RU2807998C1 - Method for spoligotyping of mycobacterium tuberculosis complex using dna amplification in immobilized phase and biological microchip for its implementation - Google Patents
Method for spoligotyping of mycobacterium tuberculosis complex using dna amplification in immobilized phase and biological microchip for its implementation Download PDFInfo
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Abstract
Description
Область техники, к которой относится изобретениеField of technology to which the invention relates
Изобретение относится к молекулярной биологии, микробиологии и медицине и обеспечивает внутривидовое и межвидовое генетическое типирование микобактерий туберкулезного комплекса.The invention relates to molecular biology, microbiology and medicine and provides intraspecific and interspecific genetic typing of mycobacteria of the tuberculosis complex.
Уровень техники State of the art
Туберкулез (ТБ) остается одним из наиболее опасных инфекционных заболеваний, уносящих миллионы жизней в год (Floyd, K., Glaziou, P., Zumla, A., Raviglione, М. (2018) The global tuberculosis epidemic and progress in care, prevention, and research: an overview in year 3 of the End ТВ era. Lancet Respir Med, 6, 299-314). Из-за быстрого трансконтинентального распространения инфекции и особенно частой коинфекции с ВИЧ (Adhikari, N., Bhattarai, R.B., Basnet, R., Joshi, L.R., Tinkari, B.S., Thapa, A., Joshi, B. (2022) Prevalence and associated risk factors for tuberculosis among people living with HIV in Nepal. PloS one, 17, e0262720; Ashenafi, S., Rao Muwa, J., Mily, A., Snail, J., Zewdie, M., Chanyalew, M, Rehn, A., Rahman, S., Aseffa, G., Bekele, A., Aderaye, G., Lema, В., Svensson, M., Brighenti, S. (2022) Immunosuppressive features of the microenvironment in lymph nodes granulomas from tuberculosis and HIV co-infected patients. Am J Path S0002-9440(22)00012-8) число новых случаев имеет тенденцию к увеличению как в промышленно развитых странах, так и в менее развитых странах (Daley, C.L. (2019) The Global Fight Against Tuberculosis. Thorac Surg Clin, 29, 19-25).Tuberculosis (TB) remains one of the most dangerous infectious diseases, claiming millions of lives per year (Floyd, K., Glaziou, P., Zumla, A., Raviglione, M. (2018) The global tuberculosis epidemic and progress in care, prevention , and research: an overview in year 3 of the End TV era. Lancet Respir Med, 6, 299-314). Due to the rapid transcontinental spread of infection and especially frequent co-infection with HIV (Adhikari, N., Bhattarai, R.B., Basnet, R., Joshi, L.R., Tinkari, B.S., Thapa, A., Joshi, B. (2022) Prevalence and associated risk factors for tuberculosis among people living with HIV in Nepal. PloS one, 17, e0262720; Ashenafi, S., Rao Muwa, J., Mily, A., Snail, J., Zewdie, M., Chanyalew, M. Rehn, A., Rahman, S., Aseffa, G., Bekele, A., Aderaye, G., Lema, V., Svensson, M., Brighenti, S. (2022) Immunosuppressive features of the microenvironment in lymph nodes granulomas from tuberculosis and HIV co-infected patients. Am J Path S0002-9440(22)00012-8) the number of new cases is trending upward in both industrialized countries and less developed countries (Daley, C. L. (2019) The Global Fight Against Tuberculosis. Thorac Surg Clin, 29, 19-25).
Помимо своевременной диагностики, ключевыми факторами в борьбе с туберкулезом являются быстрое выявление возбудителя, тестирование на лекарственную чувствительность, немедленное ограничение передачи и адекватное медикаментозное лечение. Эпидемиологический анализ, основанный на молекулярном генотипировании патогена, является мощным инструментом в идентификации источников инфекции и путей передачи, которые позволяют принимать соответствующие меры (Driscoll, J.R. (2009) Spoligotyping for molecular epidemiology of the Mycobacterium tuberculosis complex. Meth Mol Biol (Clifton, N.J.), 551, 117-128; Kamerbeek, J., Schouls, L., Kolk, A., Van Agterveld, M., Van Soolingen, D., Kuijper, S., Bunschoten, A., Molhuizen, H., Shaw, R., Goyal, M, Van Embden, J. (1997) Simultaneous detection and strain differentiation of Mycobacterium tuberculosis for diagnosis and epidemiology. J Clin Microbiol, 35, 907-914; Tejashree, A., Mahesh, P.A., Krishna Karthik, M., Nirmala Azeem, A., Reddy, R., Ravichandra, C, Nagaraja, S.B. (2022) Era of ТВ elimination: Growing need to understand diversities of Mycobacterium tuberculosis lineages!. Indian J Tuberc, 69, 79-84).In addition to timely diagnosis, the key factors in the control of tuberculosis are rapid identification of the pathogen, drug susceptibility testing, immediate limitation of transmission and adequate drug treatment. Epidemiological analysis based on molecular genotyping of the pathogen is a powerful tool in identifying sources of infection and transmission routes that allow appropriate action to be taken (Driscoll, J.R. (2009) Spoligotyping for molecular epidemiology of the Mycobacterium tuberculosis complex. Meth Mol Biol (Clifton, N.J.) , 551, 117-128; Kamerbeek, J., Schouls, L., Kolk, A., Van Agterveld, M., Van Soolingen, D., Kuijper, S., Bunschoten, A., Molhuizen, H., Shaw , R., Goyal, M, Van Embden, J. (1997) Simultaneous detection and strain differentiation of Mycobacterium tuberculosis for diagnosis and epidemiology. J Clin Microbiol, 35, 907-914; Tejashree, A., Mahesh, P.A., Krishna Karthik , M., Nirmala Azeem, A., Reddy, R., Ravichandra, C, Nagaraja, S. B. (2022) Era of TV elimination: Growing need to understand diversities of Mycobacterium tuberculosis lineages!. Indian J Tuberc, 69, 79-84 ).
Однако, если существующие экспресс-методики определения антибиотикоустойчивости, основанные на ПЦР, связаны с анализом определенной генной мишени (при известном молекулярно-генетическом механизме устойчивости), то ситуация с внутривидовым типированием не столь однозначна в силу отсутствия универсального метода, хорошо применимого для большинства туберкулезных штаммов. Принимая во внимание региональные особенности эндемичных штаммов, принадлежащих к различным эволюционным типам, что отражается на их молекулярно-биологических свойствах, часто приходится выбирать оптимальный подход типирования для каждого географического региона (Diriba, В., Berkessa, Т., Mamo, G., Tedla, Y., Ameni, G. (2013) Spoligotyping of multidrug-resistant Mycobacterium tuberculosis isolates in Ethiopia. Int J Tuberc Lung Dis, 17, 246-250; Haile, В., Tafess, K., Zewude, A., Yenew, В., Siu, G., Ameni, G. (2019) Spoligotyping and drug sensitivity of Mycobacterium tuberculosis isolated from pulmonary tuberculosis patients in the Arsi Zone of southeastern Ethiopia. New Microbes New Infect, 33,100620; Ramazanzadeh, R., Shakib, P., Rouhi, S., Mohammadi, В., Mohajeri, P., Borji, S. (2020) Molecular epidemiology of Mycobacterium tuberculosis isolates in Iran using spoligotyping. New Microbes New Infect, 38, 100767).However, if the existing rapid methods for determining antibiotic resistance, based on PCR, are associated with the analysis of a specific gene target (with a known molecular genetic mechanism of resistance), then the situation with intraspecific typing is not so clear due to the lack of a universal method that is well applicable for most tuberculosis strains . Taking into account the regional characteristics of endemic strains belonging to different evolutionary types, which is reflected in their molecular biological properties, it is often necessary to choose the optimal typing approach for each geographical region (Diriba, V., Berkessa, T., Mamo, G., Tedla , Y., Ameni, G. (2013) Spoligotyping of multidrug-resistant Mycobacterium tuberculosis isolates in Ethiopia. Int J Tuberc Lung Dis, 17, 246-250; Haile, V., Tafess, K., Zewude, A., Yenew , V., Siu, G., Ameni, G. (2019) Spoligotyping and drug sensitivity of Mycobacterium tuberculosis isolated from pulmonary tuberculosis patients in the Arsi Zone of southeastern Ethiopia. New Microbes New Infect, 33,100620; Ramazanzadeh, R., Shakib, P., Rouhi, S., Mohammadi, V., Mohajeri, P., Borji, S. (2020) Molecular epidemiology of Mycobacterium tuberculosis isolates in Iran using spoligotyping. New Microbes New Infect, 38, 100767).
Сравнению возможностей различных методов типирования посвящено довольно много работ (Ei, P.W., Aung, W.W., Lee, J.S., Choi, G.E., Chang, C.L. (2016) Molecular Strain Typing of Mycobacterium tuberculosis: a Review of Frequently Used Methods. J Korean Med Sci, 31,1673-1683; Masoud, K., Araj, G.F., Reslan, L., Fadlallah, S., Wehbe, M., Itani, L., Avedissian, A., Dbaibo, G., Saade, A., Refregier, G., Sola, C, Matar, G.M. (2020) Spoligotyping of Mycobacterium tuberculosis isolates using Luminex®-based method in Lebanon. J Infect Dev Ctries, 14, 878-885; Ravansalar, H., Tadayon, K., Ghazvini, K. (2016) Molecular typing methods used in studies of Mycobacterium tuberculosis in Iran: a systematic review. Iran J Microbiol, 8, 338-346). Кроме того, предложены комбинации нескольких методов, которые, по мнению авторов, позволяют повысить уровень дифференцировки и осуществлять более качественный эпидемиологический мониторинг (Parreiras, P.M., Andrade, G.I., Nascimento, Т., Oelemann, M.C., Gomes, H.M., Alencar, A.P., Assis, R.A., Mota, P.M., Pereira, M.A., Lobato, F.C., Lage, A.P., Suffys, P.N. (2012) Spoligotyping and variable number tandem repeat analysis of Mycobacterium bovis isolates from cattle in Brazil. Mem Inst Oswaldo Cruz, 107, 64-73; Shi, J., Zheng, D., Zhu, Y., Ma, X., Wang, S., Li, H., Xing, J. (2018) Role of MIRU-VNTR and spoligotyping in assessing the genetic diversity of Mycobacterium tuberculosis in Henan Province, China. BMC Infect Dis, 18, 447). В настоящее время широко распространенными являются методы, основанные на фингерпринтинге различных геномных мишеней, таких как IS 6110 (Dekhil, N., Skhairia, M.A., Mhenni, В., Ben Fradj, S., Warren, R., Mardassi, H. (2018) Automated IS6110-based fingerprinting of Mycobacterium tuberculosis: Reaching unprecedented discriminatory power and versatility. PloS One, 13, e0197913), PGRS (Yang, Z.H., Ijaz, K, Bates, J.H., Eisenach, K.D., Cave, M.D. (2000) Spoligotyping and polymorphic GC-rich repetitive sequence fingerprinting of Mycobacterium tuberculosis strains having few copies of IS6110. J Clin Microbiol, 38, 3572-3576), DR (Driscoll, J.R. (2009) Spoligotyping for molecular epidemiology of the Mycobacterium tuberculosis complex. Meth Mol Biol (Clifton, N.J.), 551, 117-128; Tejashree, A., Mahesh, P.A., Krishna Karthik, M., Nirmala Azeem, A., Reddy, R., Ravichandra, C, Nagaraja, S.B. (2022) Era of ТВ elimination: Growing need to understand diversities of Mycobacterium tuberculosis lineages!. Indian J Tuberc, 69, 79-84), 16S rDNA (Cheung, M.K., Lam, W.Y., Fung, W.Y., Law, P.T., Au, C.H., Nong, W., Kam, K.M., Kwan, H.S., Tsui, S.K. (2013) Sputum microbiota in tuberculosis as revealed by 16S rRNA pyrosequencing. PloS One, 8, e54574) и другие подходы; однако, основным их недостатком является необходимость использования сложного оборудования для анализа получаемых электрофорезных профилей, а также необходимость в предварительном подращивании культуры (Merker, М., Kohl, Т.А., Niemann, S., Supply, P. (2017) The Evolution of Strain Typing in the Mycobacterium tuberculosis Complex. Adv Exp Med Biol, 1019, 43-78).Quite a lot of work has been devoted to comparing the capabilities of different typing methods (Ei, P.W., Aung, W.W., Lee, J.S., Choi, G.E., Chang, C.L. (2016) Molecular Strain Typing of Mycobacterium tuberculosis: a Review of Frequently Used Methods. J Korean Med Sci , 31,1673-1683; Masoud, K., Araj, G.F., Reslan, L., Fadlallah, S., Wehbe, M., Itani, L., Avedissian, A., Dbaibo, G., Saade, A. , Refregier, G., Sola, C, Matar, G. M. (2020) Spoligotyping of Mycobacterium tuberculosis isolates using Luminex®-based method in Lebanon. J Infect Dev Ctries, 14, 878-885; Ravansalar, H., Tadayon, K. , Ghazvini, K. (2016) Molecular typing methods used in studies of Mycobacterium tuberculosis in Iran: a systematic review. Iran J Microbiol, 8, 338-346). In addition, combinations of several methods have been proposed, which, according to the authors, make it possible to increase the level of differentiation and carry out better epidemiological monitoring (Parreiras, P.M., Andrade, G.I., Nascimento, T., Oelemann, M.C., Gomes, H.M., Alencar, A.P., Assis, R. A., Mota, P. M., Pereira, M. A., Lobato, F. C., Lage, A. P., Suffys, P. N. (2012) Spoligotyping and variable number tandem repeat analysis of Mycobacterium bovis isolates from cattle in Brazil. Mem Inst Oswaldo Cruz, 107, 64 -73; Shi, J., Zheng, D., Zhu, Y., Ma, X., Wang, S., Li, H., Xing, J. (2018) Role of MIRU-VNTR and spoligotyping in assessing the genetic diversity of Mycobacterium tuberculosis in Henan Province, China. BMC Infect Dis, 18, 447). Currently, methods based on fingerprinting of various genomic targets, such as IS 6110, are widespread (Dekhil, N., Skhairia, M.A., Mhenni, V., Ben Fradj, S., Warren, R., Mardassi, H. ( 2018) Automated IS6110-based fingerprinting of Mycobacterium tuberculosis: Reaching unprecedented discriminatory power and versatility. PloS One, 13, e0197913), PGRS (Yang, Z.H., Ijaz, K, Bates, J.H., Eisenach, K.D., Cave, M.D. (2000) Spoligotyping and polymorphic GC-rich repetitive sequence fingerprinting of Mycobacterium tuberculosis strains having few copies of IS6110. J Clin Microbiol, 38, 3572-3576), DR (Driscoll, J.R. (2009) Spoligotyping for molecular epidemiology of the Mycobacterium tuberculosis complex. Meth Mol Biol (Clifton, N.J.), 551, 117-128; Tejashree, A., Mahesh, P. A., Krishna Karthik, M., Nirmala Azeem, A., Reddy, R., Ravichandra, C., Nagaraja, S. B. (2022) Era of TV elimination: Growing need to understand diversities of Mycobacterium tuberculosis lineages!. Indian J Tuberc, 69, 79-84), 16S rDNA (Cheung, M.K., Lam, W.Y., Fung, W.Y., Law, P.T., Au, C.H., Nong, W., Kam, K.M., Kwan, H.S., Tsui, S.K. (2013) Sputum microbiota in tuberculosis as revealed by 16S rRNA pyrosequencing. PloS One, 8, e54574) and other approaches; however, their main disadvantage is the need to use sophisticated equipment to analyze the resulting electrophoretic profiles, as well as the need for preliminary cultivation of the culture (Merker, M., Kohl, T.A., Niemann, S., Supply, P. (2017) The Evolution of Strain Typing in the Mycobacterium tuberculosis Complex. Adv Exp Med Biol, 1019, 43-78).
Это одна из причин, обусловивших распространение новых, основанных на ПЦР, методов типирования. Одним из таких методов является т.н. сполиготипирование (от. англ. spacer oligonucleotide typing). Метод основан на анализе локуса прямых повторов микобактериального генома, описанного в 1991 г. Hermans и соавторами (Hermans, P.W.M., van Soolingen, D., Bik, E.M., de Haas, P.E.W., Dale, J.W., van Embden, J.D.A. (1991) The insertion element IS987 from Mycobacterium bovis BCG is located in a hot spot integration region for insertion elements in Mycobacterium tuberculosis complex strains. Infect Immun, 59, 2695-2705). Данный локус характеризуется наличием многократно повторяющихся участков длиной 36 н.п.идентичной последовательности, между которыми расположены уникальные спейсерные последовательности длиной от 24 до 41 н.п. Различные штаммы характеризуются наличием разного числа таких спейсерных последовательностей, что позволяет делать заключения о степени их родства и, соответственно, проводить эпидемиологические исследования. Изначально предложенный для внутривидового типирования М. tuberculosis (Groenen, Р.М.А., Bunschoten, А.Е., van Soolingen, D., van Embden, J.D.A. (1993) Nature of DNA polymorphism in the direct repeat cluster of Mycobacterium tuberculosis; application of strain differentiation by a novel typing method. Mol Microbiol, 10, 1057-1065), метод, основанный на гибридизации ПЦР-продуктов со специфичными к спейсерным участкам зондами, затем успешно распространен и на бовисные штаммы (Aranaz, A., Liebana, Е., Mateos, A., Dominguez, L., Vidal, D., Domingo, M., Gonzolez, О., Rodriguez-Ferri, E.F., Bunschoten, A.E., Van Embden, J.D., Cousins, D. (1996) Spacer oligonucleotide typing of Mycobacterium bovis strains from cattle and other animals: a tool for studying epidemiology of tuberculosis. J Clin Microbiol, 34, 2734-2740; Cousins, D., Williams, S., Liebana, E., Aranaz, A., Bunschoten, A., Van Embden, J., Ellis, T. (1998) Evaluation of four DNA typing techniques in epidemiological investigations of bovine tuberculosis. J Clin Microbiol, 36, 168-178). К настоящему времени предпринимаются не только попытки стандартизации методики сполиготипирования (Dale, J.W., Brittain, D., Cataldi, A.A., Cousins, D., Crawford, J.T., Driscoll, J., Heersma, H., Lillebaek, Т., Quitugua, Т., Rastogi, N., Skuce, R.A., Sola, C, Van Soolingen, D., Vincent, V. (2001) Spacer oligonucleotide typing of bacteria of the Mycobacterium tuberculosis complex: recommendations for standardized nomenclature. Int J Tub Lung Dis, 5, 216-219), но и составлены обширные базы данных (Demay, С, Liens, В., Burguiere, Т., Hill, V., Couvin, D., Millet, J., Mokrousov, I., Sola, C, Zozio, Т., Rastogi, N. (2012) SITVITWEB - a publicly available international multimarker database for studying Mycobacterium tuberculosis genetic diversity and molecular epidemiology. Infect Genet Evol, 12, 755-766; Filliol, I., Driscoll, J.R., van Soolingen, D., et al. (2003) Snapshot of moving and expanding clones of Mycobacterium tuberculosis and their global distribution assessed by spoligotyping in an International Study. J Clin Microbiol, 41, 1963-1970). Стандартизированные базы данных содержат информацию о сорока трех (43) спейсерных последовательностях. Несмотря на отмечаемую многими авторами в ряде случаев меньшую дискриминирующую способность по сравнению с другими общепринятыми методами типирования (Bauer, J., Andersen, А.В., Kremer, K., Miorner, H. (1999) Usefulness of spoligotyping to discriminate IS6110 low-copy-number Mycobacterium tuberculosis complex strains cultured in Denmark. J Clin Microbiol, 37, 2602-2606; Kamerbeek, J., Schouls, L., Kolk, A., Van Agterveld, M., Van Soolingen, D., Kuijper, S., Bunschoten, A., Molhuizen, H., Shaw, R., Goyal, M., Van Embden, J. (1997) Simultaneous detection and strain differentiation of Mycobacterium tuberculosis for diagnosis and epidemiology. J Clin Microbiol, 35, 907-914; Kremer, K., Van Soolingen, D., Frothingham, R., Haas, W.H., Hermans, P.W.M., Martin, C, Palittapongarnpim, P., Plikaytis, B.B., Riley, L.W., Yakrus, M.A., Musser, J.M., Van Embden, J.D.A. (1999) Comparison of methods based on different molecular epidemiological markers for typing of Mycobacterium tuberculosis complex strains: interlaboratory study of discriminatory power and reproducibility. J Clin Microbiol, 37, 2607-2618; Wilson, S.M., Goss, S., Drobnievski, F. (1998) Evaluation of strategies for molecular fingerprinting for use in the routine work of a mycobacterium reference unit. J Clin Microbiol, 36, 3385-3388), отмечена дешевизна метода, простота его выполнения и интерпретации получаемых гибридизационных профилей (Cousins, D., Williams, S., Liebana, E., Aranaz, A., Bunschoten, A., Van Embden, J., Ellis, T. (1998) Evaluation of four DNA typing techniques in epidemiological investigations of bovine tuberculosis. J Clin Microbiol, 36, 168-178; Kremer, K., Van Soolingen, D., Frothingham, R., Haas, W.H., Hermans, P.W.M., Martin, C, Palittapongarnpim, P., Plikaytis, B.B., Riley, L.W., Yakrus, M.A., Musser, J.M., Van Embden, J.D.A. (1999) Comparison of methods based on different molecular epidemiological markers for typing of Mycobacterium tuberculosis complex strains: interlaboratory study of discriminatory power and reproducibility. J Clin Microbiol, 37, 2607-2618; Sola, C, Filliol, I., Legrand, E., Rastogi, N. (2000) Recent developments of spoligotyping as applied to the study of epidemiology, biodiversity and molecular phylogeny of the Mycobacterium tuberculosis complex. Pathol Biol (Paris), 48, 921-932). Это позволяет использовать сполиготипирование для первичной эпидемиологической оценки, затем, при необходимости, дополняя его методами с большей разрешающей способностью (Almeida, S.M., Malaspina, А.С., Leite, С, Saad, М. (2019) Usefulness of 3'- 5' IS6110-RFLP genotyping and spoligotyping of Mycobacterium tuberculosis isolated in a tertiary hospital: a retrospective study detecting unsuspected epidemiological events. Rev Inst Med Trop Sao Paulo, 61, e51, Merker, M., Kohl, T.A., Niemann, S., Supply, P. (2017) The Evolution of Strain Typing in the Mycobacterium tuberculosis Complex. Adv Exp Med Biol, 1019, 43-78). Другой подход заключается в использовании двух различных методов, основанных на ПЦР, одним из которых являлось бы сполиготипирование (Filliol, I., Ferdinand, S., Negroni, L., Sola, C, Rastogi, N. (2000) Molecular typing of Mycobacterium tuberculosis based on variable number of tandem DNA repeats used alone and in association with spoligotyping. J Clin Microbiol, 38,2520-2524). По своей разрешающей способности такой "тандем" приближается к традиционному фингерпринтингу по IS 6110. Ценность метода сполиготипирования заключается также в возможности одновременной детекции возбудителя, причем анализ может выполняться непосредственно из клинического образца без предварительного подращивания культуры, а также способность дискриминировать туберкулезные и бовисные штаммы, что представляет серьезные трудности при использовании общепринятых культуральных методов (Roring, S., Hughes, M.S., Skuce, R.A., Neill, S.D. (2000) Simultaneous detection and strain differentiation of Mycobacterium bovis directly from bovine tissue specimens by spoligotyping. Vet Microbiol, 74, 227-236).This is one of the reasons that led to the spread of new PCR-based typing methods. One of these methods is the so-called. spoligotyping (from the English spacer oligonucleotide typing). The method is based on the analysis of the direct repeat locus of the mycobacterial genome, described in 1991 by Hermans et al. insertion element IS987 from Mycobacterium bovis BCG is located in a hot spot integration region for insertion elements in Mycobacterium tuberculosis complex strains. Infect Immun, 59, 2695-2705). This locus is characterized by the presence of multiple repeating sections of 36 bp in length of an identical sequence, between which are located unique spacer sequences ranging from 24 to 41 bp in length. Different strains are characterized by the presence of different numbers of such spacer sequences, which allows one to draw conclusions about the degree of their relationship and, accordingly, conduct epidemiological studies. Originally proposed for intraspecific typing of M. tuberculosis (Groenen, R. M. A., Bunschoten, A. E., van Soolingen, D., van Embden, J. D. A. (1993) Nature of DNA polymorphism in the direct repeat cluster of Mycobacterium tuberculosis ; application of strain differentiation by a novel typing method. Mol Microbiol, 10, 1057-1065), a method based on hybridization of PCR products with probes specific to spacer regions, then successfully extended to bovis strains (Aranaz, A., Liebana , E., Mateos, A., Dominguez, L., Vidal, D., Domingo, M., Gonzolez, O., Rodriguez-Ferri, E. F., Bunschoten, A. E., Van Embden, J. D., Cousins, D. (1996 ) Spacer oligonucleotide typing of Mycobacterium bovis strains from cattle and other animals: a tool for studying epidemiology of tuberculosis. J Clin Microbiol, 34, 2734-2740; Cousins, D., Williams, S., Liebana, E., Aranaz, A. ., Bunschoten, A., Van Embden, J., Ellis, T. (1998) Evaluation of four DNA typing techniques in epidemiological investigations of bovine tuberculosis. J Clin Microbiol, 36, 168-178). To date, not only attempts have been made to standardize the spoligotyping technique (Dale, J.W., Brittain, D., Cataldi, A.A., Cousins, D., Crawford, J.T., Driscoll, J., Heersma, H., Lillebaek, T., Quitugua, T., Rastogi, N., Skuce, R.A., Sola, C., Van Soolingen, D., Vincent, V. (2001) Spacer oligonucleotide typing of bacteria of the Mycobacterium tuberculosis complex: recommendations for standardized nomenclature. Int J Tub Lung Dis , 5, 216-219), but also extensive databases have been compiled (Demay, S., Liens, V., Burguiere, T., Hill, V., Couvin, D., Millet, J., Mokrousov, I., Sola , C, Zozio, T., Rastogi, N. (2012) SITVITWEB - a publicly available international multimarker database for studying Mycobacterium tuberculosis genetic diversity and molecular epidemiology. Infect Genet Evol, 12, 755-766; Filliol, I., Driscoll, J.R., van Soolingen, D., et al (2003) Snapshot of moving and expanding clones of Mycobacterium tuberculosis and their global distribution assessed by spoligotyping in an International Study. J Clin Microbiol, 41, 1963-1970). Standardized databases contain information on forty-three (43) spacer sequences. Despite the lower discriminatory ability noted by many authors in some cases compared to other generally accepted typing methods (Bauer, J., Andersen, A.V., Kremer, K., Miorner, H. (1999) Usefulness of spoligotyping to discriminate IS6110 low -copy-number Mycobacterium tuberculosis complex strains cultured in Denmark. J Clin Microbiol, 37, 2602-2606; Kamerbeek, J., Schouls, L., Kolk, A., Van Agterveld, M., Van Soolingen, D., Kuijper , S., Bunschoten, A., Molhuizen, H., Shaw, R., Goyal, M., Van Embden, J. (1997) Simultaneous detection and strain differentiation of Mycobacterium tuberculosis for diagnosis and epidemiology. , 907-914; Kremer, K., Van Soolingen, D., Frothingham, R., Haas, W.H., Hermans, P.W.M., Martin, C., Palittapongarnpim, P., Plikaytis, B.B., Riley, L.W., Yakrus, M.A., Musser, J. M., Van Embden, J. D. A. (1999) Comparison of methods based on different molecular epidemiological markers for typing of Mycobacterium tuberculosis complex strains: interlaboratory study of discriminatory power and reproducibility. J Clin Microbiol, 37, 2607-2618; Wilson, S.M., Goss, S., Drobnievski, F. (1998) Evaluation of strategies for molecular fingerprinting for use in the routine work of a mycobacterium reference unit. J Clin Microbiol, 36, 3385-3388), noted the low cost of the method, ease of its implementation and interpretation of the resulting hybridization profiles (Cousins, D., Williams, S., Liebana, E., Aranaz, A., Bunschoten, A., Van Embden, J., Ellis, T. (1998) Evaluation of four DNA typing techniques in epidemiological investigations of bovine tuberculosis. J Clin Microbiol, 36, 168-178; Kremer, K., Van Soolingen, D., Frothingham, R. , Haas, W.H., Hermans, P.W.M., Martin, C, Palittapongarnpim, P., Plikaytis, B.B., Riley, L.W., Yakrus, M.A., Musser, J.M., Van Embden, J.D.A. (1999) Comparison of methods based on different molecular epidemiological markers for typing of Mycobacterium tuberculosis complex strains: interlaboratory study of discriminatory power and reproducibility. J Clin Microbiol, 37, 2607-2618; Sola, C, Filliol, I., Legrand, E., Rastogi, N. (2000) Recent developments of spoligotyping as applied to the study of epidemiology, biodiversity and molecular phylogeny of the Mycobacterium tuberculosis complex. Pathol Biol (Paris), 48, 921-932). This allows spoligotyping to be used for initial epidemiological assessment, then, if necessary, complemented by methods with higher resolution (Almeida, S.M., Malaspina, A.S., Leite, C, Saad, M. (2019) Usefulness of 3'-5 'IS6110-RFLP genotyping and spoligotyping of Mycobacterium tuberculosis isolated in a tertiary hospital: a retrospective study detecting unsuspected epidemiological events. Rev Inst Med Trop Sao Paulo, 61, e51, Merker, M., Kohl, T.A., Niemann, S., Supply , P. (2017) The Evolution of Strain Typing in the Mycobacterium tuberculosis Complex. Adv Exp Med Biol, 1019, 43-78). Another approach would be to use two different PCR-based methods, one of which would be spoligotyping (Filliol, I., Ferdinand, S., Negroni, L., Sola, C, Rastogi, N. (2000) Molecular typing of Mycobacterium tuberculosis based on variable number of tandem DNA repeats used alone and in association with spoligotyping. J Clin Microbiol, 38,2520-2524). In terms of its resolution, such a “tandem” approaches traditional fingerprinting according to IS 6110. The value of the spoligotyping method also lies in the possibility of simultaneous detection of the pathogen, and the analysis can be performed directly from a clinical sample without prior cultivation of the culture, as well as the ability to discriminate between tuberculosis and bovis strains, which presents serious difficulties when using conventional culture methods (Roring, S., Hughes, M.S., Skuce, R.A., Neill, S.D. (2000) Simultaneous detection and strain differentiation of Mycobacterium bovis directly from bovine tissue specimens by spoligotyping. Vet Microbiol, 74, 227 -236).
Одним из недостатков предложенного в работе (Kamerbeek, J., Schouls, L., Kolk, A., Van Agterveld, M., Van Soolingen, D., Kuijper, S., Bunschoten, A., Molhuizen, H., Shaw, R., Goyal, M., Van Embden, J. (1997) Simultaneous detection and strain differentiation of Mycobacterium tuberculosis for diagnosis and epidemiology. J Clin Microbiol, 35, 907-914) варианта метода некоторыми исследователями указывается сравнительно большое время анализа - он занимает 1-2 рабочих дня, что на сегодняшнее время нельзя считать удовлетворительным.One of the disadvantages of what was proposed in the work (Kamerbeek, J., Schouls, L., Kolk, A., Van Agterveld, M., Van Soolingen, D., Kuijper, S., Bunschoten, A., Molhuizen, H., Shaw , R., Goyal, M., Van Embden, J. (1997) Simultaneous detection and strain differentiation of Mycobacterium tuberculosis for diagnosis and epidemiology. J Clin Microbiol, 35, 907-914) version of the method, some researchers indicate a relatively long analysis time - it takes 1-2 business days, which today cannot be considered satisfactory.
Изобретение, описанное в патентной заявке RU 2237090 С1 «Способ генетического типирования штаммов микобактерий туберкулезного комплекса», связано с типированием по полиморфным локусам с применением мультиплексной системы праймеров. Способ позволяет устанавливать идентичность геномов микобактерий туберкулезного комплекса в эпидемиологических исследованиях и клинической медицине. Недостатком способа является сложность мультипраймерной схемы (пять пар праймеров, находящихся в общем реакционном объеме), возможные затруднения с интерпретацией результатов, а также недостаточная стандартизация по сравнению с другими методами типирования. Способ не использует биологический микрочип.The invention described in patent application RU 2237090 C1 “Method for genetic typing of strains of Mycobacterium tuberculosis complex” is associated with typing at polymorphic loci using a multiplex primer system. The method makes it possible to establish the identity of the genomes of Mycobacterium tuberculosis complex in epidemiological studies and clinical medicine. The disadvantage of this method is the complexity of the multiprimer design (five pairs of primers located in the total reaction volume), possible difficulties with the interpretation of the results, as well as insufficient standardization compared to other typing methods. The method does not use a biological microchip.
Изобретение RU 2689801 С1 «Способ детекции штаммов Mycobacterium bovis BCG в формате реального времени» описывает детекцию штаммов Mycobacterium bovis BCG путем лабораторного выявления геномной делеции RD1. Недостатком является невозможность типирования как Mycobacterium bovis, так и других представителей комплекса МБТ. Способ не использует биологический микрочип.Invention RU 2689801 C1 “Method for detecting Mycobacterium bovis BCG strains in real time” describes the detection of Mycobacterium bovis BCG strains by laboratory detection of the RD1 genomic deletion. The disadvantage is the impossibility of typing both Mycobacterium bovis and other representatives of the MBT complex. The method does not use a biological microchip.
Изобретение CN 102094077 A "Method for real-time detection of beijing genotype mycobacterium tuberculosis" (КНР) относится к области медицинских тестов и обеспечивает набор для быстрого определения генотипа штамма Mycobacterium tuberculosis. Набор предназначен для детекции последовательности генетического элемента IS6110. Метод не позволяет проводить эпидемиологические исследования представителей микобактериального комплекса МБТ.The invention CN 102094077 A "Method for real-time detection of beijing genotype of mycobacterium tuberculosis" (China) relates to the field of medical tests and provides a kit for quickly determining the genotype of a Mycobacterium tuberculosis strain. The kit is designed to detect the sequence of the genetic element IS6110. The method does not allow for epidemiological studies of representatives of the mycobacterial MBT complex.
Применение метода "direct on-chip PCR" - непосредственная амплификация на биологическом микрочипе - позволяет решить проблему генотипирования параллельно с миниатюризацией процесса.The use of the "direct on-chip PCR" method - direct amplification on a biological microchip - makes it possible to solve the problem of genotyping in parallel with the miniaturization of the process.
В настоящем изобретении описан метод сполиготипирования штаммов М tuberculosis и М. bovis (потенциально - и других представителей комплекса М. tuberculosis), использующий ПЦР на биологическом микрочипе с иммобилизованными праймерами, высокоспецифичньгми к DR-локусу (локус прямых повторов) микобактериального генома.The present invention describes a method for spoligotyping of M tuberculosis and M. bovis strains (potentially other representatives of the M. tuberculosis complex), using PCR on a biological microchip with immobilized primers that are highly specific to the DR locus (direct repeat locus) of the mycobacterial genome.
Раскрытие сущности изобретенияDisclosure of the invention
Изобретение, основанное на применение ПЦР на биологическом микрочипе (биочип), включает набор иммобилизованных в гелевых ячейках биочипа праймеров, специфичных к сорока трем спейсерным участкам DR-локуса, а также свободные праймеры, комплементарные прямому повтору: прямой DR-B1 и обратный флуоресцентно меченный DR-A1 (Фигура 1). Полный набор праймеров конструировали с учетом общепринятых требований (разброс расчетных температур плавления не более 2-3°С, отсутствие высокостабильных вторичных структур, высокая специфичность к выбранному региону). Кроме того, каждый иммобилизованный праймер проверяли на отсутствие дуплексов с праймерами DR-A1 и DR-B1 и анализировали на отсутствие сродства к другим локусам микобактериального и других геномов, последовательности которых доступны для BLAST-анализа. При расчете температур плавления дуплексов принимали во внимание влияние гелей биочипа на стабильность образующихся дуплексов. Поэтому реальная температура амплификации была понижена до 60°С.The invention, based on the use of PCR on a biological microchip (biochip), includes a set of primers immobilized in the gel cells of the biochip, specific to forty-three spacer regions of the DR locus, as well as free primers complementary to the direct repeat: forward DR-B1 and reverse fluorescently labeled DR -A1 (Figure 1). The complete set of primers was designed taking into account generally accepted requirements (dispersion of calculated melting temperatures no more than 2-3°C, absence of highly stable secondary structures, high specificity to the selected region). In addition, each immobilized primer was checked for the absence of duplexes with primers DR-A1 and DR-B1 and analyzed for the lack of affinity for other loci of the mycobacterial and other genomes, the sequences of which are available for BLAST analysis. When calculating the melting temperatures of duplexes, we took into account the influence of biochip gels on the stability of the resulting duplexes. Therefore, the actual amplification temperature was lowered to 60°C.
В случае, когда длина спейсерного участка не позволяла выполнить последнее условие (обеспечение необходимой стабильности дуплекса), праймер конструировали таким образом, что его 5'-концевая область заходила в правую часть прямого повтора. Последовательности праймеров указаны в Перечне последовательностей. Иммобилизованные праймеры находятся в отдельных гелевых ячейках биочипа.In cases where the length of the spacer region did not allow the last condition to be met (providing the necessary stability of the duplex), the primer was designed in such a way that its 5'-terminal region included the right side of the direct repeat. Primer sequences are listed in the Sequence List. Immobilized primers are located in separate gel cells of the biochip.
Структура самого биочипа определяется требованием миниатюризации, а именно уменьшением линейных размеров для удобства считывания получаемого ПЦР-профиля на портативном анализаторе биочипов. В результате биочип представляет собой упорядоченную матрицу 10x5 мм, элементами которой являются ячейки, содержащие праймеры, специфичные к сорока трем описанным в литературе и внесенным в базы данных прямым повторам. Сорок три олигонуклеотида расположены в порядке возрастания номера сверху вниз и слева направо, т.е. левый вертикальный столбец содержит первый десяток праймеров и т.д. (Фигура 2). Биочипы изготавливали по сополимеризационной технологии, когда праймер для иммобилизации вносится на стеклянную подложку биочипа вместе с раствором акриламида, из которого формируется ячейка.The structure of the biochip itself is determined by the requirement of miniaturization, namely, a reduction in linear dimensions for ease of reading the resulting PCR profile on a portable biochip analyzer. As a result, the biochip is an ordered 10x5 mm matrix, the elements of which are cells containing primers specific to forty-three direct repeats described in the literature and included in databases. Forty-three oligonucleotides are arranged in ascending number order from top to bottom and left to right, i.e. the left vertical column contains the first ten primers, etc. (Figure 2). The biochips were manufactured using copolymerization technology, when a primer for immobilization is applied to the glass substrate of the biochip along with an acrylamide solution from which the cell is formed.
Немаловажным этапом оптимизации метода явился выбор акриламидного геля, свойства которого позволяли проводить ПЦР с наибольшей эффективностью. Особые требования предъявляли к размеру пор в образующемся полимере, а также к его стабильности в процессе амплификации. Оптимизация условий используемой в методе ПЦР позволила получить интенсивные сигналы амплификации при высокой воспроизводимости результатов.An important step in optimizing the method was the selection of an acrylamide gel, the properties of which made it possible to carry out PCR with the greatest efficiency. Special requirements were placed on the pore size in the resulting polymer, as well as on its stability during the amplification process. Optimization of the conditions used in the PCR method made it possible to obtain intense amplification signals with high reproducibility of results.
Процедура амплификации заканчивается инкубированием ПЦР-смеси 10 мин. при 72°С - для гибридизации полученных флуоресцентно меченных ПЦР-продуктов с экстендированными (достроенными) иммобилизованными праймерами (Фигура 1). Непосредственно после этого осуществляется отмывка биочипа для удаления неспецифичных сигналов. Это позволяет резко увеличить дискриминацию между сигналами ячеек с экстендированными (достроенными) праймерами и ячеек, содержащих праймеры, специфичные к спейсерам, отсутствующим в анализируемом образце. При этом сигналы последних сравнимы с сигналами пустых гелевых ячеек. Отношение положительных и отрицательных нормализованных сигналов превышает один порядок, что позволяет проводить визуальную интерпретацию результата (Фигура 2).The amplification procedure ends with incubation of the PCR mixture for 10 minutes. at 72°C - for hybridization of the resulting fluorescently labeled PCR products with extended (completed) immobilized primers (Figure 1). Immediately after this, the biochip is washed to remove nonspecific signals. This makes it possible to sharply increase the discrimination between the signals of cells with extended (completed) primers and cells containing primers specific to spacers that are absent in the analyzed sample. In this case, the signals of the latter are comparable to the signals of empty gel cells. The ratio of positive and negative normalized signals exceeds one order of magnitude, which allows for visual interpretation of the result (Figure 2).
Детекция флуоресцентных сигналов ячеек осуществляется с использованием флуоресцентного анализатора биочипов. Анализ ПЦР-профиля проводят как визуально (Фигура 2), так и с помощью оригинального программного обеспечения "Analyze". В последнем случае появляется возможность количественной обработки интенсивностей флуоресцентных сигналов и нахождения значений нормализованных сигналов для автоматической программной интерпретации получаемых ПЦР-профилей. Результаты представляют в общепринятом для сполиготипирования виде - используя закрашенные области схемы для обозначения положительных сигналов (Таблица 1).Detection of fluorescent signals from cells is carried out using a fluorescent biochip analyzer. Analysis of the PCR profile is carried out both visually (Figure 2) and using the original "Analyze" software. In the latter case, it becomes possible to quantitatively process fluorescent signal intensities and find the values of normalized signals for automatic software interpretation of the resulting PCR profiles. The results are presented in the form generally accepted for spoligotyping - using shaded areas of the diagram to indicate positive signals (Table 1).
Заключительным этапом разработки метода явился сравнительный анализ результатов, полученных в ходе данной работы, и результатов стандартного сполиготипирования гибридизацией на мембранах. В Таблице 1 показаны профили, полученные стандартным и предлагаемым методами (обнаружено 100%-е совпадение профилей).The final stage of method development was a comparative analysis of the results obtained in this work and the results of standard spoligotyping by hybridization on membranes. Table 1 shows the profiles obtained by the standard and proposed methods (100% coincidence of the profiles was found).
Краткое описание фигур и таблицBrief description of figures and tables
Фигура 1. Схема сполиготипирования на биочипе (микрочипе, биологическом микрочипе). Прямые повторы идентичной последовательности, присутствующие в области прямых повторов (DR), амплифицируются с помощью праймеров DR-A1 и DR-B1. Праймер DR-A1 несет флуоресцентную метку и используется с 10-кратным молярным избытком по сравнению с праймеровм DR-B1; следовательно, ПЦР в растворе на биочипе дает преимущественно одноцепочечный флуоресцентно меченый продукт.Иммобилизованные праймеры, комплементарные 43 уникальным спейсерам, пронумерованы (для простоты показаны только 3 из них). В результате ПЦР образуются очень стабильные дуплексы между удлиненными иммобилизованными праймерами и флуоресцентно меченными одноцепочечными продуктами амплификации.Figure 1. Scheme of spoligotyping on a biochip (microchip, biological microchip). Direct repeats of identical sequence present in the direct repeat region (DR) are amplified using primers DR-A1 and DR-B1. Primer DR-A1 carries a fluorescent tag and is used in a 10-fold molar excess compared to primer DR-B1; therefore, solution PCR on a biochip produces predominantly a single-stranded fluorescently labeled product. Immobilized primers complementary to the 43 unique spacers are numbered (for simplicity, only 3 of them are shown). As a result of PCR, very stable duplexes are formed between extended immobilized primers and fluorescently labeled single-stranded amplification products.
Фигура 2. Примеры профилей флуоресценции после проведения «ПЦР на чипе». Сполиготипы, полученные с использованием двух образцов ДНК, М. tuberculosis H37Rv (А) и М. bovis BCG М-34 (В). Раскладка биочипа (сетка) разделена на колонки по десять ячеек, номера колонок указаны вверху. Пятая колонка включает в себя последние три ячейки из 43.Figure 2. Examples of fluorescence profiles after “PCR on a chip”. Spoligotypes obtained using two DNA samples, M. tuberculosis H37Rv (A) and M. bovis BCG M-34 (B). The biochip layout (grid) is divided into columns of ten cells, column numbers are indicated at the top. The fifth column includes the last three cells of the 43.
Таблица 1. Результаты сполиготипирования, полученные традиционным (стандартным) методам гибридизации и методом ПЦР на биологическом микрочипе. Закрашенные серым цветом ячейки указывают на наличие соответствующих прямых повторов в ДНК исследуемого образца.Table 1. Spoligotyping results obtained using traditional (standard) hybridization methods and PCR on a biological microchip. Cells shaded in gray indicate the presence of corresponding direct repeats in the DNA of the sample under study.
Осуществление изобретенияCarrying out the invention
Изобретение нацелено на внутри- и межвидовое типирование бактериальных штаммов комплекса МБТ.The invention is aimed at intra- and interspecific typing of bacterial strains of the MBT complex.
Ниже описаны основные этапы разработки способа, изготовления биологического микрочипа, синтеза олигонуклеотидных праймеров, а также осуществления способа.The main stages of developing the method, manufacturing a biological microchip, synthesizing oligonucleotide primers, and implementing the method are described below.
Праймеры. Конструирование праймеров осуществляли с использованием программного обеспечения Oligo 5 (Molecular Biology Insights, Cascade, Colo.). Праймеры были синтезированы на синтезаторе ABI-394 DNA/RNA synthesizer (Applied Biosystems, Foster City, Calif.). Для иммобилизации праймеры модифицировали в автоматическом режиме присоединением 5'-Amino-Modifier С6 (Glen Research, Sterling, Va.). Праймеры были очищены методом ВЭЖХ на обращенно-фазовых колонках Cis Nucleosil (Sigma, St. Louis, Mo.). Праймер DR-A1 содержал флуоресцентную метку Texas Red® (Molecular Probes, Inc., Eugene, Oreg.), которая вводилась в соответствии с рекомендациями производителя.Primers. Primer design was performed using Oligo 5 software (Molecular Biology Insights, Cascade, Colo.). Primers were synthesized on an ABI-394 DNA/RNA synthesizer (Applied Biosystems, Foster City, Calif.). For immobilization, primers were automatically modified by adding 5'-Amino-Modifier C6 (Glen Research, Sterling, Va.). The primers were purified by HPLC on Cis Nucleosil reverse phase columns (Sigma, St. Louis, Mo.). Primer DR-A1 contained the fluorescent tag Texas Red® (Molecular Probes, Inc., Eugene, Oreg.), which was introduced according to the manufacturer's recommendations.
DR-микрочип. Микрочипы изготавливались по методике сополимеризации, описанной ранее (Rubina, A.Y., Pan'kov, S.V., Dementieva, ЕЛ., Pen'kov, D.N., Butygin, A.V., Vasiliskov V.A., Chudinov, A.V., Mikheikin, A.L., Mikhailovich, V.M., Mirzabekov, A.D. (2004) Hydrogel drop microchips with immobilized DNA: properties and methods for large-scale production. Anal Biochem, 325, 92-106). Олигонуклеотиды вносились на стеклянную подложку вместе с остальными компонентами, формирующими ячейки. Сополимеризацию осуществляли методом фотополимеризации при λ = 260 нм в течение 30 мин. Диаметр полусферических ячеек равнялся 300 микрон при периоде 500 микрон. Размер активной области микрочипа составлял 4.8×2.3 мм.DR microchip. Microchips were manufactured using a copolymerization technique described previously (Rubina, A.Y., Pan'kov, S.V., Dementieva, E.L., Pen'kov, D.N., Butygin, A.V., Vasiliskov V.A., Chudinov, A.V., Mikheikin, A.L., Mikhailovich, V.M., Mirzabekov , A.D. (2004) Hydrogel drop microchips with immobilized DNA: properties and methods for large-scale production. Anal Biochem, 325, 92-106). Oligonucleotides were introduced onto a glass substrate along with the other components forming the cells. Copolymerization was carried out by photopolymerization at λ = 260 nm for 30 min. The diameter of the hemispherical cells was 300 microns with a period of 500 microns. The size of the active region of the microchip was 4.8×2.3 mm.
ПЦР на DR-микрочипе. В 30 мкл реакционной смеси содержались: ПЦР-буфер (10 мМ KCl, 10 мМ Tris-HCl, рН 8.3), 3 мМ MgCl2, 200 мкМ каждого dNTP, 4 Ед фрагмента Стоффеля AmpliTaq ДНК-полимеразы (Perkin Elmer, USA). В ПЦР-смесь вносили праймеры: 2 пмоль прямого DR-B1 (SEQ ID NO: 1) и 20 пмоль обратного флуоресцентно меченного DR-A1 (SEQ ID NO: 2). Температурный режим реакции был, как описано ниже: 95°С - 3 мин., 40 циклов амплификации (30 с при 95°С, 40 с при 60°С и 40 с при 72°С), 72°С - 10 мин. ПЦР проводили в камере In situ Frame (Eppendorf Sci. Inc.) на амплификаторе TGradient (Whatman Biometra, Germany). После снятия реакционной камеры чип промывался трижды по 2 мин 0.1М раствором NaCl при 72°С, затем быстро ополаскивался бидистиллированной водой для удаления солевых разводов и высушивался в потоке воздуха.PCR on DR microchip. 30 μl of the reaction mixture contained: PCR buffer (10 mM KCl, 10 mM Tris-HCl, pH 8.3), 3 mM MgCl2 , 200 μM each dNTP, 4 U of Stoffel fragment of AmpliTaq DNA polymerase (Perkin Elmer, USA). Primers were added to the PCR mixture: 2 pmol of forward DR-B1 (SEQ ID NO: 1) and 20 pmol of reverse fluorescently labeled DR-A1 (SEQ ID NO: 2). The reaction temperature was as described below: 95°C for 3 min, 40 amplification cycles (30 s at 95°C, 40 s at 60°C and 40 s at 72°C), 72°C for 10 min. PCR was carried out in an In situ Frame chamber (Eppendorf Sci. Inc.) using a TGradient thermal cycler (Whatman Biometra, Germany). After removing the reaction chamber, the chip was washed three times for 2 min each with a 0.1 M NaCl solution at 72°C, then quickly rinsed with bidistilled water to remove salt stains and dried in a stream of air.
Измерение флуоресцентного сигнала. Анализ ПЦР-профиля осуществляли на сухом микрочипе, используя портативный анализатор флуоресценции (чип-детектор). Изображение передавалось через ПЗС-камеру на компьютер для дальнейшей обработки. Измерение интенсивностей сигналов осуществляли программой "Analyze". Сохраняли как изображение ПЦР-профиля в формате pdf, так и файл с числовыми значениями сигналов.Fluorescence signal measurement. Analysis of the PCR profile was carried out on a dry microchip using a portable fluorescence analyzer (chip detector). The image was transferred through a CCD camera to a computer for further processing. Signal intensities were measured using the "Analyze" program. Both an image of the PCR profile in pdf format and a file with numerical signal values were saved.
Далее изобретение будет проиллюстрировано примерами, которые предназначены для обеспечения лучшего понимания сущности заявленного изобретения, но не должны рассматриваться как ограничивающие данное изобретение.The invention will now be illustrated by examples, which are intended to provide a better understanding of the claimed invention, but should not be construed as limiting the invention.
Пример 1. Специфичные к локусу прямых повторов (DR-локусу) олигонуклеотидные праймеры, используемые в способе.Example 1. Direct repeat locus (DR locus) specific oligonucleotide primers used in the method.
Праймеры для проведения полимеразной цепной реакции на биологическом микрочипе синтезируют на автоматическом синтезаторе 394 DNA/RNA synthesizer (Applied Biosystems, США). Последовательности праймеров DR-A1 (SEQ ID NO: 1), DR-A1 (SEQ ID NO: 2), а также список иммобилизованных праймеров (SEQ ID NO: 3-45) представлен в Перечне последовательностей.Primers for carrying out a polymerase chain reaction on a biological microchip are synthesized using an automatic synthesizer 394 DNA/RNA synthesizer (Applied Biosystems, USA). The sequences of primers DR-A1 (SEQ ID NO: 1), DR-A1 (SEQ ID NO: 2), as well as a list of immobilized primers (SEQ ID NO: 3-45) are presented in the Sequence Listing.
Пример 2. Проведение полимеразной цепной реакции для амплификации локуса прямых повторов (DR-локуса) с помощью полимеразной цепной реакции на биологическом микрочипе.Example 2. Carrying out polymerase chain reaction to amplify the direct repeat locus (DR locus) using polymerase chain reaction on a biological microchip.
К 29 мкл реакционной смеси вносят 1 мкл образца геномной ДНК анализируемого бактериального штамма.To 29 μl of the reaction mixture add 1 μl of a sample of genomic DNA of the bacterial strain being analyzed.
Мультиплексная ПЦР в иммобилизованной фазе.Multiplex PCR in the immobilized phase.
Для проведения ПЦР используют Taq-полимеразу ("Thermo Scientific") в буфере, соответствующем набору для Taq-полимеразы.To carry out PCR, Taq polymerase (Thermo Scientific) is used in a buffer corresponding to the Taq polymerase kit.
1. К аликвоте смеси, содержащей реакционный буфер и природные dNTP (по 200 мкМ каждого) добавляют праймеры (10 мкМ обратного DR-A1 (SEQ ID NO: 1) и 1.0 мкМ прямого DR-B1 (SEQ ID NO: 2)).1. Primers (10 µM reverse DR-A1 (SEQ ID NO: 1) and 1.0 µM forward DR-B1 (SEQ ID NO: 2)) are added to an aliquot of the mixture containing the reaction buffer and natural dNTPs (200 µM each).
2. Смесь помещают на биологический микрочип и герметизируют с помощью Frame-Seal 25 mkl ("Bio-Rad", США).2. The mixture is placed on a biological microchip and sealed using Frame-Seal 25 mkl (Bio-Rad, USA).
3. Проводят ПЦР на биологическом микрочипе с иммобилизованными 43 праймерами на ДНК-амплификаторе для in situ ПЦР TGradient Thermocycler ("Biometra", США) при следующих параметрах: 95°С в течение 3 мин (начальная денатурация); 36 циклов амплификации (20 с при 95°С, 30 с при 60°С и 40 с при 72°С) завершающая инкубация в течение 5 мин при 72°С.3. PCR is carried out on a biological microchip with immobilized 43 primers on a DNA amplifier for in situ PCR TGradient Thermocycler (Biometra, USA) with the following parameters: 95°C for 3 min (initial denaturation); 36 amplification cycles (20 s at 95°C, 30 s at 60°C and 40 s at 72°C) with a final incubation of 5 min at 72°C.
4. Осуществляют промывку биологического микрочипа дистиллированной водой в течение 1 минуты при комнатной температуре, после чего биологический микрочип высушивают в струе воздуха.4. The biological microchip is washed with distilled water for 1 minute at room temperature, after which the biological microchip is dried in a stream of air.
5. Производят анализ полученного на биочипе ПЦР-профиля и интерпретацию результатов анализа с помощью "Чип-детектора".5. The PCR profile obtained on the biochip is analyzed and the analysis results are interpreted using a “Chip Detector”.
Пример 3. Интерпретация результатов сполиготипирования (определения сполиготипа).Example 3. Interpretation of spoligotyping results (spoligotype determination).
Считывание флуоресцентного сигнала с биочипа после удлинения иммобилизованных праймеров осуществляют с применением анализатора "Чип-детектор" (ИМБ, Россия). Интенсивность полученных сигналов определяют с помощью программного обеспечения "Analyse" ver. 1.0 (ИМБ, Россия) или "ImaGeWare" ver. 3.50 (ИМБ, Россия).Reading of the fluorescent signal from the biochip after extension of the immobilized primers is carried out using a “Chip Detector” analyzer (IMB, Russia). The intensity of the received signals is determined using the software "Analyse" ver. 1.0 (IMB, Russia) or "ImaGeWare" ver. 3.50 (IMB, Russia).
Интерпретацию наличия в образце того или иного набора спейсеров в локусе прямых повторов осуществляют визуально либо полуавтоматически с помощью программного обеспечения "Analyse" ver. 1.0 (ИМБ, Россия) или "ImaGeWare" ver. 3.50 (ИМБ, Россия).Interpretation of the presence of a particular set of spacers in the direct repeat locus in a sample is carried out visually or semi-automatically using the "Analyse" software ver. 1.0 (IMB, Russia) or "ImaGeWare" ver. 3.50 (IMB, Russia).
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</INSDFeature> </INSDFeature>
<INSDFeature> <INSDFeature>
<INSDFeature_key>misc_feature</INSDFeature_key> <INSDFeature_key>misc_feature</INSDFeature_key>
<INSDFeature_location>1..32</INSDFeature_location> <INSDFeature_location>1..32</INSDFeature_location>
<INSDFeature_quals> <INSDFeature_quals>
<INSDQualifier id="q30"> <INSDQualifier id="q30">
<INSDQualifier_name>product</INSDQualifier_name> <INSDQualifier_name>product</INSDQualifier_name>
<INSDQualifier_value>immobilized primer</INSDQualifier_value> <INSDQualifier_value>immobilized primer</INSDQualifier_value>
<NonEnglishQualifier_value>иммобилизованный <NonEnglishQualifier_value>immobilized
праймер</NonEnglishQualifier_value>primer</NonEnglishQualifier_value>
</INSDQualifier> </INSDQualifier>
<INSDQualifier id="q29"> <INSDQualifier id="q29">
<INSDQualifier_name>standard_name</INSDQualifier_name> <INSDQualifier_name>standard_name</INSDQualifier_name>
<INSDQualifier_value>#4</INSDQualifier_value> <INSDQualifier_value>#4</INSDQualifier_value>
<NonEnglishQualifier_value>№4</NonEnglishQualifier_value> <NonEnglishQualifier_value>№4</NonEnglishQualifier_value>
</INSDQualifier> </INSDQualifier>
</INSDFeature_quals> </INSDFeature_quals>
</INSDFeature> </INSDFeature>
<INSDFeature> <INSDFeature>
<INSDFeature_key>misc_feature</INSDFeature_key> <INSDFeature_key>misc_feature</INSDFeature_key>
<INSDFeature_location><1</INSDFeature_location> <INSDFeature_location><1</INSDFeature_location>
<INSDFeature_quals> <INSDFeature_quals>
<INSDQualifier id="q31"> <INSDQualifier id="q31">
<INSDQualifier_name>function</INSDQualifier_name> <INSDQualifier_name>function</INSDQualifier_name>
<INSDQualifier_value>NH2-group</INSDQualifier_value> <INSDQualifier_value>NH2-group</INSDQualifier_value>
<NonEnglishQualifier_value>аминогруппа</NonEnglishQualifier_val <NonEnglishQualifier_value>amino group</NonEnglishQualifier_val
ue>ue>
</INSDQualifier> </INSDQualifier>
</INSDFeature_quals> </INSDFeature_quals>
</INSDFeature> </INSDFeature>
</INSDSeq_feature-table> </INSDSeq_feature-table>
<INSDSeq_sequence>ctcagtagcacgtcatacgccgaccaatcatc</INSDSeq_sequenc <INSDSeq_sequence>ctcagtagcacgtcatacgccgaccaatcatc</INSDSeq_sequenc
e>e>
</INSDSeq> </INSDSeq>
</SequenceData> </SequenceData>
<SequenceData sequenceIDNumber="7"> <SequenceData sequenceIDNumber="7">
<INSDSeq> <INSDSeq>
<INSDSeq_length>30</INSDSeq_length> <INSDSeq_length>30</INSDSeq_length>
<INSDSeq_moltype>DNA</INSDSeq_moltype> <INSDSeq_moltype>DNA</INSDSeq_moltype>
<INSDSeq_division>PAT</INSDSeq_division> <INSDSeq_division>PAT</INSDSeq_division>
<INSDSeq_feature-table> <INSDSeq_feature-table>
<INSDFeature> <INSDFeature>
<INSDFeature_key>source</INSDFeature_key> <INSDFeature_key>source</INSDFeature_key>
<INSDFeature_location>1..30</INSDFeature_location> <INSDFeature_location>1..30</INSDFeature_location>
<INSDFeature_quals> <INSDFeature_quals>
<INSDQualifier> <INSDQualifier>
<INSDQualifier_name>mol_type</INSDQualifier_name> <INSDQualifier_name>mol_type</INSDQualifier_name>
<INSDQualifier_value>other DNA</INSDQualifier_value> <INSDQualifier_value>other DNA</INSDQualifier_value>
</INSDQualifier> </INSDQualifier>
<INSDQualifier id="q33"> <INSDQualifier id="q33">
<INSDQualifier_name>organism</INSDQualifier_name> <INSDQualifier_name>organism</INSDQualifier_name>
<INSDQualifier_value>synthetic construct</INSDQualifier_value> <INSDQualifier_value>synthetic construct</INSDQualifier_value>
</INSDQualifier> </INSDQualifier>
</INSDFeature_quals> </INSDFeature_quals>
</INSDFeature> </INSDFeature>
<INSDFeature> <INSDFeature>
<INSDFeature_key>misc_feature</INSDFeature_key> <INSDFeature_key>misc_feature</INSDFeature_key>
<INSDFeature_location>1..30</INSDFeature_location> <INSDFeature_location>1..30</INSDFeature_location>
<INSDFeature_quals> <INSDFeature_quals>
<INSDQualifier id="q35"> <INSDQualifier id="q35">
<INSDQualifier_name>product</INSDQualifier_name> <INSDQualifier_name>product</INSDQualifier_name>
<INSDQualifier_value>immobilized primer</INSDQualifier_value> <INSDQualifier_value>immobilized primer</INSDQualifier_value>
<NonEnglishQualifier_value>иммобилизованный <NonEnglishQualifier_value>immobilized
праймер</NonEnglishQualifier_value>primer</NonEnglishQualifier_value>
</INSDQualifier> </INSDQualifier>
<INSDQualifier id="q34"> <INSDQualifier id="q34">
<INSDQualifier_name>standard_name</INSDQualifier_name> <INSDQualifier_name>standard_name</INSDQualifier_name>
<INSDQualifier_value>#5</INSDQualifier_value> <INSDQualifier_value>#5</INSDQualifier_value>
<NonEnglishQualifier_value>№5</NonEnglishQualifier_value> <NonEnglishQualifier_value>№5</NonEnglishQualifier_value>
</INSDQualifier> </INSDQualifier>
</INSDFeature_quals> </INSDFeature_quals>
</INSDFeature> </INSDFeature>
<INSDFeature> <INSDFeature>
<INSDFeature_key>misc_feature</INSDFeature_key> <INSDFeature_key>misc_feature</INSDFeature_key>
<INSDFeature_location><1</INSDFeature_location> <INSDFeature_location><1</INSDFeature_location>
<INSDFeature_quals> <INSDFeature_quals>
<INSDQualifier id="q36"> <INSDQualifier id="q36">
<INSDQualifier_name>function</INSDQualifier_name> <INSDQualifier_name>function</INSDQualifier_name>
<INSDQualifier_value>NH2-group</INSDQualifier_value> <INSDQualifier_value>NH2-group</INSDQualifier_value>
<NonEnglishQualifier_value>аминогруппа</NonEnglishQualifier_val <NonEnglishQualifier_value>amino group</NonEnglishQualifier_val
ue>ue>
</INSDQualifier> </INSDQualifier>
</INSDFeature_quals> </INSDFeature_quals>
</INSDFeature> </INSDFeature>
</INSDSeq_feature-table> </INSDSeq_feature-table>
<INSDSeq_sequence>ttttctgaccacttgtgcgggattagcggg</INSDSeq_sequence> <INSDSeq_sequence>ttttctgaccacttgtgcgggattagcggg</INSDSeq_sequence>
</INSDSeq> </INSDSeq>
</SequenceData> </SequenceData>
<SequenceData sequenceIDNumber="8"> <SequenceData sequenceIDNumber="8">
<INSDSeq> <INSDSeq>
<INSDSeq_length>30</INSDSeq_length> <INSDSeq_length>30</INSDSeq_length>
<INSDSeq_moltype>DNA</INSDSeq_moltype> <INSDSeq_moltype>DNA</INSDSeq_moltype>
<INSDSeq_division>PAT</INSDSeq_division> <INSDSeq_division>PAT</INSDSeq_division>
<INSDSeq_feature-table> <INSDSeq_feature-table>
<INSDFeature> <INSDFeature>
<INSDFeature_key>source</INSDFeature_key> <INSDFeature_key>source</INSDFeature_key>
<INSDFeature_location>1..30</INSDFeature_location> <INSDFeature_location>1..30</INSDFeature_location>
<INSDFeature_quals> <INSDFeature_quals>
<INSDQualifier> <INSDQualifier>
<INSDQualifier_name>mol_type</INSDQualifier_name> <INSDQualifier_name>mol_type</INSDQualifier_name>
<INSDQualifier_value>other DNA</INSDQualifier_value> <INSDQualifier_value>other DNA</INSDQualifier_value>
</INSDQualifier> </INSDQualifier>
<INSDQualifier id="q38"> <INSDQualifier id="q38">
<INSDQualifier_name>organism</INSDQualifier_name> <INSDQualifier_name>organism</INSDQualifier_name>
<INSDQualifier_value>synthetic construct</INSDQualifier_value> <INSDQualifier_value>synthetic construct</INSDQualifier_value>
</INSDQualifier> </INSDQualifier>
</INSDFeature_quals> </INSDFeature_quals>
</INSDFeature> </INSDFeature>
<INSDFeature> <INSDFeature>
<INSDFeature_key>misc_feature</INSDFeature_key> <INSDFeature_key>misc_feature</INSDFeature_key>
<INSDFeature_location>1..30</INSDFeature_location> <INSDFeature_location>1..30</INSDFeature_location>
<INSDFeature_quals> <INSDFeature_quals>
<INSDQualifier id="q40"> <INSDQualifier id="q40">
<INSDQualifier_name>product</INSDQualifier_name> <INSDQualifier_name>product</INSDQualifier_name>
<INSDQualifier_value>immobilized primer</INSDQualifier_value> <INSDQualifier_value>immobilized primer</INSDQualifier_value>
<NonEnglishQualifier_value>иммобилизованный <NonEnglishQualifier_value>immobilized
праймер</NonEnglishQualifier_value>primer</NonEnglishQualifier_value>
</INSDQualifier> </INSDQualifier>
<INSDQualifier id="q39"> <INSDQualifier id="q39">
<INSDQualifier_name>standard_name</INSDQualifier_name> <INSDQualifier_name>standard_name</INSDQualifier_name>
<INSDQualifier_value>#6</INSDQualifier_value> <INSDQualifier_value>#6</INSDQualifier_value>
<NonEnglishQualifier_value>№6</NonEnglishQualifier_value> <NonEnglishQualifier_value>№6</NonEnglishQualifier_value>
</INSDQualifier> </INSDQualifier>
</INSDFeature_quals> </INSDFeature_quals>
</INSDFeature> </INSDFeature>
<INSDFeature> <INSDFeature>
<INSDFeature_key>misc_feature</INSDFeature_key> <INSDFeature_key>misc_feature</INSDFeature_key>
<INSDFeature_location><1</INSDFeature_location> <INSDFeature_location><1</INSDFeature_location>
<INSDFeature_quals> <INSDFeature_quals>
<INSDQualifier id="q41"> <INSDQualifier id="q41">
<INSDQualifier_name>function</INSDQualifier_name> <INSDQualifier_name>function</INSDQualifier_name>
<INSDQualifier_value>NH2-group</INSDQualifier_value> <INSDQualifier_value>NH2-group</INSDQualifier_value>
<NonEnglishQualifier_value>аминогруппа</NonEnglishQualifier_val <NonEnglishQualifier_value>amino group</NonEnglishQualifier_val
ue>ue>
</INSDQualifier> </INSDQualifier>
</INSDFeature_quals> </INSDFeature_quals>
</INSDFeature> </INSDFeature>
</INSDSeq_feature-table> </INSDSeq_feature-table>
<INSDSeq_sequence>ccaatgcgtcgtcatttccggcttcaattt</INSDSeq_sequence> <INSDSeq_sequence>ccaatgcgtcgtcatttccggcttcaattt</INSDSeq_sequence>
</INSDSeq> </INSDSeq>
</SequenceData> </SequenceData>
<SequenceData sequenceIDNumber="9"> <SequenceData sequenceIDNumber="9">
<INSDSeq> <INSDSeq>
<INSDSeq_length>27</INSDSeq_length> <INSDSeq_length>27</INSDSeq_length>
<INSDSeq_moltype>DNA</INSDSeq_moltype> <INSDSeq_moltype>DNA</INSDSeq_moltype>
<INSDSeq_division>PAT</INSDSeq_division> <INSDSeq_division>PAT</INSDSeq_division>
<INSDSeq_feature-table> <INSDSeq_feature-table>
<INSDFeature> <INSDFeature>
<INSDFeature_key>source</INSDFeature_key> <INSDFeature_key>source</INSDFeature_key>
<INSDFeature_location>1..27</INSDFeature_location> <INSDFeature_location>1..27</INSDFeature_location>
<INSDFeature_quals> <INSDFeature_quals>
<INSDQualifier> <INSDQualifier>
<INSDQualifier_name>mol_type</INSDQualifier_name> <INSDQualifier_name>mol_type</INSDQualifier_name>
<INSDQualifier_value>other DNA</INSDQualifier_value> <INSDQualifier_value>other DNA</INSDQualifier_value>
</INSDQualifier> </INSDQualifier>
<INSDQualifier id="q43"> <INSDQualifier id="q43">
<INSDQualifier_name>organism</INSDQualifier_name> <INSDQualifier_name>organism</INSDQualifier_name>
<INSDQualifier_value>synthetic construct</INSDQualifier_value> <INSDQualifier_value>synthetic construct</INSDQualifier_value>
</INSDQualifier> </INSDQualifier>
</INSDFeature_quals> </INSDFeature_quals>
</INSDFeature> </INSDFeature>
<INSDFeature> <INSDFeature>
<INSDFeature_key>misc_feature</INSDFeature_key> <INSDFeature_key>misc_feature</INSDFeature_key>
<INSDFeature_location>1..27</INSDFeature_location> <INSDFeature_location>1..27</INSDFeature_location>
<INSDFeature_quals> <INSDFeature_quals>
<INSDQualifier id="q45"> <INSDQualifier id="q45">
<INSDQualifier_name>product</INSDQualifier_name> <INSDQualifier_name>product</INSDQualifier_name>
<INSDQualifier_value>immobilized primer</INSDQualifier_value> <INSDQualifier_value>immobilized primer</INSDQualifier_value>
<NonEnglishQualifier_value>иммобилизованный <NonEnglishQualifier_value>immobilized
праймер</NonEnglishQualifier_value>primer</NonEnglishQualifier_value>
</INSDQualifier> </INSDQualifier>
<INSDQualifier id="q44"> <INSDQualifier id="q44">
<INSDQualifier_name>standard_name</INSDQualifier_name> <INSDQualifier_name>standard_name</INSDQualifier_name>
<INSDQualifier_value>#7</INSDQualifier_value> <INSDQualifier_value>#7</INSDQualifier_value>
<NonEnglishQualifier_value>#7</NonEnglishQualifier_value> <NonEnglishQualifier_value>#7</NonEnglishQualifier_value>
</INSDQualifier> </INSDQualifier>
</INSDFeature_quals> </INSDFeature_quals>
</INSDFeature> </INSDFeature>
<INSDFeature> <INSDFeature>
<INSDFeature_key>misc_feature</INSDFeature_key> <INSDFeature_key>misc_feature</INSDFeature_key>
<INSDFeature_location><1</INSDFeature_location> <INSDFeature_location><1</INSDFeature_location>
<INSDFeature_quals> <INSDFeature_quals>
<INSDQualifier id="q46"> <INSDQualifier id="q46">
<INSDQualifier_name>function</INSDQualifier_name> <INSDQualifier_name>function</INSDQualifier_name>
<INSDQualifier_value>NH2-group</INSDQualifier_value> <INSDQualifier_value>NH2-group</INSDQualifier_value>
<NonEnglishQualifier_value>аминогруппа</NonEnglishQualifier_val <NonEnglishQualifier_value>amino group</NonEnglishQualifier_val
ue>ue>
</INSDQualifier> </INSDQualifier>
</INSDFeature_quals> </INSDFeature_quals>
</INSDFeature> </INSDFeature>
</INSDSeq_feature-table> </INSDSeq_feature-table>
<INSDSeq_sequence>ctgaggagagcgagtactcggggctgc</INSDSeq_sequence> <INSDSeq_sequence>ctgaggagagcgagtactcggggctgc</INSDSeq_sequence>
</INSDSeq> </INSDSeq>
</SequenceData> </SequenceData>
<SequenceData sequenceIDNumber="10"> <SequenceData sequenceIDNumber="10">
<INSDSeq> <INSDSeq>
<INSDSeq_length>21</INSDSeq_length> <INSDSeq_length>21</INSDSeq_length>
<INSDSeq_moltype>DNA</INSDSeq_moltype> <INSDSeq_moltype>DNA</INSDSeq_moltype>
<INSDSeq_division>PAT</INSDSeq_division> <INSDSeq_division>PAT</INSDSeq_division>
<INSDSeq_feature-table> <INSDSeq_feature-table>
<INSDFeature> <INSDFeature>
<INSDFeature_key>source</INSDFeature_key> <INSDFeature_key>source</INSDFeature_key>
<INSDFeature_location>1..21</INSDFeature_location> <INSDFeature_location>1..21</INSDFeature_location>
<INSDFeature_quals> <INSDFeature_quals>
<INSDQualifier> <INSDQualifier>
<INSDQualifier_name>mol_type</INSDQualifier_name> <INSDQualifier_name>mol_type</INSDQualifier_name>
<INSDQualifier_value>other DNA</INSDQualifier_value> <INSDQualifier_value>other DNA</INSDQualifier_value>
</INSDQualifier> </INSDQualifier>
<INSDQualifier id="q48"> <INSDQualifier id="q48">
<INSDQualifier_name>organism</INSDQualifier_name> <INSDQualifier_name>organism</INSDQualifier_name>
<INSDQualifier_value>synthetic construct</INSDQualifier_value> <INSDQualifier_value>synthetic construct</INSDQualifier_value>
</INSDQualifier> </INSDQualifier>
</INSDFeature_quals> </INSDFeature_quals>
</INSDFeature> </INSDFeature>
<INSDFeature> <INSDFeature>
<INSDFeature_key>misc_feature</INSDFeature_key> <INSDFeature_key>misc_feature</INSDFeature_key>
<INSDFeature_location>1..21</INSDFeature_location> <INSDFeature_location>1..21</INSDFeature_location>
<INSDFeature_quals> <INSDFeature_quals>
<INSDQualifier id="q50"> <INSDQualifier id="q50">
<INSDQualifier_name>product</INSDQualifier_name> <INSDQualifier_name>product</INSDQualifier_name>
<INSDQualifier_value>immobilized primer</INSDQualifier_value> <INSDQualifier_value>immobilized primer</INSDQualifier_value>
<NonEnglishQualifier_value>иммобилизованный <NonEnglishQualifier_value>immobilized
праймер</NonEnglishQualifier_value>primer</NonEnglishQualifier_value>
</INSDQualifier> </INSDQualifier>
<INSDQualifier id="q49"> <INSDQualifier id="q49">
<INSDQualifier_name>standard_name</INSDQualifier_name> <INSDQualifier_name>standard_name</INSDQualifier_name>
<INSDQualifier_value>#8</INSDQualifier_value> <INSDQualifier_value>#8</INSDQualifier_value>
<NonEnglishQualifier_value>№8</NonEnglishQualifier_value> <NonEnglishQualifier_value>№8</NonEnglishQualifier_value>
</INSDQualifier> </INSDQualifier>
</INSDFeature_quals> </INSDFeature_quals>
</INSDFeature> </INSDFeature>
<INSDFeature> <INSDFeature>
<INSDFeature_key>misc_feature</INSDFeature_key> <INSDFeature_key>misc_feature</INSDFeature_key>
<INSDFeature_location><1</INSDFeature_location> <INSDFeature_location><1</INSDFeature_location>
<INSDFeature_quals> <INSDFeature_quals>
<INSDQualifier id="q51"> <INSDQualifier id="q51">
<INSDQualifier_name>function</INSDQualifier_name> <INSDQualifier_name>function</INSDQualifier_name>
<INSDQualifier_value>NH2-group</INSDQualifier_value> <INSDQualifier_value>NH2-group</INSDQualifier_value>
<NonEnglishQualifier_value>аминогруппа</NonEnglishQualifier_val <NonEnglishQualifier_value>amino group</NonEnglishQualifier_val
ue>ue>
</INSDQualifier> </INSDQualifier>
</INSDFeature_quals> </INSDFeature_quals>
</INSDFeature> </INSDFeature>
</INSDSeq_feature-table> </INSDSeq_feature-table>
<INSDSeq_sequence>gaaaccgcccccagcctcgcc</INSDSeq_sequence> <INSDSeq_sequence>gaaaccgcccccagcctcgcc</INSDSeq_sequence>
</INSDSeq> </INSDSeq>
</SequenceData> </SequenceData>
<SequenceData sequenceIDNumber="11"> <SequenceData sequenceIDNumber="11">
<INSDSeq> <INSDSeq>
<INSDSeq_length>27</INSDSeq_length> <INSDSeq_length>27</INSDSeq_length>
<INSDSeq_moltype>DNA</INSDSeq_moltype> <INSDSeq_moltype>DNA</INSDSeq_moltype>
<INSDSeq_division>PAT</INSDSeq_division> <INSDSeq_division>PAT</INSDSeq_division>
<INSDSeq_feature-table> <INSDSeq_feature-table>
<INSDFeature> <INSDFeature>
<INSDFeature_key>source</INSDFeature_key> <INSDFeature_key>source</INSDFeature_key>
<INSDFeature_location>1..27</INSDFeature_location> <INSDFeature_location>1..27</INSDFeature_location>
<INSDFeature_quals> <INSDFeature_quals>
<INSDQualifier> <INSDQualifier>
<INSDQualifier_name>mol_type</INSDQualifier_name> <INSDQualifier_name>mol_type</INSDQualifier_name>
<INSDQualifier_value>other DNA</INSDQualifier_value> <INSDQualifier_value>other DNA</INSDQualifier_value>
</INSDQualifier> </INSDQualifier>
<INSDQualifier id="q53"> <INSDQualifier id="q53">
<INSDQualifier_name>organism</INSDQualifier_name> <INSDQualifier_name>organism</INSDQualifier_name>
<INSDQualifier_value>synthetic construct</INSDQualifier_value> <INSDQualifier_value>synthetic construct</INSDQualifier_value>
</INSDQualifier> </INSDQualifier>
</INSDFeature_quals> </INSDFeature_quals>
</INSDFeature> </INSDFeature>
<INSDFeature> <INSDFeature>
<INSDFeature_key>misc_feature</INSDFeature_key> <INSDFeature_key>misc_feature</INSDFeature_key>
<INSDFeature_location>1..27</INSDFeature_location> <INSDFeature_location>1..27</INSDFeature_location>
<INSDFeature_quals> <INSDFeature_quals>
<INSDQualifier id="q55"> <INSDQualifier id="q55">
<INSDQualifier_name>product</INSDQualifier_name> <INSDQualifier_name>product</INSDQualifier_name>
<INSDQualifier_value>immobilized primer</INSDQualifier_value> <INSDQualifier_value>immobilized primer</INSDQualifier_value>
<NonEnglishQualifier_value>иммобилизованный <NonEnglishQualifier_value>immobilized
праймер</NonEnglishQualifier_value>primer</NonEnglishQualifier_value>
</INSDQualifier> </INSDQualifier>
<INSDQualifier id="q54"> <INSDQualifier id="q54">
<INSDQualifier_name>standard_name</INSDQualifier_name> <INSDQualifier_name>standard_name</INSDQualifier_name>
<INSDQualifier_value>#9</INSDQualifier_value> <INSDQualifier_value>#9</INSDQualifier_value>
<NonEnglishQualifier_value>№9</NonEnglishQualifier_value> <NonEnglishQualifier_value>№9</NonEnglishQualifier_value>
</INSDQualifier> </INSDQualifier>
</INSDFeature_quals> </INSDFeature_quals>
</INSDFeature> </INSDFeature>
<INSDFeature> <INSDFeature>
<INSDFeature_key>misc_feature</INSDFeature_key> <INSDFeature_key>misc_feature</INSDFeature_key>
<INSDFeature_location><1</INSDFeature_location> <INSDFeature_location><1</INSDFeature_location>
<INSDFeature_quals> <INSDFeature_quals>
<INSDQualifier id="q56"> <INSDQualifier id="q56">
<INSDQualifier_name>function</INSDQualifier_name> <INSDQualifier_name>function</INSDQualifier_name>
<INSDQualifier_value>NH2-group</INSDQualifier_value> <INSDQualifier_value>NH2-group</INSDQualifier_value>
<NonEnglishQualifier_value>NH2-group</NonEnglishQualifier_value <NonEnglishQualifier_value>NH2-group</NonEnglishQualifier_value
>>
</INSDQualifier> </INSDQualifier>
</INSDFeature_quals> </INSDFeature_quals>
</INSDFeature> </INSDFeature>
</INSDSeq_feature-table> </INSDSeq_feature-table>
<INSDSeq_sequence>tcccatgtgctgacagcggattcgcat</INSDSeq_sequence> <INSDSeq_sequence>tcccatgtgctgacagcggattcgcat</INSDSeq_sequence>
</INSDSeq> </INSDSeq>
</SequenceData> </SequenceData>
<SequenceData sequenceIDNumber="12"> <SequenceData sequenceIDNumber="12">
<INSDSeq> <INSDSeq>
<INSDSeq_length>27</INSDSeq_length> <INSDSeq_length>27</INSDSeq_length>
<INSDSeq_moltype>DNA</INSDSeq_moltype> <INSDSeq_moltype>DNA</INSDSeq_moltype>
<INSDSeq_division>PAT</INSDSeq_division> <INSDSeq_division>PAT</INSDSeq_division>
<INSDSeq_feature-table> <INSDSeq_feature-table>
<INSDFeature> <INSDFeature>
<INSDFeature_key>source</INSDFeature_key> <INSDFeature_key>source</INSDFeature_key>
<INSDFeature_location>1..27</INSDFeature_location> <INSDFeature_location>1..27</INSDFeature_location>
<INSDFeature_quals> <INSDFeature_quals>
<INSDQualifier> <INSDQualifier>
<INSDQualifier_name>mol_type</INSDQualifier_name> <INSDQualifier_name>mol_type</INSDQualifier_name>
<INSDQualifier_value>other DNA</INSDQualifier_value> <INSDQualifier_value>other DNA</INSDQualifier_value>
</INSDQualifier> </INSDQualifier>
<INSDQualifier id="q58"> <INSDQualifier id="q58">
<INSDQualifier_name>organism</INSDQualifier_name> <INSDQualifier_name>organism</INSDQualifier_name>
<INSDQualifier_value>synthetic construct</INSDQualifier_value> <INSDQualifier_value>synthetic construct</INSDQualifier_value>
</INSDQualifier> </INSDQualifier>
</INSDFeature_quals> </INSDFeature_quals>
</INSDFeature> </INSDFeature>
<INSDFeature> <INSDFeature>
<INSDFeature_key>misc_feature</INSDFeature_key> <INSDFeature_key>misc_feature</INSDFeature_key>
<INSDFeature_location>1..27</INSDFeature_location> <INSDFeature_location>1..27</INSDFeature_location>
<INSDFeature_quals> <INSDFeature_quals>
<INSDQualifier id="q60"> <INSDQualifier id="q60">
<INSDQualifier_name>product</INSDQualifier_name> <INSDQualifier_name>product</INSDQualifier_name>
<INSDQualifier_value>immobilized primer</INSDQualifier_value> <INSDQualifier_value>immobilized primer</INSDQualifier_value>
<NonEnglishQualifier_value>иммобилизованный <NonEnglishQualifier_value>immobilized
праймер</NonEnglishQualifier_value>primer</NonEnglishQualifier_value>
</INSDQualifier> </INSDQualifier>
<INSDQualifier id="q59"> <INSDQualifier id="q59">
<INSDQualifier_name>standard_name</INSDQualifier_name> <INSDQualifier_name>standard_name</INSDQualifier_name>
<INSDQualifier_value>#10</INSDQualifier_value> <INSDQualifier_value>#10</INSDQualifier_value>
<NonEnglishQualifier_value>№10</NonEnglishQualifier_value> <NonEnglishQualifier_value>№10</NonEnglishQualifier_value>
</INSDQualifier> </INSDQualifier>
</INSDFeature_quals> </INSDFeature_quals>
</INSDFeature> </INSDFeature>
<INSDFeature> <INSDFeature>
<INSDFeature_key>misc_feature</INSDFeature_key> <INSDFeature_key>misc_feature</INSDFeature_key>
<INSDFeature_location><1</INSDFeature_location> <INSDFeature_location><1</INSDFeature_location>
<INSDFeature_quals> <INSDFeature_quals>
<INSDQualifier id="q61"> <INSDQualifier id="q61">
<INSDQualifier_name>function</INSDQualifier_name> <INSDQualifier_name>function</INSDQualifier_name>
<INSDQualifier_value>NH2-group</INSDQualifier_value> <INSDQualifier_value>NH2-group</INSDQualifier_value>
<NonEnglishQualifier_value>аминогруппа</NonEnglishQualifier_val <NonEnglishQualifier_value>amino group</NonEnglishQualifier_val
ue>ue>
</INSDQualifier> </INSDQualifier>
</INSDFeature_quals> </INSDFeature_quals>
</INSDFeature> </INSDFeature>
</INSDSeq_feature-table> </INSDSeq_feature-table>
<INSDSeq_sequence>gcgaaatacaggctccacgagaccaca</INSDSeq_sequence> <INSDSeq_sequence>gcgaaatacaggctccacgagaccaca</INSDSeq_sequence>
</INSDSeq> </INSDSeq>
</SequenceData> </SequenceData>
<SequenceData sequenceIDNumber="13"> <SequenceData sequenceIDNumber="13">
<INSDSeq> <INSDSeq>
<INSDSeq_length>22</INSDSeq_length> <INSDSeq_length>22</INSDSeq_length>
<INSDSeq_moltype>DNA</INSDSeq_moltype> <INSDSeq_moltype>DNA</INSDSeq_moltype>
<INSDSeq_division>PAT</INSDSeq_division> <INSDSeq_division>PAT</INSDSeq_division>
<INSDSeq_feature-table> <INSDSeq_feature-table>
<INSDFeature> <INSDFeature>
<INSDFeature_key>source</INSDFeature_key> <INSDFeature_key>source</INSDFeature_key>
<INSDFeature_location>1..22</INSDFeature_location> <INSDFeature_location>1..22</INSDFeature_location>
<INSDFeature_quals> <INSDFeature_quals>
<INSDQualifier> <INSDQualifier>
<INSDQualifier_name>mol_type</INSDQualifier_name> <INSDQualifier_name>mol_type</INSDQualifier_name>
<INSDQualifier_value>other DNA</INSDQualifier_value> <INSDQualifier_value>other DNA</INSDQualifier_value>
</INSDQualifier> </INSDQualifier>
<INSDQualifier id="q63"> <INSDQualifier id="q63">
<INSDQualifier_name>organism</INSDQualifier_name> <INSDQualifier_name>organism</INSDQualifier_name>
<INSDQualifier_value>synthetic construct</INSDQualifier_value> <INSDQualifier_value>synthetic construct</INSDQualifier_value>
</INSDQualifier> </INSDQualifier>
</INSDFeature_quals> </INSDFeature_quals>
</INSDFeature> </INSDFeature>
<INSDFeature> <INSDFeature>
<INSDFeature_key>misc_feature</INSDFeature_key> <INSDFeature_key>misc_feature</INSDFeature_key>
<INSDFeature_location>1..22</INSDFeature_location> <INSDFeature_location>1..22</INSDFeature_location>
<INSDFeature_quals> <INSDFeature_quals>
<INSDQualifier id="q65"> <INSDQualifier id="q65">
<INSDQualifier_name>product</INSDQualifier_name> <INSDQualifier_name>product</INSDQualifier_name>
<INSDQualifier_value>immobilized primer</INSDQualifier_value> <INSDQualifier_value>immobilized primer</INSDQualifier_value>
<NonEnglishQualifier_value>иммобилизованный <NonEnglishQualifier_value>immobilized
праймер</NonEnglishQualifier_value>primer</NonEnglishQualifier_value>
</INSDQualifier> </INSDQualifier>
<INSDQualifier id="q64"> <INSDQualifier id="q64">
<INSDQualifier_name>standard_name</INSDQualifier_name> <INSDQualifier_name>standard_name</INSDQualifier_name>
<INSDQualifier_value>#11</INSDQualifier_value> <INSDQualifier_value>#11</INSDQualifier_value>
<NonEnglishQualifier_value>№11</NonEnglishQualifier_value> <NonEnglishQualifier_value>№11</NonEnglishQualifier_value>
</INSDQualifier> </INSDQualifier>
</INSDFeature_quals> </INSDFeature_quals>
</INSDFeature> </INSDFeature>
<INSDFeature> <INSDFeature>
<INSDFeature_key>misc_feature</INSDFeature_key> <INSDFeature_key>misc_feature</INSDFeature_key>
<INSDFeature_location><1</INSDFeature_location> <INSDFeature_location><1</INSDFeature_location>
<INSDFeature_quals> <INSDFeature_quals>
<INSDQualifier id="q66"> <INSDQualifier id="q66">
<INSDQualifier_name>function</INSDQualifier_name> <INSDQualifier_name>function</INSDQualifier_name>
<INSDQualifier_value>NH2-group</INSDQualifier_value> <INSDQualifier_value>NH2-group</INSDQualifier_value>
<NonEnglishQualifier_value>аминогруппа</NonEnglishQualifier_val <NonEnglishQualifier_value>amino group</NonEnglishQualifier_val
ue>ue>
</INSDQualifier> </INSDQualifier>
</INSDFeature_quals> </INSDFeature_quals>
</INSDFeature> </INSDFeature>
</INSDSeq_feature-table> </INSDSeq_feature-table>
<INSDSeq_sequence>gagcaacggcggcggcaacctg</INSDSeq_sequence> <INSDSeq_sequence>gagcaacggcggcggcaacctg</INSDSeq_sequence>
</INSDSeq> </INSDSeq>
</SequenceData> </SequenceData>
<SequenceData sequenceIDNumber="14"> <SequenceData sequenceIDNumber="14">
<INSDSeq> <INSDSeq>
<INSDSeq_length>22</INSDSeq_length> <INSDSeq_length>22</INSDSeq_length>
<INSDSeq_moltype>DNA</INSDSeq_moltype> <INSDSeq_moltype>DNA</INSDSeq_moltype>
<INSDSeq_division>PAT</INSDSeq_division> <INSDSeq_division>PAT</INSDSeq_division>
<INSDSeq_feature-table> <INSDSeq_feature-table>
<INSDFeature> <INSDFeature>
<INSDFeature_key>source</INSDFeature_key> <INSDFeature_key>source</INSDFeature_key>
<INSDFeature_location>1..22</INSDFeature_location> <INSDFeature_location>1..22</INSDFeature_location>
<INSDFeature_quals> <INSDFeature_quals>
<INSDQualifier> <INSDQualifier>
<INSDQualifier_name>mol_type</INSDQualifier_name> <INSDQualifier_name>mol_type</INSDQualifier_name>
<INSDQualifier_value>other DNA</INSDQualifier_value> <INSDQualifier_value>other DNA</INSDQualifier_value>
</INSDQualifier> </INSDQualifier>
<INSDQualifier id="q68"> <INSDQualifier id="q68">
<INSDQualifier_name>organism</INSDQualifier_name> <INSDQualifier_name>organism</INSDQualifier_name>
<INSDQualifier_value>synthetic construct</INSDQualifier_value> <INSDQualifier_value>synthetic construct</INSDQualifier_value>
</INSDQualifier> </INSDQualifier>
</INSDFeature_quals> </INSDFeature_quals>
</INSDFeature> </INSDFeature>
<INSDFeature> <INSDFeature>
<INSDFeature_key>misc_feature</INSDFeature_key> <INSDFeature_key>misc_feature</INSDFeature_key>
<INSDFeature_location>1..22</INSDFeature_location> <INSDFeature_location>1..22</INSDFeature_location>
<INSDFeature_quals> <INSDFeature_quals>
<INSDQualifier id="q70"> <INSDQualifier id="q70">
<INSDQualifier_name>product</INSDQualifier_name> <INSDQualifier_name>product</INSDQualifier_name>
<INSDQualifier_value>immobilized primer</INSDQualifier_value> <INSDQualifier_value>immobilized primer</INSDQualifier_value>
<NonEnglishQualifier_value>иммобилизованный <NonEnglishQualifier_value>immobilized
праймер</NonEnglishQualifier_value>primer</NonEnglishQualifier_value>
</INSDQualifier> </INSDQualifier>
<INSDQualifier id="q69"> <INSDQualifier id="q69">
<INSDQualifier_name>standard_name</INSDQualifier_name> <INSDQualifier_name>standard_name</INSDQualifier_name>
<INSDQualifier_value>#12</INSDQualifier_value> <INSDQualifier_value>#12</INSDQualifier_value>
<NonEnglishQualifier_value>№12</NonEnglishQualifier_value> <NonEnglishQualifier_value>№12</NonEnglishQualifier_value>
</INSDQualifier> </INSDQualifier>
</INSDFeature_quals> </INSDFeature_quals>
</INSDFeature> </INSDFeature>
<INSDFeature> <INSDFeature>
<INSDFeature_key>misc_feature</INSDFeature_key> <INSDFeature_key>misc_feature</INSDFeature_key>
<INSDFeature_location><1</INSDFeature_location> <INSDFeature_location><1</INSDFeature_location>
<INSDFeature_quals> <INSDFeature_quals>
<INSDQualifier id="q71"> <INSDQualifier id="q71">
<INSDQualifier_name>function</INSDQualifier_name> <INSDQualifier_name>function</INSDQualifier_name>
<INSDQualifier_value>NH2-group</INSDQualifier_value> <INSDQualifier_value>NH2-group</INSDQualifier_value>
<NonEnglishQualifier_value>аминогруппа</NonEnglishQualifier_val <NonEnglishQualifier_value>amino group</NonEnglishQualifier_val
ue>ue>
</INSDQualifier> </INSDQualifier>
</INSDFeature_quals> </INSDFeature_quals>
</INSDFeature> </INSDFeature>
</INSDSeq_feature-table> </INSDSeq_feature-table>
<INSDSeq_sequence>cgcccggggagatgctgtccga</INSDSeq_sequence> <INSDSeq_sequence>cgcccggggagatgctgtccga</INSDSeq_sequence>
</INSDSeq> </INSDSeq>
</SequenceData> </SequenceData>
<SequenceData sequenceIDNumber="15"> <SequenceData sequenceIDNumber="15">
<INSDSeq> <INSDSeq>
<INSDSeq_length>28</INSDSeq_length> <INSDSeq_length>28</INSDSeq_length>
<INSDSeq_moltype>DNA</INSDSeq_moltype> <INSDSeq_moltype>DNA</INSDSeq_moltype>
<INSDSeq_division>PAT</INSDSeq_division> <INSDSeq_division>PAT</INSDSeq_division>
<INSDSeq_feature-table> <INSDSeq_feature-table>
<INSDFeature> <INSDFeature>
<INSDFeature_key>source</INSDFeature_key> <INSDFeature_key>source</INSDFeature_key>
<INSDFeature_location>1..28</INSDFeature_location> <INSDFeature_location>1..28</INSDFeature_location>
<INSDFeature_quals> <INSDFeature_quals>
<INSDQualifier> <INSDQualifier>
<INSDQualifier_name>mol_type</INSDQualifier_name> <INSDQualifier_name>mol_type</INSDQualifier_name>
<INSDQualifier_value>other DNA</INSDQualifier_value> <INSDQualifier_value>other DNA</INSDQualifier_value>
</INSDQualifier> </INSDQualifier>
<INSDQualifier id="q73"> <INSDQualifier id="q73">
<INSDQualifier_name>organism</INSDQualifier_name> <INSDQualifier_name>organism</INSDQualifier_name>
<INSDQualifier_value>synthetic construct</INSDQualifier_value> <INSDQualifier_value>synthetic construct</INSDQualifier_value>
</INSDQualifier> </INSDQualifier>
</INSDFeature_quals> </INSDFeature_quals>
</INSDFeature> </INSDFeature>
<INSDFeature> <INSDFeature>
<INSDFeature_key>misc_feature</INSDFeature_key> <INSDFeature_key>misc_feature</INSDFeature_key>
<INSDFeature_location>1..28</INSDFeature_location> <INSDFeature_location>1..28</INSDFeature_location>
<INSDFeature_quals> <INSDFeature_quals>
<INSDQualifier id="q75"> <INSDQualifier id="q75">
<INSDQualifier_name>product</INSDQualifier_name> <INSDQualifier_name>product</INSDQualifier_name>
<INSDQualifier_value>immobilized primer</INSDQualifier_value> <INSDQualifier_value>immobilized primer</INSDQualifier_value>
<NonEnglishQualifier_value>иммобилизованный <NonEnglishQualifier_value>immobilized
праймер</NonEnglishQualifier_value>primer</NonEnglishQualifier_value>
</INSDQualifier> </INSDQualifier>
<INSDQualifier id="q74"> <INSDQualifier id="q74">
<INSDQualifier_name>standard_name</INSDQualifier_name> <INSDQualifier_name>standard_name</INSDQualifier_name>
<INSDQualifier_value>#13</INSDQualifier_value> <INSDQualifier_value>#13</INSDQualifier_value>
<NonEnglishQualifier_value>№13</NonEnglishQualifier_value> <NonEnglishQualifier_value>№13</NonEnglishQualifier_value>
</INSDQualifier> </INSDQualifier>
</INSDFeature_quals> </INSDFeature_quals>
</INSDFeature> </INSDFeature>
<INSDFeature> <INSDFeature>
<INSDFeature_key>misc_feature</INSDFeature_key> <INSDFeature_key>misc_feature</INSDFeature_key>
<INSDFeature_location><1</INSDFeature_location> <INSDFeature_location><1</INSDFeature_location>
<INSDFeature_quals> <INSDFeature_quals>
<INSDQualifier id="q76"> <INSDQualifier id="q76">
<INSDQualifier_name>function</INSDQualifier_name> <INSDQualifier_name>function</INSDQualifier_name>
<INSDQualifier_value>NH2-group</INSDQualifier_value> <INSDQualifier_value>NH2-group</INSDQualifier_value>
<NonEnglishQualifier_value>аминогруппа</NonEnglishQualifier_val <NonEnglishQualifier_value>amino group</NonEnglishQualifier_val
ue>ue>
</INSDQualifier> </INSDQualifier>
</INSDFeature_quals> </INSDFeature_quals>
</INSDFeature> </INSDFeature>
</INSDSeq_feature-table> </INSDSeq_feature-table>
<INSDSeq_sequence>gaccatcattgccattccctctccccac</INSDSeq_sequence> <INSDSeq_sequence>gaccatcattgccattccctctccccac</INSDSeq_sequence>
</INSDSeq> </INSDSeq>
</SequenceData> </SequenceData>
<SequenceData sequenceIDNumber="16"> <SequenceData sequenceIDNumber="16">
<INSDSeq> <INSDSeq>
<INSDSeq_length>27</INSDSeq_length> <INSDSeq_length>27</INSDSeq_length>
<INSDSeq_moltype>DNA</INSDSeq_moltype> <INSDSeq_moltype>DNA</INSDSeq_moltype>
<INSDSeq_division>PAT</INSDSeq_division> <INSDSeq_division>PAT</INSDSeq_division>
<INSDSeq_feature-table> <INSDSeq_feature-table>
<INSDFeature> <INSDFeature>
<INSDFeature_key>source</INSDFeature_key> <INSDFeature_key>source</INSDFeature_key>
<INSDFeature_location>1..27</INSDFeature_location> <INSDFeature_location>1..27</INSDFeature_location>
<INSDFeature_quals> <INSDFeature_quals>
<INSDQualifier> <INSDQualifier>
<INSDQualifier_name>mol_type</INSDQualifier_name> <INSDQualifier_name>mol_type</INSDQualifier_name>
<INSDQualifier_value>other DNA</INSDQualifier_value> <INSDQualifier_value>other DNA</INSDQualifier_value>
</INSDQualifier> </INSDQualifier>
<INSDQualifier id="q78"> <INSDQualifier id="q78">
<INSDQualifier_name>organism</INSDQualifier_name> <INSDQualifier_name>organism</INSDQualifier_name>
<INSDQualifier_value>synthetic construct</INSDQualifier_value> <INSDQualifier_value>synthetic construct</INSDQualifier_value>
</INSDQualifier> </INSDQualifier>
</INSDFeature_quals> </INSDFeature_quals>
</INSDFeature> </INSDFeature>
<INSDFeature> <INSDFeature>
<INSDFeature_key>misc_feature</INSDFeature_key> <INSDFeature_key>misc_feature</INSDFeature_key>
<INSDFeature_location>1..27</INSDFeature_location> <INSDFeature_location>1..27</INSDFeature_location>
<INSDFeature_quals> <INSDFeature_quals>
<INSDQualifier id="q80"> <INSDQualifier id="q80">
<INSDQualifier_name>product</INSDQualifier_name> <INSDQualifier_name>product</INSDQualifier_name>
<INSDQualifier_value>immobilized primer</INSDQualifier_value> <INSDQualifier_value>immobilized primer</INSDQualifier_value>
<NonEnglishQualifier_value>иммобилизованный <NonEnglishQualifier_value>immobilized
праймер</NonEnglishQualifier_value>primer</NonEnglishQualifier_value>
</INSDQualifier> </INSDQualifier>
<INSDQualifier id="q79"> <INSDQualifier id="q79">
<INSDQualifier_name>standard_name</INSDQualifier_name> <INSDQualifier_name>standard_name</INSDQualifier_name>
<INSDQualifier_value>#14</INSDQualifier_value> <INSDQualifier_value>#14</INSDQualifier_value>
<NonEnglishQualifier_value>№14</NonEnglishQualifier_value> <NonEnglishQualifier_value>№14</NonEnglishQualifier_value>
</INSDQualifier> </INSDQualifier>
</INSDFeature_quals> </INSDFeature_quals>
</INSDFeature> </INSDFeature>
<INSDFeature> <INSDFeature>
<INSDFeature_key>misc_feature</INSDFeature_key> <INSDFeature_key>misc_feature</INSDFeature_key>
<INSDFeature_location><1</INSDFeature_location> <INSDFeature_location><1</INSDFeature_location>
<INSDFeature_quals> <INSDFeature_quals>
<INSDQualifier id="q81"> <INSDQualifier id="q81">
<INSDQualifier_name>function</INSDQualifier_name> <INSDQualifier_name>function</INSDQualifier_name>
<INSDQualifier_value>NH2-group</INSDQualifier_value> <INSDQualifier_value>NH2-group</INSDQualifier_value>
<NonEnglishQualifier_value>аминогруппа</NonEnglishQualifier_val <NonEnglishQualifier_value>amino group</NonEnglishQualifier_val
ue>ue>
</INSDQualifier> </INSDQualifier>
</INSDFeature_quals> </INSDFeature_quals>
</INSDFeature> </INSDFeature>
</INSDSeq_feature-table> </INSDSeq_feature-table>
<INSDSeq_sequence>gcgaaatacagccacgacacgaccaca</INSDSeq_sequence> <INSDSeq_sequence>gcgaaatacagccacgacacgaccaca</INSDSeq_sequence>
</INSDSeq> </INSDSeq>
</SequenceData> </SequenceData>
<SequenceData sequenceIDNumber="17"> <SequenceData sequenceIDNumber="17">
<INSDSeq> <INSDSeq>
<INSDSeq_length>32</INSDSeq_length> <INSDSeq_length>32</INSDSeq_length>
<INSDSeq_moltype>DNA</INSDSeq_moltype> <INSDSeq_moltype>DNA</INSDSeq_moltype>
<INSDSeq_division>PAT</INSDSeq_division> <INSDSeq_division>PAT</INSDSeq_division>
<INSDSeq_feature-table> <INSDSeq_feature-table>
<INSDFeature> <INSDFeature>
<INSDFeature_key>source</INSDFeature_key> <INSDFeature_key>source</INSDFeature_key>
<INSDFeature_location>1..32</INSDFeature_location> <INSDFeature_location>1..32</INSDFeature_location>
<INSDFeature_quals> <INSDFeature_quals>
<INSDQualifier> <INSDQualifier>
<INSDQualifier_name>mol_type</INSDQualifier_name> <INSDQualifier_name>mol_type</INSDQualifier_name>
<INSDQualifier_value>other DNA</INSDQualifier_value> <INSDQualifier_value>other DNA</INSDQualifier_value>
</INSDQualifier> </INSDQualifier>
<INSDQualifier id="q83"> <INSDQualifier id="q83">
<INSDQualifier_name>organism</INSDQualifier_name> <INSDQualifier_name>organism</INSDQualifier_name>
<INSDQualifier_value>synthetic construct</INSDQualifier_value> <INSDQualifier_value>synthetic construct</INSDQualifier_value>
</INSDQualifier> </INSDQualifier>
</INSDFeature_quals> </INSDFeature_quals>
</INSDFeature> </INSDFeature>
<INSDFeature> <INSDFeature>
<INSDFeature_key>misc_feature</INSDFeature_key> <INSDFeature_key>misc_feature</INSDFeature_key>
<INSDFeature_location>1..32</INSDFeature_location> <INSDFeature_location>1..32</INSDFeature_location>
<INSDFeature_quals> <INSDFeature_quals>
<INSDQualifier id="q85"> <INSDQualifier id="q85">
<INSDQualifier_name>product</INSDQualifier_name> <INSDQualifier_name>product</INSDQualifier_name>
<INSDQualifier_value>immobilized primer</INSDQualifier_value> <INSDQualifier_value>immobilized primer</INSDQualifier_value>
<NonEnglishQualifier_value>иммобилизованный <NonEnglishQualifier_value>immobilized
праймер</NonEnglishQualifier_value>primer</NonEnglishQualifier_value>
</INSDQualifier> </INSDQualifier>
<INSDQualifier id="q84"> <INSDQualifier id="q84">
<INSDQualifier_name>standard_name</INSDQualifier_name> <INSDQualifier_name>standard_name</INSDQualifier_name>
<INSDQualifier_value>#15</INSDQualifier_value> <INSDQualifier_value>#15</INSDQualifier_value>
<NonEnglishQualifier_value>№15</NonEnglishQualifier_value> <NonEnglishQualifier_value>№15</NonEnglishQualifier_value>
</INSDQualifier> </INSDQualifier>
</INSDFeature_quals> </INSDFeature_quals>
</INSDFeature> </INSDFeature>
<INSDFeature> <INSDFeature>
<INSDFeature_key>misc_feature</INSDFeature_key> <INSDFeature_key>misc_feature</INSDFeature_key>
<INSDFeature_location><1</INSDFeature_location> <INSDFeature_location><1</INSDFeature_location>
<INSDFeature_quals> <INSDFeature_quals>
<INSDQualifier id="q86"> <INSDQualifier id="q86">
<INSDQualifier_name>function</INSDQualifier_name> <INSDQualifier_name>function</INSDQualifier_name>
<INSDQualifier_value>NH2-group</INSDQualifier_value> <INSDQualifier_value>NH2-group</INSDQualifier_value>
<NonEnglishQualifier_value>аминогруппа</NonEnglishQualifier_val <NonEnglishQualifier_value>amino group</NonEnglishQualifier_val
ue>ue>
</INSDQualifier> </INSDQualifier>
</INSDFeature_quals> </INSDFeature_quals>
</INSDFeature> </INSDFeature>
</INSDSeq_feature-table> </INSDSeq_feature-table>
<INSDSeq_sequence>cggaaaccttgaataacgcgcagtgaatttcg</INSDSeq_sequenc <INSDSeq_sequence>cggaaaccttgaataacgcgcagtgaatttcg</INSDSeq_sequenc
e>e>
</INSDSeq> </INSDSeq>
</SequenceData> </SequenceData>
<SequenceData sequenceIDNumber="18"> <SequenceData sequenceIDNumber="18">
<INSDSeq> <INSDSeq>
<INSDSeq_length>27</INSDSeq_length> <INSDSeq_length>27</INSDSeq_length>
<INSDSeq_moltype>DNA</INSDSeq_moltype> <INSDSeq_moltype>DNA</INSDSeq_moltype>
<INSDSeq_division>PAT</INSDSeq_division> <INSDSeq_division>PAT</INSDSeq_division>
<INSDSeq_feature-table> <INSDSeq_feature-table>
<INSDFeature> <INSDFeature>
<INSDFeature_key>source</INSDFeature_key> <INSDFeature_key>source</INSDFeature_key>
<INSDFeature_location>1..27</INSDFeature_location> <INSDFeature_location>1..27</INSDFeature_location>
<INSDFeature_quals> <INSDFeature_quals>
<INSDQualifier> <INSDQualifier>
<INSDQualifier_name>mol_type</INSDQualifier_name> <INSDQualifier_name>mol_type</INSDQualifier_name>
<INSDQualifier_value>other DNA</INSDQualifier_value> <INSDQualifier_value>other DNA</INSDQualifier_value>
</INSDQualifier> </INSDQualifier>
<INSDQualifier id="q88"> <INSDQualifier id="q88">
<INSDQualifier_name>organism</INSDQualifier_name> <INSDQualifier_name>organism</INSDQualifier_name>
<INSDQualifier_value>synthetic construct</INSDQualifier_value> <INSDQualifier_value>synthetic construct</INSDQualifier_value>
</INSDQualifier> </INSDQualifier>
</INSDFeature_quals> </INSDFeature_quals>
</INSDFeature> </INSDFeature>
<INSDFeature> <INSDFeature>
<INSDFeature_key>misc_feature</INSDFeature_key> <INSDFeature_key>misc_feature</INSDFeature_key>
<INSDFeature_location>1..27</INSDFeature_location> <INSDFeature_location>1..27</INSDFeature_location>
<INSDFeature_quals> <INSDFeature_quals>
<INSDQualifier id="q90"> <INSDQualifier id="q90">
<INSDQualifier_name>product</INSDQualifier_name> <INSDQualifier_name>product</INSDQualifier_name>
<INSDQualifier_value>immobilized primer</INSDQualifier_value> <INSDQualifier_value>immobilized primer</INSDQualifier_value>
<NonEnglishQualifier_value>иммобилизованный <NonEnglishQualifier_value>immobilized
праймер</NonEnglishQualifier_value>primer</NonEnglishQualifier_value>
</INSDQualifier> </INSDQualifier>
<INSDQualifier id="q89"> <INSDQualifier id="q89">
<INSDQualifier_name>standard_name</INSDQualifier_name> <INSDQualifier_name>standard_name</INSDQualifier_name>
<INSDQualifier_value>#16</INSDQualifier_value> <INSDQualifier_value>#16</INSDQualifier_value>
<NonEnglishQualifier_value>№16</NonEnglishQualifier_value> <NonEnglishQualifier_value>№16</NonEnglishQualifier_value>
</INSDQualifier> </INSDQualifier>
</INSDFeature_quals> </INSDFeature_quals>
</INSDFeature> </INSDFeature>
<INSDFeature> <INSDFeature>
<INSDFeature_key>misc_feature</INSDFeature_key> <INSDFeature_key>misc_feature</INSDFeature_key>
<INSDFeature_location><1</INSDFeature_location> <INSDFeature_location><1</INSDFeature_location>
<INSDFeature_quals> <INSDFeature_quals>
<INSDQualifier id="q91"> <INSDQualifier id="q91">
<INSDQualifier_name>function</INSDQualifier_name> <INSDQualifier_name>function</INSDQualifier_name>
<INSDQualifier_value>NH2-group</INSDQualifier_value> <INSDQualifier_value>NH2-group</INSDQualifier_value>
<NonEnglishQualifier_value>аминогруппа</NonEnglishQualifier_val <NonEnglishQualifier_value>amino group</NonEnglishQualifier_val
ue>ue>
</INSDQualifier> </INSDQualifier>
</INSDFeature_quals> </INSDFeature_quals>
</INSDFeature> </INSDFeature>
</INSDSeq_feature-table> </INSDSeq_feature-table>
<INSDSeq_sequence>gcgaacaggctccacgacacgaccaca</INSDSeq_sequence> <INSDSeq_sequence>gcgaacaggctccacgacacgaccaca</INSDSeq_sequence>
</INSDSeq> </INSDSeq>
</SequenceData> </SequenceData>
<SequenceData sequenceIDNumber="19"> <SequenceData sequenceIDNumber="19">
<INSDSeq> <INSDSeq>
<INSDSeq_length>20</INSDSeq_length> <INSDSeq_length>20</INSDSeq_length>
<INSDSeq_moltype>DNA</INSDSeq_moltype> <INSDSeq_moltype>DNA</INSDSeq_moltype>
<INSDSeq_division>PAT</INSDSeq_division> <INSDSeq_division>PAT</INSDSeq_division>
<INSDSeq_feature-table> <INSDSeq_feature-table>
<INSDFeature> <INSDFeature>
<INSDFeature_key>source</INSDFeature_key> <INSDFeature_key>source</INSDFeature_key>
<INSDFeature_location>1..20</INSDFeature_location> <INSDFeature_location>1..20</INSDFeature_location>
<INSDFeature_quals> <INSDFeature_quals>
<INSDQualifier> <INSDQualifier>
<INSDQualifier_name>mol_type</INSDQualifier_name> <INSDQualifier_name>mol_type</INSDQualifier_name>
<INSDQualifier_value>other DNA</INSDQualifier_value> <INSDQualifier_value>other DNA</INSDQualifier_value>
</INSDQualifier> </INSDQualifier>
<INSDQualifier id="q93"> <INSDQualifier id="q93">
<INSDQualifier_name>organism</INSDQualifier_name> <INSDQualifier_name>organism</INSDQualifier_name>
<INSDQualifier_value>synthetic construct</INSDQualifier_value> <INSDQualifier_value>synthetic construct</INSDQualifier_value>
</INSDQualifier> </INSDQualifier>
</INSDFeature_quals> </INSDFeature_quals>
</INSDFeature> </INSDFeature>
<INSDFeature> <INSDFeature>
<INSDFeature_key>misc_feature</INSDFeature_key> <INSDFeature_key>misc_feature</INSDFeature_key>
<INSDFeature_location>1..20</INSDFeature_location> <INSDFeature_location>1..20</INSDFeature_location>
<INSDFeature_quals> <INSDFeature_quals>
<INSDQualifier id="q95"> <INSDQualifier id="q95">
<INSDQualifier_name>product</INSDQualifier_name> <INSDQualifier_name>product</INSDQualifier_name>
<INSDQualifier_value>immobilized primer</INSDQualifier_value> <INSDQualifier_value>immobilized primer</INSDQualifier_value>
<NonEnglishQualifier_value>иммобилизованный <NonEnglishQualifier_value>immobilized
праймер</NonEnglishQualifier_value>primer</NonEnglishQualifier_value>
</INSDQualifier> </INSDQualifier>
<INSDQualifier id="q94"> <INSDQualifier id="q94">
<INSDQualifier_name>standard_name</INSDQualifier_name> <INSDQualifier_name>standard_name</INSDQualifier_name>
<INSDQualifier_value>#17</INSDQualifier_value> <INSDQualifier_value>#17</INSDQualifier_value>
<NonEnglishQualifier_value>№17</NonEnglishQualifier_value> <NonEnglishQualifier_value>№17</NonEnglishQualifier_value>
</INSDQualifier> </INSDQualifier>
</INSDFeature_quals> </INSDFeature_quals>
</INSDFeature> </INSDFeature>
<INSDFeature> <INSDFeature>
<INSDFeature_key>misc_feature</INSDFeature_key> <INSDFeature_key>misc_feature</INSDFeature_key>
<INSDFeature_location><1</INSDFeature_location> <INSDFeature_location><1</INSDFeature_location>
<INSDFeature_quals> <INSDFeature_quals>
<INSDQualifier id="q96"> <INSDQualifier id="q96">
<INSDQualifier_name>function</INSDQualifier_name> <INSDQualifier_name>function</INSDQualifier_name>
<INSDQualifier_value>NH2-group</INSDQualifier_value> <INSDQualifier_value>NH2-group</INSDQualifier_value>
<NonEnglishQualifier_value>аминогруппа</NonEnglishQualifier_val <NonEnglishQualifier_value>amino group</NonEnglishQualifier_val
ue>ue>
</INSDQualifier> </INSDQualifier>
</INSDFeature_quals> </INSDFeature_quals>
</INSDFeature> </INSDFeature>
</INSDSeq_feature-table> </INSDSeq_feature-table>
<INSDSeq_sequence>cggcccgcgcggatgactcc</INSDSeq_sequence> <INSDSeq_sequence>cggcccgcgcggatgactcc</INSDSeq_sequence>
</INSDSeq> </INSDSeq>
</SequenceData> </SequenceData>
<SequenceData sequenceIDNumber="20"> <SequenceData sequenceIDNumber="20">
<INSDSeq> <INSDSeq>
<INSDSeq_length>23</INSDSeq_length> <INSDSeq_length>23</INSDSeq_length>
<INSDSeq_moltype>DNA</INSDSeq_moltype> <INSDSeq_moltype>DNA</INSDSeq_moltype>
<INSDSeq_division>PAT</INSDSeq_division> <INSDSeq_division>PAT</INSDSeq_division>
<INSDSeq_feature-table> <INSDSeq_feature-table>
<INSDFeature> <INSDFeature>
<INSDFeature_key>source</INSDFeature_key> <INSDFeature_key>source</INSDFeature_key>
<INSDFeature_location>1..23</INSDFeature_location> <INSDFeature_location>1..23</INSDFeature_location>
<INSDFeature_quals> <INSDFeature_quals>
<INSDQualifier> <INSDQualifier>
<INSDQualifier_name>mol_type</INSDQualifier_name> <INSDQualifier_name>mol_type</INSDQualifier_name>
<INSDQualifier_value>other DNA</INSDQualifier_value> <INSDQualifier_value>other DNA</INSDQualifier_value>
</INSDQualifier> </INSDQualifier>
<INSDQualifier id="q98"> <INSDQualifier id="q98">
<INSDQualifier_name>organism</INSDQualifier_name> <INSDQualifier_name>organism</INSDQualifier_name>
<INSDQualifier_value>synthetic construct</INSDQualifier_value> <INSDQualifier_value>synthetic construct</INSDQualifier_value>
</INSDQualifier> </INSDQualifier>
</INSDFeature_quals> </INSDFeature_quals>
</INSDFeature> </INSDFeature>
<INSDFeature> <INSDFeature>
<INSDFeature_key>misc_feature</INSDFeature_key> <INSDFeature_key>misc_feature</INSDFeature_key>
<INSDFeature_location>1..23</INSDFeature_location> <INSDFeature_location>1..23</INSDFeature_location>
<INSDFeature_quals> <INSDFeature_quals>
<INSDQualifier id="q100"> <INSDQualifier id="q100">
<INSDQualifier_name>product</INSDQualifier_name> <INSDQualifier_name>product</INSDQualifier_name>
<INSDQualifier_value>immobilized primer</INSDQualifier_value> <INSDQualifier_value>immobilized primer</INSDQualifier_value>
<NonEnglishQualifier_value>иммобилизованный <NonEnglishQualifier_value>immobilized
праймер</NonEnglishQualifier_value>primer</NonEnglishQualifier_value>
</INSDQualifier> </INSDQualifier>
<INSDQualifier id="q99"> <INSDQualifier id="q99">
<INSDQualifier_name>standard_name</INSDQualifier_name> <INSDQualifier_name>standard_name</INSDQualifier_name>
<INSDQualifier_value>#18</INSDQualifier_value> <INSDQualifier_value>#18</INSDQualifier_value>
<NonEnglishQualifier_value>№18</NonEnglishQualifier_value> <NonEnglishQualifier_value>№18</NonEnglishQualifier_value>
</INSDQualifier> </INSDQualifier>
</INSDFeature_quals> </INSDFeature_quals>
</INSDFeature> </INSDFeature>
<INSDFeature> <INSDFeature>
<INSDFeature_key>misc_feature</INSDFeature_key> <INSDFeature_key>misc_feature</INSDFeature_key>
<INSDFeature_location><1</INSDFeature_location> <INSDFeature_location><1</INSDFeature_location>
<INSDFeature_quals> <INSDFeature_quals>
<INSDQualifier id="q101"> <INSDQualifier id="q101">
<INSDQualifier_name>function</INSDQualifier_name> <INSDQualifier_name>function</INSDQualifier_name>
<INSDQualifier_value>NH2-group</INSDQualifier_value> <INSDQualifier_value>NH2-group</INSDQualifier_value>
<NonEnglishQualifier_value>аминогруппа</NonEnglishQualifier_val <NonEnglishQualifier_value>amino group</NonEnglishQualifier_val
ue>ue>
</INSDQualifier> </INSDQualifier>
</INSDFeature_quals> </INSDFeature_quals>
</INSDFeature> </INSDFeature>
</INSDSeq_feature-table> </INSDSeq_feature-table>
<INSDSeq_sequence>gggcgagctgcagatggtccggg</INSDSeq_sequence> <INSDSeq_sequence>gggcgagctgcagatggtccggg</INSDSeq_sequence>
</INSDSeq> </INSDSeq>
</SequenceData> </SequenceData>
<SequenceData sequenceIDNumber="21"> <SequenceData sequenceIDNumber="21">
<INSDSeq> <INSDSeq>
<INSDSeq_length>28</INSDSeq_length> <INSDSeq_length>28</INSDSeq_length>
<INSDSeq_moltype>DNA</INSDSeq_moltype> <INSDSeq_moltype>DNA</INSDSeq_moltype>
<INSDSeq_division>PAT</INSDSeq_division> <INSDSeq_division>PAT</INSDSeq_division>
<INSDSeq_feature-table> <INSDSeq_feature-table>
<INSDFeature> <INSDFeature>
<INSDFeature_key>source</INSDFeature_key> <INSDFeature_key>source</INSDFeature_key>
<INSDFeature_location>1..28</INSDFeature_location> <INSDFeature_location>1..28</INSDFeature_location>
<INSDFeature_quals> <INSDFeature_quals>
<INSDQualifier> <INSDQualifier>
<INSDQualifier_name>mol_type</INSDQualifier_name> <INSDQualifier_name>mol_type</INSDQualifier_name>
<INSDQualifier_value>other DNA</INSDQualifier_value> <INSDQualifier_value>other DNA</INSDQualifier_value>
</INSDQualifier> </INSDQualifier>
<INSDQualifier id="q103"> <INSDQualifier id="q103">
<INSDQualifier_name>organism</INSDQualifier_name> <INSDQualifier_name>organism</INSDQualifier_name>
<INSDQualifier_value>synthetic construct</INSDQualifier_value> <INSDQualifier_value>synthetic construct</INSDQualifier_value>
</INSDQualifier> </INSDQualifier>
</INSDFeature_quals> </INSDFeature_quals>
</INSDFeature> </INSDFeature>
<INSDFeature> <INSDFeature>
<INSDFeature_key>misc_feature</INSDFeature_key> <INSDFeature_key>misc_feature</INSDFeature_key>
<INSDFeature_location>1..28</INSDFeature_location> <INSDFeature_location>1..28</INSDFeature_location>
<INSDFeature_quals> <INSDFeature_quals>
<INSDQualifier id="q105"> <INSDQualifier id="q105">
<INSDQualifier_name>product</INSDQualifier_name> <INSDQualifier_name>product</INSDQualifier_name>
<INSDQualifier_value>immobilized primer</INSDQualifier_value> <INSDQualifier_value>immobilized primer</INSDQualifier_value>
<NonEnglishQualifier_value>иммобилизованный <NonEnglishQualifier_value>immobilized
праймер</NonEnglishQualifier_value>primer</NonEnglishQualifier_value>
</INSDQualifier> </INSDQualifier>
<INSDQualifier id="q104"> <INSDQualifier id="q104">
<INSDQualifier_name>standard_name</INSDQualifier_name> <INSDQualifier_name>standard_name</INSDQualifier_name>
<INSDQualifier_value>#19</INSDQualifier_value> <INSDQualifier_value>#19</INSDQualifier_value>
<NonEnglishQualifier_value>№19</NonEnglishQualifier_value> <NonEnglishQualifier_value>№19</NonEnglishQualifier_value>
</INSDQualifier> </INSDQualifier>
</INSDFeature_quals> </INSDFeature_quals>
</INSDFeature> </INSDFeature>
<INSDFeature> <INSDFeature>
<INSDFeature_key>misc_feature</INSDFeature_key> <INSDFeature_key>misc_feature</INSDFeature_key>
<INSDFeature_location><1</INSDFeature_location> <INSDFeature_location><1</INSDFeature_location>
<INSDFeature_quals> <INSDFeature_quals>
<INSDQualifier id="q106"> <INSDQualifier id="q106">
<INSDQualifier_name>function</INSDQualifier_name> <INSDQualifier_name>function</INSDQualifier_name>
<INSDQualifier_value>NH2-group</INSDQualifier_value> <INSDQualifier_value>NH2-group</INSDQualifier_value>
<NonEnglishQualifier_value>аминогруппа</NonEnglishQualifier_val <NonEnglishQualifier_value>amino group</NonEnglishQualifier_val
ue>ue>
</INSDQualifier> </INSDQualifier>
</INSDFeature_quals> </INSDFeature_quals>
</INSDFeature> </INSDFeature>
</INSDSeq_feature-table> </INSDSeq_feature-table>
<INSDSeq_sequence>cgctaactggcttggcgctgatcctggt</INSDSeq_sequence> <INSDSeq_sequence>cgctaactggcttggcgctgatcctggt</INSDSeq_sequence>
</INSDSeq> </INSDSeq>
</SequenceData> </SequenceData>
<SequenceData sequenceIDNumber="22"> <SequenceData sequenceIDNumber="22">
<INSDSeq> <INSDSeq>
<INSDSeq_length>29</INSDSeq_length> <INSDSeq_length>29</INSDSeq_length>
<INSDSeq_moltype>DNA</INSDSeq_moltype> <INSDSeq_moltype>DNA</INSDSeq_moltype>
<INSDSeq_division>PAT</INSDSeq_division> <INSDSeq_division>PAT</INSDSeq_division>
<INSDSeq_feature-table> <INSDSeq_feature-table>
<INSDFeature> <INSDFeature>
<INSDFeature_key>source</INSDFeature_key> <INSDFeature_key>source</INSDFeature_key>
<INSDFeature_location>1..29</INSDFeature_location> <INSDFeature_location>1..29</INSDFeature_location>
<INSDFeature_quals> <INSDFeature_quals>
<INSDQualifier> <INSDQualifier>
<INSDQualifier_name>mol_type</INSDQualifier_name> <INSDQualifier_name>mol_type</INSDQualifier_name>
<INSDQualifier_value>other DNA</INSDQualifier_value> <INSDQualifier_value>other DNA</INSDQualifier_value>
</INSDQualifier> </INSDQualifier>
<INSDQualifier id="q108"> <INSDQualifier id="q108">
<INSDQualifier_name>organism</INSDQualifier_name> <INSDQualifier_name>organism</INSDQualifier_name>
<INSDQualifier_value>synthetic construct</INSDQualifier_value> <INSDQualifier_value>synthetic construct</INSDQualifier_value>
</INSDQualifier> </INSDQualifier>
</INSDFeature_quals> </INSDFeature_quals>
</INSDFeature> </INSDFeature>
<INSDFeature> <INSDFeature>
<INSDFeature_key>misc_feature</INSDFeature_key> <INSDFeature_key>misc_feature</INSDFeature_key>
<INSDFeature_location>1..29</INSDFeature_location> <INSDFeature_location>1..29</INSDFeature_location>
<INSDFeature_quals> <INSDFeature_quals>
<INSDQualifier id="q110"> <INSDQualifier id="q110">
<INSDQualifier_name>product</INSDQualifier_name> <INSDQualifier_name>product</INSDQualifier_name>
<INSDQualifier_value>immobilized primer</INSDQualifier_value> <INSDQualifier_value>immobilized primer</INSDQualifier_value>
<NonEnglishQualifier_value>иммобилизованный <NonEnglishQualifier_value>immobilized
праймер</NonEnglishQualifier_value>primer</NonEnglishQualifier_value>
</INSDQualifier> </INSDQualifier>
<INSDQualifier id="q109"> <INSDQualifier id="q109">
<INSDQualifier_name>standard_name</INSDQualifier_name> <INSDQualifier_name>standard_name</INSDQualifier_name>
<INSDQualifier_value>#20</INSDQualifier_value> <INSDQualifier_value>#20</INSDQualifier_value>
<NonEnglishQualifier_value>№20</NonEnglishQualifier_value> <NonEnglishQualifier_value>№20</NonEnglishQualifier_value>
</INSDQualifier> </INSDQualifier>
</INSDFeature_quals> </INSDFeature_quals>
</INSDFeature> </INSDFeature>
<INSDFeature> <INSDFeature>
<INSDFeature_key>misc_feature</INSDFeature_key> <INSDFeature_key>misc_feature</INSDFeature_key>
<INSDFeature_location><1</INSDFeature_location> <INSDFeature_location><1</INSDFeature_location>
<INSDFeature_quals> <INSDFeature_quals>
<INSDQualifier id="q111"> <INSDQualifier id="q111">
<INSDQualifier_name>function</INSDQualifier_name> <INSDQualifier_name>function</INSDQualifier_name>
<INSDQualifier_value>NH2-group</INSDQualifier_value> <INSDQualifier_value>NH2-group</INSDQualifier_value>
<NonEnglishQualifier_value>аминогруппа</NonEnglishQualifier_val <NonEnglishQualifier_value>amino group</NonEnglishQualifier_val
ue>ue>
</INSDQualifier> </INSDQualifier>
</INSDFeature_quals> </INSDFeature_quals>
</INSDFeature> </INSDFeature>
</INSDSeq_feature-table> </INSDSeq_feature-table>
<INSDSeq_sequence>tccacatcgatttccttgacctcgccagg</INSDSeq_sequence> <INSDSeq_sequence>tccacatcgatttccttgacctcgccagg</INSDSeq_sequence>
</INSDSeq> </INSDSeq>
</SequenceData> </SequenceData>
<SequenceData sequenceIDNumber="23"> <SequenceData sequenceIDNumber="23">
<INSDSeq> <INSDSeq>
<INSDSeq_length>25</INSDSeq_length> <INSDSeq_length>25</INSDSeq_length>
<INSDSeq_moltype>DNA</INSDSeq_moltype> <INSDSeq_moltype>DNA</INSDSeq_moltype>
<INSDSeq_division>PAT</INSDSeq_division> <INSDSeq_division>PAT</INSDSeq_division>
<INSDSeq_feature-table> <INSDSeq_feature-table>
<INSDFeature> <INSDFeature>
<INSDFeature_key>source</INSDFeature_key> <INSDFeature_key>source</INSDFeature_key>
<INSDFeature_location>1..25</INSDFeature_location> <INSDFeature_location>1..25</INSDFeature_location>
<INSDFeature_quals> <INSDFeature_quals>
<INSDQualifier> <INSDQualifier>
<INSDQualifier_name>mol_type</INSDQualifier_name> <INSDQualifier_name>mol_type</INSDQualifier_name>
<INSDQualifier_value>other DNA</INSDQualifier_value> <INSDQualifier_value>other DNA</INSDQualifier_value>
</INSDQualifier> </INSDQualifier>
<INSDQualifier id="q113"> <INSDQualifier id="q113">
<INSDQualifier_name>organism</INSDQualifier_name> <INSDQualifier_name>organism</INSDQualifier_name>
<INSDQualifier_value>synthetic construct</INSDQualifier_value> <INSDQualifier_value>synthetic construct</INSDQualifier_value>
</INSDQualifier> </INSDQualifier>
</INSDFeature_quals> </INSDFeature_quals>
</INSDFeature> </INSDFeature>
<INSDFeature> <INSDFeature>
<INSDFeature_key>misc_feature</INSDFeature_key> <INSDFeature_key>misc_feature</INSDFeature_key>
<INSDFeature_location>1..25</INSDFeature_location> <INSDFeature_location>1..25</INSDFeature_location>
<INSDFeature_quals> <INSDFeature_quals>
<INSDQualifier id="q115"> <INSDQualifier id="q115">
<INSDQualifier_name>product</INSDQualifier_name> <INSDQualifier_name>product</INSDQualifier_name>
<INSDQualifier_value>immobilized primer</INSDQualifier_value> <INSDQualifier_value>immobilized primer</INSDQualifier_value>
<NonEnglishQualifier_value>иммобилизованный <NonEnglishQualifier_value>immobilized
праймер</NonEnglishQualifier_value>primer</NonEnglishQualifier_value>
</INSDQualifier> </INSDQualifier>
<INSDQualifier id="q114"> <INSDQualifier id="q114">
<INSDQualifier_name>standard_name</INSDQualifier_name> <INSDQualifier_name>standard_name</INSDQualifier_name>
<INSDQualifier_value>#21</INSDQualifier_value> <INSDQualifier_value>#21</INSDQualifier_value>
<NonEnglishQualifier_value>№21</NonEnglishQualifier_value> <NonEnglishQualifier_value>№21</NonEnglishQualifier_value>
</INSDQualifier> </INSDQualifier>
</INSDFeature_quals> </INSDFeature_quals>
</INSDFeature> </INSDFeature>
<INSDFeature> <INSDFeature>
<INSDFeature_key>misc_feature</INSDFeature_key> <INSDFeature_key>misc_feature</INSDFeature_key>
<INSDFeature_location><1</INSDFeature_location> <INSDFeature_location><1</INSDFeature_location>
<INSDFeature_quals> <INSDFeature_quals>
<INSDQualifier id="q116"> <INSDQualifier id="q116">
<INSDQualifier_name>function</INSDQualifier_name> <INSDQualifier_name>function</INSDQualifier_name>
<INSDQualifier_value>NH2-group</INSDQualifier_value> <INSDQualifier_value>NH2-group</INSDQualifier_value>
<NonEnglishQualifier_value>аминогруппа</NonEnglishQualifier_val <NonEnglishQualifier_value>amino group</NonEnglishQualifier_val
ue>ue>
</INSDQualifier> </INSDQualifier>
</INSDFeature_quals> </INSDFeature_quals>
</INSDFeature> </INSDFeature>
</INSDSeq_feature-table> </INSDSeq_feature-table>
<INSDSeq_sequence>gcatacaggctcccgacacgaccaa</INSDSeq_sequence> <INSDSeq_sequence>gcatacaggctcccgacacgaccaa</INSDSeq_sequence>
</INSDSeq> </INSDSeq>
</SequenceData> </SequenceData>
<SequenceData sequenceIDNumber="24"> <SequenceData sequenceIDNumber="24">
<INSDSeq> <INSDSeq>
<INSDSeq_length>26</INSDSeq_length> <INSDSeq_length>26</INSDSeq_length>
<INSDSeq_moltype>DNA</INSDSeq_moltype> <INSDSeq_moltype>DNA</INSDSeq_moltype>
<INSDSeq_division>PAT</INSDSeq_division> <INSDSeq_division>PAT</INSDSeq_division>
<INSDSeq_feature-table> <INSDSeq_feature-table>
<INSDFeature> <INSDFeature>
<INSDFeature_key>source</INSDFeature_key> <INSDFeature_key>source</INSDFeature_key>
<INSDFeature_location>1..26</INSDFeature_location> <INSDFeature_location>1..26</INSDFeature_location>
<INSDFeature_quals> <INSDFeature_quals>
<INSDQualifier> <INSDQualifier>
<INSDQualifier_name>mol_type</INSDQualifier_name> <INSDQualifier_name>mol_type</INSDQualifier_name>
<INSDQualifier_value>other DNA</INSDQualifier_value> <INSDQualifier_value>other DNA</INSDQualifier_value>
</INSDQualifier> </INSDQualifier>
<INSDQualifier id="q118"> <INSDQualifier id="q118">
<INSDQualifier_name>organism</INSDQualifier_name> <INSDQualifier_name>organism</INSDQualifier_name>
<INSDQualifier_value>synthetic construct</INSDQualifier_value> <INSDQualifier_value>synthetic construct</INSDQualifier_value>
</INSDQualifier> </INSDQualifier>
</INSDFeature_quals> </INSDFeature_quals>
</INSDFeature> </INSDFeature>
<INSDFeature> <INSDFeature>
<INSDFeature_key>misc_feature</INSDFeature_key> <INSDFeature_key>misc_feature</INSDFeature_key>
<INSDFeature_location>1..26</INSDFeature_location> <INSDFeature_location>1..26</INSDFeature_location>
<INSDFeature_quals> <INSDFeature_quals>
<INSDQualifier id="q120"> <INSDQualifier id="q120">
<INSDQualifier_name>product</INSDQualifier_name> <INSDQualifier_name>product</INSDQualifier_name>
<INSDQualifier_value>immobilized primer</INSDQualifier_value> <INSDQualifier_value>immobilized primer</INSDQualifier_value>
<NonEnglishQualifier_value>иммобилизованный <NonEnglishQualifier_value>immobilized
праймер</NonEnglishQualifier_value>primer</NonEnglishQualifier_value>
</INSDQualifier> </INSDQualifier>
<INSDQualifier id="q119"> <INSDQualifier id="q119">
<INSDQualifier_name>standard_name</INSDQualifier_name> <INSDQualifier_name>standard_name</INSDQualifier_name>
<INSDQualifier_value>#22</INSDQualifier_value> <INSDQualifier_value>#22</INSDQualifier_value>
<NonEnglishQualifier_value>№22</NonEnglishQualifier_value> <NonEnglishQualifier_value>№22</NonEnglishQualifier_value>
</INSDQualifier> </INSDQualifier>
</INSDFeature_quals> </INSDFeature_quals>
</INSDFeature> </INSDFeature>
<INSDFeature> <INSDFeature>
<INSDFeature_key>misc_feature</INSDFeature_key> <INSDFeature_key>misc_feature</INSDFeature_key>
<INSDFeature_location><1</INSDFeature_location> <INSDFeature_location><1</INSDFeature_location>
<INSDFeature_quals> <INSDFeature_quals>
<INSDQualifier id="q121"> <INSDQualifier id="q121">
<INSDQualifier_name>function</INSDQualifier_name> <INSDQualifier_name>function</INSDQualifier_name>
<INSDQualifier_value>NH2-group</INSDQualifier_value> <INSDQualifier_value>NH2-group</INSDQualifier_value>
<NonEnglishQualifier_value>аминогруппа</NonEnglishQualifier_val <NonEnglishQualifier_value>amino group</NonEnglishQualifier_val
ue>ue>
</INSDQualifier> </INSDQualifier>
</INSDFeature_quals> </INSDFeature_quals>
</INSDFeature> </INSDFeature>
</INSDSeq_feature-table> </INSDSeq_feature-table>
<INSDSeq_sequence>gtgtcaccgcagacggcacgattgag</INSDSeq_sequence> <INSDSeq_sequence>gtgtcaccgcagacggcacgattgag</INSDSeq_sequence>
</INSDSeq> </INSDSeq>
</SequenceData> </SequenceData>
<SequenceData sequenceIDNumber="25"> <SequenceData sequenceIDNumber="25">
<INSDSeq> <INSDSeq>
<INSDSeq_length>26</INSDSeq_length> <INSDSeq_length>26</INSDSeq_length>
<INSDSeq_moltype>DNA</INSDSeq_moltype> <INSDSeq_moltype>DNA</INSDSeq_moltype>
<INSDSeq_division>PAT</INSDSeq_division> <INSDSeq_division>PAT</INSDSeq_division>
<INSDSeq_feature-table> <INSDSeq_feature-table>
<INSDFeature> <INSDFeature>
<INSDFeature_key>source</INSDFeature_key> <INSDFeature_key>source</INSDFeature_key>
<INSDFeature_location>1..26</INSDFeature_location> <INSDFeature_location>1..26</INSDFeature_location>
<INSDFeature_quals> <INSDFeature_quals>
<INSDQualifier> <INSDQualifier>
<INSDQualifier_name>mol_type</INSDQualifier_name> <INSDQualifier_name>mol_type</INSDQualifier_name>
<INSDQualifier_value>other DNA</INSDQualifier_value> <INSDQualifier_value>other DNA</INSDQualifier_value>
</INSDQualifier> </INSDQualifier>
<INSDQualifier id="q123"> <INSDQualifier id="q123">
<INSDQualifier_name>organism</INSDQualifier_name> <INSDQualifier_name>organism</INSDQualifier_name>
<INSDQualifier_value>synthetic construct</INSDQualifier_value> <INSDQualifier_value>synthetic construct</INSDQualifier_value>
</INSDQualifier> </INSDQualifier>
</INSDFeature_quals> </INSDFeature_quals>
</INSDFeature> </INSDFeature>
<INSDFeature> <INSDFeature>
<INSDFeature_key>misc_feature</INSDFeature_key> <INSDFeature_key>misc_feature</INSDFeature_key>
<INSDFeature_location>1..26</INSDFeature_location> <INSDFeature_location>1..26</INSDFeature_location>
<INSDFeature_quals> <INSDFeature_quals>
<INSDQualifier id="q125"> <INSDQualifier id="q125">
<INSDQualifier_name>product</INSDQualifier_name> <INSDQualifier_name>product</INSDQualifier_name>
<INSDQualifier_value>immobilized primer</INSDQualifier_value> <INSDQualifier_value>immobilized primer</INSDQualifier_value>
<NonEnglishQualifier_value>иммобилизованный <NonEnglishQualifier_value>immobilized
праймер</NonEnglishQualifier_value>primer</NonEnglishQualifier_value>
</INSDQualifier> </INSDQualifier>
<INSDQualifier id="q124"> <INSDQualifier id="q124">
<INSDQualifier_name>standard_name</INSDQualifier_name> <INSDQualifier_name>standard_name</INSDQualifier_name>
<INSDQualifier_value>#23</INSDQualifier_value> <INSDQualifier_value>#23</INSDQualifier_value>
<NonEnglishQualifier_value>№23</NonEnglishQualifier_value> <NonEnglishQualifier_value>№23</NonEnglishQualifier_value>
</INSDQualifier> </INSDQualifier>
</INSDFeature_quals> </INSDFeature_quals>
</INSDFeature> </INSDFeature>
<INSDFeature> <INSDFeature>
<INSDFeature_key>misc_feature</INSDFeature_key> <INSDFeature_key>misc_feature</INSDFeature_key>
<INSDFeature_location><1</INSDFeature_location> <INSDFeature_location><1</INSDFeature_location>
<INSDFeature_quals> <INSDFeature_quals>
<INSDQualifier id="q126"> <INSDQualifier id="q126">
<INSDQualifier_name>function</INSDQualifier_name> <INSDQualifier_name>function</INSDQualifier_name>
<INSDQualifier_value>NH2-group</INSDQualifier_value> <INSDQualifier_value>NH2-group</INSDQualifier_value>
<NonEnglishQualifier_value>аминогруппа</NonEnglishQualifier_val <NonEnglishQualifier_value>amino group</NonEnglishQualifier_val
ue>ue>
</INSDQualifier> </INSDQualifier>
</INSDFeature_quals> </INSDFeature_quals>
</INSDFeature> </INSDFeature>
</INSDSeq_feature-table> </INSDSeq_feature-table>
<INSDSeq_sequence>agcatcgctgatgcggtccagctcgt</INSDSeq_sequence> <INSDSeq_sequence>agcatcgctgatgcggtccagctcgt</INSDSeq_sequence>
</INSDSeq> </INSDSeq>
</SequenceData> </SequenceData>
<SequenceData sequenceIDNumber="26"> <SequenceData sequenceIDNumber="26">
<INSDSeq> <INSDSeq>
<INSDSeq_length>27</INSDSeq_length> <INSDSeq_length>27</INSDSeq_length>
<INSDSeq_moltype>DNA</INSDSeq_moltype> <INSDSeq_moltype>DNA</INSDSeq_moltype>
<INSDSeq_division>PAT</INSDSeq_division> <INSDSeq_division>PAT</INSDSeq_division>
<INSDSeq_feature-table> <INSDSeq_feature-table>
<INSDFeature> <INSDFeature>
<INSDFeature_key>source</INSDFeature_key> <INSDFeature_key>source</INSDFeature_key>
<INSDFeature_location>1..27</INSDFeature_location> <INSDFeature_location>1..27</INSDFeature_location>
<INSDFeature_quals> <INSDFeature_quals>
<INSDQualifier> <INSDQualifier>
<INSDQualifier_name>mol_type</INSDQualifier_name> <INSDQualifier_name>mol_type</INSDQualifier_name>
<INSDQualifier_value>other DNA</INSDQualifier_value> <INSDQualifier_value>other DNA</INSDQualifier_value>
</INSDQualifier> </INSDQualifier>
<INSDQualifier id="q128"> <INSDQualifier id="q128">
<INSDQualifier_name>organism</INSDQualifier_name> <INSDQualifier_name>organism</INSDQualifier_name>
<INSDQualifier_value>synthetic construct</INSDQualifier_value> <INSDQualifier_value>synthetic construct</INSDQualifier_value>
</INSDQualifier> </INSDQualifier>
</INSDFeature_quals> </INSDFeature_quals>
</INSDFeature> </INSDFeature>
<INSDFeature> <INSDFeature>
<INSDFeature_key>misc_feature</INSDFeature_key> <INSDFeature_key>misc_feature</INSDFeature_key>
<INSDFeature_location>1..27</INSDFeature_location> <INSDFeature_location>1..27</INSDFeature_location>
<INSDFeature_quals> <INSDFeature_quals>
<INSDQualifier id="q130"> <INSDQualifier id="q130">
<INSDQualifier_name>product</INSDQualifier_name> <INSDQualifier_name>product</INSDQualifier_name>
<INSDQualifier_value>immobilized primer</INSDQualifier_value> <INSDQualifier_value>immobilized primer</INSDQualifier_value>
<NonEnglishQualifier_value>иммобилизованный <NonEnglishQualifier_value>immobilized
праймер</NonEnglishQualifier_value>primer</NonEnglishQualifier_value>
</INSDQualifier> </INSDQualifier>
<INSDQualifier id="q129"> <INSDQualifier id="q129">
<INSDQualifier_name>standard_name</INSDQualifier_name> <INSDQualifier_name>standard_name</INSDQualifier_name>
<INSDQualifier_value>#24</INSDQualifier_value> <INSDQualifier_value>#24</INSDQualifier_value>
<NonEnglishQualifier_value>№24</NonEnglishQualifier_value> <NonEnglishQualifier_value>№24</NonEnglishQualifier_value>
</INSDQualifier> </INSDQualifier>
</INSDFeature_quals> </INSDFeature_quals>
</INSDFeature> </INSDFeature>
<INSDFeature> <INSDFeature>
<INSDFeature_key>misc_feature</INSDFeature_key> <INSDFeature_key>misc_feature</INSDFeature_key>
<INSDFeature_location><1</INSDFeature_location> <INSDFeature_location><1</INSDFeature_location>
<INSDFeature_quals> <INSDFeature_quals>
<INSDQualifier id="q131"> <INSDQualifier id="q131">
<INSDQualifier_name>function</INSDQualifier_name> <INSDQualifier_name>function</INSDQualifier_name>
<INSDQualifier_value>NH2-group</INSDQualifier_value> <INSDQualifier_value>NH2-group</INSDQualifier_value>
<NonEnglishQualifier_value>аминогруппа</NonEnglishQualifier_val <NonEnglishQualifier_value>amino group</NonEnglishQualifier_val
ue>ue>
</INSDQualifier> </INSDQualifier>
</INSDFeature_quals> </INSDFeature_quals>
</INSDFeature> </INSDFeature>
</INSDSeq_feature-table> </INSDSeq_feature-table>
<INSDSeq_sequence>ggaaatcaggctccacgcacgaccaca</INSDSeq_sequence> <INSDSeq_sequence>ggaaatcaggctccacgcacgaccaca</INSDSeq_sequence>
</INSDSeq> </INSDSeq>
</SequenceData> </SequenceData>
<SequenceData sequenceIDNumber="27"> <SequenceData sequenceIDNumber="27">
<INSDSeq> <INSDSeq>
<INSDSeq_length>25</INSDSeq_length> <INSDSeq_length>25</INSDSeq_length>
<INSDSeq_moltype>DNA</INSDSeq_moltype> <INSDSeq_moltype>DNA</INSDSeq_moltype>
<INSDSeq_division>PAT</INSDSeq_division> <INSDSeq_division>PAT</INSDSeq_division>
<INSDSeq_feature-table> <INSDSeq_feature-table>
<INSDFeature> <INSDFeature>
<INSDFeature_key>source</INSDFeature_key> <INSDFeature_key>source</INSDFeature_key>
<INSDFeature_location>1..25</INSDFeature_location> <INSDFeature_location>1..25</INSDFeature_location>
<INSDFeature_quals> <INSDFeature_quals>
<INSDQualifier> <INSDQualifier>
<INSDQualifier_name>mol_type</INSDQualifier_name> <INSDQualifier_name>mol_type</INSDQualifier_name>
<INSDQualifier_value>other DNA</INSDQualifier_value> <INSDQualifier_value>other DNA</INSDQualifier_value>
</INSDQualifier> </INSDQualifier>
<INSDQualifier id="q133"> <INSDQualifier id="q133">
<INSDQualifier_name>organism</INSDQualifier_name> <INSDQualifier_name>organism</INSDQualifier_name>
<INSDQualifier_value>synthetic construct</INSDQualifier_value> <INSDQualifier_value>synthetic construct</INSDQualifier_value>
</INSDQualifier> </INSDQualifier>
</INSDFeature_quals> </INSDFeature_quals>
</INSDFeature> </INSDFeature>
<INSDFeature> <INSDFeature>
<INSDFeature_key>misc_feature</INSDFeature_key> <INSDFeature_key>misc_feature</INSDFeature_key>
<INSDFeature_location>1..25</INSDFeature_location> <INSDFeature_location>1..25</INSDFeature_location>
<INSDFeature_quals> <INSDFeature_quals>
<INSDQualifier id="q135"> <INSDQualifier id="q135">
<INSDQualifier_name>product</INSDQualifier_name> <INSDQualifier_name>product</INSDQualifier_name>
<INSDQualifier_value>immobilized primer</INSDQualifier_value> <INSDQualifier_value>immobilized primer</INSDQualifier_value>
<NonEnglishQualifier_value>иммобилизованный <NonEnglishQualifier_value>immobilized
праймер</NonEnglishQualifier_value>primer</NonEnglishQualifier_value>
</INSDQualifier> </INSDQualifier>
<INSDQualifier id="q134"> <INSDQualifier id="q134">
<INSDQualifier_name>standard_name</INSDQualifier_name> <INSDQualifier_name>standard_name</INSDQualifier_name>
<INSDQualifier_value>#25</INSDQualifier_value> <INSDQualifier_value>#25</INSDQualifier_value>
<NonEnglishQualifier_value>№25</NonEnglishQualifier_value> <NonEnglishQualifier_value>№25</NonEnglishQualifier_value>
</INSDQualifier> </INSDQualifier>
</INSDFeature_quals> </INSDFeature_quals>
</INSDFeature> </INSDFeature>
<INSDFeature> <INSDFeature>
<INSDFeature_key>misc_feature</INSDFeature_key> <INSDFeature_key>misc_feature</INSDFeature_key>
<INSDFeature_location><1</INSDFeature_location> <INSDFeature_location><1</INSDFeature_location>
<INSDFeature_quals> <INSDFeature_quals>
<INSDQualifier id="q136"> <INSDQualifier id="q136">
<INSDQualifier_name>function</INSDQualifier_name> <INSDQualifier_name>function</INSDQualifier_name>
<INSDQualifier_value>NH2-group</INSDQualifier_value> <INSDQualifier_value>NH2-group</INSDQualifier_value>
<NonEnglishQualifier_value>аминогруппа</NonEnglishQualifier_val <NonEnglishQualifier_value>amino group</NonEnglishQualifier_val
ue>ue>
</INSDQualifier> </INSDQualifier>
</INSDFeature_quals> </INSDFeature_quals>
</INSDFeature> </INSDFeature>
</INSDSeq_feature-table> </INSDSeq_feature-table>
<INSDSeq_sequence>gatcagcgaccaccgcaccctgtca</INSDSeq_sequence> <INSDSeq_sequence>gatcagcgaccaccgcaccctgtca</INSDSeq_sequence>
</INSDSeq> </INSDSeq>
</SequenceData> </SequenceData>
<SequenceData sequenceIDNumber="28"> <SequenceData sequenceIDNumber="28">
<INSDSeq> <INSDSeq>
<INSDSeq_length>28</INSDSeq_length> <INSDSeq_length>28</INSDSeq_length>
<INSDSeq_moltype>DNA</INSDSeq_moltype> <INSDSeq_moltype>DNA</INSDSeq_moltype>
<INSDSeq_division>PAT</INSDSeq_division> <INSDSeq_division>PAT</INSDSeq_division>
<INSDSeq_feature-table> <INSDSeq_feature-table>
<INSDFeature> <INSDFeature>
<INSDFeature_key>source</INSDFeature_key> <INSDFeature_key>source</INSDFeature_key>
<INSDFeature_location>1..28</INSDFeature_location> <INSDFeature_location>1..28</INSDFeature_location>
<INSDFeature_quals> <INSDFeature_quals>
<INSDQualifier> <INSDQualifier>
<INSDQualifier_name>mol_type</INSDQualifier_name> <INSDQualifier_name>mol_type</INSDQualifier_name>
<INSDQualifier_value>other DNA</INSDQualifier_value> <INSDQualifier_value>other DNA</INSDQualifier_value>
</INSDQualifier> </INSDQualifier>
<INSDQualifier id="q138"> <INSDQualifier id="q138">
<INSDQualifier_name>organism</INSDQualifier_name> <INSDQualifier_name>organism</INSDQualifier_name>
<INSDQualifier_value>synthetic construct</INSDQualifier_value> <INSDQualifier_value>synthetic construct</INSDQualifier_value>
</INSDQualifier> </INSDQualifier>
</INSDFeature_quals> </INSDFeature_quals>
</INSDFeature> </INSDFeature>
<INSDFeature> <INSDFeature>
<INSDFeature_key>misc_feature</INSDFeature_key> <INSDFeature_key>misc_feature</INSDFeature_key>
<INSDFeature_location>1..28</INSDFeature_location> <INSDFeature_location>1..28</INSDFeature_location>
<INSDFeature_quals> <INSDFeature_quals>
<INSDQualifier id="q140"> <INSDQualifier id="q140">
<INSDQualifier_name>product</INSDQualifier_name> <INSDQualifier_name>product</INSDQualifier_name>
<INSDQualifier_value>immobilized primer</INSDQualifier_value> <INSDQualifier_value>immobilized primer</INSDQualifier_value>
<NonEnglishQualifier_value>иммобилизованный <NonEnglishQualifier_value>immobilized
праймер</NonEnglishQualifier_value>primer</NonEnglishQualifier_value>
</INSDQualifier> </INSDQualifier>
<INSDQualifier id="q139"> <INSDQualifier id="q139">
<INSDQualifier_name>standard_name</INSDQualifier_name> <INSDQualifier_name>standard_name</INSDQualifier_name>
<INSDQualifier_value>#26</INSDQualifier_value> <INSDQualifier_value>#26</INSDQualifier_value>
<NonEnglishQualifier_value>№26</NonEnglishQualifier_value> <NonEnglishQualifier_value>№26</NonEnglishQualifier_value>
</INSDQualifier> </INSDQualifier>
</INSDFeature_quals> </INSDFeature_quals>
</INSDFeature> </INSDFeature>
<INSDFeature> <INSDFeature>
<INSDFeature_key>misc_feature</INSDFeature_key> <INSDFeature_key>misc_feature</INSDFeature_key>
<INSDFeature_location><1</INSDFeature_location> <INSDFeature_location><1</INSDFeature_location>
<INSDFeature_quals> <INSDFeature_quals>
<INSDQualifier id="q141"> <INSDQualifier id="q141">
<INSDQualifier_name>function</INSDQualifier_name> <INSDQualifier_name>function</INSDQualifier_name>
<INSDQualifier_value>NH2-group</INSDQualifier_value> <INSDQualifier_value>NH2-group</INSDQualifier_value>
<NonEnglishQualifier_value>аминогруппа</NonEnglishQualifier_val <NonEnglishQualifier_value>amino group</NonEnglishQualifier_val
ue>ue>
</INSDQualifier> </INSDQualifier>
</INSDFeature_quals> </INSDFeature_quals>
</INSDFeature> </INSDFeature>
</INSDSeq_feature-table> </INSDSeq_feature-table>
<INSDSeq_sequence>gaaaccttgaatacgcgcagtgaattcg</INSDSeq_sequence> <INSDSeq_sequence>gaaaccttgaatacgcgcagtgaattcg</INSDSeq_sequence>
</INSDSeq> </INSDSeq>
</SequenceData> </SequenceData>
<SequenceData sequenceIDNumber="29"> <SequenceData sequenceIDNumber="29">
<INSDSeq> <INSDSeq>
<INSDSeq_length>30</INSDSeq_length> <INSDSeq_length>30</INSDSeq_length>
<INSDSeq_moltype>DNA</INSDSeq_moltype> <INSDSeq_moltype>DNA</INSDSeq_moltype>
<INSDSeq_division>PAT</INSDSeq_division> <INSDSeq_division>PAT</INSDSeq_division>
<INSDSeq_feature-table> <INSDSeq_feature-table>
<INSDFeature> <INSDFeature>
<INSDFeature_key>source</INSDFeature_key> <INSDFeature_key>source</INSDFeature_key>
<INSDFeature_location>1..30</INSDFeature_location> <INSDFeature_location>1..30</INSDFeature_location>
<INSDFeature_quals> <INSDFeature_quals>
<INSDQualifier> <INSDQualifier>
<INSDQualifier_name>mol_type</INSDQualifier_name> <INSDQualifier_name>mol_type</INSDQualifier_name>
<INSDQualifier_value>other DNA</INSDQualifier_value> <INSDQualifier_value>other DNA</INSDQualifier_value>
</INSDQualifier> </INSDQualifier>
<INSDQualifier id="q143"> <INSDQualifier id="q143">
<INSDQualifier_name>organism</INSDQualifier_name> <INSDQualifier_name>organism</INSDQualifier_name>
<INSDQualifier_value>synthetic construct</INSDQualifier_value> <INSDQualifier_value>synthetic construct</INSDQualifier_value>
</INSDQualifier> </INSDQualifier>
</INSDFeature_quals> </INSDFeature_quals>
</INSDFeature> </INSDFeature>
<INSDFeature> <INSDFeature>
<INSDFeature_key>misc_feature</INSDFeature_key> <INSDFeature_key>misc_feature</INSDFeature_key>
<INSDFeature_location>1..30</INSDFeature_location> <INSDFeature_location>1..30</INSDFeature_location>
<INSDFeature_quals> <INSDFeature_quals>
<INSDQualifier id="q145"> <INSDQualifier id="q145">
<INSDQualifier_name>product</INSDQualifier_name> <INSDQualifier_name>product</INSDQualifier_name>
<INSDQualifier_value>immobilized primer</INSDQualifier_value> <INSDQualifier_value>immobilized primer</INSDQualifier_value>
<NonEnglishQualifier_value>иммобилизованный <NonEnglishQualifier_value>immobilized
праймер</NonEnglishQualifier_value>primer</NonEnglishQualifier_value>
</INSDQualifier> </INSDQualifier>
<INSDQualifier id="q144"> <INSDQualifier id="q144">
<INSDQualifier_name>standard_name</INSDQualifier_name> <INSDQualifier_name>standard_name</INSDQualifier_name>
<INSDQualifier_value>#27</INSDQualifier_value> <INSDQualifier_value>#27</INSDQualifier_value>
<NonEnglishQualifier_value>№27</NonEnglishQualifier_value> <NonEnglishQualifier_value>№27</NonEnglishQualifier_value>
</INSDQualifier> </INSDQualifier>
</INSDFeature_quals> </INSDFeature_quals>
</INSDFeature> </INSDFeature>
<INSDFeature> <INSDFeature>
<INSDFeature_key>misc_feature</INSDFeature_key> <INSDFeature_key>misc_feature</INSDFeature_key>
<INSDFeature_location><1</INSDFeature_location> <INSDFeature_location><1</INSDFeature_location>
<INSDFeature_quals> <INSDFeature_quals>
<INSDQualifier id="q146"> <INSDQualifier id="q146">
<INSDQualifier_name>function</INSDQualifier_name> <INSDQualifier_name>function</INSDQualifier_name>
<INSDQualifier_value>NH2-group</INSDQualifier_value> <INSDQualifier_value>NH2-group</INSDQualifier_value>
<NonEnglishQualifier_value>аминогруппа</NonEnglishQualifier_val <NonEnglishQualifier_value>amino group</NonEnglishQualifier_val
ue>ue>
</INSDQualifier> </INSDQualifier>
</INSDFeature_quals> </INSDFeature_quals>
</INSDFeature> </INSDFeature>
</INSDSeq_feature-table> </INSDSeq_feature-table>
<INSDSeq_sequence>cggaaacttgaataagcgcagtgaatttcg</INSDSeq_sequence> <INSDSeq_sequence>cggaaacttgaataagcgcagtgaatttcg</INSDSeq_sequence>
</INSDSeq> </INSDSeq>
</SequenceData> </SequenceData>
<SequenceData sequenceIDNumber="30"> <SequenceData sequenceIDNumber="30">
<INSDSeq> <INSDSeq>
<INSDSeq_length>26</INSDSeq_length> <INSDSeq_length>26</INSDSeq_length>
<INSDSeq_moltype>DNA</INSDSeq_moltype> <INSDSeq_moltype>DNA</INSDSeq_moltype>
<INSDSeq_division>PAT</INSDSeq_division> <INSDSeq_division>PAT</INSDSeq_division>
<INSDSeq_feature-table> <INSDSeq_feature-table>
<INSDFeature> <INSDFeature>
<INSDFeature_key>source</INSDFeature_key> <INSDFeature_key>source</INSDFeature_key>
<INSDFeature_location>1..26</INSDFeature_location> <INSDFeature_location>1..26</INSDFeature_location>
<INSDFeature_quals> <INSDFeature_quals>
<INSDQualifier> <INSDQualifier>
<INSDQualifier_name>mol_type</INSDQualifier_name> <INSDQualifier_name>mol_type</INSDQualifier_name>
<INSDQualifier_value>other DNA</INSDQualifier_value> <INSDQualifier_value>other DNA</INSDQualifier_value>
</INSDQualifier> </INSDQualifier>
<INSDQualifier id="q148"> <INSDQualifier id="q148">
<INSDQualifier_name>organism</INSDQualifier_name> <INSDQualifier_name>organism</INSDQualifier_name>
<INSDQualifier_value>synthetic construct</INSDQualifier_value> <INSDQualifier_value>synthetic construct</INSDQualifier_value>
</INSDQualifier> </INSDQualifier>
</INSDFeature_quals> </INSDFeature_quals>
</INSDFeature> </INSDFeature>
<INSDFeature> <INSDFeature>
<INSDFeature_key>misc_feature</INSDFeature_key> <INSDFeature_key>misc_feature</INSDFeature_key>
<INSDFeature_location>1..26</INSDFeature_location> <INSDFeature_location>1..26</INSDFeature_location>
<INSDFeature_quals> <INSDFeature_quals>
<INSDQualifier id="q150"> <INSDQualifier id="q150">
<INSDQualifier_name>product</INSDQualifier_name> <INSDQualifier_name>product</INSDQualifier_name>
<INSDQualifier_value>immobilized primer</INSDQualifier_value> <INSDQualifier_value>immobilized primer</INSDQualifier_value>
<NonEnglishQualifier_value>иммобилизованный <NonEnglishQualifier_value>immobilized
праймер</NonEnglishQualifier_value>primer</NonEnglishQualifier_value>
</INSDQualifier> </INSDQualifier>
<INSDQualifier id="q149"> <INSDQualifier id="q149">
<INSDQualifier_name>standard_name</INSDQualifier_name> <INSDQualifier_name>standard_name</INSDQualifier_name>
<INSDQualifier_value>#28</INSDQualifier_value> <INSDQualifier_value>#28</INSDQualifier_value>
<NonEnglishQualifier_value>№28</NonEnglishQualifier_value> <NonEnglishQualifier_value>№28</NonEnglishQualifier_value>
</INSDQualifier> </INSDQualifier>
</INSDFeature_quals> </INSDFeature_quals>
</INSDFeature> </INSDFeature>
<INSDFeature> <INSDFeature>
<INSDFeature_key>misc_feature</INSDFeature_key> <INSDFeature_key>misc_feature</INSDFeature_key>
<INSDFeature_location><1</INSDFeature_location> <INSDFeature_location><1</INSDFeature_location>
<INSDFeature_quals> <INSDFeature_quals>
<INSDQualifier id="q151"> <INSDQualifier id="q151">
<INSDQualifier_name>function</INSDQualifier_name> <INSDQualifier_name>function</INSDQualifier_name>
<INSDQualifier_value>NH2-group</INSDQualifier_value> <INSDQualifier_value>NH2-group</INSDQualifier_value>
<NonEnglishQualifier_value>аминогруппа</NonEnglishQualifier_val <NonEnglishQualifier_value>amino group</NonEnglishQualifier_val
ue>ue>
</INSDQualifier> </INSDQualifier>
</INSDFeature_quals> </INSDFeature_quals>
</INSDFeature> </INSDFeature>
</INSDSeq_feature-table> </INSDSeq_feature-table>
<INSDSeq_sequence>tgccccggcgtttagcgatcacaaca</INSDSeq_sequence> <INSDSeq_sequence>tgccccggcgtttagcgatcacaaca</INSDSeq_sequence>
</INSDSeq> </INSDSeq>
</SequenceData> </SequenceData>
<SequenceData sequenceIDNumber="31"> <SequenceData sequenceIDNumber="31">
<INSDSeq> <INSDSeq>
<INSDSeq_length>30</INSDSeq_length> <INSDSeq_length>30</INSDSeq_length>
<INSDSeq_moltype>DNA</INSDSeq_moltype> <INSDSeq_moltype>DNA</INSDSeq_moltype>
<INSDSeq_division>PAT</INSDSeq_division> <INSDSeq_division>PAT</INSDSeq_division>
<INSDSeq_feature-table> <INSDSeq_feature-table>
<INSDFeature> <INSDFeature>
<INSDFeature_key>source</INSDFeature_key> <INSDFeature_key>source</INSDFeature_key>
<INSDFeature_location>1..30</INSDFeature_location> <INSDFeature_location>1..30</INSDFeature_location>
<INSDFeature_quals> <INSDFeature_quals>
<INSDQualifier> <INSDQualifier>
<INSDQualifier_name>mol_type</INSDQualifier_name> <INSDQualifier_name>mol_type</INSDQualifier_name>
<INSDQualifier_value>other DNA</INSDQualifier_value> <INSDQualifier_value>other DNA</INSDQualifier_value>
</INSDQualifier> </INSDQualifier>
<INSDQualifier id="q153"> <INSDQualifier id="q153">
<INSDQualifier_name>organism</INSDQualifier_name> <INSDQualifier_name>organism</INSDQualifier_name>
<INSDQualifier_value>synthetic construct</INSDQualifier_value> <INSDQualifier_value>synthetic construct</INSDQualifier_value>
</INSDQualifier> </INSDQualifier>
</INSDFeature_quals> </INSDFeature_quals>
</INSDFeature> </INSDFeature>
<INSDFeature> <INSDFeature>
<INSDFeature_key>misc_feature</INSDFeature_key> <INSDFeature_key>misc_feature</INSDFeature_key>
<INSDFeature_location>1..30</INSDFeature_location> <INSDFeature_location>1..30</INSDFeature_location>
<INSDFeature_quals> <INSDFeature_quals>
<INSDQualifier id="q155"> <INSDQualifier id="q155">
<INSDQualifier_name>product</INSDQualifier_name> <INSDQualifier_name>product</INSDQualifier_name>
<INSDQualifier_value>immobilized primer</INSDQualifier_value> <INSDQualifier_value>immobilized primer</INSDQualifier_value>
<NonEnglishQualifier_value>иммобилизованный <NonEnglishQualifier_value>immobilized
праймер</NonEnglishQualifier_value>primer</NonEnglishQualifier_value>
</INSDQualifier> </INSDQualifier>
<INSDQualifier id="q154"> <INSDQualifier id="q154">
<INSDQualifier_name>standard_name</INSDQualifier_name> <INSDQualifier_name>standard_name</INSDQualifier_name>
<INSDQualifier_value>#29</INSDQualifier_value> <INSDQualifier_value>#29</INSDQualifier_value>
<NonEnglishQualifier_value>№29</NonEnglishQualifier_value> <NonEnglishQualifier_value>№29</NonEnglishQualifier_value>
</INSDQualifier> </INSDQualifier>
</INSDFeature_quals> </INSDFeature_quals>
</INSDFeature> </INSDFeature>
<INSDFeature> <INSDFeature>
<INSDFeature_key>misc_feature</INSDFeature_key> <INSDFeature_key>misc_feature</INSDFeature_key>
<INSDFeature_location><1</INSDFeature_location> <INSDFeature_location><1</INSDFeature_location>
<INSDFeature_quals> <INSDFeature_quals>
<INSDQualifier id="q156"> <INSDQualifier id="q156">
<INSDQualifier_name>function</INSDQualifier_name> <INSDQualifier_name>function</INSDQualifier_name>
<INSDQualifier_value>NH2-group</INSDQualifier_value> <INSDQualifier_value>NH2-group</INSDQualifier_value>
<NonEnglishQualifier_value>аминогруппа</NonEnglishQualifier_val <NonEnglishQualifier_value>amino group</NonEnglishQualifier_val
ue>ue>
</INSDQualifier> </INSDQualifier>
</INSDFeature_quals> </INSDFeature_quals>
</INSDFeature> </INSDFeature>
</INSDSeq_feature-table> </INSDSeq_feature-table>
<INSDSeq_sequence>gcgaaatacaggctccacgacacgaccaca</INSDSeq_sequence> <INSDSeq_sequence>gcgaaatacaggctccacgacacgaccaca</INSDSeq_sequence>
</INSDSeq> </INSDSeq>
</SequenceData> </SequenceData>
<SequenceData sequenceIDNumber="32"> <SequenceData sequenceIDNumber="32">
<INSDSeq> <INSDSeq>
<INSDSeq_length>21</INSDSeq_length> <INSDSeq_length>21</INSDSeq_length>
<INSDSeq_moltype>DNA</INSDSeq_moltype> <INSDSeq_moltype>DNA</INSDSeq_moltype>
<INSDSeq_division>PAT</INSDSeq_division> <INSDSeq_division>PAT</INSDSeq_division>
<INSDSeq_feature-table> <INSDSeq_feature-table>
<INSDFeature> <INSDFeature>
<INSDFeature_key>source</INSDFeature_key> <INSDFeature_key>source</INSDFeature_key>
<INSDFeature_location>1..21</INSDFeature_location> <INSDFeature_location>1..21</INSDFeature_location>
<INSDFeature_quals> <INSDFeature_quals>
<INSDQualifier> <INSDQualifier>
<INSDQualifier_name>mol_type</INSDQualifier_name> <INSDQualifier_name>mol_type</INSDQualifier_name>
<INSDQualifier_value>other DNA</INSDQualifier_value> <INSDQualifier_value>other DNA</INSDQualifier_value>
</INSDQualifier> </INSDQualifier>
<INSDQualifier id="q158"> <INSDQualifier id="q158">
<INSDQualifier_name>organism</INSDQualifier_name> <INSDQualifier_name>organism</INSDQualifier_name>
<INSDQualifier_value>synthetic construct</INSDQualifier_value> <INSDQualifier_value>synthetic construct</INSDQualifier_value>
</INSDQualifier> </INSDQualifier>
</INSDFeature_quals> </INSDFeature_quals>
</INSDFeature> </INSDFeature>
<INSDFeature> <INSDFeature>
<INSDFeature_key>misc_feature</INSDFeature_key> <INSDFeature_key>misc_feature</INSDFeature_key>
<INSDFeature_location>1..21</INSDFeature_location> <INSDFeature_location>1..21</INSDFeature_location>
<INSDFeature_quals> <INSDFeature_quals>
<INSDQualifier id="q160"> <INSDQualifier id="q160">
<INSDQualifier_name>product</INSDQualifier_name> <INSDQualifier_name>product</INSDQualifier_name>
<INSDQualifier_value>immobilized primer</INSDQualifier_value> <INSDQualifier_value>immobilized primer</INSDQualifier_value>
<NonEnglishQualifier_value>иммобилизованный <NonEnglishQualifier_value>immobilized
праймер</NonEnglishQualifier_value>primer</NonEnglishQualifier_value>
</INSDQualifier> </INSDQualifier>
<INSDQualifier id="q159"> <INSDQualifier id="q159">
<INSDQualifier_name>standard_name</INSDQualifier_name> <INSDQualifier_name>standard_name</INSDQualifier_name>
<INSDQualifier_value>#30</INSDQualifier_value> <INSDQualifier_value>#30</INSDQualifier_value>
<NonEnglishQualifier_value>№30</NonEnglishQualifier_value> <NonEnglishQualifier_value>№30</NonEnglishQualifier_value>
</INSDQualifier> </INSDQualifier>
</INSDFeature_quals> </INSDFeature_quals>
</INSDFeature> </INSDFeature>
<INSDFeature> <INSDFeature>
<INSDFeature_key>misc_feature</INSDFeature_key> <INSDFeature_key>misc_feature</INSDFeature_key>
<INSDFeature_location><1</INSDFeature_location> <INSDFeature_location><1</INSDFeature_location>
<INSDFeature_quals> <INSDFeature_quals>
<INSDQualifier id="q161"> <INSDQualifier id="q161">
<INSDQualifier_name>function</INSDQualifier_name> <INSDQualifier_name>function</INSDQualifier_name>
<INSDQualifier_value>NH2-group</INSDQualifier_value> <INSDQualifier_value>NH2-group</INSDQualifier_value>
<NonEnglishQualifier_value>аминогруппа</NonEnglishQualifier_val <NonEnglishQualifier_value>amino group</NonEnglishQualifier_val
ue>ue>
</INSDQualifier> </INSDQualifier>
</INSDFeature_quals> </INSDFeature_quals>
</INSDFeature> </INSDFeature>
</INSDSeq_feature-table> </INSDSeq_feature-table>
<INSDSeq_sequence>gccccgcgcccttttccagcc</INSDSeq_sequence> <INSDSeq_sequence>gccccgcgcccttttccagcc</INSDSeq_sequence>
</INSDSeq> </INSDSeq>
</SequenceData> </SequenceData>
<SequenceData sequenceIDNumber="33"> <SequenceData sequenceIDNumber="33">
<INSDSeq> <INSDSeq>
<INSDSeq_length>26</INSDSeq_length> <INSDSeq_length>26</INSDSeq_length>
<INSDSeq_moltype>DNA</INSDSeq_moltype> <INSDSeq_moltype>DNA</INSDSeq_moltype>
<INSDSeq_division>PAT</INSDSeq_division> <INSDSeq_division>PAT</INSDSeq_division>
<INSDSeq_feature-table> <INSDSeq_feature-table>
<INSDFeature> <INSDFeature>
<INSDFeature_key>source</INSDFeature_key> <INSDFeature_key>source</INSDFeature_key>
<INSDFeature_location>1..26</INSDFeature_location> <INSDFeature_location>1..26</INSDFeature_location>
<INSDFeature_quals> <INSDFeature_quals>
<INSDQualifier> <INSDQualifier>
<INSDQualifier_name>mol_type</INSDQualifier_name> <INSDQualifier_name>mol_type</INSDQualifier_name>
<INSDQualifier_value>other DNA</INSDQualifier_value> <INSDQualifier_value>other DNA</INSDQualifier_value>
</INSDQualifier> </INSDQualifier>
<INSDQualifier id="q163"> <INSDQualifier id="q163">
<INSDQualifier_name>organism</INSDQualifier_name> <INSDQualifier_name>organism</INSDQualifier_name>
<INSDQualifier_value>synthetic construct</INSDQualifier_value> <INSDQualifier_value>synthetic construct</INSDQualifier_value>
</INSDQualifier> </INSDQualifier>
</INSDFeature_quals> </INSDFeature_quals>
</INSDFeature> </INSDFeature>
<INSDFeature> <INSDFeature>
<INSDFeature_key>misc_feature</INSDFeature_key> <INSDFeature_key>misc_feature</INSDFeature_key>
<INSDFeature_location>1..26</INSDFeature_location> <INSDFeature_location>1..26</INSDFeature_location>
<INSDFeature_quals> <INSDFeature_quals>
<INSDQualifier id="q165"> <INSDQualifier id="q165">
<INSDQualifier_name>product</INSDQualifier_name> <INSDQualifier_name>product</INSDQualifier_name>
<INSDQualifier_value>immobilized primer</INSDQualifier_value> <INSDQualifier_value>immobilized primer</INSDQualifier_value>
<NonEnglishQualifier_value>иммобилизованный <NonEnglishQualifier_value>immobilized
праймер</NonEnglishQualifier_value>primer</NonEnglishQualifier_value>
</INSDQualifier> </INSDQualifier>
<INSDQualifier id="q164"> <INSDQualifier id="q164">
<INSDQualifier_name>standard_name</INSDQualifier_name> <INSDQualifier_name>standard_name</INSDQualifier_name>
<INSDQualifier_value>#31</INSDQualifier_value> <INSDQualifier_value>#31</INSDQualifier_value>
<NonEnglishQualifier_value>№31</NonEnglishQualifier_value> <NonEnglishQualifier_value>№31</NonEnglishQualifier_value>
</INSDQualifier> </INSDQualifier>
</INSDFeature_quals> </INSDFeature_quals>
</INSDFeature> </INSDFeature>
<INSDFeature> <INSDFeature>
<INSDFeature_key>misc_feature</INSDFeature_key> <INSDFeature_key>misc_feature</INSDFeature_key>
<INSDFeature_location><1</INSDFeature_location> <INSDFeature_location><1</INSDFeature_location>
<INSDFeature_quals> <INSDFeature_quals>
<INSDQualifier id="q166"> <INSDQualifier id="q166">
<INSDQualifier_name>function</INSDQualifier_name> <INSDQualifier_name>function</INSDQualifier_name>
<INSDQualifier_value>NH2-group</INSDQualifier_value> <INSDQualifier_value>NH2-group</INSDQualifier_value>
<NonEnglishQualifier_value>аминогруппа</NonEnglishQualifier_val <NonEnglishQualifier_value>amino group</NonEnglishQualifier_val
ue>ue>
</INSDQualifier> </INSDQualifier>
</INSDFeature_quals> </INSDFeature_quals>
</INSDFeature> </INSDFeature>
</INSDSeq_feature-table> </INSDSeq_feature-table>
<INSDSeq_sequence>cggaccttgtaacgcgcagaatttcg</INSDSeq_sequence> <INSDSeq_sequence>cggaccttgtaacgcgcagaatttcg</INSDSeq_sequence>
</INSDSeq> </INSDSeq>
</SequenceData> </SequenceData>
<SequenceData sequenceIDNumber="34"> <SequenceData sequenceIDNumber="34">
<INSDSeq> <INSDSeq>
<INSDSeq_length>34</INSDSeq_length> <INSDSeq_length>34</INSDSeq_length>
<INSDSeq_moltype>DNA</INSDSeq_moltype> <INSDSeq_moltype>DNA</INSDSeq_moltype>
<INSDSeq_division>PAT</INSDSeq_division> <INSDSeq_division>PAT</INSDSeq_division>
<INSDSeq_feature-table> <INSDSeq_feature-table>
<INSDFeature> <INSDFeature>
<INSDFeature_key>source</INSDFeature_key> <INSDFeature_key>source</INSDFeature_key>
<INSDFeature_location>1..34</INSDFeature_location> <INSDFeature_location>1..34</INSDFeature_location>
<INSDFeature_quals> <INSDFeature_quals>
<INSDQualifier> <INSDQualifier>
<INSDQualifier_name>mol_type</INSDQualifier_name> <INSDQualifier_name>mol_type</INSDQualifier_name>
<INSDQualifier_value>other DNA</INSDQualifier_value> <INSDQualifier_value>other DNA</INSDQualifier_value>
</INSDQualifier> </INSDQualifier>
<INSDQualifier id="q168"> <INSDQualifier id="q168">
<INSDQualifier_name>organism</INSDQualifier_name> <INSDQualifier_name>organism</INSDQualifier_name>
<INSDQualifier_value>synthetic construct</INSDQualifier_value> <INSDQualifier_value>synthetic construct</INSDQualifier_value>
</INSDQualifier> </INSDQualifier>
</INSDFeature_quals> </INSDFeature_quals>
</INSDFeature> </INSDFeature>
<INSDFeature> <INSDFeature>
<INSDFeature_key>misc_feature</INSDFeature_key> <INSDFeature_key>misc_feature</INSDFeature_key>
<INSDFeature_location>1..34</INSDFeature_location> <INSDFeature_location>1..34</INSDFeature_location>
<INSDFeature_quals> <INSDFeature_quals>
<INSDQualifier id="q170"> <INSDQualifier id="q170">
<INSDQualifier_name>product</INSDQualifier_name> <INSDQualifier_name>product</INSDQualifier_name>
<INSDQualifier_value>immobilized primer</INSDQualifier_value> <INSDQualifier_value>immobilized primer</INSDQualifier_value>
<NonEnglishQualifier_value>иммобилизованный <NonEnglishQualifier_value>immobilized
праймер</NonEnglishQualifier_value>primer</NonEnglishQualifier_value>
</INSDQualifier> </INSDQualifier>
<INSDQualifier id="q169"> <INSDQualifier id="q169">
<INSDQualifier_name>standard_name</INSDQualifier_name> <INSDQualifier_name>standard_name</INSDQualifier_name>
<INSDQualifier_value>#32</INSDQualifier_value> <INSDQualifier_value>#32</INSDQualifier_value>
<NonEnglishQualifier_value>№32</NonEnglishQualifier_value> <NonEnglishQualifier_value>№32</NonEnglishQualifier_value>
</INSDQualifier> </INSDQualifier>
</INSDFeature_quals> </INSDFeature_quals>
</INSDFeature> </INSDFeature>
<INSDFeature> <INSDFeature>
<INSDFeature_key>misc_feature</INSDFeature_key> <INSDFeature_key>misc_feature</INSDFeature_key>
<INSDFeature_location><1</INSDFeature_location> <INSDFeature_location><1</INSDFeature_location>
<INSDFeature_quals> <INSDFeature_quals>
<INSDQualifier id="q171"> <INSDQualifier id="q171">
<INSDQualifier_name>function</INSDQualifier_name> <INSDQualifier_name>function</INSDQualifier_name>
<INSDQualifier_value>NH2-group</INSDQualifier_value> <INSDQualifier_value>NH2-group</INSDQualifier_value>
<NonEnglishQualifier_value>аминогруппа</NonEnglishQualifier_val <NonEnglishQualifier_value>amino group</NonEnglishQualifier_val
ue>ue>
</INSDQualifier> </INSDQualifier>
</INSDFeature_quals> </INSDFeature_quals>
</INSDFeature> </INSDFeature>
</INSDSeq_feature-table> </INSDSeq_feature-table>
<INSDSeq_sequence>tatcactcccgaccaaataggtatcggcgtgttc</INSDSeq_seque <INSDSeq_sequence>tatcactcccgaccaaataggtatcggcgtgttc</INSDSeq_seque
nce>nce>
</INSDSeq> </INSDSeq>
</SequenceData> </SequenceData>
<SequenceData sequenceIDNumber="35"> <SequenceData sequenceIDNumber="35">
<INSDSeq> <INSDSeq>
<INSDSeq_length>28</INSDSeq_length> <INSDSeq_length>28</INSDSeq_length>
<INSDSeq_moltype>DNA</INSDSeq_moltype> <INSDSeq_moltype>DNA</INSDSeq_moltype>
<INSDSeq_division>PAT</INSDSeq_division> <INSDSeq_division>PAT</INSDSeq_division>
<INSDSeq_feature-table> <INSDSeq_feature-table>
<INSDFeature> <INSDFeature>
<INSDFeature_key>source</INSDFeature_key> <INSDFeature_key>source</INSDFeature_key>
<INSDFeature_location>1..28</INSDFeature_location> <INSDFeature_location>1..28</INSDFeature_location>
<INSDFeature_quals> <INSDFeature_quals>
<INSDQualifier> <INSDQualifier>
<INSDQualifier_name>mol_type</INSDQualifier_name> <INSDQualifier_name>mol_type</INSDQualifier_name>
<INSDQualifier_value>other DNA</INSDQualifier_value> <INSDQualifier_value>other DNA</INSDQualifier_value>
</INSDQualifier> </INSDQualifier>
<INSDQualifier id="q173"> <INSDQualifier id="q173">
<INSDQualifier_name>organism</INSDQualifier_name> <INSDQualifier_name>organism</INSDQualifier_name>
<INSDQualifier_value>synthetic construct</INSDQualifier_value> <INSDQualifier_value>synthetic construct</INSDQualifier_value>
</INSDQualifier> </INSDQualifier>
</INSDFeature_quals> </INSDFeature_quals>
</INSDFeature> </INSDFeature>
<INSDFeature> <INSDFeature>
<INSDFeature_key>misc_feature</INSDFeature_key> <INSDFeature_key>misc_feature</INSDFeature_key>
<INSDFeature_location>1..28</INSDFeature_location> <INSDFeature_location>1..28</INSDFeature_location>
<INSDFeature_quals> <INSDFeature_quals>
<INSDQualifier id="q175"> <INSDQualifier id="q175">
<INSDQualifier_name>product</INSDQualifier_name> <INSDQualifier_name>product</INSDQualifier_name>
<INSDQualifier_value>immobilized primer</INSDQualifier_value> <INSDQualifier_value>immobilized primer</INSDQualifier_value>
<NonEnglishQualifier_value>иммобилизованный <NonEnglishQualifier_value>immobilized
праймер</NonEnglishQualifier_value>primer</NonEnglishQualifier_value>
</INSDQualifier> </INSDQualifier>
<INSDQualifier id="q174"> <INSDQualifier id="q174">
<INSDQualifier_name>standard_name</INSDQualifier_name> <INSDQualifier_name>standard_name</INSDQualifier_name>
<INSDQualifier_value>#33</INSDQualifier_value> <INSDQualifier_value>#33</INSDQualifier_value>
<NonEnglishQualifier_value>№33</NonEnglishQualifier_value> <NonEnglishQualifier_value>№33</NonEnglishQualifier_value>
</INSDQualifier> </INSDQualifier>
</INSDFeature_quals> </INSDFeature_quals>
</INSDFeature> </INSDFeature>
<INSDFeature> <INSDFeature>
<INSDFeature_key>misc_feature</INSDFeature_key> <INSDFeature_key>misc_feature</INSDFeature_key>
<INSDFeature_location><1</INSDFeature_location> <INSDFeature_location><1</INSDFeature_location>
<INSDFeature_quals> <INSDFeature_quals>
<INSDQualifier id="q176"> <INSDQualifier id="q176">
<INSDQualifier_name>function</INSDQualifier_name> <INSDQualifier_name>function</INSDQualifier_name>
<INSDQualifier_value>NH2-group</INSDQualifier_value> <INSDQualifier_value>NH2-group</INSDQualifier_value>
<NonEnglishQualifier_value>аминогруппа</NonEnglishQualifier_val <NonEnglishQualifier_value>amino group</NonEnglishQualifier_val
ue>ue>
</INSDQualifier> </INSDQualifier>
</INSDFeature_quals> </INSDFeature_quals>
</INSDFeature> </INSDFeature>
</INSDSeq_feature-table> </INSDSeq_feature-table>
<INSDSeq_sequence>gacatgacgacggtgccgcacttgacgc</INSDSeq_sequence> <INSDSeq_sequence>gacatgacgacggtgccgcacttgacgc</INSDSeq_sequence>
</INSDSeq> </INSDSeq>
</SequenceData> </SequenceData>
<SequenceData sequenceIDNumber="36"> <SequenceData sequenceIDNumber="36">
<INSDSeq> <INSDSeq>
<INSDSeq_length>26</INSDSeq_length> <INSDSeq_length>26</INSDSeq_length>
<INSDSeq_moltype>DNA</INSDSeq_moltype> <INSDSeq_moltype>DNA</INSDSeq_moltype>
<INSDSeq_division>PAT</INSDSeq_division> <INSDSeq_division>PAT</INSDSeq_division>
<INSDSeq_feature-table> <INSDSeq_feature-table>
<INSDFeature> <INSDFeature>
<INSDFeature_key>source</INSDFeature_key> <INSDFeature_key>source</INSDFeature_key>
<INSDFeature_location>1..26</INSDFeature_location> <INSDFeature_location>1..26</INSDFeature_location>
<INSDFeature_quals> <INSDFeature_quals>
<INSDQualifier> <INSDQualifier>
<INSDQualifier_name>mol_type</INSDQualifier_name> <INSDQualifier_name>mol_type</INSDQualifier_name>
<INSDQualifier_value>other DNA</INSDQualifier_value> <INSDQualifier_value>other DNA</INSDQualifier_value>
</INSDQualifier> </INSDQualifier>
<INSDQualifier id="q178"> <INSDQualifier id="q178">
<INSDQualifier_name>organism</INSDQualifier_name> <INSDQualifier_name>organism</INSDQualifier_name>
<INSDQualifier_value>synthetic construct</INSDQualifier_value> <INSDQualifier_value>synthetic construct</INSDQualifier_value>
</INSDQualifier> </INSDQualifier>
</INSDFeature_quals> </INSDFeature_quals>
</INSDFeature> </INSDFeature>
<INSDFeature> <INSDFeature>
<INSDFeature_key>misc_feature</INSDFeature_key> <INSDFeature_key>misc_feature</INSDFeature_key>
<INSDFeature_location>1..26</INSDFeature_location> <INSDFeature_location>1..26</INSDFeature_location>
<INSDFeature_quals> <INSDFeature_quals>
<INSDQualifier id="q180"> <INSDQualifier id="q180">
<INSDQualifier_name>product</INSDQualifier_name> <INSDQualifier_name>product</INSDQualifier_name>
<INSDQualifier_value>immobilized primer</INSDQualifier_value> <INSDQualifier_value>immobilized primer</INSDQualifier_value>
<NonEnglishQualifier_value>иммобилизованный <NonEnglishQualifier_value>immobilized
праймер</NonEnglishQualifier_value>primer</NonEnglishQualifier_value>
</INSDQualifier> </INSDQualifier>
<INSDQualifier id="q179"> <INSDQualifier id="q179">
<INSDQualifier_name>standard_name</INSDQualifier_name> <INSDQualifier_name>standard_name</INSDQualifier_name>
<INSDQualifier_value>#34</INSDQualifier_value> <INSDQualifier_value>#34</INSDQualifier_value>
<NonEnglishQualifier_value>№34</NonEnglishQualifier_value> <NonEnglishQualifier_value>№34</NonEnglishQualifier_value>
</INSDQualifier> </INSDQualifier>
</INSDFeature_quals> </INSDFeature_quals>
</INSDFeature> </INSDFeature>
<INSDFeature> <INSDFeature>
<INSDFeature_key>misc_feature</INSDFeature_key> <INSDFeature_key>misc_feature</INSDFeature_key>
<INSDFeature_location><1</INSDFeature_location> <INSDFeature_location><1</INSDFeature_location>
<INSDFeature_quals> <INSDFeature_quals>
<INSDQualifier id="q181"> <INSDQualifier id="q181">
<INSDQualifier_name>function</INSDQualifier_name> <INSDQualifier_name>function</INSDQualifier_name>
<INSDQualifier_value>NH2-group</INSDQualifier_value> <INSDQualifier_value>NH2-group</INSDQualifier_value>
<NonEnglishQualifier_value>аминогруппа</NonEnglishQualifier_val <NonEnglishQualifier_value>amino group</NonEnglishQualifier_val
ue>ue>
</INSDQualifier> </INSDQualifier>
</INSDFeature_quals> </INSDFeature_quals>
</INSDFeature> </INSDFeature>
</INSDSeq_feature-table> </INSDSeq_feature-table>
<INSDSeq_sequence>agtcacctcgcccacaccgtcgaagc</INSDSeq_sequence> <INSDSeq_sequence>agtcacctcgcccacaccgtcgaagc</INSDSeq_sequence>
</INSDSeq> </INSDSeq>
</SequenceData> </SequenceData>
<SequenceData sequenceIDNumber="37"> <SequenceData sequenceIDNumber="37">
<INSDSeq> <INSDSeq>
<INSDSeq_length>29</INSDSeq_length> <INSDSeq_length>29</INSDSeq_length>
<INSDSeq_moltype>DNA</INSDSeq_moltype> <INSDSeq_moltype>DNA</INSDSeq_moltype>
<INSDSeq_division>PAT</INSDSeq_division> <INSDSeq_division>PAT</INSDSeq_division>
<INSDSeq_feature-table> <INSDSeq_feature-table>
<INSDFeature> <INSDFeature>
<INSDFeature_key>source</INSDFeature_key> <INSDFeature_key>source</INSDFeature_key>
<INSDFeature_location>1..29</INSDFeature_location> <INSDFeature_location>1..29</INSDFeature_location>
<INSDFeature_quals> <INSDFeature_quals>
<INSDQualifier> <INSDQualifier>
<INSDQualifier_name>mol_type</INSDQualifier_name> <INSDQualifier_name>mol_type</INSDQualifier_name>
<INSDQualifier_value>other DNA</INSDQualifier_value> <INSDQualifier_value>other DNA</INSDQualifier_value>
</INSDQualifier> </INSDQualifier>
<INSDQualifier id="q183"> <INSDQualifier id="q183">
<INSDQualifier_name>organism</INSDQualifier_name> <INSDQualifier_name>organism</INSDQualifier_name>
<INSDQualifier_value>synthetic construct</INSDQualifier_value> <INSDQualifier_value>synthetic construct</INSDQualifier_value>
</INSDQualifier> </INSDQualifier>
</INSDFeature_quals> </INSDFeature_quals>
</INSDFeature> </INSDFeature>
<INSDFeature> <INSDFeature>
<INSDFeature_key>misc_feature</INSDFeature_key> <INSDFeature_key>misc_feature</INSDFeature_key>
<INSDFeature_location>1..29</INSDFeature_location> <INSDFeature_location>1..29</INSDFeature_location>
<INSDFeature_quals> <INSDFeature_quals>
<INSDQualifier id="q185"> <INSDQualifier id="q185">
<INSDQualifier_name>product</INSDQualifier_name> <INSDQualifier_name>product</INSDQualifier_name>
<INSDQualifier_value>immobilized primer</INSDQualifier_value> <INSDQualifier_value>immobilized primer</INSDQualifier_value>
<NonEnglishQualifier_value>иммобилизованный <NonEnglishQualifier_value>immobilized
праймер</NonEnglishQualifier_value>primer</NonEnglishQualifier_value>
</INSDQualifier> </INSDQualifier>
<INSDQualifier id="q184"> <INSDQualifier id="q184">
<INSDQualifier_name>standard_name</INSDQualifier_name> <INSDQualifier_name>standard_name</INSDQualifier_name>
<INSDQualifier_value>#35</INSDQualifier_value> <INSDQualifier_value>#35</INSDQualifier_value>
<NonEnglishQualifier_value>№35</NonEnglishQualifier_value> <NonEnglishQualifier_value>№35</NonEnglishQualifier_value>
</INSDQualifier> </INSDQualifier>
</INSDFeature_quals> </INSDFeature_quals>
</INSDFeature> </INSDFeature>
<INSDFeature> <INSDFeature>
<INSDFeature_key>misc_feature</INSDFeature_key> <INSDFeature_key>misc_feature</INSDFeature_key>
<INSDFeature_location><1</INSDFeature_location> <INSDFeature_location><1</INSDFeature_location>
<INSDFeature_quals> <INSDFeature_quals>
<INSDQualifier id="q186"> <INSDQualifier id="q186">
<INSDQualifier_name>function</INSDQualifier_name> <INSDQualifier_name>function</INSDQualifier_name>
<INSDQualifier_value>NH2-group</INSDQualifier_value> <INSDQualifier_value>NH2-group</INSDQualifier_value>
<NonEnglishQualifier_value>аминогруппа</NonEnglishQualifier_val <NonEnglishQualifier_value>amino group</NonEnglishQualifier_val
ue>ue>
</INSDQualifier> </INSDQualifier>
</INSDFeature_quals> </INSDFeature_quals>
</INSDFeature> </INSDFeature>
</INSDSeq_feature-table> </INSDSeq_feature-table>
<INSDSeq_sequence>aagtccgtacgctcgaaacgcttccaacg</INSDSeq_sequence> <INSDSeq_sequence>aagtccgtacgctcgaaacgcttccaacg</INSDSeq_sequence>
</INSDSeq> </INSDSeq>
</SequenceData> </SequenceData>
<SequenceData sequenceIDNumber="38"> <SequenceData sequenceIDNumber="38">
<INSDSeq> <INSDSeq>
<INSDSeq_length>26</INSDSeq_length> <INSDSeq_length>26</INSDSeq_length>
<INSDSeq_moltype>DNA</INSDSeq_moltype> <INSDSeq_moltype>DNA</INSDSeq_moltype>
<INSDSeq_division>PAT</INSDSeq_division> <INSDSeq_division>PAT</INSDSeq_division>
<INSDSeq_feature-table> <INSDSeq_feature-table>
<INSDFeature> <INSDFeature>
<INSDFeature_key>source</INSDFeature_key> <INSDFeature_key>source</INSDFeature_key>
<INSDFeature_location>1..26</INSDFeature_location> <INSDFeature_location>1..26</INSDFeature_location>
<INSDFeature_quals> <INSDFeature_quals>
<INSDQualifier> <INSDQualifier>
<INSDQualifier_name>mol_type</INSDQualifier_name> <INSDQualifier_name>mol_type</INSDQualifier_name>
<INSDQualifier_value>other DNA</INSDQualifier_value> <INSDQualifier_value>other DNA</INSDQualifier_value>
</INSDQualifier> </INSDQualifier>
<INSDQualifier id="q188"> <INSDQualifier id="q188">
<INSDQualifier_name>organism</INSDQualifier_name> <INSDQualifier_name>organism</INSDQualifier_name>
<INSDQualifier_value>synthetic construct</INSDQualifier_value> <INSDQualifier_value>synthetic construct</INSDQualifier_value>
</INSDQualifier> </INSDQualifier>
</INSDFeature_quals> </INSDFeature_quals>
</INSDFeature> </INSDFeature>
<INSDFeature> <INSDFeature>
<INSDFeature_key>misc_feature</INSDFeature_key> <INSDFeature_key>misc_feature</INSDFeature_key>
<INSDFeature_location>1..26</INSDFeature_location> <INSDFeature_location>1..26</INSDFeature_location>
<INSDFeature_quals> <INSDFeature_quals>
<INSDQualifier id="q190"> <INSDQualifier id="q190">
<INSDQualifier_name>product</INSDQualifier_name> <INSDQualifier_name>product</INSDQualifier_name>
<INSDQualifier_value>immobilized primer</INSDQualifier_value> <INSDQualifier_value>immobilized primer</INSDQualifier_value>
<NonEnglishQualifier_value>иммобилизованный <NonEnglishQualifier_value>immobilized
праймер</NonEnglishQualifier_value>primer</NonEnglishQualifier_value>
</INSDQualifier> </INSDQualifier>
<INSDQualifier id="q189"> <INSDQualifier id="q189">
<INSDQualifier_name>standard_name</INSDQualifier_name> <INSDQualifier_name>standard_name</INSDQualifier_name>
<INSDQualifier_value>#36</INSDQualifier_value> <INSDQualifier_value>#36</INSDQualifier_value>
<NonEnglishQualifier_value>№36</NonEnglishQualifier_value> <NonEnglishQualifier_value>№36</NonEnglishQualifier_value>
</INSDQualifier> </INSDQualifier>
</INSDFeature_quals> </INSDFeature_quals>
</INSDFeature> </INSDFeature>
<INSDFeature> <INSDFeature>
<INSDFeature_key>misc_feature</INSDFeature_key> <INSDFeature_key>misc_feature</INSDFeature_key>
<INSDFeature_location><1</INSDFeature_location> <INSDFeature_location><1</INSDFeature_location>
<INSDFeature_quals> <INSDFeature_quals>
<INSDQualifier id="q191"> <INSDQualifier id="q191">
<INSDQualifier_name>function</INSDQualifier_name> <INSDQualifier_name>function</INSDQualifier_name>
<INSDQualifier_value>NH2-group</INSDQualifier_value> <INSDQualifier_value>NH2-group</INSDQualifier_value>
<NonEnglishQualifier_value>аминогруппа</NonEnglishQualifier_val <NonEnglishQualifier_value>amino group</NonEnglishQualifier_val
ue>ue>
</INSDQualifier> </INSDQualifier>
</INSDFeature_quals> </INSDFeature_quals>
</INSDFeature> </INSDFeature>
</INSDSeq_feature-table> </INSDSeq_feature-table>
<INSDSeq_sequence>tcgaaatccagcaccacatccgcagc</INSDSeq_sequence> <INSDSeq_sequence>tcgaaatccagcaccacatccgcagc</INSDSeq_sequence>
</INSDSeq> </INSDSeq>
</SequenceData> </SequenceData>
<SequenceData sequenceIDNumber="39"> <SequenceData sequenceIDNumber="39">
<INSDSeq> <INSDSeq>
<INSDSeq_length>25</INSDSeq_length> <INSDSeq_length>25</INSDSeq_length>
<INSDSeq_moltype>DNA</INSDSeq_moltype> <INSDSeq_moltype>DNA</INSDSeq_moltype>
<INSDSeq_division>PAT</INSDSeq_division> <INSDSeq_division>PAT</INSDSeq_division>
<INSDSeq_feature-table> <INSDSeq_feature-table>
<INSDFeature> <INSDFeature>
<INSDFeature_key>source</INSDFeature_key> <INSDFeature_key>source</INSDFeature_key>
<INSDFeature_location>1..25</INSDFeature_location> <INSDFeature_location>1..25</INSDFeature_location>
<INSDFeature_quals> <INSDFeature_quals>
<INSDQualifier> <INSDQualifier>
<INSDQualifier_name>mol_type</INSDQualifier_name> <INSDQualifier_name>mol_type</INSDQualifier_name>
<INSDQualifier_value>other DNA</INSDQualifier_value> <INSDQualifier_value>other DNA</INSDQualifier_value>
</INSDQualifier> </INSDQualifier>
<INSDQualifier id="q193"> <INSDQualifier id="q193">
<INSDQualifier_name>organism</INSDQualifier_name> <INSDQualifier_name>organism</INSDQualifier_name>
<INSDQualifier_value>synthetic construct</INSDQualifier_value> <INSDQualifier_value>synthetic construct</INSDQualifier_value>
</INSDQualifier> </INSDQualifier>
</INSDFeature_quals> </INSDFeature_quals>
</INSDFeature> </INSDFeature>
<INSDFeature> <INSDFeature>
<INSDFeature_key>misc_feature</INSDFeature_key> <INSDFeature_key>misc_feature</INSDFeature_key>
<INSDFeature_location>1..25</INSDFeature_location> <INSDFeature_location>1..25</INSDFeature_location>
<INSDFeature_quals> <INSDFeature_quals>
<INSDQualifier id="q195"> <INSDQualifier id="q195">
<INSDQualifier_name>product</INSDQualifier_name> <INSDQualifier_name>product</INSDQualifier_name>
<INSDQualifier_value>immobilized primer</INSDQualifier_value> <INSDQualifier_value>immobilized primer</INSDQualifier_value>
<NonEnglishQualifier_value>иммобилизованный <NonEnglishQualifier_value>immobilized
праймер</NonEnglishQualifier_value>primer</NonEnglishQualifier_value>
</INSDQualifier> </INSDQualifier>
<INSDQualifier id="q194"> <INSDQualifier id="q194">
<INSDQualifier_name>standard_name</INSDQualifier_name> <INSDQualifier_name>standard_name</INSDQualifier_name>
<INSDQualifier_value>#37</INSDQualifier_value> <INSDQualifier_value>#37</INSDQualifier_value>
<NonEnglishQualifier_value>№37</NonEnglishQualifier_value> <NonEnglishQualifier_value>№37</NonEnglishQualifier_value>
</INSDQualifier> </INSDQualifier>
</INSDFeature_quals> </INSDFeature_quals>
</INSDFeature> </INSDFeature>
<INSDFeature> <INSDFeature>
<INSDFeature_key>misc_feature</INSDFeature_key> <INSDFeature_key>misc_feature</INSDFeature_key>
<INSDFeature_location><1</INSDFeature_location> <INSDFeature_location><1</INSDFeature_location>
<INSDFeature_quals> <INSDFeature_quals>
<INSDQualifier id="q196"> <INSDQualifier id="q196">
<INSDQualifier_name>function</INSDQualifier_name> <INSDQualifier_name>function</INSDQualifier_name>
<INSDQualifier_value>NH2-group</INSDQualifier_value> <INSDQualifier_value>NH2-group</INSDQualifier_value>
<NonEnglishQualifier_value>аминогруппа</NonEnglishQualifier_val <NonEnglishQualifier_value>amino group</NonEnglishQualifier_val
ue>ue>
</INSDQualifier> </INSDQualifier>
</INSDFeature_quals> </INSDFeature_quals>
</INSDFeature> </INSDFeature>
</INSDSeq_feature-table> </INSDSeq_feature-table>
<INSDSeq_sequence>gcgcgaactcgtccacagtccccct</INSDSeq_sequence> <INSDSeq_sequence>gcgcgaactcgtccacagtccccct</INSDSeq_sequence>
</INSDSeq> </INSDSeq>
</SequenceData> </SequenceData>
<SequenceData sequenceIDNumber="40"> <SequenceData sequenceIDNumber="40">
<INSDSeq> <INSDSeq>
<INSDSeq_length>23</INSDSeq_length> <INSDSeq_length>23</INSDSeq_length>
<INSDSeq_moltype>DNA</INSDSeq_moltype> <INSDSeq_moltype>DNA</INSDSeq_moltype>
<INSDSeq_division>PAT</INSDSeq_division> <INSDSeq_division>PAT</INSDSeq_division>
<INSDSeq_feature-table> <INSDSeq_feature-table>
<INSDFeature> <INSDFeature>
<INSDFeature_key>source</INSDFeature_key> <INSDFeature_key>source</INSDFeature_key>
<INSDFeature_location>1..23</INSDFeature_location> <INSDFeature_location>1..23</INSDFeature_location>
<INSDFeature_quals> <INSDFeature_quals>
<INSDQualifier> <INSDQualifier>
<INSDQualifier_name>mol_type</INSDQualifier_name> <INSDQualifier_name>mol_type</INSDQualifier_name>
<INSDQualifier_value>other DNA</INSDQualifier_value> <INSDQualifier_value>other DNA</INSDQualifier_value>
</INSDQualifier> </INSDQualifier>
<INSDQualifier id="q198"> <INSDQualifier id="q198">
<INSDQualifier_name>organism</INSDQualifier_name> <INSDQualifier_name>organism</INSDQualifier_name>
<INSDQualifier_value>synthetic construct</INSDQualifier_value> <INSDQualifier_value>synthetic construct</INSDQualifier_value>
</INSDQualifier> </INSDQualifier>
</INSDFeature_quals> </INSDFeature_quals>
</INSDFeature> </INSDFeature>
<INSDFeature> <INSDFeature>
<INSDFeature_key>misc_feature</INSDFeature_key> <INSDFeature_key>misc_feature</INSDFeature_key>
<INSDFeature_location>1..23</INSDFeature_location> <INSDFeature_location>1..23</INSDFeature_location>
<INSDFeature_quals> <INSDFeature_quals>
<INSDQualifier id="q200"> <INSDQualifier id="q200">
<INSDQualifier_name>product</INSDQualifier_name> <INSDQualifier_name>product</INSDQualifier_name>
<INSDQualifier_value>immobilized primer</INSDQualifier_value> <INSDQualifier_value>immobilized primer</INSDQualifier_value>
<NonEnglishQualifier_value>иммобилизованный <NonEnglishQualifier_value>immobilized
праймер</NonEnglishQualifier_value>primer</NonEnglishQualifier_value>
</INSDQualifier> </INSDQualifier>
<INSDQualifier id="q199"> <INSDQualifier id="q199">
<INSDQualifier_name>standard_name</INSDQualifier_name> <INSDQualifier_name>standard_name</INSDQualifier_name>
<INSDQualifier_value>#38</INSDQualifier_value> <INSDQualifier_value>#38</INSDQualifier_value>
<NonEnglishQualifier_value>№38</NonEnglishQualifier_value> <NonEnglishQualifier_value>№38</NonEnglishQualifier_value>
</INSDQualifier> </INSDQualifier>
</INSDFeature_quals> </INSDFeature_quals>
</INSDFeature> </INSDFeature>
<INSDFeature> <INSDFeature>
<INSDFeature_key>misc_feature</INSDFeature_key> <INSDFeature_key>misc_feature</INSDFeature_key>
<INSDFeature_location><1</INSDFeature_location> <INSDFeature_location><1</INSDFeature_location>
<INSDFeature_quals> <INSDFeature_quals>
<INSDQualifier id="q201"> <INSDQualifier id="q201">
<INSDQualifier_name>function</INSDQualifier_name> <INSDQualifier_name>function</INSDQualifier_name>
<INSDQualifier_value>NH2-group</INSDQualifier_value> <INSDQualifier_value>NH2-group</INSDQualifier_value>
<NonEnglishQualifier_value>аминогруппа</NonEnglishQualifier_val <NonEnglishQualifier_value>amino group</NonEnglishQualifier_val
ue>ue>
</INSDQualifier> </INSDQualifier>
</INSDFeature_quals> </INSDFeature_quals>
</INSDFeature> </INSDFeature>
</INSDSeq_feature-table> </INSDSeq_feature-table>
<INSDSeq_sequence>gtggatggcggatgcgttgtgcg</INSDSeq_sequence> <INSDSeq_sequence>gtggatggcggatgcgttgtgcg</INSDSeq_sequence>
</INSDSeq> </INSDSeq>
</SequenceData> </SequenceData>
<SequenceData sequenceIDNumber="41"> <SequenceData sequenceIDNumber="41">
<INSDSeq> <INSDSeq>
<INSDSeq_length>27</INSDSeq_length> <INSDSeq_length>27</INSDSeq_length>
<INSDSeq_moltype>DNA</INSDSeq_moltype> <INSDSeq_moltype>DNA</INSDSeq_moltype>
<INSDSeq_division>PAT</INSDSeq_division> <INSDSeq_division>PAT</INSDSeq_division>
<INSDSeq_feature-table> <INSDSeq_feature-table>
<INSDFeature> <INSDFeature>
<INSDFeature_key>source</INSDFeature_key> <INSDFeature_key>source</INSDFeature_key>
<INSDFeature_location>1..27</INSDFeature_location> <INSDFeature_location>1..27</INSDFeature_location>
<INSDFeature_quals> <INSDFeature_quals>
<INSDQualifier> <INSDQualifier>
<INSDQualifier_name>mol_type</INSDQualifier_name> <INSDQualifier_name>mol_type</INSDQualifier_name>
<INSDQualifier_value>other DNA</INSDQualifier_value> <INSDQualifier_value>other DNA</INSDQualifier_value>
</INSDQualifier> </INSDQualifier>
<INSDQualifier id="q203"> <INSDQualifier id="q203">
<INSDQualifier_name>organism</INSDQualifier_name> <INSDQualifier_name>organism</INSDQualifier_name>
<INSDQualifier_value>synthetic construct</INSDQualifier_value> <INSDQualifier_value>synthetic construct</INSDQualifier_value>
</INSDQualifier> </INSDQualifier>
</INSDFeature_quals> </INSDFeature_quals>
</INSDFeature> </INSDFeature>
<INSDFeature> <INSDFeature>
<INSDFeature_key>misc_feature</INSDFeature_key> <INSDFeature_key>misc_feature</INSDFeature_key>
<INSDFeature_location>1..27</INSDFeature_location> <INSDFeature_location>1..27</INSDFeature_location>
<INSDFeature_quals> <INSDFeature_quals>
<INSDQualifier id="q205"> <INSDQualifier id="q205">
<INSDQualifier_name>product</INSDQualifier_name> <INSDQualifier_name>product</INSDQualifier_name>
<INSDQualifier_value>immobilized primer</INSDQualifier_value> <INSDQualifier_value>immobilized primer</INSDQualifier_value>
<NonEnglishQualifier_value>иммобилизованный <NonEnglishQualifier_value>immobilized
праймер</NonEnglishQualifier_value>primer</NonEnglishQualifier_value>
</INSDQualifier> </INSDQualifier>
<INSDQualifier id="q204"> <INSDQualifier id="q204">
<INSDQualifier_name>standard_name</INSDQualifier_name> <INSDQualifier_name>standard_name</INSDQualifier_name>
<INSDQualifier_value>#39</INSDQualifier_value> <INSDQualifier_value>#39</INSDQualifier_value>
<NonEnglishQualifier_value>№39</NonEnglishQualifier_value> <NonEnglishQualifier_value>№39</NonEnglishQualifier_value>
</INSDQualifier> </INSDQualifier>
</INSDFeature_quals> </INSDFeature_quals>
</INSDFeature> </INSDFeature>
<INSDFeature> <INSDFeature>
<INSDFeature_key>misc_feature</INSDFeature_key> <INSDFeature_key>misc_feature</INSDFeature_key>
<INSDFeature_location><1</INSDFeature_location> <INSDFeature_location><1</INSDFeature_location>
<INSDFeature_quals> <INSDFeature_quals>
<INSDQualifier id="q206"> <INSDQualifier id="q206">
<INSDQualifier_name>function</INSDQualifier_name> <INSDQualifier_name>function</INSDQualifier_name>
<INSDQualifier_value>NH2-group</INSDQualifier_value> <INSDQualifier_value>NH2-group</INSDQualifier_value>
<NonEnglishQualifier_value>аминогруппа</NonEnglishQualifier_val <NonEnglishQualifier_value>amino group</NonEnglishQualifier_val
ue>ue>
</INSDQualifier> </INSDQualifier>
</INSDFeature_quals> </INSDFeature_quals>
</INSDFeature> </INSDFeature>
</INSDSeq_feature-table> </INSDSeq_feature-table>
<INSDSeq_sequence>gacgatggccagtaaatcggcgtgggt</INSDSeq_sequence> <INSDSeq_sequence>gacgatggccagtaaatcggcgtgggt</INSDSeq_sequence>
</INSDSeq> </INSDSeq>
</SequenceData> </SequenceData>
<SequenceData sequenceIDNumber="42"> <SequenceData sequenceIDNumber="42">
<INSDSeq> <INSDSeq>
<INSDSeq_length>30</INSDSeq_length> <INSDSeq_length>30</INSDSeq_length>
<INSDSeq_moltype>DNA</INSDSeq_moltype> <INSDSeq_moltype>DNA</INSDSeq_moltype>
<INSDSeq_division>PAT</INSDSeq_division> <INSDSeq_division>PAT</INSDSeq_division>
<INSDSeq_feature-table> <INSDSeq_feature-table>
<INSDFeature> <INSDFeature>
<INSDFeature_key>source</INSDFeature_key> <INSDFeature_key>source</INSDFeature_key>
<INSDFeature_location>1..30</INSDFeature_location> <INSDFeature_location>1..30</INSDFeature_location>
<INSDFeature_quals> <INSDFeature_quals>
<INSDQualifier> <INSDQualifier>
<INSDQualifier_name>mol_type</INSDQualifier_name> <INSDQualifier_name>mol_type</INSDQualifier_name>
<INSDQualifier_value>other DNA</INSDQualifier_value> <INSDQualifier_value>other DNA</INSDQualifier_value>
</INSDQualifier> </INSDQualifier>
<INSDQualifier id="q208"> <INSDQualifier id="q208">
<INSDQualifier_name>organism</INSDQualifier_name> <INSDQualifier_name>organism</INSDQualifier_name>
<INSDQualifier_value>synthetic construct</INSDQualifier_value> <INSDQualifier_value>synthetic construct</INSDQualifier_value>
</INSDQualifier> </INSDQualifier>
</INSDFeature_quals> </INSDFeature_quals>
</INSDFeature> </INSDFeature>
<INSDFeature> <INSDFeature>
<INSDFeature_key>misc_feature</INSDFeature_key> <INSDFeature_key>misc_feature</INSDFeature_key>
<INSDFeature_location>1..30</INSDFeature_location> <INSDFeature_location>1..30</INSDFeature_location>
<INSDFeature_quals> <INSDFeature_quals>
<INSDQualifier id="q210"> <INSDQualifier id="q210">
<INSDQualifier_name>product</INSDQualifier_name> <INSDQualifier_name>product</INSDQualifier_name>
<INSDQualifier_value>immobilized primer</INSDQualifier_value> <INSDQualifier_value>immobilized primer</INSDQualifier_value>
<NonEnglishQualifier_value>иммобилизованный <NonEnglishQualifier_value>immobilized
праймер</NonEnglishQualifier_value>primer</NonEnglishQualifier_value>
</INSDQualifier> </INSDQualifier>
<INSDQualifier id="q209"> <INSDQualifier id="q209">
<INSDQualifier_name>standard_name</INSDQualifier_name> <INSDQualifier_name>standard_name</INSDQualifier_name>
<INSDQualifier_value>#40</INSDQualifier_value> <INSDQualifier_value>#40</INSDQualifier_value>
<NonEnglishQualifier_value>№40</NonEnglishQualifier_value> <NonEnglishQualifier_value>№40</NonEnglishQualifier_value>
</INSDQualifier> </INSDQualifier>
</INSDFeature_quals> </INSDFeature_quals>
</INSDFeature> </INSDFeature>
<INSDFeature> <INSDFeature>
<INSDFeature_key>misc_feature</INSDFeature_key> <INSDFeature_key>misc_feature</INSDFeature_key>
<INSDFeature_location><1</INSDFeature_location> <INSDFeature_location><1</INSDFeature_location>
<INSDFeature_quals> <INSDFeature_quals>
<INSDQualifier id="q211"> <INSDQualifier id="q211">
<INSDQualifier_name>function</INSDQualifier_name> <INSDQualifier_name>function</INSDQualifier_name>
<INSDQualifier_value>NH2-group</INSDQualifier_value> <INSDQualifier_value>NH2-group</INSDQualifier_value>
<NonEnglishQualifier_value>аминогруппа</NonEnglishQualifier_val <NonEnglishQualifier_value>amino group</NonEnglishQualifier_val
ue>ue>
</INSDQualifier> </INSDQualifier>
</INSDFeature_quals> </INSDFeature_quals>
</INSDFeature> </INSDFeature>
</INSDSeq_feature-table> </INSDSeq_feature-table>
<INSDSeq_sequence>ccatctgtgcctcatacaggtccagtgccc</INSDSeq_sequence> <INSDSeq_sequence>ccatctgtgcctcatacaggtccagtgccc</INSDSeq_sequence>
</INSDSeq> </INSDSeq>
</SequenceData> </SequenceData>
<SequenceData sequenceIDNumber="43"> <SequenceData sequenceIDNumber="43">
<INSDSeq> <INSDSeq>
<INSDSeq_length>32</INSDSeq_length> <INSDSeq_length>32</INSDSeq_length>
<INSDSeq_moltype>DNA</INSDSeq_moltype> <INSDSeq_moltype>DNA</INSDSeq_moltype>
<INSDSeq_division>PAT</INSDSeq_division> <INSDSeq_division>PAT</INSDSeq_division>
<INSDSeq_feature-table> <INSDSeq_feature-table>
<INSDFeature> <INSDFeature>
<INSDFeature_key>source</INSDFeature_key> <INSDFeature_key>source</INSDFeature_key>
<INSDFeature_location>1..32</INSDFeature_location> <INSDFeature_location>1..32</INSDFeature_location>
<INSDFeature_quals> <INSDFeature_quals>
<INSDQualifier> <INSDQualifier>
<INSDQualifier_name>mol_type</INSDQualifier_name> <INSDQualifier_name>mol_type</INSDQualifier_name>
<INSDQualifier_value>other DNA</INSDQualifier_value> <INSDQualifier_value>other DNA</INSDQualifier_value>
</INSDQualifier> </INSDQualifier>
<INSDQualifier id="q213"> <INSDQualifier id="q213">
<INSDQualifier_name>organism</INSDQualifier_name> <INSDQualifier_name>organism</INSDQualifier_name>
<INSDQualifier_value>synthetic construct</INSDQualifier_value> <INSDQualifier_value>synthetic construct</INSDQualifier_value>
</INSDQualifier> </INSDQualifier>
</INSDFeature_quals> </INSDFeature_quals>
</INSDFeature> </INSDFeature>
<INSDFeature> <INSDFeature>
<INSDFeature_key>misc_feature</INSDFeature_key> <INSDFeature_key>misc_feature</INSDFeature_key>
<INSDFeature_location>1..32</INSDFeature_location> <INSDFeature_location>1..32</INSDFeature_location>
<INSDFeature_quals> <INSDFeature_quals>
<INSDQualifier id="q215"> <INSDQualifier id="q215">
<INSDQualifier_name>product</INSDQualifier_name> <INSDQualifier_name>product</INSDQualifier_name>
<INSDQualifier_value>immobilized primer</INSDQualifier_value> <INSDQualifier_value>immobilized primer</INSDQualifier_value>
<NonEnglishQualifier_value>иммобилизованный <NonEnglishQualifier_value>immobilized
праймер</NonEnglishQualifier_value>primer</NonEnglishQualifier_value>
</INSDQualifier> </INSDQualifier>
<INSDQualifier id="q214"> <INSDQualifier id="q214">
<INSDQualifier_name>standard_name</INSDQualifier_name> <INSDQualifier_name>standard_name</INSDQualifier_name>
<INSDQualifier_value>#41</INSDQualifier_value> <INSDQualifier_value>#41</INSDQualifier_value>
<NonEnglishQualifier_value>№41</NonEnglishQualifier_value> <NonEnglishQualifier_value>№41</NonEnglishQualifier_value>
</INSDQualifier> </INSDQualifier>
</INSDFeature_quals> </INSDFeature_quals>
</INSDFeature> </INSDFeature>
<INSDFeature> <INSDFeature>
<INSDFeature_key>misc_feature</INSDFeature_key> <INSDFeature_key>misc_feature</INSDFeature_key>
<INSDFeature_location><1</INSDFeature_location> <INSDFeature_location><1</INSDFeature_location>
<INSDFeature_quals> <INSDFeature_quals>
<INSDQualifier id="q216"> <INSDQualifier id="q216">
<INSDQualifier_name>function</INSDQualifier_name> <INSDQualifier_name>function</INSDQualifier_name>
<INSDQualifier_value>NH2-group</INSDQualifier_value> <INSDQualifier_value>NH2-group</INSDQualifier_value>
<NonEnglishQualifier_value>аминогруппа</NonEnglishQualifier_val <NonEnglishQualifier_value>amino group</NonEnglishQualifier_val
ue>ue>
</INSDQualifier> </INSDQualifier>
</INSDFeature_quals> </INSDFeature_quals>
</INSDFeature> </INSDFeature>
</INSDSeq_feature-table> </INSDSeq_feature-table>
<INSDSeq_sequence>tgacggcaaggagctttccggcttctatcagg</INSDSeq_sequenc <INSDSeq_sequence>tgacggcaaggagctttccggcttctatcagg</INSDSeq_sequenc
e>e>
</INSDSeq> </INSDSeq>
</SequenceData> </SequenceData>
<SequenceData sequenceIDNumber="44"> <SequenceData sequenceIDNumber="44">
<INSDSeq> <INSDSeq>
<INSDSeq_length>26</INSDSeq_length> <INSDSeq_length>26</INSDSeq_length>
<INSDSeq_moltype>DNA</INSDSeq_moltype> <INSDSeq_moltype>DNA</INSDSeq_moltype>
<INSDSeq_division>PAT</INSDSeq_division> <INSDSeq_division>PAT</INSDSeq_division>
<INSDSeq_feature-table> <INSDSeq_feature-table>
<INSDFeature> <INSDFeature>
<INSDFeature_key>source</INSDFeature_key> <INSDFeature_key>source</INSDFeature_key>
<INSDFeature_location>1..26</INSDFeature_location> <INSDFeature_location>1..26</INSDFeature_location>
<INSDFeature_quals> <INSDFeature_quals>
<INSDQualifier> <INSDQualifier>
<INSDQualifier_name>mol_type</INSDQualifier_name> <INSDQualifier_name>mol_type</INSDQualifier_name>
<INSDQualifier_value>other DNA</INSDQualifier_value> <INSDQualifier_value>other DNA</INSDQualifier_value>
</INSDQualifier> </INSDQualifier>
<INSDQualifier id="q218"> <INSDQualifier id="q218">
<INSDQualifier_name>organism</INSDQualifier_name> <INSDQualifier_name>organism</INSDQualifier_name>
<INSDQualifier_value>synthetic construct</INSDQualifier_value> <INSDQualifier_value>synthetic construct</INSDQualifier_value>
</INSDQualifier> </INSDQualifier>
</INSDFeature_quals> </INSDFeature_quals>
</INSDFeature> </INSDFeature>
<INSDFeature> <INSDFeature>
<INSDFeature_key>misc_feature</INSDFeature_key> <INSDFeature_key>misc_feature</INSDFeature_key>
<INSDFeature_location>1..26</INSDFeature_location> <INSDFeature_location>1..26</INSDFeature_location>
<INSDFeature_quals> <INSDFeature_quals>
<INSDQualifier id="q220"> <INSDQualifier id="q220">
<INSDQualifier_name>product</INSDQualifier_name> <INSDQualifier_name>product</INSDQualifier_name>
<INSDQualifier_value>immobilized primer</INSDQualifier_value> <INSDQualifier_value>immobilized primer</INSDQualifier_value>
<NonEnglishQualifier_value>иммобилизованный <NonEnglishQualifier_value>immobilized
праймер</NonEnglishQualifier_value>primer</NonEnglishQualifier_value>
</INSDQualifier> </INSDQualifier>
<INSDQualifier id="q219"> <INSDQualifier id="q219">
<INSDQualifier_name>standard_name</INSDQualifier_name> <INSDQualifier_name>standard_name</INSDQualifier_name>
<INSDQualifier_value>#42</INSDQualifier_value> <INSDQualifier_value>#42</INSDQualifier_value>
<NonEnglishQualifier_value>№42</NonEnglishQualifier_value> <NonEnglishQualifier_value>№42</NonEnglishQualifier_value>
</INSDQualifier> </INSDQualifier>
</INSDFeature_quals> </INSDFeature_quals>
</INSDFeature> </INSDFeature>
<INSDFeature> <INSDFeature>
<INSDFeature_key>misc_feature</INSDFeature_key> <INSDFeature_key>misc_feature</INSDFeature_key>
<INSDFeature_location><1</INSDFeature_location> <INSDFeature_location><1</INSDFeature_location>
<INSDFeature_quals> <INSDFeature_quals>
<INSDQualifier id="q221"> <INSDQualifier id="q221">
<INSDQualifier_name>function</INSDQualifier_name> <INSDQualifier_name>function</INSDQualifier_name>
<INSDQualifier_value>NH2-group</INSDQualifier_value> <INSDQualifier_value>NH2-group</INSDQualifier_value>
<NonEnglishQualifier_value>аминогруппа</NonEnglishQualifier_val <NonEnglishQualifier_value>amino group</NonEnglishQualifier_val
ue>ue>
</INSDQualifier> </INSDQualifier>
</INSDFeature_quals> </INSDFeature_quals>
</INSDFeature> </INSDFeature>
</INSDSeq_feature-table> </INSDSeq_feature-table>
<INSDSeq_sequence>ggacatggacgagcgcgactatcggg</INSDSeq_sequence> <INSDSeq_sequence>ggacatggacgagcgcgactatcggg</INSDSeq_sequence>
</INSDSeq> </INSDSeq>
</SequenceData> </SequenceData>
<SequenceData sequenceIDNumber="45"> <SequenceData sequenceIDNumber="45">
<INSDSeq> <INSDSeq>
<INSDSeq_length>23</INSDSeq_length> <INSDSeq_length>23</INSDSeq_length>
<INSDSeq_moltype>DNA</INSDSeq_moltype> <INSDSeq_moltype>DNA</INSDSeq_moltype>
<INSDSeq_division>PAT</INSDSeq_division> <INSDSeq_division>PAT</INSDSeq_division>
<INSDSeq_feature-table> <INSDSeq_feature-table>
<INSDFeature> <INSDFeature>
<INSDFeature_key>source</INSDFeature_key> <INSDFeature_key>source</INSDFeature_key>
<INSDFeature_location>1..23</INSDFeature_location> <INSDFeature_location>1..23</INSDFeature_location>
<INSDFeature_quals> <INSDFeature_quals>
<INSDQualifier> <INSDQualifier>
<INSDQualifier_name>mol_type</INSDQualifier_name> <INSDQualifier_name>mol_type</INSDQualifier_name>
<INSDQualifier_value>other DNA</INSDQualifier_value> <INSDQualifier_value>other DNA</INSDQualifier_value>
</INSDQualifier> </INSDQualifier>
<INSDQualifier id="q223"> <INSDQualifier id="q223">
<INSDQualifier_name>organism</INSDQualifier_name> <INSDQualifier_name>organism</INSDQualifier_name>
<INSDQualifier_value>synthetic construct</INSDQualifier_value> <INSDQualifier_value>synthetic construct</INSDQualifier_value>
</INSDQualifier> </INSDQualifier>
</INSDFeature_quals> </INSDFeature_quals>
</INSDFeature> </INSDFeature>
<INSDFeature> <INSDFeature>
<INSDFeature_key>misc_feature</INSDFeature_key> <INSDFeature_key>misc_feature</INSDFeature_key>
<INSDFeature_location>1..23</INSDFeature_location> <INSDFeature_location>1..23</INSDFeature_location>
<INSDFeature_quals> <INSDFeature_quals>
<INSDQualifier id="q225"> <INSDQualifier id="q225">
<INSDQualifier_name>product</INSDQualifier_name> <INSDQualifier_name>product</INSDQualifier_name>
<INSDQualifier_value>immobilized primer</INSDQualifier_value> <INSDQualifier_value>immobilized primer</INSDQualifier_value>
<NonEnglishQualifier_value>иммобилизованный <NonEnglishQualifier_value>immobilized
праймер</NonEnglishQualifier_value>primer</NonEnglishQualifier_value>
</INSDQualifier> </INSDQualifier>
<INSDQualifier id="q224"> <INSDQualifier id="q224">
<INSDQualifier_name>standard_name</INSDQualifier_name> <INSDQualifier_name>standard_name</INSDQualifier_name>
<INSDQualifier_value>#43</INSDQualifier_value> <INSDQualifier_value>#43</INSDQualifier_value>
<NonEnglishQualifier_value>№43</NonEnglishQualifier_value> <NonEnglishQualifier_value>№43</NonEnglishQualifier_value>
</INSDQualifier> </INSDQualifier>
</INSDFeature_quals> </INSDFeature_quals>
</INSDFeature> </INSDFeature>
<INSDFeature> <INSDFeature>
<INSDFeature_key>misc_feature</INSDFeature_key> <INSDFeature_key>misc_feature</INSDFeature_key>
<INSDFeature_location><1</INSDFeature_location> <INSDFeature_location><1</INSDFeature_location>
<INSDFeature_quals> <INSDFeature_quals>
<INSDQualifier id="q226"> <INSDQualifier id="q226">
<INSDQualifier_name>function</INSDQualifier_name> <INSDQualifier_name>function</INSDQualifier_name>
<INSDQualifier_value>NH2-group</INSDQualifier_value> <INSDQualifier_value>NH2-group</INSDQualifier_value>
<NonEnglishQualifier_value>аминогруппа</NonEnglishQualifier_val <NonEnglishQualifier_value>amino group</NonEnglishQualifier_val
ue>ue>
</INSDQualifier> </INSDQualifier>
</INSDFeature_quals> </INSDFeature_quals>
</INSDFeature> </INSDFeature>
</INSDSeq_feature-table> </INSDSeq_feature-table>
<INSDSeq_sequence>tggacgcagaatcgcaccgggtg</INSDSeq_sequence> <INSDSeq_sequence>tggacgcagaatcgcaccgggtg</INSDSeq_sequence>
</INSDSeq> </INSDSeq>
</SequenceData> </SequenceData>
</ST26SequenceListing></ST26SequenceListing>
<---<---
Claims (6)
Publications (1)
| Publication Number | Publication Date |
|---|---|
| RU2807998C1 true RU2807998C1 (en) | 2023-11-21 |
Family
ID=
Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| RU2405836C2 (en) * | 2008-05-12 | 2010-12-10 | Федеральное государственное учреждение "Санкт-Петербургский научно-исследовательский институт фтизиопульмонологии Федерального агентства по высокотехнологичной медицинской помощи" | Method for mycobacterium tuberculosis beijing genotype detection |
| CN108977503A (en) * | 2018-06-25 | 2018-12-11 | 河南省疾病预防控制中心 | A kind of mycobacterium tuberculosis spacer oligonucleotide typing method of oligonucleotide probe hybridization |
Patent Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| RU2405836C2 (en) * | 2008-05-12 | 2010-12-10 | Федеральное государственное учреждение "Санкт-Петербургский научно-исследовательский институт фтизиопульмонологии Федерального агентства по высокотехнологичной медицинской помощи" | Method for mycobacterium tuberculosis beijing genotype detection |
| CN108977503A (en) * | 2018-06-25 | 2018-12-11 | 河南省疾病预防控制中心 | A kind of mycobacterium tuberculosis spacer oligonucleotide typing method of oligonucleotide probe hybridization |
Non-Patent Citations (1)
| Title |
|---|
| Bespyatykh JA, Zimenkov DV, Shitikov EA, Kulagina EV, Lapa SA, Gryadunov DA, Ilina EN, Govorun VM. 2014. Spoligotyping of Mycobacterium tuberculosis complex isolates using hydrogel oligonucleotide microarrays. Infect Genet Evol 26:41-46. doi: 10.1016/j.meegid.2014.04.024. * |
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