CN110662835B - Engineering minimized SaCas9 CRISPR/Cas systems for gene editing and transcriptional regulation optimized by enhanced guide RNAs - Google Patents

Engineering minimized SaCas9 CRISPR/Cas systems for gene editing and transcriptional regulation optimized by enhanced guide RNAs Download PDF

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CN110662835B
CN110662835B CN201780091204.2A CN201780091204A CN110662835B CN 110662835 B CN110662835 B CN 110662835B CN 201780091204 A CN201780091204 A CN 201780091204A CN 110662835 B CN110662835 B CN 110662835B
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谢震
马大程
彭曙光
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Abstract

Programmable and precise modulation of Cas9 function by utilizing a set of compact Cas9 derivatives generated by deletion of conserved HNH and/or REC-C domains based on structural information across multiple class 2 CRISPR effectors is provided. Also provided are new strategies for engineering dimeric gRNA-directed nucleases by fragmenting mini-dscas 9 and fusing the fokl domain immediately after the cleavage point to increase target DNA cleavage efficiency and potentially reduce off-target effects due to the closer proximity of the dimeric fokl nuclease to the target sequence. By combining an optimized compact gRNA expression cassette with a reduced size SaCas9 derivative, the entire CRISPR/Cas system with different effector domains for transactivation, DNA cleavage and base editing is loaded into a single AAV virus. Such use of only one AAV-CRISPR/Cas9 system is particularly attractive in biomedical applications where safe and effective in vivo delivery is required.

Description

Engineering minimized SaCas9 CRISPR/Cas systems for gene editing and transcriptional regulation optimized by enhanced guide RNAs
Technical Field
The present application relates to the field of biology, in particular to directed modification of genetic material. More particularly, the claimed invention relates to improvements to Cas9 CRISPR-associated proteins and related products thereof.
Background
CRISPR-associated protein 9 (Cas 9) found in streptococcus pyogenes (Streptococcus pyogenes) is a multidomain protein that has been widely used for genome editing and transcriptional control of mammalian cells due to its excellent modularity and versatility. Due to size limitations, the in vivo delivery of synthetic gene lines is limited, especially for smaller delivery systems with payload capacity nearly equal to the entire Cas9 complex.
Disclosure of Invention
Technical problem
Various strategies have been developed to engineer modular and stratified gene circuits in mammalian cells by modulating dCas9 and gRNA expression. Transcriptional control in mammalian cells can be achieved by direct fusion of the transcriptional regulatory domain with nuclease-inactivated Cas9 (dCas 9). Alternatively, multiple transcriptional regulatory domains can be recruited to dCas9 by labeling dCas9 with a repeat peptide scaffold or by fusing repeat RNA motifs to homologous grnas. However, biomedical applications of CRISPR/Cas systems require exploration of new platforms for engineering mammalian synthetic lines that integrate and process multiple endogenous inputs. In addition, the application of CRISPR/Cas therapy lines is also challenging due to the limited cargo size of existing viral delivery vectors.
Solution to the problem
The split Cas9 system is generally useful to bypass packaging limitations of viral delivery vectors and in the claimed invention dCas9 is split and reconstituted in human cells. One of the challenges of therapeutic applications is to find an optimal delivery system that can carry all CRISPR/Cas9 components to a desired organ or cell population for gene manipulation. When incorporated into a viral delivery system, the use of CRISPR/Cas systems with maximum potential is greatly limited by their physical dimensions. When used for synthetic biological purposes in high value delivery systems with site-specific integration (e.g., adeno-associated virus/AAV), the entire cas9 complex resembles a computer manipulation system, which occupies 95% of the available memory, leaving only a small portion for synthetic biological programming purposes. By dividing CRISPR/CAS9 into multiple smaller regions and delivering each region in separate viral delivery vectors, this retains powerful gene manipulation functions while greatly increasing the space for cell programming purposes. The claimed invention shows substantial improvements over existing CAS9 delivery techniques and includes other enhancements to gene control and programming.
Although a variety of viral delivery systems have been employed and have met with varying success, the implementation of systems that rely on alternative viral systems may lead to an undesirably strong immune response. The use of recombinant adeno-associated virus (rAAV) has high gene transfer efficiency and very low immune response. Unfortunately, the packaging capacity is limited to 4.7kb to 5kb, optimizing Cas9 size relative to humans over 4.2kb and adding promoter sequences up to the limit of over 5kb is problematic. With intein-mediated splitting of Cas9, the intein acts as a protein intron and is excised from the sequence and linked to the remaining flanking region (the extein) by peptide bonds without scarring. Regarding the selection of the cleavage sites, special attention is paid to the surface-exposed cleavage sites due to the spatial requirements of protein splicing. Such a system allows the coding sequence of Cas9 to be distributed over a dual-vector or multi-vector system and reconstructed post-translationally.
The claimed invention expands the accessibility of synthetic biology through targeted specific diagnostic and therapeutic applications via improved and higher level gene line delivery enhancement of the gene line. The presently claimed embodiments overcome the existing size limitations by optimal splitting of Cas9, allowing higher levels of synthetic gene lines to be accommodated by smaller delivery systems.
The presently claimed invention utilizes a reduced size staphylococcus aureus Cas9 variant (mini-SaCas 9) that retains DNA binding activity by deleting a conserved functional domain. In a preferred illustrative embodiment, a fokl nuclease domain is fused to the middle of split mini-SaCas9 to trigger efficient DNA cleavage. In another illustrative embodiment, the gene editing system is small enough to be housed in a single AAV comprising mini-SaCas9 fused to a reduced-size transactivation domain and an optimized compact gRNA expression cassette with effective transactivation activity. The claimed invention highlights a practical approach to generating a single AAV-CRISPR/Cas9 system with different effector domains for in vivo applications.
To bypass AAV payload restrictions, 4.2kb Cas9 (SpCas 9) from streptococcus pyogenes (Streptococcus pyogenes) was split and packaged with guide RNA (gRNA) expression units into two separate AAV, which allowed functional reconstitution of full length SpCas9 in vivo. Another strategy is to find natural class 2 CRISPR effectors of reduced size, such as 3.2-kb SaCas9 and-3-kb CasX identified in uncultured organisms by using metagenomic datasets. To further reduce the transgene size, the expression of a tRNA: gRNA fusion transcript that is cleaved by endogenous tRNAse Z to produce active gRNA can be driven using a 70-bp glutamine tRNA in place of the 250bp RNA polymerase III promoter. These efforts have facilitated the construction of only one AAV delivery vector for in vivo applications of CRISPR/Cas technology.
Recent structural studies of SpCas9, saCas9 and amino acid coccus sp. Cpf1 (AsCpf 1) elucidated the function of the conserved domains (including HNH/NUC and RuvC nuclease domains that cleave complementary and non-complementary DNA strands, respectively), recognition (REC) domains and pre-spacer adjacent motif (PAM) interaction (PI) domains in these 2 classes of CRISPR effectors. Interestingly, essentially the complete DNA binding activity or half-cleavage activity was retained by the truncated SpCas9 mutants lacking HNH or REC2 domains. These results highlight the possibility of minimizing Cas9 (mini-Cas 9) to further size down wild-type Cas9 to have only DNA binding activity and no DNA cleavage activity, allowing the accommodation of other DNA templates, effector domains, and control elements in a single AAV vector.
Advantageous effects of the invention
Such CRISPR/Cas9 systems have particular utility in biomedical applications where viral delivery vectors with limited cargo sizes are preferred. Contemplated variants include combinations of split Cas9/dCas9 systems with rAAV delivery systems, where the activity of Cas9/dCas9 can be controlled to edit and regulate endogenous genes in vivo.
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The accompanying drawings are included to better illustrate illustrative embodiments of the claimed invention.
Figure 1 is a graph of EBFP2 transcriptional activation assay of compact Cas9 derivatives fused to VPR domains.
FIG. 2 is a graph of dSpCas9, mini-dSpCas9-1, and mini-dSpCas9-2 domain organization and corresponding gene activation efficiencies.
Fig. 3 is a schematic diagram of domain organization of dscas 9 and its derivatives and the resulting gene activation efficiencies.
FIG. 4 is a graph of the organization of the dASCpf1 and mini-AsCpf1-1 domains and the corresponding gene activation efficiencies.
FIG. 5 is a schematic representation of optimized gRNA and corresponding gene activation efficiency.
FIG. 6 is a diagram of the use of glutamine (Glu) tRNA as a promoter for expression optimized gRNA-2.
FIG. 7 is a diagram of domain organization including segmented dSaCas9 fused to FokI domains and segmented mini-dSaCas9-4 dSaCas9 variants.
FIG. 8 is a graph of EYFP reconstitution assay demonstrating DNA cleavage efficiency and a graphical representation of dimeric FokI nuclease fused to split dSaCas9 and split minidSaCas9-4 versus DNA cleavage.
FIG. 9 is a graph of DNA cleavage efficiency for split dSaCas9 variants with spacer lengths in the range of 12-bp to 24-bp.
FIG. 10 is a schematic representation of the transcriptional activation domains of VPR, VTR1, VTR2 and VTR3 and their corresponding gene activation efficiencies assessed using the EBFP2 reporting system.
FIG. 11 is a schematic of a repeated array of SpyTag and MoonTag for transcriptional activation and corresponding gene activation efficiency data.
Figure 12 is a schematic of transcriptional activation by using a single AAV loaded with a compact CRISPR/Cas9 system and a graphical representation of EBFP2 activation efficiency.
FIG. 13 is a schematic of packaging mini-Cas9, effector domains, gRNA expression cassettes, and other parts in a single AAV vector for transcriptional activation, DNA cleavage, and base editing.
Fig. 14 is a schematic representation of miniscas 9-3 and split SaCas9 and the corresponding miniscas 9-3 and split SaCas9 activation efficiencies.
FIG. 15 is a gene sequence diagram of the CCR5 gene showing the spacer and corresponding gel agarose map data.
Detailed Description
Rational design of the compact CRISPR/Cas9 system is further detailed in the following embodiments as further detailed in the corresponding figures.
Fig. 1 is a graph of EBFP2 transcriptional activation assay of compact Cas9 derivatives fused to VPR domains (101). Constitutively expressed mKate2 was used as transfection control. In a first illustrative example, the reporting system is used in cultured human embryonic kidney 293 (HEK 293) cells. Two mini-dSpCas9 were generated by deleting the C-terminal region (REC-C,. DELTA.501-710) and HNH domain (,. DELTA.777-891) of the REC1 domain that was not necessary for the DNA binding activity of nuclease-inactivated Cas9 (dCAS 9), respectively, and the two mini-dSpCas9 were fused to VP64-p65-Rta (VPR) transactivation domains, respectively.
FIG. 2 is a graph of dSpCas9, mini-dSpCas9-1, and mini-dSpCas9-2 domain organization (201) and a graphical representation of the corresponding gene activation efficiencies (203). In this illustrative example, both mini-dSpCas9:VPR variants retain more than 50% of transactivation capacity as compared to dSpCas 9:VPR.
Fig. 3 is a schematic representation of domain organization of dscas 9 and its derivatives (301) and a diagram describing the efficiency of gene activation (303). Recently, wild-type SaCas9 has been engineered to recognize altered PAM ("NNNRRT"). Based on the SaCas9 mutant, the disclosed embodiments construct mini-SaCas9-1:VPR, which retains only-2.5% of the transactivation activity of dSaCas9:VPR, by replacing the conserved REC-C domain (Δ234-444) with a "GGGGSGGGG" linker (GS-linker). Although the GS linker is widely used as a flexible linker, it may still deform the SaCas9 structure. Inspired by a computational scheme recently called search, the existing structure between discontinuous Cas9 fragments is searched using the neighbor residue search (ARS) scheme. Substitution of the GS linker with a "KRRRRHR" (R linker) from the SaCas9 BH domain suitable for filling in the REC-C deletion gap by using the ARS protocol results in an 11-fold increase in transactivation capacity of mini-SaCas 9-2. As an enhanced example of the claimed invention, by using SAR protocols, a "GSK linker" derived from a putative gene (accession No. O67859) in liquid aeolian (Aquifex aeolicus) was used, which fits into the deletion gap (Δ479-649) of the HNH domain. By deleting both REC-C domain and HNH domain a 2-kb mini-SaCas9-3 was produced which retained only 0.6% transactivation activity relative to the control.
FIG. 4 is a graph of the organization of the dASCpf1 and mini-AsCpf1-1 domains (401) and the corresponding gene activation efficiencies (403). To evaluate the DNA cleavage efficiency of the mini-SaCas9 variants in this illustrative example, a report reconstitution assay was used, wherein DNA cleavage can trigger reconstitution of an activity-Enhanced Yellow Fluorescent Protein (EYFP) reporter gene from an inactive form. Deletion of REC-C or both REC-C and HNH domains resulted in background EYFP expression, indicating that domain deletion abrogated DNA cleavage activity of mini-SaCas9 variants. In this illustrative example, the transactivation capacity of dASCpf1:VPR was retained at 46% by deletion of the REC2 domain (. DELTA.324-525), indicating that the deletion strategy was applicable to unique class 2 CRISPR effectors.
FIG. 5 is a schematic representation of optimized gRNA (501) and a corresponding representation of gene activation efficiency (503). A-U flip or U-G conversion can be introduced to disrupt the putative RNA Pol III terminator sequence in the first stem loop of the gRNA scaffold, thereby enhancing the efficiency of dCAS 9-mediated DNA labeling. Putative RNA Pol III terminator sequences are shown in boxes with A-U and G-C mutations marked with shading at the third and fourth positions. Representative scatter plots of flow cytometry data are also shown on the right. Constitutively expressed mKate2 was used as transfection control. In a preferred illustrative embodiment, the transactivation efficiency of miniSaCas9-3 is increased to about 100-fold by introducing one or two point mutations in the putative RNA Pol III terminator sequence.
FIG. 6 is a diagram of glutamine (Glu) tRNA (601) used as a promoter for expression of optimized gRNA-2. In this illustrative example, HEK293 cells are co-transfected with plasmids expressing mini-SaCas9-3:VPR and gRNA-2 driven by Glu RNA or U6 promoter. A representative flow cytometry scatter plot (603) is shown on the right. Constitutively expressed iRFP was used as transfection control. Data are shown as mean ± SEM fold change of EBFP2 fluorescence measured using flow cytometry in three independent replicates 48 hours after transfection into HEK293 cells. In this illustrative example, a transactivation activity of 50% is obtained when glutamine tRNA is used to drive gRNA expression instead of the U6 promoter.
Improving the DNA cleavage specificity of the CRISPR/Cas9 system is crucial for future clinical applications. Dimerization of hybrid proteins in which the fokl nuclease domain is fused to the N-terminus of dsbacas 9 but not to the C-terminus improves DNA cleavage specificity in the PAM-out orientation. In addition, truncated grnas with shorter target complementarity regions reduce off-target cleavage efficiency.
In the following illustrative examples, the effect of compact SaCas9 derivatives on DNA cleavage and base editing is further disclosed.
FIG. 7 is a diagram of the domain organization of the dSaCas9 variant (701) comprising segmented dSaCas9 and segmented mini-dSaCas9-4 fused to the FokI domain and further comprising Fok1 fused to the N-terminus of dSaCas9 and mini-dSaCas 9-4.
FIG. 8 is a graph of EYFP reconstitution assay demonstrating DNA cleavage efficiency (801) and DNA cleavage by dimeric FokI nucleases fused to either split dSaCas9 or split miniSaCas 9-4803). The EYFP reconstitution assay evaluates the DNA cleavage efficiency of the dscas 9 derivative fused to fokl and two truncated grnas each comprising an 18-nt sequence complementary to the target. In one illustrative embodiment, no DNA cleavage activity is detected when FokI is fused to the N-terminus of mini-SaCas 9-2. One possible reason is that the fokl in this protein structure may be located distally to the target DNA. However, since the predicted distance between the N-and C-termini of the FokI nuclease domain is
Figure BDA0002287271310000061
This makes finding a suitable insertion location in the middle of dscas 9 challenging. In an alternative embodiment, dscas 9 is split at residue 733 and fokl is fused after the split point using a triplicate G-linker ("GGGGS"). In another illustrative embodiment, the first four residues (Delta734-737) of the C-terminal fragment that could interfere with the reconstruction of the two split fragments are removed. The split dscas 9 or split mini-dscas 9-4 without HNH domain resulted in about 8% to 30% of wild-type SaCas9 induced EYFP expression level, with a spacer in the PAM outer direction in the range of 12bp to 24 bp.
FIG. 9 is a graphical representation of the DNA cleavage efficiency of split dSaCas9 variants with spacer lengths of 12-bp to 24-bp (901). Division of spacer length from 12-bp to 24-bp dSaCas9:FokI, division of mini-dSaCas9-4:FokI, and DNA cleavage efficiency of Fok1 fusion with the N-terminus of dSaCas9 and mini-dSacas 9-4. Each bar shows the mean fold change in EYFP fluorescence measured by flow cytometry 48 hours after HEK293 cell transfection (mean ± SEM; n=3).
In the following illustrative embodiments, the construction of a compact CRISPR/Cas system for transcriptional activation is described in further detail.
FIG. 10 is a schematic representation of the transcriptional activation domains of VPR, VTR1, VTR2 and VTR3 (1001), and a graphical representation of the corresponding gene activation efficiencies assessed by using the EBFP2 reporting system (1003). In this illustrative example, a compact transcription activator based on dCas9-VPR was designed. The entire P65 comprises a DNA binding domain at the N-terminus and two transactivation domains at the C-terminus (TA 1 and TA 2). However, only TA2 and part TA1 are contained in the three-part VPR domain. To reduce the size of dCas9-VPR, mini-SaCas9-3:vtr1 can be constructed by replacing the P65 domain in VPR with TA1 and TA2 domains. In another illustrative embodiment, the P65 domain in VPR is constructed with two repeating TA1 domains and is referred to as VTR2. The VTR1 domain and VTR2 domain retain 45% and 21% of the transactivation efficiency of the VPR domain, respectively. Also described is VTR3, which is about 200bp shorter than VTR2 and VTR1, making it the best illustrative embodiment.
FIG. 11 is a schematic representation of a repetition of SpyTag and MoonTag arrays for transcriptional activation (1101), and a graphical representation of corresponding gene activation efficiency data (1103). In an illustrative embodiment, the corresponding gene activation efficiency is assessed by using an EBFP2 reporting system using optimized gRNA-2. The data are shown as mean fold change in EBFP2 fluorescence measured with flow cytometry 72 hours after transfection into HEK293 cells (mean ± SEM; n=3). Recently, a 13 residue peptide tag (SpyTag) derived from streptococcus pyogenes fibronectin binding protein (FbaB) has been shown to form a covalent bond with a 116 residue binding partner known as SpyCatcher.
In an illustrative example, a repetitive peptide array of smaller size than the SunTag system was constructed by fusing four tandem repeats of SpyTag to the C-terminus of mini-SaCas9-3 and SpyCatcher to the VPR domain, allowing spontaneous assembly of the VPR transactivation scaffold in cells. The SpyTag system induced 100-fold enhanced expression of the blue fluorescent protein 2 (EBFP 2) reporter gene compared to the negative control. In the illustrative example, a putative protein (accession number WP 054278706) with 60% sequence similarity to FbaB was found from streptococcus seal (Streptococcus phocae) by searching for homologs of SpyTag and SpyCatcher. In the illustrative embodiment, the protein is split similar to SpyTag and Spycatcher. As a direct and expected result of the illustrative examples, a similar scaffold system, known as the MoonTag system, is disclosed herein and is achieved by fusing four tandem repeats of a 13-residue MoonTag with mini-SaCas9-3 and complexing the MoonCatcher and VPR domains. Although the Moontag system is not orthogonal to the SpyTag system, the Moontag system increases the efficiency of activating EBFP2 expression by a factor of 5.
Fig. 12 is a schematic representation of transcriptional activation by using a single AAV loaded with a compact CRISPR/Cas9 system (1201), and a graphical representation of EBFP2 activation efficiency (1203). In the illustrative example, transcriptional activation by using a single AAV loaded with a compact CRISPR/Cas9 system comprising mini-SaCas9-3:vtr1 and U6 driven optimized gRNA-2 is depicted. Plasmid DNA encoding the EBFP2 reporter gene and the mKate2 control gene was introduced into HEK293 cells by transient transfection followed by AAV infection. The lower panel shows the activation efficiency of EBFP2 and representative microscopic image after AAV transfection encoding mini-SaCas9-3:vtr1 (1205). The scale bar in the image represents 200 μm. Data are shown as mean fold change in EBFP2 fluorescence measured using flow cytometry 72 hours after transfection and AAV infection (mean ± SEM; n=3). In this illustrative embodiment, a single AAV virus encoding constitutively expressed mini-SaCas9-3:VTR1 and optimized gRNA targeting the TRE promoter is produced. The TRE-driven EBFP2 reporter gene was introduced into HEK293 cells by transient transfection. In an illustrative embodiment, AAV infection activates EBFP2 expression up to-130 fold compared to negative controls in HEK293 cells, which is also more efficient than AAV viruses loaded with mini-SaCas 9-3:vp64.
FIG. 13 is a schematic representation of packaging mini-Cas9, effector domains, gRNA expression cassettes, and other parts in a single AAV vector for transcriptional activation, DNA cleavage, and base editing (1301). By combining the illustrative examples with a broad working example, a set of compact Cas9 derivatives was engineered by deleting conserved HNH and/or REC-C domains based on structural information across multiple class 2 CRISPR effectors. In addition, a new strategy for engineering dimeric gRNA-directed nucleases by partitioning mini-dSaCas9 and fusing the FokI domain just behind the partitioning point is disclosed, which can increase target DNA cleavage efficiency and potentially reduce off-target efficiency due to the closer proximity of the dimeric FokI nuclease to the target sequence. By combining an optimized compact gRNA expression cassette with a reduced size SaCas9 derivative, practical methods of loading an entire CRISPR/Cas system with different effector domains for transactivation, DNA cleavage and base editing into a single AAV virus are disclosed. Such use of only one AAV-CRISPR/Cas9 system is particularly attractive in biomedical applications where safe and effective in vivo delivery is required.
Fig. 14 is a schematic representation of miniscas 9-3 and split SaCas9 (1401) and graphical data of the respective miniscas 9-3 and split SaCas9 activation efficiencies (1403).
FIG. 15 is a graphical representation of the gene sequence of the CCR5 gene showing the spacer (1501), and gel electrophoresis graphical data showing the relative percentage of cleavage of the CCR5 gene (1503).
In the description, numerous specific details are set forth in order to provide a thorough understanding of embodiments of the invention. However, it will be apparent to one of ordinary skill in the art that the specific details need not be employed to practice the present embodiments. In other instances, well-known materials or methods have not been described in detail in order to avoid obscuring embodiments of the present invention.
Reference throughout this specification to "one embodiment," "an embodiment," "one example," or "an example" means that a particular feature, structure, or characteristic described in connection with the embodiment or example is included in at least one embodiment of the present invention. Thus, the appearances of the phrases "in one embodiment," "in an embodiment," "one example," or "an example" in various places throughout this specification are not necessarily all referring to the same embodiment or example. Furthermore, the particular features, structures, or characteristics may be combined in any suitable combination and/or sub-combination in one or more embodiments or examples. Additionally, it should be understood that the drawings provided herewith are for explanation purposes to persons ordinarily skilled in the art and that the drawings are not necessarily drawn to scale.
As used herein, the terms "comprises," "comprising," "includes," "including," "having," or any other variation thereof, are intended to cover a non-exclusive inclusion. For example, a process, article, or apparatus that comprises a list of elements is not necessarily limited to only those elements but may include other elements not expressly listed or inherent to such process, article, or apparatus.
In addition, any examples or examples set forth herein should not be construed as in any way limiting, restricting, or expressing definitions of any term or terms used therewith. Rather, these examples or illustrations should be considered as being described with respect to one particular embodiment and are merely exemplary. One of ordinary skill in the art will understand that one or more terms used with these examples or illustrations will encompass other embodiments that may not be presented with them or elsewhere in the specification, and that all such embodiments are intended to be included within the scope of the one or more terms. Languages that specify such non-limiting examples or examples include, but are not limited to: "for example," such as, "" as, "and" in one embodiment.
INDUSTRIAL APPLICABILITY
The claimed invention has industrial applicability in biomedical and industrial biotechnology applications. Because of the higher specificity, the improved Cas9 system provides greater gene editing and regulatory control capabilities than conventional Cas9 systems.
Free text of sequence Listing
The present application contains a sequence listing that has been submitted in ASCII format and is incorporated herein by reference in its entirety. The ASCII copy was created on 28 th 2017, named ZX2seqlist_ST25.Txt, 225KB in size.
The sequences to which this patent relates are as follows.
Sequence listing
Seq ID 1gRNAa TACGTTCTCTATCACTGATA
Seq ID 2gRNAb TACGTTCTCTATCACTGATA
Seq ID 3crRNAa CTCCCTATCAGTGATAGAGAACG
Seq ID 4gRNAc CGTTCTCTATCACTGATA
Seq ID 5gRNAd ACTAGAAATTCACCGAGC
Seq ID 6gRNA-15bp CTCACTCAACAGTGATAGAGA
Seq ID 7gRNA-12bp TTGCTCACTCAACAGTGATAG
Seq ID 8
dSpCas9:VPR
MPKKKRKVGGGSPGMDKKYSIGLAIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKKNLIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEESFLVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIKFRGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRLENLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQIGDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVRQQLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLLRKQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARGNSRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEYFTVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFDSVEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEERLKTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFMQLIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGRHKPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLYLYYLQNGRDMYVDQELDINRLSDYDVDAIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVPSEEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKHVAQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYLNAVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKTEITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSKESILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGITIMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNELALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILADANLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVLDATLIHQSITGLYETRIDLSQLGGDTSRADPKKKRKVEASGSGRADALDDFDLDMLGSDALDDFDLDMLGSDALDDFDLDMLGSDALDDFDLDMLINSRSQYLPDTDDRHRIEEKRKRTYETFKSIMKKSPFSGPTDPRPPPRRIAVPSRSSASVPKPAPQPYPFTSSLSTINYDEFPTMVFPSGQISQASALAPAPPQVLPQAPAPAPAPAMVSALAQAPAPVPVLAPGPPQAVAPPAPKPTQAGEGTLSEALLQLQFDDEDLGALLGNSTDPAVFTDLASVDNSEFQQLLNQGIPVAPHTTEPMLMEYPEAITRLVTGAQRPPDPAPAPLGAPGLPNGLLSGDEDFSSIADMDFSALLGSGSGSRDSREGMFLPKPEAGSAISDVFEGREVCQPKRIRPFHPPGSPWANRPLPASLAPTPTGPVHEPVGSLTPAPVPQPLDPAPAVTPEASHLLEDPDEETSQAVKALREMADTVIPQKEEAAICGQMDLSHPPPRGHLDELTTTLESMTEDLNLDSPLTPELNEILDTFLNDECLLHAMHISTGLSIFDTSLF
Seq ID 9
mini-dSpCas9-1:VPR
MPKKKRKVGGGSPGMDKKYSIGLAIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKKNLIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEESFLVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIKFRGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRLENLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQIGDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVRQQLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLLRKQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARGNSRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGRHKPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLYLYYLQNGRDMYVDQELDINRLSDYDVDAIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVPSEEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKHVAQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYLNAVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKTEITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSKESILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGITIMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNELALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILADANLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVLDATLIHQSITGLYETRIDLSQLGGDTSRADPKKKRKVEASGSGRADALDDFDLDMLGSDALDDFDLDMLGSDALDDFDLDMLGSDALDDFDLDMLINSRSQYLPDTDDRHRIEEKRKRTYETFKSIMKKSPFSGPTDPRPPPRRIAVPSRSSASVPKPAPQPYPFTSSLSTINYDEFPTMVFPSGQISQASALAPAPPQVLPQAPAPAPAPAMVSALAQAPAPVPVLAPGPPQAVAPPAPKPTQAGEGTLSEALLQLQFDDEDLGALLGNSTDPAVFTDLASVDNSEFQQLLNQGIPVAPHTTEPMLMEYPEAITRLVTGAQRPPDPAPAPLGAPGLPNGLLSGDEDFSSIADMDFSALLGSGSGSRDSREGMFLPKPEAGSAISDVFEGREVCQPKRIRPFHPPGSPWANRPLPASLAPTPTGPVHEPVGSLTPAPVPQPLDPAPAVTPEASHLLEDPDEETSQAVKALREMADTVIPQKEEAAICGQMDLSHPPPRGHLDELTTTLESMTEDLNLDSPLTPELNEILDTFLNDECLLHAMHISTGLSIFDTSLF
Seq ID 10
mini-dSpCas9-2:VPR
MPKKKRKVGGGSPGMDKKYSIGLAIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKKNLIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEESFLVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIKFRGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRLENLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQIGDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVRQQLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLLRKQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARGNSRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEYFTVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFDSVEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEERLKTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFMQLIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGRHKPENIVIEMARENQTTQKGQKNITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKHVAQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYLNAVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKTEITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSKESILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGITIMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNELALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILADANLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVLDATLIHQSITGLYETRIDLSQLGGDTSRADPKKKRKVEASGSGRADALDDFDLDMLGSDALDDFDLDMLGSDALDDFDLDMLGSDALDDFDLDMLINSRSQYLPDTDDRHRIEEKRKRTYETFKSIMKKSPFSGPTDPRPPPRRIAVPSRSSASVPKPAPQPYPFTSSLSTINYDEFPTMVFPSGQISQASALAPAPPQVLPQAPAPAPAPAMVSALAQAPAPVPVLAPGPPQAVAPPAPKPTQAGEGTLSEALLQLQFDDEDLGALLGNSTDPAVFTDLASVDNSEFQQLLNQGIPVAPHTTEPMLMEYPEAITRLVTGAQRPPDPAPAPLGAPGLPNGLLSGDEDFSSIADMDFSALLGSGSGSRDSREGMFLPKPEAGSAISDVFEGREVCQPKRIRPFHPPGSPWANRPLPASLAPTPTGPVHEPVGSLTPAPVPQPLDPAPAVTPEASHLLEDPDEETSQAVKALREMADTVIPQKEEAAICGQMDLSHPPPRGHLDELTTTLESMTEDLNLDSPLTPELNEILDTFLNDECLLHAMHISTGLSIFDTSLF
Seq ID 11
dSaCas9:VPR
MPKKKRKVGGGSPGKRNYILGLAIGITSVGYGIIDYETRDVIDAGVRLFKEANVENNEGRRSKRGARRLKRRRRHRIQRVKKLLFDYNLLTDHSELSGINPYEARVKGLSQKLSEEEFSAALLHLAKRRGVHNVNEVEEDTGNELSTKEQISRNSKALEEKYVAELQLERLKKDGEVRGSINRFKTSDYVKEAKQLLKVQKAYHQLDQSFIDTYIDLLETRRTYYEGPGEGSPFGWKDIKEWYEMLMGHCTYFPEELRSVKYAYNADLYNALNDLNNLVITRDENEKLEYYEKFQIIENVFKQKKKPTLKQIAKEILVNEEDIKGYRVTSTGKPEFTNLKVYHDIKDITARKEIIENAELLDQIAKILTIYQSSEDIQEELTNLNSELTQEEIEQISNLKGYTGTHNLSLKAINLILDELWHTNDNQIAIFNRLKLVPKKVDLSQQKEIPTTLVDDFILSPVVKRSFIQSIKVINAIIKKYGLPNDIIIELAREKNSKDAQKMINEMQKRNRQTNERIEEIIRTTGKENAKYLIEKIKLHDMQEGKCLYSLEAIPLEDLLNNPFNYEVDHIIPRSVSFDNSFNNKVLVKQEEASKKGNRTPFQYLSSSDSKISYETFKKHILNLAKGKGRISKTKKEYLLEERDINRFSVQKDFINRNLVDTRYATRGLMNLLRSYFRVNNLDVKVKSINGGFTSFLRRKWKFKKERNKGYKHHAEDALIIANADFIFKEWKKLDKAKKVMENQMFEEKQAESMPEIETEQEYKEIFITPHQIKHIKDFKDYKYSHRVDKKPNRKLINDTLYSTRKDDKGNTLIVNNLNGLYDKDNDKLKKLINKSPEKLLMYHHDPQTYQKLKLIMEQYGDEKNPLYKYYEETGNYLTKYSKKDNGPVIKKIKYYGNKLNAHLDITDDYPNSRNKVVKLSLKPYRFDVYLDNGVYKFVTVKNLDVIKKENYYEVNSKCYEEAKKLKKISNQAEFIASFYKNDLIKINGELYRVIGVNNDLLNRIEVNMIDITYREYLENMNDKRPPHIIKTIASKTQSIKKYSTDILGNLYEVKSKKHPQIIKKGTSRADPKKKRKVEASGSGRADALDDFDLDMLGSDALDDFDLDMLGSDALDDFDLDMLGSDALDDFDLDMLINSRSQYLPDTDDRHRIEEKRKRTYETFKSIMKKSPFSGPTDPRPPPRRIAVPSRSSASVPKPAPQPYPFTSSLSTINYDEFPTMVFPSGQISQASALAPAPPQVLPQAPAPAPAPAMVSALAQAPAPVPVLAPGPPQAVAPPAPKPTQAGEGTLSEALLQLQFDDEDLGALLGNSTDPAVFTDLASVDNSEFQQLLNQGIPVAPHTTEPMLMEYPEAITRLVTGAQRPPDPAPAPLGAPGLPNGLLSGDEDFSSIADMDFSALLGSGSGSRDSREGMFLPKPEAGSAISDVFEGREVCQPKRIRPFHPPGSPWANRPLPASLAPTPTGPVHEPVGSLTPAPVPQPLDPAPAVTPEASHLLEDPDEETSQAVKALREMADTVIPQKEEAAICGQMDLSHPPPRGHLDELTTTLESMTEDLNLDSPLTPELNEILDTFLNDECLLHAMHISTGLSIFDTSLF
Seq ID 12
mini-SaCas9-1:VPR
MPKKKRKVGGGSPGKRNYILGLDIGITSVGYGIIDYETRDVIDAGVRLFKEANVENNEGRRSKRGARRLKRRRRHRIQRVKKLLFDYNLLTDHSELSGINPYEARVKGLSQKLSEEEFSAALLHLAKRRGVHNVNEVEEDTGNELSTKEQISRNSKALEEKYVAELQLERLKKDGEVRGSINRFKTSDYVKEAKQLLKVQKAYHQLDQSFIDTYIDLLETRRTYYEGPGEGSPFGWKDIKEWYEMLGGGSGGGILSPVVKRSFIQSIKVINAIIKKYGLPNDIIIELAREKNSKDAQKMINEMQKRNRQTNERIEEIIRTTGKENAKYLIEKIKLHDMQEGKCLYSLEAIPLEDLLNNPFNYEVDHIIPRSVSFDNSFNNKVLVKQEENSKKGNRTPFQYLSSSDSKISYETFKKHILNLAKGKGRISKTKKEYLLEERDINRFSVQKDFINRNLVDTRYATRGLMNLLRSYFRVNNLDVKVKSINGGFTSFLRRKWKFKKERNKGYKHHAEDALIIANADFIFKEWKKLDKAKKVMENQMFEEKQAESMPEIETEQEYKEIFITPHQIKHIKDFKDYKYSHRVDKKPNRKLINDTLYSTRKDDKGNTLIVNNLNGLYDKDNDKLKKLINKSPEKLLMYHHDPQTYQKLKLIMEQYGDEKNPLYKYYEETGNYLTKYSKKDNGPVIKKIKYYGNKLNAHLDITDDYPNSRNKVVKLSLKPYRFDVYLDNGVYKFVTVKNLDVIKKENYYEVNSKCYEEAKKLKKISNQAEFIASFYKNDLIKINGELYRVIGVNNDLLNRIEVNMIDITYREYLENMNDKRPPHIIKTIASKTQSIKKYSTDILGNLYEVKSKKHPQIIKKGTSRADPKKKRKVEASGSGRADALDDFDLDMLGSDALDDFDLDMLGSDALDDFDLDMLGSDALDDFDLDMLINSRSQYLPDTDDRHRIEEKRKRTYETFKSIMKKSPFSGPTDPRPPPRRIAVPSRSSASVPKPAPQPYPFTSSLSTINYDEFPTMVFPSGQISQASALAPAPPQVLPQAPAPAPAPAMVSALAQAPAPVPVLAPGPPQAVAPPAPKPTQAGEGTLSEALLQLQFDDEDLGALLGNSTDPAVFTDLASVDNSEFQQLLNQGIPVAPHTTEPMLMEYPEAITRLVTGAQRPPDPAPAPLGAPGLPNGLLSGDEDFSSIADMDFSALLGSGSGSRDSREGMFLPKPEAGSAISDVFEGREVCQPKRIRPFHPPGSPWANRPLPASLAPTPTGPVHEPVGSLTPAPVPQPLDPAPAVTPEASHLLEDPDEETSQAVKALREMADTVIPQKEEAAICGQMDLSHPPPRGHLDELTTTLESMTEDLNLDSPLTPELNEILDTFLNDECLLHAMHISTGLSIFDTSLF
Seq ID 13
mini-SaCas9-2:VPR
MPKKKRKVGGGSPGKRNYILGLDIGITSVGYGIIDYETRDVIDAGVRLFKEANVENNEGRRSKRGARRLKRRRRHRIQRVKKLLFDYNLLTDHSELSGINPYEARVKGLSQKLSEEEFSAALLHLAKRRGVHNVNEVEEDTGNELSTKEQISRNSKALEEKYVAELQLERLKKDGEVRGSINRFKTSDYVKEAKQLLKVQKAYHQLDQSFIDTYIDLLETRRTYYEGPGEGSPFGWKDIKEWYEMLKRRRRHRILSPVVKRSFIQSIKVINAIIKKYGLPNDIIIELAREKNSKDAQKMINEMQKRNRQTNERIEEIIRTTGKENAKYLIEKIKLHDMQEGKCLYSLEAIPLEDLLNNPFNYEVDHIIPRSVSFDNSFNNKVLVKQEENSKKGNRTPFQYLSSSDSKISYETFKKHILNLAKGKGRISKTKKEYLLEERDINRFSVQKDFINRNLVDTRYATRGLMNLLRSYFRVNNLDVKVKSINGGFTSFLRRKWKFKKERNKGYKHHAEDALIIANADFIFKEWKKLDKAKKVMENQMFEEKQAESMPEIETEQEYKEIFITPHQIKHIKDFKDYKYSHRVDKKPNRKLINDTLYSTRKDDKGNTLIVNNLNGLYDKDNDKLKKLINKSPEKLLMYHHDPQTYQKLKLIMEQYGDEKNPLYKYYEETGNYLTKYSKKDNGPVIKKIKYYGNKLNAHLDITDDYPNSRNKVVKLSLKPYRFDVYLDNGVYKFVTVKNLDVIKKENYYEVNSKCYEEAKKLKKISNQAEFIASFYKNDLIKINGELYRVIGVNNDLLNRIEVNMIDITYREYLENMNDKRPPHIIKTIASKTQSIKKYSTDILGNLYEVKSKKHPQIIKKGTSRADPKKKRKVEASGSGRADALDDFDLDMLGSDALDDFDLDMLGSDALDDFDLDMLGSDALDDFDLDMLINSRSQYLPDTDDRHRIEEKRKRTYETFKSIMKKSPFSGPTDPRPPPRRIAVPSRSSASVPKPAPQPYPFTSSLSTINYDEFPTMVFPSGQISQASALAPAPPQVLPQAPAPAPAPAMVSALAQAPAPVPVLAPGPPQAVAPPAPKPTQAGEGTLSEALLQLQFDDEDLGALLGNSTDPAVFTDLASVDNSEFQQLLNQGIPVAPHTTEPMLMEYPEAITRLVTGAQRPPDPAPAPLGAPGLPNGLLSGDEDFSSIADMDFSALLGSGSGSRDSREGMFLPKPEAGSAISDVFEGREVCQPKRIRPFHPPGSPWANRPLPASLAPTPTGPVHEPVGSLTPAPVPQPLDPAPAVTPEASHLLEDPDEETSQAVKALREMADTVIPQKEEAAICGQMDLSHPPPRGHLDELTTTLESMTEDLNLDSPLTPELNEILDTFLNDECLLHAMHISTGLSIFDTSLF
Seq ID 14
mini-SaCas9-3:VPR
MPKKKRKVGGGSPGKRNYILGLDIGITSVGYGIIDYETRDVIDAGVRLFKEANVENNEGRRSKRGARRLKRRRRHRIQRVKKLLFDYNLLTDHSELSGINPYEARVKGLSQKLSEEEFSAALLHLAKRRGVHNVNEVEEDTGNELSTKEQISRNSKALEEKYVAELQLERLKKDGEVRGSINRFKTSDYVKEAKQLLKVQKAYHQLDQSFIDTYIDLLETRRTYYEGPGEGSPFGWKDIKEWYEMLKRRRRHRILSPVVKRSFIQSIKVINAIIKKYGLPNDIIIELGSKRYATRGLMNLLRSYFRVNNLDVKVKSINGGFTSFLRRKWKFKKERNKGYKHHAEDALIIANADFIFKEWKKLDKAKKVMENQMFEEKQAESMPEIETEQEYKEIFITPHQIKHIKDFKDYKYSHRVDKKPNRKLINDTLYSTRKDDKGNTLIVNNLNGLYDKDNDKLKKLINKSPEKLLMYHHDPQTYQKLKLIMEQYGDEKNPLYKYYEETGNYLTKYSKKDNGPVIKKIKYYGNKLNAHLDITDDYPNSRNKVVKLSLKPYRFDVYLDNGVYKFVTVKNLDVIKKENYYEVNSKCYEEAKKLKKISNQAEFIASFYKNDLIKINGELYRVIGVNNDLLNRIEVNMIDITYREYLENMNDKRPPHIIKTIASKTQSIKKYSTDILGNLYEVKSKKHPQIIKKGTSRADPKKKRKVEASGSGRADALDDFDLDMLGSDALDDFDLDMLGSDALDDFDLDMLGSDALDDFDLDMLINSRSQYLPDTDDRHRIEEKRKRTYETFKSIMKKSPFSGPTDPRPPPRRIAVPSRSSASVPKPAPQPYPFTSSLSTINYDEFPTMVFPSGQISQASALAPAPPQVLPQAPAPAPAPAMVSALAQAPAPVPVLAPGPPQAVAPPAPKPTQAGEGTLSEALLQLQFDDEDLGALLGNSTDPAVFTDLASVDNSEFQQLLNQGIPVAPHTTEPMLMEYPEAITRLVTGAQRPPDPAPAPLGAPGLPNGLLSGDEDFSSIADMDFSALLGSGSGSRDSREGMFLPKPEAGSAISDVFEGREVCQPKRIRPFHPPGSPWANRPLPASLAPTPTGPVHEPVGSLTPAPVPQPLDPAPAVTPEASHLLEDPDEETSQAVKALREMADTVIPQKEEAAICGQMDLSHPPPRGHLDELTTTLESMTEDLNLDSPLTPELNEILDTFLNDECLLHAMHISTGLSIFDTSLF
Seq ID 15
dAsCpf1:VPR
MPKKKRKVGGGSPGMTQFEGFTNLYQVSKTLRFELIPQGKTLKHIQEQGFIEEDKARNDHYKELKPIIDRIYKTYADQCLQLVQLDWENLSAAIDSYRKEKTEETRNALIEEQATYRNAIHDYFIGRTDNLTDAINKRHAEIYKGLFKAELFNGKVLKQLGTVTTTEHENALLRSFDKFTTYFSGFYENRKNVFSAEDISTAIPHRIVQDNFPKFKENCHIFTRLITAVPSLREHFENVKKAIGIFVSTSIEEVFSFPFYNQLLTQTQIDLYNQLLGGISREAGTEKIKGLNEVLNLAIQKNDETAHIIASLPHRFIPLFKQILSDRNTLSFILEEFKSDEEVIQSFCKYKTLLRNENVLETAEALFNELNSIDLTHIFISHKKLETISSALCDHWDTLRNALYERRISELTGKITKSAKEKVQRSLKHEDINLQEIISAAGKELSEAFKQKTSEILSHAHAALDQPLPTTLKKQEEKEILKSQLDSLLGLYHLLDWFAVDESNEVDPEFSARLTGIKLEMEPSLSFYNKARNYATKKPYSVEKFKLNFQMPTLASGWDVNKEKNNGAILFVKNGLYYLGIMPKQKGRYKALSFEPTEKTSEGFDKMYYDYFPDAAKMIPKCSTQLKAVTAHFQTHTTPILLSNNFIEPLEITKEIYDLNNPEKEPKKFQTAYAKKTGDQKGYREALCKWIDFTRDFLSKYTKTTSIDLSSLRPSSQYKDLGEYYAELNPLLYHISFQRIAEKEIMDAVETGKLYLFQIYNKDFAKGHHGKPNLHTLYWTGLFSPENLAKTSIKLNGQAELFYRPKSRMKRMAHRLGEKMLNKKLKDQKTPIPDTLYQELYDYVNHRLSHDLSDEARALLPNVITKEVSHEIIKDRRFTSDKFFFHVPITLNYQAANSPSKFNQRVNAYLKEHPETPIIGIARGERNLIYITVIDSTGKILEQRSLNTIQQFDYQKKLDNREKERVAARQAWSVVGTIKDLKQGYLSQVIHEIVDLMIHYQAVVVLENLNFGFKSKRTGIAEKAVYQQFEKMLIDKLNCLVLKDYPAEKVGGVLNPYQLTDQFTSFAKMGTQSGFLFYVPAPYTSKIDPLTGFVDPFVWKTIKNHESRKHFLEGFDFLHYDVKTGDFILHFKMNRNLSFQRGLPGFMPAWDIVFEKNETQFDAKGTPFIAGKRIVPVIENHRFTGRYRDLYPANELIALLEEKGIVFRDGSNILPKLLENDDSHAIDTMVALIRSVLQMRNSNAATGEDYINSPVRDLNGVCFDSRFQNPEWPMDADANGAYHIALKGQLLLNHLKESKDLKLQNGISNQDWLAYIQELRNTSRADPKKKRKVEASGSGRADALDDFDLDMLGSDALDDFDLDMLGSDALDDFDLDMLGSDALDDFDLDMLINSRSQYLPDTDDRHRIEEKRKRTYETFKSIMKKSPFSGPTDPRPPPRRIAVPSRSSASVPKPAPQPYPFTSSLSTINYDEFPTMVFPSGQISQASALAPAPPQVLPQAPAPAPAPAMVSALAQAPAPVPVLAPGPPQAVAPPAPKPTQAGEGTLSEALLQLQFDDEDLGALLGNSTDPAVFTDLASVDNSEFQQLLNQGIPVAPHTTEPMLMEYPEAITRLVTGAQRPPDPAPAPLGAPGLPNGLLSGDEDFSSIADMDFSALLGSGSGSRDSREGMFLPKPEAGSAISDVFEGREVCQPKRIRPFHPPGSPWANRPLPASLAPTPTGPVHEPVGSLTPAPVPQPLDPAPAVTPEASHLLEDPDEETSQAVKALREMADTVIPQKEEAAICGQMDLSHPPPRGHLDELTTTLESMTEDLNLDSPLTPELNEILDTFLNDECLLHAMHISTGLSIFDTSLF
Seq ID 16
mini-AsCpf1-1:VPR
MPKKKRKVGGGSPGMTQFEGFTNLYQVSKTLRFELIPQGKTLKHIQEQGFIEEDKARNDHYKELKPIIDRIYKTYADQCLQLVQLDWENLSAAIDSYRKEKTEETRNALIEEQATYRNAIHDYFIGRTDNLTDAINKRHAEIYKGLFKAELFNGKVLKQLGTVTTTEHENALLRSFDKFTTYFSGFYENRKNVFSAEDISTAIPHRIVQDNFPKFKENCHIFTRLITAVPSLREHFENVKKAIGIFVSTSIEEVFSFPFYNQLLTQTQIDLYNQLLGGISREAGTEKIKGLNEVLNLAIQKNDETAHIIASLPHRFIPLFKQILSDRNTLSFILEEFGGGSYSVEKFKLNFQMPTLASGWDVNKEKNNGAILFVKNGLYYLGIMPKQKGRYKALSFEPTEKTSEGFDKMYYDYFPDAAKMIPKCSTQLKAVTAHFQTHTTPILLSNNFIEPLEITKEIYDLNNPEKEPKKFQTAYAKKTGDQKGYREALCKWIDFTRDFLSKYTKTTSIDLSSLRPSSQYKDLGEYYAELNPLLYHISFQRIAEKEIMDAVETGKLYLFQIYNKDFAKGHHGKPNLHTLYWTGLFSPENLAKTSIKLNGQAELFYRPKSRMKRMAHRLGEKMLNKKLKDQKTPIPDTLYQELYDYVNHRLSHDLSDEARALLPNVITKEVSHEIIKDRRFTSDKFFFHVPITLNYQAANSPSKFNQRVNAYLKEHPETPIIGIDRGERNLIYITVIDSTGKILEQRSLNTIQQFDYQKKLDNREKERVAARQAWSVVGTIKDLKQGYLSQVIHEIVDLMIHYQAVVVLENLNFGFKSKRTGIAEKAVYQQFEKMLIDKLNCLVLKDYPAEKVGGVLNPYQLTDQFTSFAKMGTQSGFLFYVPAPYTSKIDPLTGFVDPFVWKTIKNHESRKHFLEGFDFLHYDVKTGDFILHFKMNRNLSFQRGLPGFMPAWDIVFEKNETQFDAKGTPFIAGKRIVPVIENHRFTGRYRDLYPANELIALLEEKGIVFRDGSNILPKLLENDDSHAIDTMVALIRSVLQMRNSNAATGEDYINSPVRDLNGVCFDSRFQNPEWPMDADANGAYHIALKGQLLLNHLKESKDLKLQNGISNQDWLAYIQELRNTSRADPKKKRKVEASGSGRADALDDFDLDMLGSDALDDFDLDMLGSDALDDFDLDMLGSDALDDFDLDMLINSRSQYLPDTDDRHRIEEKRKRTYETFKSIMKKSPFSGPTDPRPPPRRIAVPSRSSASVPKPAPQPYPFTSSLSTINYDEFPTMVFPSGQISQASALAPAPPQVLPQAPAPAPAPAMVSALAQAPAPVPVLAPGPPQAVAPPAPKPTQAGEGTLSEALLQLQFDDEDLGALLGNSTDPAVFTDLASVDNSEFQQLLNQGIPVAPHTTEPMLMEYPEAITRLVTGAQRPPDPAPAPLGAPGLPNGLLSGDEDFSSIADMDFSALLGSGSGSRDSREGMFLPKPEAGSAISDVFEGREVCQPKRIRPFHPPGSPWANRPLPASLAPTPTGPVHEPVGSLTPAPVPQPLDPAPAVTPEASHLLEDPDEETSQAVKALREMADTVIPQKEEAAICGQMDLSHPPPRGHLDELTTTLESMTEDLNLDSPLTPELNEILDTFLNDECLLHAMHISTGLSIFDTSLF
Seq ID 17
dSaCas9N:FokI
MPKKKRKVGGGSPGKRNYILGLAIGITSVGYGIIDYETRDVIDAGVRLFKEANVENNEGRRSKRGARRLKRRRRHRIQRVKKLLFDYNLLTDHSELSGINPYEARVKGLSQKLSEEEFSAALLHLAKRRGVHNVNEVEEDTGNELSTKEQISRNSKALEEKYVAELQLERLKKDGEVRGSINRFKTSDYVKEAKQLLKVQKAYHQLDQSFIDTYIDLLETRRTYYEGPGEGSPFGWKDIKEWYEMLMGHCTYFPEELRSVKYAYNADLYNALNDLNNLVITRDENEKLEYYEKFQIIENVFKQKKKPTLKQIAKEILVNEEDIKGYRVTSTGKPEFTNLKVYHDIKDITARKEIIENAELLDQIAKILTIYQSSEDIQEELTNLNSELTQEEIEQISNLKGYTGTHNLSLKAINLILDELWHTNDNQIAIFNRLKLVPKKVDLSQQKEIPTTLVDDFILSPVVKRSFIQSIKVINAIIKKYGLPNDIIIELAREKNSKDAQKMINEMQKRNRQTNERIEEIIRTTGKENAKYLIEKIKLHDMQEGKCLYSLEAIPLEDLLNNPFNYEVDHIIPRSVSFDNSFNNKVLVKQEEASKKGNRTPFQYLSSSDSKISYETFKKHILNLAKGKGRISKTKKEYLLEERDINRFSVQKDFINRNLVDTRYATRGLMNLLRSYFRVNNLDVKVKSINGGFTSFLRRKWKFKKERNKGYKHHAEDALIIANADFIFKEWKKLDKAKKVMENQMFGGGSGGGSGGGSLVKSELEEKKSELRHKLKYVPHEYIELIEIARNSTQDRILEMKVMEFFMKVYGYRGKHLGGSRKPDGAIYTVGSPIDYGVIVDTKAYSGGYNLPIGQADEMQRYVEENQTRNKHINPNEWWKVYPSSVTEFKFLFVSGHFKGNYKAQLTRLNHITNCNGAVLSVEELLIGGEMIKAGTLTLEEVRRKFNNGEINFTSGGGSKRPAATKKAGQAKKKKSR
Seq ID 18
dSaCas9C
MPKKKRKVGGGSPGAESMPEIETEQEYKEIFITPHQIKHIKDFKDYKYSHRVDKKPNRKLINDTLYSTRKDDKGNTLIVNNLNGLYDKDNDKLKKLINKSPEKLLMYHHDPQTYQKLKLIMEQYGDEKNPLYKYYEETGNYLTKYSKKDNGPVIKKIKYYGNKLNAHLDITDDYPNSRNKVVKLSLKPYRFDVYLDNGVYKFVTVKNLDVIKKENYYEVNSKCYEEAKKLKKISNQAEFIASFYKNDLIKINGELYRVIGVNNDLLNRIEVNMIDITYREYLENMNDKRPPHIIKTIASKTQSIKKYSTDILGNLYEVKSKKHPQIIKKGTSGGGSKRPAATKKAGQAKKKKSR
Seq ID 19
mini-dSaCas9-4N:FokI
MPKKKRKVGGGSPGKRNYILGLAIGITSVGYGIIDYETRDVIDAGVRLFKEANVENNEGRRSKRGARRLKRRRRHRIQRVKKLLFDYNLLTDHSELSGINPYEARVKGLSQKLSEEEFSAALLHLAKRRGVHNVNEVEEDTGNELSTKEQISRNSKALEEKYVAELQLERLKKDGEVRGSINRFKTSDYVKEAKQLLKVQKAYHQLDQSFIDTYIDLLETRRTYYEGPGEGSPFGWKDIKEWYEMLMGHCTYFPEELRSVKYAYNADLYNALNDLNNLVITRDENEKLEYYEKFQIIENVFKQKKKPTLKQIAKEILVNEEDIKGYRVTSTGKPEFTNLKVYHDIKDITARKEIIENAELLDQIAKILTIYQSSEDIQEELTNLNSELTQEEIEQISNLKGYTGTHNLSLKAINLILDELWHTNDNQIAIFNRLKLVPKKVDLSQQKEIPTTLVDDFILSPVVKRSFIQSIKVINAIIKKYGLPNDIIIELGSKRYATRGLMNLLRSYFRVNNLDVKVKSINGGFTSFLRRKWKFKKERNKGYKHHAEDALIIANADFIFKEWKKLDKAKKVMENQMFGGGSGGGSGGGSLVKSELEEKKSELRHKLKYVPHEYIELIEIARNSTQDRILEMKVMEFFMKVYGYRGKHLGGSRKPDGAIYTVGSPIDYGVIVDTKAYSGGYNLPIGQADEMQRYVEENQTRNKHINPNEWWKVYPSSVTEFKFLFVSGHFKGNYKAQLTRLNHITNCNGAVLSVEELLIGGEMIKAGTLTLEEVRRKFNNGEINFTSGGGSKRPAATKKAGQAKKKKSR
Seq ID 20
AID:dSaCas9:UGI
MPKKKRKVGGGSPGMSSETGPVAVDPTLRRRIEPHEFEVFFDPRELRKETCLLYEINWGGRHSIWRHTSQNTNKHVEVNFIEKFTTERYFCPNTRCSITWFLSWSPCGECSRAITEFLSRYPHVTLFIYIARLYHHADPRNRQGLRDLISSGVTIQIMTEQESGYCWRNFVNYSPSNEAHWPRYPHLWVRLYVLELYCIILGLPPCLNILRRKQPQLTFFTIALQSCHYQRLPPHILWATGLKSGSETPGTSESATPEKRNYILGLAIGITSVGYGIIDYETRDVIDAGVRLFKEANVENNEGRRSKRGARRLKRRRRHRIQRVKKLLFDYNLLTDHSELSGINPYEARVKGLSQKLSEEEFSAALLHLAKRRGVHNVNEVEEDTGNELSTKEQISRNSKALEEKYVAELQLERLKKDGEVRGSINRFKTSDYVKEAKQLLKVQKAYHQLDQSFIDTYIDLLETRRTYYEGPGEGSPFGWKDIKEWYEMLMGHCTYFPEELRSVKYAYNADLYNALNDLNNLVITRDENEKLEYYEKFQIIENVFKQKKKPTLKQIAKEILVNEEDIKGYRVTSTGKPEFTNLKVYHDIKDITARKEIIENAELLDQIAKILTIYQSSEDIQEELTNLNSELTQEEIEQISNLKGYTGTHNLSLKAINLILDELWHTNDNQIAIFNRLKLVPKKVDLSQQKEIPTTLVDDFILSPVVKRSFIQSIKVINAIIKKYGLPNDIIIELAREKNSKDAQKMINEMQKRNRQTNERIEEIIRTTGKENAKYLIEKIKLHDMQEGKCLYSLEAIPLEDLLNNPFNYEVDHIIPRSVSFDNSFNNKVLVKQEEASKKGNRTPFQYLSSSDSKISYETFKKHILNLAKGKGRISKTKKEYLLEERDINRFSVQKDFINRNLVDTRYATRGLMNLLRSYFRVNNLDVKVKSINGGFTSFLRRKWKFKKERNKGYKHHAEDALIIANADFIFKEWKKLDKAKKVMENQMFEEKQAESMPEIETEQEYKEIFITPHQIKHIKDFKDYKYSHRVDKKPNRKLINDTLYSTRKDDKGNTLIVNNLNGLYDKDNDKLKKLINKSPEKLLMYHHDPQTYQKLKLIMEQYGDEKNPLYKYYEETGNYLTKYSKKDNGPVIKKIKYYGNKLNAHLDITDDYPNSRNKVVKLSLKPYRFDVYLDNGVYKFVTVKNLDVIKKENYYEVNSKCYEEAKKLKKISNQAEFIASFYKNDLIKINGELYRVIGVNNDLLNRIEVNMIDITYREYLENMNDKRPPHIIKTIASKTQSIKKYSTDILGNLYEVKSKKHPQIIKKGSGGSTNLSDIIEKETGKQLVIQESILMLPEEVEEVIGNKPESDILVHTAYDESTDENVMLLTSDAPEYKPWALVIQDSNGENKIKMLSGGSPPKKKRKV
Seq ID 21
AID:mini-dSaCas9-5:UGI
MPKKKRKVGGGSPGMSSETGPVAVDPTLRRRIEPHEFEVFFDPRELRKETCLLYEINWGGRHSIWRHTSQNTNKHVEVNFIEKFTTERYFCPNTRCSITWFLSWSPCGECSRAITEFLSRYPHVTLFIYIARLYHHADPRNRQGLRDLISSGVTIQIMTEQESGYCWRNFVNYSPSNEAHWPRYPHLWVRLYVLELYCIILGLPPCLNILRRKQPQLTFFTIALQSCHYQRLPPHILWATGLKSGSETPGTSESATPEKRNYILGLAIGITSVGYGIIDYETRDVIDAGVRLFKEANVENNEGRRSKRGARRLKRRRRHRIQRVKKLLFDYNLLTDHSELSGINPYEARVKGLSQKLSEEEFSAALLHLAKRRGVHNVNEVEEDTGNELSTKEQISRNSKALEEKYVAELQLERLKKDGEVRGSINRFKTSDYVKEAKQLLKVQKAYHQLDQSFIDTYIDLLETRRTYYEGPGEGSPFGWKDIKEWYEMLMGHCTYFPEELRSVKYAYNADLYNALNDLNNLVITRDENEKLEYYEKFQIIENVFKQKKKPTLKQIAKEILVNEEDIKGYRVTSTGKPEFTNLKVYHDIKDITARKEIIENAELLDQIAKILTIYQSSEDIQEELTNLNSELTQEEIEQISNLKGYTGTHNLSLKAINLILDELWHTNDNQIAIFNRLKLVPKKVDLSQQKEIPTTLVDDFILSPVVKRSFIQSIKVINAIIKKYGLPNDIIIELGSKRYATRGLMNLLRSYFRVNNLDVKVKSINGGFTSFLRRKWKFKKERNKGYKHHAEDALIIANADFIFKEWKKLDKAKKVMENQMFEEKQAESMPEIETEQEYKEIFITPHQIKHIKDFKDYKYSHRVDKKPNRKLINDTLYSTRKDDKGNTLIVNNLNGLYDKDNDKLKKLINKSPEKLLMYHHDPQTYQKLKLIMEQYGDEKNPLYKYYEETGNYLTKYSKKDNGPVIKKIKYYGNKLNAHLDITDDYPNSRNKVVKLSLKPYRFDVYLDNGVYKFVTVKNLDVIKKENYYEVNSKCYEEAKKLKKISNQAEFIASFYKNDLIKINGELYRVIGVNNDLLNRIEVNMIDITYREYLENMNDKRPPHIIKTIASKTQSIKKYSTDILGNLYEVKSKKHPQIIKKGSGGSTNLSDIIEKETGKQLVIQESILMLPEEVEEVIGNKPESDILVHTAYDESTDENVMLLTSDAPEYKPWALVIQDSNGENKIKMLSGGSPPKKKRKV
Seq ID 22
mini-dSaCas9-6N:AID
MPKKKRKVGGGSPGKRNYILGLAIGITSVGYGIIDYETRDVIDAGVRLFKEANVENNEGRRSKRGARRLKRRRRHRIQRVKKLLFDYNLLTDHSELSGINPYEARVKGLSQKLSEEEFSAALLHLAKRRGVHNVNEVEEDTGNELSTKEQISRNSKALEEKYVAELQLERLKKDGEVRGSINRFKTSDYVKEAKQLLKVQKAYHQLDQSFIDTYIDLLETRRTYYEGPGEGSPFGWKDIKEWYEMLMGHCTYFPEELRSVKYAYNADLYNALNDLNNLVITRDENEKLEYYEKFQIIENVFKQKKKPTLKQIAKEILVNEEDIKGYRVTSTGKPEFTNLKVYHDIKDITARKEIIENAELLDQIAKILTIYQSSEDIQEELTNLNSELTQEEIEQISNLKGYTGTHNLSLKAINLILDELWHTNDNQIAIFNRLKLVPKKVDLSQQKEIPTTLVDDFILSPVVKRSFIQSIKVINAIIKKYGLPNDIIIELAREKGGGSGGGSMSSETGPVAVDPTLRRRIEPHEFEVFFDPRELRKETCLLYEINWGGRHSIWRHTSQNTNKHVEVNFIEKFTTERYFCPNTRCSITWFLSWSPCGECSRAITEFLSRYPHVTLFIYIARLYHHADPRNRQGLRDLISSGVTIQIMTEQESGYCWRNFVNYSPSNEAHWPRYPHLWVRLYVLELYCIILGLPPCLNILRRKQPQLTFFTIALQSCHYQRLPPHILWATGLKSGSETPGTSESATPETSGGGSKRPAATKKAGQAKKKKSR
Seq ID 23
mini-dSaCas9-6C:UGI
MPKKKRKVGGGSPGTRYATRGLMNLLRSYFRVNNLDVKVKSINGGFTSFLRRKWKFKKERNKGYKHHAEDALIIANADFIFKEWKKLDKAKKVMENQMFEEKQAESMPEIETEQEYKEIFITPHQIKHIKDFKDYKYSHRVDKKPNRKLINDTLYSTRKDDKGNTLIVNNLNGLYDKDNDKLKKLINKSPEKLLMYHHDPQTYQKLKLIMEQYGDEKNPLYKYYEETGNYLTKYSKKDNGPVIKKIKYYGNKLNAHLDITDDYPNSRNKVVKLSLKPYRFDVYLDNGVYKFVTVKNLDVIKKENYYEVNSKCYEEAKKLKKISNQAEFIASFYKNDLIKINGELYRVIGVNNDLLNRIEVNMIDITYREYLENMNDKRPPHIIKTIASKTQSIKKYSTDILGNLYEVKSKKHPQIIKKGSGGSTNLSDIIEKETGKQLVIQESILMLPEEVEEVIGNKPESDILVHTAYDESTDENVMLLTSDAPEYKPWALVIQDSNGENKIKMLSGGSPPKKKRKV
Seq ID 24
VTR1
GSGRADALDDFDLDMLGSDALDDFDLDMLGSDALDDFDLDMLGSDALDDFDLDMLINSALLQLQFDDEDLGALLGNSTDPAVFTDLASVDNSEFQQLLNQGIPVAPHTTEPMLMEYPEAITRLVTGAQRPPDPAPAPLGAPGLPNGLLSGDEDFSSIADMDFSALLSQISSGSGSGSRDSREGMFLPKPEAGSAISDVFEGREVCQPKRIRPFHPPGSPWANRPLPASLAPTPTGPVHEPVGSLTPAPVPQPLDPAPAVTPEASHLLEDPDEETSQAVKALREMADTVIPQKEEAAICGQMDLSHPPPRGHLDELTTTLESMTEDLNLDSPLTPELNEILDTFLNDECLLHAMHISTGLSIFDTSLFTSGGGSKRPAATKKAGQAKKKKSR
Seq ID 25
VTR2
GSGRADALDDFDLDMLGSDALDDFDLDMLGSDALDDFDLDMLGSDALDDFDLDMLINSRLVTGAQRPPDPAPAPLGAPGLPNGLLSGDEDFSSIADMDFSALLSQISSGGGSRLVTGAQRPPDPAPAPLGAPGLPNGLLSGDEDFSSIADMDFSALLSQISSGSGSGSRDSREGMFLPKPEAGSAISDVFEGREVCQPKRIRPFHPPGSPWANRPLPASLAPTPTGPVHEPVGSLTPAPVPQPLDPAPAVTPEASHLLEDPDEETSQAVKALREMADTVIPQKEEAAICGQMDLSHPPPRGHLDELTTTLESMTEDLNLDSPLTPELNEILDTFLNDECLLHAMHISTGLSIFDTSLFTSGGGSKRPAATKKAGQAKKKKSR
Seq ID 26
SpyCatcher:VPR
MPKKKRKVGGGSPGGAMVDTLSGLSSEQGQSGDMTIEEDSATHIKFSKRDEDGKELAGATMELRDSSGKTISTWISDGQVKDFYLYPGKYTFVETAAPDGYEVATAITFTVNEQGQVTVNGKATKGDAHITGTSRADPKKKRKVEASGSGRADALDDFDLDMLGSDALDDFDLDMLGSDALDDFDLDMLGSDALDDFDLDMLINSRSQYLPDTDDRHRIEEKRKRTYETFKSIMKKSPFSGPTDPRPPPRRIAVPSRSSASVPKPAPQPYPFTSSLSTINYDEFPTMVFPSGQISQASALAPAPPQVLPQAPAPAPAPAMVSALAQAPAPVPVLAPGPPQAVAPPAPKPTQAGEGTLSEALLQLQFDDEDLGALLGNSTDPAVFTDLASVDNSEFQQLLNQGIPVAPHTTEPMLMEYPEAITRLVTGAQRPPDPAPAPLGAPGLPNGLLSGDEDFSSIADMDFSALLGSGSGSRDSREGMFLPKPEAGSAISDVFEGREVCQPKRIRPFHPPGSPWANRPLPASLAPTPTGPVHEPVGSLTPAPVPQPLDPAPAVTPEASHLLEDPDEETSQAVKALREMADTVIPQKEEAAICGQMDLSHPPPRGHLDELTTTLESMTEDLNLDSPLTPELNEILDTFLNDECLLHAMHISTGLSIFDTSLF
Seq ID 27
4x SpyTag
GGGSGGAHIVMVDAYKPTKGGSGGGGSGGAHIVMVDAYKPTKGGSGGGGSGGAHIVMVDAYKPTKGGSGGGGSGGAHIVMVDAYKPTK
Seq ID 28
MoonCatcher:VPR
MPKKKRKVGGGSPGNHVIETEQNLPNEDGQSGNIIEQEDSKTLVKFSKRDIKGNELAGATIELRDLSGKSIQSWVSDGKAKDFYLLPGSYEFVETAAPEGYQIATKIMFTISTDGRITVDGQLVTGTGSGRADALDDFDLDMLGSDALDDFDLDMLGSDALDDFDLDMLGSDALDDFDLDMLINSRSQYLPDTDDRHRIEEKRKRTYETFKSIMKKSPFSGPTDPRPPPRRIAVPSRSSASVPKPAPQPYPFTSSLSTINYDEFPTMVFPSGQISQASALAPAPPQVLPQAPAPAPAPAMVSALAQAPAPVPVLAPGPPQAVAPPAPKPTQAGEGTLSEALLQLQFDDEDLGALLGNSTDPAVFTDLASVDNSEFQQLLNQGIPVAPHTTEPMLMEYPEAITRLVTGAQRPPDPAPAPLGAPGLPNGLLSGDEDFSSIADMDFSALLGSGSGSRDSREGMFLPKPEAGSAISDVFEGREVCQPKRIRPFHPPGSPWANRPLPASLAPTPTGPVHEPVGSLTPAPVPQPLDPAPAVTPEASHLLEDPDEETSQAVKALREMADTVIPQKEEAAICGQMDLSHPPPRGHLDELTTTLESMTEDLNLDSPLTPELNEILDTFLNDECLLHAMHISTGLSIFDTSLF
Seq ID 29
4x MoonTag
GGGSGGAHIVMVDNYKPIVGGSGGAHIVMVDNYKPIVGGSGGAHIVMVDNYKPIVGGSGGGGSGGAHIVMVDNYKPIV
Seq ID 30
VTR3 sequences
GSGRADALDDFDLDMLGSDALDDFDLDMLGSDALDDFDLDMLGSDALDDFDLDMLINSRLVTGAQRPPDPAPAPLGAPGLPNGLLSGDEDFSSIADMDFSALLSQISSGGGSRLVTGAQRPPDPAPAPLGAPGLPNGLLSGDEDFSSIADMDFSALLSQISSGSGSQPLDPAPAVTPEASHLLEDPDEETSQAVKALREMADTVIPQKEEAAICGQMDLSHPPPRGHLDELTTTLESMTEDLNLDSPLTPELNEILDTFLNDECLLHAMHISTGLSIFDTSLFTS
Seq ID 31
gRNA-ccr5-1
GCTGTGTTTGCGTCTCTCCC
Seq ID 32
gRNA-ccr5-2
GGGGTGGTGACAAGTGTGAT
Seq ID 33
Mini-saCas9-4:VPR
MPKKKRKVGGGSPGGGGSKRNYILGLDIGITSVGYGIIDYETRDVIDAGVRLFKEANVENNEGRRSKRGARRLKRRRRHRIQRVKKLLFDYNLLTDHSELSGINPYEARVKGLSQKLSEEEFSAALLHLAKRRGVHNVNEVEEDTGNELSTKEQISRNSKALEEKYVAELQLERLKKDGEVRGSINRFKTSDYVKEAKQLLKVQKAYHQLDQSFIDTYIDLLETRRTYYEGPGEGSPFGWKDIKEWYEMLMGHCTYFPEELRSVKYAYNADLYNALNDLNNLVITRDENEKLEYYEKFQIIENVFKQKKKPTLKQIAKEILVNEEDIKGYRVTSTGKPEFTNLKVYHDIKDITARKEIIENAELLDQIAKILTIYQSSEDIQEELTNLNSELTQEEIEQISNLKGYTGTHNLSLKAINLILDELWHTNDNQIAIFNRLKLVPKKVDLSQQKEIPTTLVDDFILSPVVKRSFIQSIKVINAIIKKYGLPNDIIIELGSKRYATRGLMNLLRSYFRVNNLDVKVKSINGGFTSFLRRKWKFKKERNKGYKHHAEDALIIANADFIFKEWKKLDKAKKVMENQMFEEKQAESMPEIETEQEYKEIFITPHQIKHIKDFKDYKYSHRVDKKPNRKLINDTLYSTRKDDKGNTLIVNNLNGLYDKDNDKLKKLINKSPEKLLMYHHDPQTYQKLKLIMEQYGDEKNPLYKYYEETGNYLTKYSKKDNGPVIKKIKYYGNKLNAHLDITDDYPNSRNKVVKLSLKPYRFDVYLDNGVYKFVTVKNLDVIKKENYYEVNSKCYEEAKKLKKISNQAEFIASFYKNDLIKINGELYRVIGVNNDLLNRIEVNMIDITYREYLENMNDKRPPHIIKTIASKTQSIKKYSTDILGNLYEVKSKKHPQIIKKGTSRADPKKKRKVEASGSGRADALDDFDLDMLGSDALDDFDLDMLGSDALDDFDLDMLGSDALDDFDLDMLINSRSQYLPDTDDRHRIEEKRKRTYETFKSIMKKSPFSGPTDPRPPPRRIAVPSRSSASVPKPAPQPYPFTSSLSTINYDEFPTMVFPSGQISQASALAPAPPQVLPQAPAPAPAPAMVSALAQAPAPVPVLAPGPPQAVAPPAPKPTQAGEGTLSEALLQLQFDDEDLGALLGNSTDPAVFTDLASVDNSEFQQLLNQGIPVAPHTTEPMLMEYPEAITRLVTGAQRPPDPAPAPLGAPGLPNGLLSGDEDFSSIADMDFSALLGSGSGSRDSREGMFLPKPEAGSAISDVFEGREVCQPKRIRPFHPPGSPWANRPLPASLAPTPTGPVHEPVGSLTPAPVPQPLDPAPAVTPEASHLLEDPDEETSQAVKALREMADTVIPQKEEAAICGQMDLSHPPPRGHLDELTTTLESMTEDLNLDSPLTPELNEILDTFLNDECLLHAMHISTGLSIFDTSLF
Seq ID 34
Mini-saCas9-5:VPR
MPKKKRKVGGGSPGGGGSKRNYILGLDIGITSVGYGIIDYETRDVIDAGVRLFKEANVENNEGRRSKRGARRLKRRRRHRIQRVKKLLFDYNLLTDHSELSGINPYEARVKGLSQKLSEEEFSAALLHLAKRRGVHNVNEVEEDTGNELSTKEQISRNSKALEEKYVAELQLERLKKDGEVRGSINRFKTSDYVKEAKQLLKVQKAYHQLDQSFIDTYIDLLETRRTYYEGPGEGSPFGWKDIKEWYEMLMGHCTKKVDLSQQKEIPTTLVDDFILSPVVKRSFIQSIKVINAIIKKYGLPNDIIIELAREKNSKDAQKMINEMQKRNRQTNERIEEIIRTTGKENAKYLIEKIKLHDMQEGKCLYSLEAIPLEDLLNNPFNYEVDHIIPRSVSFDNSFNNKVLVKQEENSKKGNRTPFQYLSSSDSKISYETFKKHILNLAKGKGRISKTKKEYLLEERDINRFSVQKDFINRNLVDTRYATRGLMNLLRSYFRVNNLDVKVKSINGGFTSFLRRKWKFKKERNKGYKHHAEDALIIANADFIFKEWKKLDKAKKVMENQMFEEKQAESMPEIETEQEYKEIFITPHQIKHIKDFKDYKYSHRVDKKPNRKLINDTLYSTRKDDKGNTLIVNNLNGLYDKDNDKLKKLINKSPEKLLMYHHDPQTYQKLKLIMEQYGDEKNPLYKYYEETGNYLTKYSKKDNGPVIKKIKYYGNKLNAHLDITDDYPNSRNKVVKLSLKPYRFDVYLDNGVYKFVTVKNLDVIKKENYYEVNSKCYEEAKKLKKISNQAEFIASFYKNDLIKINGELYRVIGVNNDLLNRIEVNMIDITYREYLENMNDKRPPHIIKTIASKTQSIKKYSTDILGNLYEVKSKKHPQIIKKGTSRADPKKKRKVEASGSGRADALDDFDLDMLGSDALDDFDLDMLGSDALDDFDLDMLGSDALDDFDLDMLINSRSQYLPDTDDRHRIEEKRKRTYETFKSIMKKSPFSGPTDPRPPPRRIAVPSRSSASVPKPAPQPYPFTSSLSTINYDEFPTMVFPSGQISQASALAPAPPQVLPQAPAPAPAPAMVSALAQAPAPVPVLAPGPPQAVAPPAPKPTQAGEGTLSEALLQLQFDDEDLGALLGNSTDPAVFTDLASVDNSEFQQLLNQGIPVAPHTTEPMLMEYPEAITRLVTGAQRPPDPAPAPLGAPGLPNGLLSGDEDFSSIADMDFSALLGSGSGSRDSREGMFLPKPEAGSAISDVFEGREVCQPKRIRPFHPPGSPWANRPLPASLAPTPTGPVHEPVGSLTPAPVPQPLDPAPAVTPEASHLLEDPDEETSQAVKALREMADTVIPQKEEAAICGQMDLSHPPPRGHLDELTTTLESMTEDLNLDSPLTPELNEILDTFLNDECLLHAMHISTGLSIFDTSLF
Seq ID 35
GSK joint
GSK
Sequence listing
<110> university of Qinghua
<120> engineering minimized SaCas9 CRISPR/Cas systems for gene editing and transcriptional regulation optimized by enhanced guide RNA
<130> PIDC3171800PCN
<160> 34
<170> PatentIn version 3.5
<210> 1
<211> 20
<212> DNA
<213> artificial sequence
<220>
<223> synthetic constructs produced by rational design
<400> 1
tacgttctct atcactgata 20
<210> 2
<211> 20
<212> DNA
<213> artificial sequence
<220>
<223> synthetic constructs produced by rational design
<400> 2
tacgttctct atcactgata 20
<210> 3
<211> 23
<212> DNA
<213> artificial sequence
<220>
<223> synthetic constructs produced by rational design
<400> 3
ctccctatca gtgatagaga acg 23
<210> 4
<211> 18
<212> DNA
<213> artificial sequence
<220>
<223> synthetic constructs produced by rational design
<400> 4
cgttctctat cactgata 18
<210> 5
<211> 18
<212> DNA
<213> artificial sequence
<220>
<223> synthetic constructs produced by rational design
<400> 5
actagaaatt caccgagc 18
<210> 6
<211> 21
<212> DNA
<213> artificial sequence
<220>
<223> synthetic constructs produced by rational design
<400> 6
ctcactcaac agtgatagag a 21
<210> 7
<211> 21
<212> DNA
<213> artificial sequence
<220>
<223> synthetic constructs produced by rational design
<400> 7
ttgctcactc aacagtgata g 21
<210> 8
<211> 1913
<212> PRT
<213> artificial sequence
<220>
<223> synthetic constructs produced by rational design
<400> 8
Met Pro Lys Lys Lys Arg Lys Val Gly Gly Gly Ser Pro Gly Met Asp
1 5 10 15
Lys Lys Tyr Ser Ile Gly Leu Ala Ile Gly Thr Asn Ser Val Gly Trp
20 25 30
Ala Val Ile Thr Asp Glu Tyr Lys Val Pro Ser Lys Lys Phe Lys Val
35 40 45
Leu Gly Asn Thr Asp Arg His Ser Ile Lys Lys Asn Leu Ile Gly Ala
50 55 60
Leu Leu Phe Asp Ser Gly Glu Thr Ala Glu Ala Thr Arg Leu Lys Arg
65 70 75 80
Thr Ala Arg Arg Arg Tyr Thr Arg Arg Lys Asn Arg Ile Cys Tyr Leu
85 90 95
Gln Glu Ile Phe Ser Asn Glu Met Ala Lys Val Asp Asp Ser Phe Phe
100 105 110
His Arg Leu Glu Glu Ser Phe Leu Val Glu Glu Asp Lys Lys His Glu
115 120 125
Arg His Pro Ile Phe Gly Asn Ile Val Asp Glu Val Ala Tyr His Glu
130 135 140
Lys Tyr Pro Thr Ile Tyr His Leu Arg Lys Lys Leu Val Asp Ser Thr
145 150 155 160
Asp Lys Ala Asp Leu Arg Leu Ile Tyr Leu Ala Leu Ala His Met Ile
165 170 175
Lys Phe Arg Gly His Phe Leu Ile Glu Gly Asp Leu Asn Pro Asp Asn
180 185 190
Ser Asp Val Asp Lys Leu Phe Ile Gln Leu Val Gln Thr Tyr Asn Gln
195 200 205
Leu Phe Glu Glu Asn Pro Ile Asn Ala Ser Gly Val Asp Ala Lys Ala
210 215 220
Ile Leu Ser Ala Arg Leu Ser Lys Ser Arg Arg Leu Glu Asn Leu Ile
225 230 235 240
Ala Gln Leu Pro Gly Glu Lys Lys Asn Gly Leu Phe Gly Asn Leu Ile
245 250 255
Ala Leu Ser Leu Gly Leu Thr Pro Asn Phe Lys Ser Asn Phe Asp Leu
260 265 270
Ala Glu Asp Ala Lys Leu Gln Leu Ser Lys Asp Thr Tyr Asp Asp Asp
275 280 285
Leu Asp Asn Leu Leu Ala Gln Ile Gly Asp Gln Tyr Ala Asp Leu Phe
290 295 300
Leu Ala Ala Lys Asn Leu Ser Asp Ala Ile Leu Leu Ser Asp Ile Leu
305 310 315 320
Arg Val Asn Thr Glu Ile Thr Lys Ala Pro Leu Ser Ala Ser Met Ile
325 330 335
Lys Arg Tyr Asp Glu His His Gln Asp Leu Thr Leu Leu Lys Ala Leu
340 345 350
Val Arg Gln Gln Leu Pro Glu Lys Tyr Lys Glu Ile Phe Phe Asp Gln
355 360 365
Ser Lys Asn Gly Tyr Ala Gly Tyr Ile Asp Gly Gly Ala Ser Gln Glu
370 375 380
Glu Phe Tyr Lys Phe Ile Lys Pro Ile Leu Glu Lys Met Asp Gly Thr
385 390 395 400
Glu Glu Leu Leu Val Lys Leu Asn Arg Glu Asp Leu Leu Arg Lys Gln
405 410 415
Arg Thr Phe Asp Asn Gly Ser Ile Pro His Gln Ile His Leu Gly Glu
420 425 430
Leu His Ala Ile Leu Arg Arg Gln Glu Asp Phe Tyr Pro Phe Leu Lys
435 440 445
Asp Asn Arg Glu Lys Ile Glu Lys Ile Leu Thr Phe Arg Ile Pro Tyr
450 455 460
Tyr Val Gly Pro Leu Ala Arg Gly Asn Ser Arg Phe Ala Trp Met Thr
465 470 475 480
Arg Lys Ser Glu Glu Thr Ile Thr Pro Trp Asn Phe Glu Glu Val Val
485 490 495
Asp Lys Gly Ala Ser Ala Gln Ser Phe Ile Glu Arg Met Thr Asn Phe
500 505 510
Asp Lys Asn Leu Pro Asn Glu Lys Val Leu Pro Lys His Ser Leu Leu
515 520 525
Tyr Glu Tyr Phe Thr Val Tyr Asn Glu Leu Thr Lys Val Lys Tyr Val
530 535 540
Thr Glu Gly Met Arg Lys Pro Ala Phe Leu Ser Gly Glu Gln Lys Lys
545 550 555 560
Ala Ile Val Asp Leu Leu Phe Lys Thr Asn Arg Lys Val Thr Val Lys
565 570 575
Gln Leu Lys Glu Asp Tyr Phe Lys Lys Ile Glu Cys Phe Asp Ser Val
580 585 590
Glu Ile Ser Gly Val Glu Asp Arg Phe Asn Ala Ser Leu Gly Thr Tyr
595 600 605
His Asp Leu Leu Lys Ile Ile Lys Asp Lys Asp Phe Leu Asp Asn Glu
610 615 620
Glu Asn Glu Asp Ile Leu Glu Asp Ile Val Leu Thr Leu Thr Leu Phe
625 630 635 640
Glu Asp Arg Glu Met Ile Glu Glu Arg Leu Lys Thr Tyr Ala His Leu
645 650 655
Phe Asp Asp Lys Val Met Lys Gln Leu Lys Arg Arg Arg Tyr Thr Gly
660 665 670
Trp Gly Arg Leu Ser Arg Lys Leu Ile Asn Gly Ile Arg Asp Lys Gln
675 680 685
Ser Gly Lys Thr Ile Leu Asp Phe Leu Lys Ser Asp Gly Phe Ala Asn
690 695 700
Arg Asn Phe Met Gln Leu Ile His Asp Asp Ser Leu Thr Phe Lys Glu
705 710 715 720
Asp Ile Gln Lys Ala Gln Val Ser Gly Gln Gly Asp Ser Leu His Glu
725 730 735
His Ile Ala Asn Leu Ala Gly Ser Pro Ala Ile Lys Lys Gly Ile Leu
740 745 750
Gln Thr Val Lys Val Val Asp Glu Leu Val Lys Val Met Gly Arg His
755 760 765
Lys Pro Glu Asn Ile Val Ile Glu Met Ala Arg Glu Asn Gln Thr Thr
770 775 780
Gln Lys Gly Gln Lys Asn Ser Arg Glu Arg Met Lys Arg Ile Glu Glu
785 790 795 800
Gly Ile Lys Glu Leu Gly Ser Gln Ile Leu Lys Glu His Pro Val Glu
805 810 815
Asn Thr Gln Leu Gln Asn Glu Lys Leu Tyr Leu Tyr Tyr Leu Gln Asn
820 825 830
Gly Arg Asp Met Tyr Val Asp Gln Glu Leu Asp Ile Asn Arg Leu Ser
835 840 845
Asp Tyr Asp Val Asp Ala Ile Val Pro Gln Ser Phe Leu Lys Asp Asp
850 855 860
Ser Ile Asp Asn Lys Val Leu Thr Arg Ser Asp Lys Asn Arg Gly Lys
865 870 875 880
Ser Asp Asn Val Pro Ser Glu Glu Val Val Lys Lys Met Lys Asn Tyr
885 890 895
Trp Arg Gln Leu Leu Asn Ala Lys Leu Ile Thr Gln Arg Lys Phe Asp
900 905 910
Asn Leu Thr Lys Ala Glu Arg Gly Gly Leu Ser Glu Leu Asp Lys Ala
915 920 925
Gly Phe Ile Lys Arg Gln Leu Val Glu Thr Arg Gln Ile Thr Lys His
930 935 940
Val Ala Gln Ile Leu Asp Ser Arg Met Asn Thr Lys Tyr Asp Glu Asn
945 950 955 960
Asp Lys Leu Ile Arg Glu Val Lys Val Ile Thr Leu Lys Ser Lys Leu
965 970 975
Val Ser Asp Phe Arg Lys Asp Phe Gln Phe Tyr Lys Val Arg Glu Ile
980 985 990
Asn Asn Tyr His His Ala His Asp Ala Tyr Leu Asn Ala Val Val Gly
995 1000 1005
Thr Ala Leu Ile Lys Lys Tyr Pro Lys Leu Glu Ser Glu Phe Val
1010 1015 1020
Tyr Gly Asp Tyr Lys Val Tyr Asp Val Arg Lys Met Ile Ala Lys
1025 1030 1035
Ser Glu Gln Glu Ile Gly Lys Ala Thr Ala Lys Tyr Phe Phe Tyr
1040 1045 1050
Ser Asn Ile Met Asn Phe Phe Lys Thr Glu Ile Thr Leu Ala Asn
1055 1060 1065
Gly Glu Ile Arg Lys Arg Pro Leu Ile Glu Thr Asn Gly Glu Thr
1070 1075 1080
Gly Glu Ile Val Trp Asp Lys Gly Arg Asp Phe Ala Thr Val Arg
1085 1090 1095
Lys Val Leu Ser Met Pro Gln Val Asn Ile Val Lys Lys Thr Glu
1100 1105 1110
Val Gln Thr Gly Gly Phe Ser Lys Glu Ser Ile Leu Pro Lys Arg
1115 1120 1125
Asn Ser Asp Lys Leu Ile Ala Arg Lys Lys Asp Trp Asp Pro Lys
1130 1135 1140
Lys Tyr Gly Gly Phe Asp Ser Pro Thr Val Ala Tyr Ser Val Leu
1145 1150 1155
Val Val Ala Lys Val Glu Lys Gly Lys Ser Lys Lys Leu Lys Ser
1160 1165 1170
Val Lys Glu Leu Leu Gly Ile Thr Ile Met Glu Arg Ser Ser Phe
1175 1180 1185
Glu Lys Asn Pro Ile Asp Phe Leu Glu Ala Lys Gly Tyr Lys Glu
1190 1195 1200
Val Lys Lys Asp Leu Ile Ile Lys Leu Pro Lys Tyr Ser Leu Phe
1205 1210 1215
Glu Leu Glu Asn Gly Arg Lys Arg Met Leu Ala Ser Ala Gly Glu
1220 1225 1230
Leu Gln Lys Gly Asn Glu Leu Ala Leu Pro Ser Lys Tyr Val Asn
1235 1240 1245
Phe Leu Tyr Leu Ala Ser His Tyr Glu Lys Leu Lys Gly Ser Pro
1250 1255 1260
Glu Asp Asn Glu Gln Lys Gln Leu Phe Val Glu Gln His Lys His
1265 1270 1275
Tyr Leu Asp Glu Ile Ile Glu Gln Ile Ser Glu Phe Ser Lys Arg
1280 1285 1290
Val Ile Leu Ala Asp Ala Asn Leu Asp Lys Val Leu Ser Ala Tyr
1295 1300 1305
Asn Lys His Arg Asp Lys Pro Ile Arg Glu Gln Ala Glu Asn Ile
1310 1315 1320
Ile His Leu Phe Thr Leu Thr Asn Leu Gly Ala Pro Ala Ala Phe
1325 1330 1335
Lys Tyr Phe Asp Thr Thr Ile Asp Arg Lys Arg Tyr Thr Ser Thr
1340 1345 1350
Lys Glu Val Leu Asp Ala Thr Leu Ile His Gln Ser Ile Thr Gly
1355 1360 1365
Leu Tyr Glu Thr Arg Ile Asp Leu Ser Gln Leu Gly Gly Asp Thr
1370 1375 1380
Ser Arg Ala Asp Pro Lys Lys Lys Arg Lys Val Glu Ala Ser Gly
1385 1390 1395
Ser Gly Arg Ala Asp Ala Leu Asp Asp Phe Asp Leu Asp Met Leu
1400 1405 1410
Gly Ser Asp Ala Leu Asp Asp Phe Asp Leu Asp Met Leu Gly Ser
1415 1420 1425
Asp Ala Leu Asp Asp Phe Asp Leu Asp Met Leu Gly Ser Asp Ala
1430 1435 1440
Leu Asp Asp Phe Asp Leu Asp Met Leu Ile Asn Ser Arg Ser Gln
1445 1450 1455
Tyr Leu Pro Asp Thr Asp Asp Arg His Arg Ile Glu Glu Lys Arg
1460 1465 1470
Lys Arg Thr Tyr Glu Thr Phe Lys Ser Ile Met Lys Lys Ser Pro
1475 1480 1485
Phe Ser Gly Pro Thr Asp Pro Arg Pro Pro Pro Arg Arg Ile Ala
1490 1495 1500
Val Pro Ser Arg Ser Ser Ala Ser Val Pro Lys Pro Ala Pro Gln
1505 1510 1515
Pro Tyr Pro Phe Thr Ser Ser Leu Ser Thr Ile Asn Tyr Asp Glu
1520 1525 1530
Phe Pro Thr Met Val Phe Pro Ser Gly Gln Ile Ser Gln Ala Ser
1535 1540 1545
Ala Leu Ala Pro Ala Pro Pro Gln Val Leu Pro Gln Ala Pro Ala
1550 1555 1560
Pro Ala Pro Ala Pro Ala Met Val Ser Ala Leu Ala Gln Ala Pro
1565 1570 1575
Ala Pro Val Pro Val Leu Ala Pro Gly Pro Pro Gln Ala Val Ala
1580 1585 1590
Pro Pro Ala Pro Lys Pro Thr Gln Ala Gly Glu Gly Thr Leu Ser
1595 1600 1605
Glu Ala Leu Leu Gln Leu Gln Phe Asp Asp Glu Asp Leu Gly Ala
1610 1615 1620
Leu Leu Gly Asn Ser Thr Asp Pro Ala Val Phe Thr Asp Leu Ala
1625 1630 1635
Ser Val Asp Asn Ser Glu Phe Gln Gln Leu Leu Asn Gln Gly Ile
1640 1645 1650
Pro Val Ala Pro His Thr Thr Glu Pro Met Leu Met Glu Tyr Pro
1655 1660 1665
Glu Ala Ile Thr Arg Leu Val Thr Gly Ala Gln Arg Pro Pro Asp
1670 1675 1680
Pro Ala Pro Ala Pro Leu Gly Ala Pro Gly Leu Pro Asn Gly Leu
1685 1690 1695
Leu Ser Gly Asp Glu Asp Phe Ser Ser Ile Ala Asp Met Asp Phe
1700 1705 1710
Ser Ala Leu Leu Gly Ser Gly Ser Gly Ser Arg Asp Ser Arg Glu
1715 1720 1725
Gly Met Phe Leu Pro Lys Pro Glu Ala Gly Ser Ala Ile Ser Asp
1730 1735 1740
Val Phe Glu Gly Arg Glu Val Cys Gln Pro Lys Arg Ile Arg Pro
1745 1750 1755
Phe His Pro Pro Gly Ser Pro Trp Ala Asn Arg Pro Leu Pro Ala
1760 1765 1770
Ser Leu Ala Pro Thr Pro Thr Gly Pro Val His Glu Pro Val Gly
1775 1780 1785
Ser Leu Thr Pro Ala Pro Val Pro Gln Pro Leu Asp Pro Ala Pro
1790 1795 1800
Ala Val Thr Pro Glu Ala Ser His Leu Leu Glu Asp Pro Asp Glu
1805 1810 1815
Glu Thr Ser Gln Ala Val Lys Ala Leu Arg Glu Met Ala Asp Thr
1820 1825 1830
Val Ile Pro Gln Lys Glu Glu Ala Ala Ile Cys Gly Gln Met Asp
1835 1840 1845
Leu Ser His Pro Pro Pro Arg Gly His Leu Asp Glu Leu Thr Thr
1850 1855 1860
Thr Leu Glu Ser Met Thr Glu Asp Leu Asn Leu Asp Ser Pro Leu
1865 1870 1875
Thr Pro Glu Leu Asn Glu Ile Leu Asp Thr Phe Leu Asn Asp Glu
1880 1885 1890
Cys Leu Leu His Ala Met His Ile Ser Thr Gly Leu Ser Ile Phe
1895 1900 1905
Asp Thr Ser Leu Phe
1910
<210> 9
<211> 1703
<212> PRT
<213> artificial sequence
<220>
<223> synthetic constructs produced by rational design
<400> 9
Met Pro Lys Lys Lys Arg Lys Val Gly Gly Gly Ser Pro Gly Met Asp
1 5 10 15
Lys Lys Tyr Ser Ile Gly Leu Ala Ile Gly Thr Asn Ser Val Gly Trp
20 25 30
Ala Val Ile Thr Asp Glu Tyr Lys Val Pro Ser Lys Lys Phe Lys Val
35 40 45
Leu Gly Asn Thr Asp Arg His Ser Ile Lys Lys Asn Leu Ile Gly Ala
50 55 60
Leu Leu Phe Asp Ser Gly Glu Thr Ala Glu Ala Thr Arg Leu Lys Arg
65 70 75 80
Thr Ala Arg Arg Arg Tyr Thr Arg Arg Lys Asn Arg Ile Cys Tyr Leu
85 90 95
Gln Glu Ile Phe Ser Asn Glu Met Ala Lys Val Asp Asp Ser Phe Phe
100 105 110
His Arg Leu Glu Glu Ser Phe Leu Val Glu Glu Asp Lys Lys His Glu
115 120 125
Arg His Pro Ile Phe Gly Asn Ile Val Asp Glu Val Ala Tyr His Glu
130 135 140
Lys Tyr Pro Thr Ile Tyr His Leu Arg Lys Lys Leu Val Asp Ser Thr
145 150 155 160
Asp Lys Ala Asp Leu Arg Leu Ile Tyr Leu Ala Leu Ala His Met Ile
165 170 175
Lys Phe Arg Gly His Phe Leu Ile Glu Gly Asp Leu Asn Pro Asp Asn
180 185 190
Ser Asp Val Asp Lys Leu Phe Ile Gln Leu Val Gln Thr Tyr Asn Gln
195 200 205
Leu Phe Glu Glu Asn Pro Ile Asn Ala Ser Gly Val Asp Ala Lys Ala
210 215 220
Ile Leu Ser Ala Arg Leu Ser Lys Ser Arg Arg Leu Glu Asn Leu Ile
225 230 235 240
Ala Gln Leu Pro Gly Glu Lys Lys Asn Gly Leu Phe Gly Asn Leu Ile
245 250 255
Ala Leu Ser Leu Gly Leu Thr Pro Asn Phe Lys Ser Asn Phe Asp Leu
260 265 270
Ala Glu Asp Ala Lys Leu Gln Leu Ser Lys Asp Thr Tyr Asp Asp Asp
275 280 285
Leu Asp Asn Leu Leu Ala Gln Ile Gly Asp Gln Tyr Ala Asp Leu Phe
290 295 300
Leu Ala Ala Lys Asn Leu Ser Asp Ala Ile Leu Leu Ser Asp Ile Leu
305 310 315 320
Arg Val Asn Thr Glu Ile Thr Lys Ala Pro Leu Ser Ala Ser Met Ile
325 330 335
Lys Arg Tyr Asp Glu His His Gln Asp Leu Thr Leu Leu Lys Ala Leu
340 345 350
Val Arg Gln Gln Leu Pro Glu Lys Tyr Lys Glu Ile Phe Phe Asp Gln
355 360 365
Ser Lys Asn Gly Tyr Ala Gly Tyr Ile Asp Gly Gly Ala Ser Gln Glu
370 375 380
Glu Phe Tyr Lys Phe Ile Lys Pro Ile Leu Glu Lys Met Asp Gly Thr
385 390 395 400
Glu Glu Leu Leu Val Lys Leu Asn Arg Glu Asp Leu Leu Arg Lys Gln
405 410 415
Arg Thr Phe Asp Asn Gly Ser Ile Pro His Gln Ile His Leu Gly Glu
420 425 430
Leu His Ala Ile Leu Arg Arg Gln Glu Asp Phe Tyr Pro Phe Leu Lys
435 440 445
Asp Asn Arg Glu Lys Ile Glu Lys Ile Leu Thr Phe Arg Ile Pro Tyr
450 455 460
Tyr Val Gly Pro Leu Ala Arg Gly Asn Ser Arg Phe Ala Trp Met Thr
465 470 475 480
Arg Lys Ser Glu Glu Thr Ile Thr Pro Trp Asn Phe Glu Glu Val Val
485 490 495
Asp Lys Gly Ala Ser Ala Gln Ser Phe Ile Glu Arg Met Thr Asn Phe
500 505 510
Asp Lys Ala Gln Val Ser Gly Gln Gly Asp Ser Leu His Glu His Ile
515 520 525
Ala Asn Leu Ala Gly Ser Pro Ala Ile Lys Lys Gly Ile Leu Gln Thr
530 535 540
Val Lys Val Val Asp Glu Leu Val Lys Val Met Gly Arg His Lys Pro
545 550 555 560
Glu Asn Ile Val Ile Glu Met Ala Arg Glu Asn Gln Thr Thr Gln Lys
565 570 575
Gly Gln Lys Asn Ser Arg Glu Arg Met Lys Arg Ile Glu Glu Gly Ile
580 585 590
Lys Glu Leu Gly Ser Gln Ile Leu Lys Glu His Pro Val Glu Asn Thr
595 600 605
Gln Leu Gln Asn Glu Lys Leu Tyr Leu Tyr Tyr Leu Gln Asn Gly Arg
610 615 620
Asp Met Tyr Val Asp Gln Glu Leu Asp Ile Asn Arg Leu Ser Asp Tyr
625 630 635 640
Asp Val Asp Ala Ile Val Pro Gln Ser Phe Leu Lys Asp Asp Ser Ile
645 650 655
Asp Asn Lys Val Leu Thr Arg Ser Asp Lys Asn Arg Gly Lys Ser Asp
660 665 670
Asn Val Pro Ser Glu Glu Val Val Lys Lys Met Lys Asn Tyr Trp Arg
675 680 685
Gln Leu Leu Asn Ala Lys Leu Ile Thr Gln Arg Lys Phe Asp Asn Leu
690 695 700
Thr Lys Ala Glu Arg Gly Gly Leu Ser Glu Leu Asp Lys Ala Gly Phe
705 710 715 720
Ile Lys Arg Gln Leu Val Glu Thr Arg Gln Ile Thr Lys His Val Ala
725 730 735
Gln Ile Leu Asp Ser Arg Met Asn Thr Lys Tyr Asp Glu Asn Asp Lys
740 745 750
Leu Ile Arg Glu Val Lys Val Ile Thr Leu Lys Ser Lys Leu Val Ser
755 760 765
Asp Phe Arg Lys Asp Phe Gln Phe Tyr Lys Val Arg Glu Ile Asn Asn
770 775 780
Tyr His His Ala His Asp Ala Tyr Leu Asn Ala Val Val Gly Thr Ala
785 790 795 800
Leu Ile Lys Lys Tyr Pro Lys Leu Glu Ser Glu Phe Val Tyr Gly Asp
805 810 815
Tyr Lys Val Tyr Asp Val Arg Lys Met Ile Ala Lys Ser Glu Gln Glu
820 825 830
Ile Gly Lys Ala Thr Ala Lys Tyr Phe Phe Tyr Ser Asn Ile Met Asn
835 840 845
Phe Phe Lys Thr Glu Ile Thr Leu Ala Asn Gly Glu Ile Arg Lys Arg
850 855 860
Pro Leu Ile Glu Thr Asn Gly Glu Thr Gly Glu Ile Val Trp Asp Lys
865 870 875 880
Gly Arg Asp Phe Ala Thr Val Arg Lys Val Leu Ser Met Pro Gln Val
885 890 895
Asn Ile Val Lys Lys Thr Glu Val Gln Thr Gly Gly Phe Ser Lys Glu
900 905 910
Ser Ile Leu Pro Lys Arg Asn Ser Asp Lys Leu Ile Ala Arg Lys Lys
915 920 925
Asp Trp Asp Pro Lys Lys Tyr Gly Gly Phe Asp Ser Pro Thr Val Ala
930 935 940
Tyr Ser Val Leu Val Val Ala Lys Val Glu Lys Gly Lys Ser Lys Lys
945 950 955 960
Leu Lys Ser Val Lys Glu Leu Leu Gly Ile Thr Ile Met Glu Arg Ser
965 970 975
Ser Phe Glu Lys Asn Pro Ile Asp Phe Leu Glu Ala Lys Gly Tyr Lys
980 985 990
Glu Val Lys Lys Asp Leu Ile Ile Lys Leu Pro Lys Tyr Ser Leu Phe
995 1000 1005
Glu Leu Glu Asn Gly Arg Lys Arg Met Leu Ala Ser Ala Gly Glu
1010 1015 1020
Leu Gln Lys Gly Asn Glu Leu Ala Leu Pro Ser Lys Tyr Val Asn
1025 1030 1035
Phe Leu Tyr Leu Ala Ser His Tyr Glu Lys Leu Lys Gly Ser Pro
1040 1045 1050
Glu Asp Asn Glu Gln Lys Gln Leu Phe Val Glu Gln His Lys His
1055 1060 1065
Tyr Leu Asp Glu Ile Ile Glu Gln Ile Ser Glu Phe Ser Lys Arg
1070 1075 1080
Val Ile Leu Ala Asp Ala Asn Leu Asp Lys Val Leu Ser Ala Tyr
1085 1090 1095
Asn Lys His Arg Asp Lys Pro Ile Arg Glu Gln Ala Glu Asn Ile
1100 1105 1110
Ile His Leu Phe Thr Leu Thr Asn Leu Gly Ala Pro Ala Ala Phe
1115 1120 1125
Lys Tyr Phe Asp Thr Thr Ile Asp Arg Lys Arg Tyr Thr Ser Thr
1130 1135 1140
Lys Glu Val Leu Asp Ala Thr Leu Ile His Gln Ser Ile Thr Gly
1145 1150 1155
Leu Tyr Glu Thr Arg Ile Asp Leu Ser Gln Leu Gly Gly Asp Thr
1160 1165 1170
Ser Arg Ala Asp Pro Lys Lys Lys Arg Lys Val Glu Ala Ser Gly
1175 1180 1185
Ser Gly Arg Ala Asp Ala Leu Asp Asp Phe Asp Leu Asp Met Leu
1190 1195 1200
Gly Ser Asp Ala Leu Asp Asp Phe Asp Leu Asp Met Leu Gly Ser
1205 1210 1215
Asp Ala Leu Asp Asp Phe Asp Leu Asp Met Leu Gly Ser Asp Ala
1220 1225 1230
Leu Asp Asp Phe Asp Leu Asp Met Leu Ile Asn Ser Arg Ser Gln
1235 1240 1245
Tyr Leu Pro Asp Thr Asp Asp Arg His Arg Ile Glu Glu Lys Arg
1250 1255 1260
Lys Arg Thr Tyr Glu Thr Phe Lys Ser Ile Met Lys Lys Ser Pro
1265 1270 1275
Phe Ser Gly Pro Thr Asp Pro Arg Pro Pro Pro Arg Arg Ile Ala
1280 1285 1290
Val Pro Ser Arg Ser Ser Ala Ser Val Pro Lys Pro Ala Pro Gln
1295 1300 1305
Pro Tyr Pro Phe Thr Ser Ser Leu Ser Thr Ile Asn Tyr Asp Glu
1310 1315 1320
Phe Pro Thr Met Val Phe Pro Ser Gly Gln Ile Ser Gln Ala Ser
1325 1330 1335
Ala Leu Ala Pro Ala Pro Pro Gln Val Leu Pro Gln Ala Pro Ala
1340 1345 1350
Pro Ala Pro Ala Pro Ala Met Val Ser Ala Leu Ala Gln Ala Pro
1355 1360 1365
Ala Pro Val Pro Val Leu Ala Pro Gly Pro Pro Gln Ala Val Ala
1370 1375 1380
Pro Pro Ala Pro Lys Pro Thr Gln Ala Gly Glu Gly Thr Leu Ser
1385 1390 1395
Glu Ala Leu Leu Gln Leu Gln Phe Asp Asp Glu Asp Leu Gly Ala
1400 1405 1410
Leu Leu Gly Asn Ser Thr Asp Pro Ala Val Phe Thr Asp Leu Ala
1415 1420 1425
Ser Val Asp Asn Ser Glu Phe Gln Gln Leu Leu Asn Gln Gly Ile
1430 1435 1440
Pro Val Ala Pro His Thr Thr Glu Pro Met Leu Met Glu Tyr Pro
1445 1450 1455
Glu Ala Ile Thr Arg Leu Val Thr Gly Ala Gln Arg Pro Pro Asp
1460 1465 1470
Pro Ala Pro Ala Pro Leu Gly Ala Pro Gly Leu Pro Asn Gly Leu
1475 1480 1485
Leu Ser Gly Asp Glu Asp Phe Ser Ser Ile Ala Asp Met Asp Phe
1490 1495 1500
Ser Ala Leu Leu Gly Ser Gly Ser Gly Ser Arg Asp Ser Arg Glu
1505 1510 1515
Gly Met Phe Leu Pro Lys Pro Glu Ala Gly Ser Ala Ile Ser Asp
1520 1525 1530
Val Phe Glu Gly Arg Glu Val Cys Gln Pro Lys Arg Ile Arg Pro
1535 1540 1545
Phe His Pro Pro Gly Ser Pro Trp Ala Asn Arg Pro Leu Pro Ala
1550 1555 1560
Ser Leu Ala Pro Thr Pro Thr Gly Pro Val His Glu Pro Val Gly
1565 1570 1575
Ser Leu Thr Pro Ala Pro Val Pro Gln Pro Leu Asp Pro Ala Pro
1580 1585 1590
Ala Val Thr Pro Glu Ala Ser His Leu Leu Glu Asp Pro Asp Glu
1595 1600 1605
Glu Thr Ser Gln Ala Val Lys Ala Leu Arg Glu Met Ala Asp Thr
1610 1615 1620
Val Ile Pro Gln Lys Glu Glu Ala Ala Ile Cys Gly Gln Met Asp
1625 1630 1635
Leu Ser His Pro Pro Pro Arg Gly His Leu Asp Glu Leu Thr Thr
1640 1645 1650
Thr Leu Glu Ser Met Thr Glu Asp Leu Asn Leu Asp Ser Pro Leu
1655 1660 1665
Thr Pro Glu Leu Asn Glu Ile Leu Asp Thr Phe Leu Asn Asp Glu
1670 1675 1680
Cys Leu Leu His Ala Met His Ile Ser Thr Gly Leu Ser Ile Phe
1685 1690 1695
Asp Thr Ser Leu Phe
1700
<210> 10
<211> 1798
<212> PRT
<213> artificial sequence
<220>
<223> synthetic constructs produced by rational design
<400> 10
Met Pro Lys Lys Lys Arg Lys Val Gly Gly Gly Ser Pro Gly Met Asp
1 5 10 15
Lys Lys Tyr Ser Ile Gly Leu Ala Ile Gly Thr Asn Ser Val Gly Trp
20 25 30
Ala Val Ile Thr Asp Glu Tyr Lys Val Pro Ser Lys Lys Phe Lys Val
35 40 45
Leu Gly Asn Thr Asp Arg His Ser Ile Lys Lys Asn Leu Ile Gly Ala
50 55 60
Leu Leu Phe Asp Ser Gly Glu Thr Ala Glu Ala Thr Arg Leu Lys Arg
65 70 75 80
Thr Ala Arg Arg Arg Tyr Thr Arg Arg Lys Asn Arg Ile Cys Tyr Leu
85 90 95
Gln Glu Ile Phe Ser Asn Glu Met Ala Lys Val Asp Asp Ser Phe Phe
100 105 110
His Arg Leu Glu Glu Ser Phe Leu Val Glu Glu Asp Lys Lys His Glu
115 120 125
Arg His Pro Ile Phe Gly Asn Ile Val Asp Glu Val Ala Tyr His Glu
130 135 140
Lys Tyr Pro Thr Ile Tyr His Leu Arg Lys Lys Leu Val Asp Ser Thr
145 150 155 160
Asp Lys Ala Asp Leu Arg Leu Ile Tyr Leu Ala Leu Ala His Met Ile
165 170 175
Lys Phe Arg Gly His Phe Leu Ile Glu Gly Asp Leu Asn Pro Asp Asn
180 185 190
Ser Asp Val Asp Lys Leu Phe Ile Gln Leu Val Gln Thr Tyr Asn Gln
195 200 205
Leu Phe Glu Glu Asn Pro Ile Asn Ala Ser Gly Val Asp Ala Lys Ala
210 215 220
Ile Leu Ser Ala Arg Leu Ser Lys Ser Arg Arg Leu Glu Asn Leu Ile
225 230 235 240
Ala Gln Leu Pro Gly Glu Lys Lys Asn Gly Leu Phe Gly Asn Leu Ile
245 250 255
Ala Leu Ser Leu Gly Leu Thr Pro Asn Phe Lys Ser Asn Phe Asp Leu
260 265 270
Ala Glu Asp Ala Lys Leu Gln Leu Ser Lys Asp Thr Tyr Asp Asp Asp
275 280 285
Leu Asp Asn Leu Leu Ala Gln Ile Gly Asp Gln Tyr Ala Asp Leu Phe
290 295 300
Leu Ala Ala Lys Asn Leu Ser Asp Ala Ile Leu Leu Ser Asp Ile Leu
305 310 315 320
Arg Val Asn Thr Glu Ile Thr Lys Ala Pro Leu Ser Ala Ser Met Ile
325 330 335
Lys Arg Tyr Asp Glu His His Gln Asp Leu Thr Leu Leu Lys Ala Leu
340 345 350
Val Arg Gln Gln Leu Pro Glu Lys Tyr Lys Glu Ile Phe Phe Asp Gln
355 360 365
Ser Lys Asn Gly Tyr Ala Gly Tyr Ile Asp Gly Gly Ala Ser Gln Glu
370 375 380
Glu Phe Tyr Lys Phe Ile Lys Pro Ile Leu Glu Lys Met Asp Gly Thr
385 390 395 400
Glu Glu Leu Leu Val Lys Leu Asn Arg Glu Asp Leu Leu Arg Lys Gln
405 410 415
Arg Thr Phe Asp Asn Gly Ser Ile Pro His Gln Ile His Leu Gly Glu
420 425 430
Leu His Ala Ile Leu Arg Arg Gln Glu Asp Phe Tyr Pro Phe Leu Lys
435 440 445
Asp Asn Arg Glu Lys Ile Glu Lys Ile Leu Thr Phe Arg Ile Pro Tyr
450 455 460
Tyr Val Gly Pro Leu Ala Arg Gly Asn Ser Arg Phe Ala Trp Met Thr
465 470 475 480
Arg Lys Ser Glu Glu Thr Ile Thr Pro Trp Asn Phe Glu Glu Val Val
485 490 495
Asp Lys Gly Ala Ser Ala Gln Ser Phe Ile Glu Arg Met Thr Asn Phe
500 505 510
Asp Lys Asn Leu Pro Asn Glu Lys Val Leu Pro Lys His Ser Leu Leu
515 520 525
Tyr Glu Tyr Phe Thr Val Tyr Asn Glu Leu Thr Lys Val Lys Tyr Val
530 535 540
Thr Glu Gly Met Arg Lys Pro Ala Phe Leu Ser Gly Glu Gln Lys Lys
545 550 555 560
Ala Ile Val Asp Leu Leu Phe Lys Thr Asn Arg Lys Val Thr Val Lys
565 570 575
Gln Leu Lys Glu Asp Tyr Phe Lys Lys Ile Glu Cys Phe Asp Ser Val
580 585 590
Glu Ile Ser Gly Val Glu Asp Arg Phe Asn Ala Ser Leu Gly Thr Tyr
595 600 605
His Asp Leu Leu Lys Ile Ile Lys Asp Lys Asp Phe Leu Asp Asn Glu
610 615 620
Glu Asn Glu Asp Ile Leu Glu Asp Ile Val Leu Thr Leu Thr Leu Phe
625 630 635 640
Glu Asp Arg Glu Met Ile Glu Glu Arg Leu Lys Thr Tyr Ala His Leu
645 650 655
Phe Asp Asp Lys Val Met Lys Gln Leu Lys Arg Arg Arg Tyr Thr Gly
660 665 670
Trp Gly Arg Leu Ser Arg Lys Leu Ile Asn Gly Ile Arg Asp Lys Gln
675 680 685
Ser Gly Lys Thr Ile Leu Asp Phe Leu Lys Ser Asp Gly Phe Ala Asn
690 695 700
Arg Asn Phe Met Gln Leu Ile His Asp Asp Ser Leu Thr Phe Lys Glu
705 710 715 720
Asp Ile Gln Lys Ala Gln Val Ser Gly Gln Gly Asp Ser Leu His Glu
725 730 735
His Ile Ala Asn Leu Ala Gly Ser Pro Ala Ile Lys Lys Gly Ile Leu
740 745 750
Gln Thr Val Lys Val Val Asp Glu Leu Val Lys Val Met Gly Arg His
755 760 765
Lys Pro Glu Asn Ile Val Ile Glu Met Ala Arg Glu Asn Gln Thr Thr
770 775 780
Gln Lys Gly Gln Lys Asn Ile Thr Gln Arg Lys Phe Asp Asn Leu Thr
785 790 795 800
Lys Ala Glu Arg Gly Gly Leu Ser Glu Leu Asp Lys Ala Gly Phe Ile
805 810 815
Lys Arg Gln Leu Val Glu Thr Arg Gln Ile Thr Lys His Val Ala Gln
820 825 830
Ile Leu Asp Ser Arg Met Asn Thr Lys Tyr Asp Glu Asn Asp Lys Leu
835 840 845
Ile Arg Glu Val Lys Val Ile Thr Leu Lys Ser Lys Leu Val Ser Asp
850 855 860
Phe Arg Lys Asp Phe Gln Phe Tyr Lys Val Arg Glu Ile Asn Asn Tyr
865 870 875 880
His His Ala His Asp Ala Tyr Leu Asn Ala Val Val Gly Thr Ala Leu
885 890 895
Ile Lys Lys Tyr Pro Lys Leu Glu Ser Glu Phe Val Tyr Gly Asp Tyr
900 905 910
Lys Val Tyr Asp Val Arg Lys Met Ile Ala Lys Ser Glu Gln Glu Ile
915 920 925
Gly Lys Ala Thr Ala Lys Tyr Phe Phe Tyr Ser Asn Ile Met Asn Phe
930 935 940
Phe Lys Thr Glu Ile Thr Leu Ala Asn Gly Glu Ile Arg Lys Arg Pro
945 950 955 960
Leu Ile Glu Thr Asn Gly Glu Thr Gly Glu Ile Val Trp Asp Lys Gly
965 970 975
Arg Asp Phe Ala Thr Val Arg Lys Val Leu Ser Met Pro Gln Val Asn
980 985 990
Ile Val Lys Lys Thr Glu Val Gln Thr Gly Gly Phe Ser Lys Glu Ser
995 1000 1005
Ile Leu Pro Lys Arg Asn Ser Asp Lys Leu Ile Ala Arg Lys Lys
1010 1015 1020
Asp Trp Asp Pro Lys Lys Tyr Gly Gly Phe Asp Ser Pro Thr Val
1025 1030 1035
Ala Tyr Ser Val Leu Val Val Ala Lys Val Glu Lys Gly Lys Ser
1040 1045 1050
Lys Lys Leu Lys Ser Val Lys Glu Leu Leu Gly Ile Thr Ile Met
1055 1060 1065
Glu Arg Ser Ser Phe Glu Lys Asn Pro Ile Asp Phe Leu Glu Ala
1070 1075 1080
Lys Gly Tyr Lys Glu Val Lys Lys Asp Leu Ile Ile Lys Leu Pro
1085 1090 1095
Lys Tyr Ser Leu Phe Glu Leu Glu Asn Gly Arg Lys Arg Met Leu
1100 1105 1110
Ala Ser Ala Gly Glu Leu Gln Lys Gly Asn Glu Leu Ala Leu Pro
1115 1120 1125
Ser Lys Tyr Val Asn Phe Leu Tyr Leu Ala Ser His Tyr Glu Lys
1130 1135 1140
Leu Lys Gly Ser Pro Glu Asp Asn Glu Gln Lys Gln Leu Phe Val
1145 1150 1155
Glu Gln His Lys His Tyr Leu Asp Glu Ile Ile Glu Gln Ile Ser
1160 1165 1170
Glu Phe Ser Lys Arg Val Ile Leu Ala Asp Ala Asn Leu Asp Lys
1175 1180 1185
Val Leu Ser Ala Tyr Asn Lys His Arg Asp Lys Pro Ile Arg Glu
1190 1195 1200
Gln Ala Glu Asn Ile Ile His Leu Phe Thr Leu Thr Asn Leu Gly
1205 1210 1215
Ala Pro Ala Ala Phe Lys Tyr Phe Asp Thr Thr Ile Asp Arg Lys
1220 1225 1230
Arg Tyr Thr Ser Thr Lys Glu Val Leu Asp Ala Thr Leu Ile His
1235 1240 1245
Gln Ser Ile Thr Gly Leu Tyr Glu Thr Arg Ile Asp Leu Ser Gln
1250 1255 1260
Leu Gly Gly Asp Thr Ser Arg Ala Asp Pro Lys Lys Lys Arg Lys
1265 1270 1275
Val Glu Ala Ser Gly Ser Gly Arg Ala Asp Ala Leu Asp Asp Phe
1280 1285 1290
Asp Leu Asp Met Leu Gly Ser Asp Ala Leu Asp Asp Phe Asp Leu
1295 1300 1305
Asp Met Leu Gly Ser Asp Ala Leu Asp Asp Phe Asp Leu Asp Met
1310 1315 1320
Leu Gly Ser Asp Ala Leu Asp Asp Phe Asp Leu Asp Met Leu Ile
1325 1330 1335
Asn Ser Arg Ser Gln Tyr Leu Pro Asp Thr Asp Asp Arg His Arg
1340 1345 1350
Ile Glu Glu Lys Arg Lys Arg Thr Tyr Glu Thr Phe Lys Ser Ile
1355 1360 1365
Met Lys Lys Ser Pro Phe Ser Gly Pro Thr Asp Pro Arg Pro Pro
1370 1375 1380
Pro Arg Arg Ile Ala Val Pro Ser Arg Ser Ser Ala Ser Val Pro
1385 1390 1395
Lys Pro Ala Pro Gln Pro Tyr Pro Phe Thr Ser Ser Leu Ser Thr
1400 1405 1410
Ile Asn Tyr Asp Glu Phe Pro Thr Met Val Phe Pro Ser Gly Gln
1415 1420 1425
Ile Ser Gln Ala Ser Ala Leu Ala Pro Ala Pro Pro Gln Val Leu
1430 1435 1440
Pro Gln Ala Pro Ala Pro Ala Pro Ala Pro Ala Met Val Ser Ala
1445 1450 1455
Leu Ala Gln Ala Pro Ala Pro Val Pro Val Leu Ala Pro Gly Pro
1460 1465 1470
Pro Gln Ala Val Ala Pro Pro Ala Pro Lys Pro Thr Gln Ala Gly
1475 1480 1485
Glu Gly Thr Leu Ser Glu Ala Leu Leu Gln Leu Gln Phe Asp Asp
1490 1495 1500
Glu Asp Leu Gly Ala Leu Leu Gly Asn Ser Thr Asp Pro Ala Val
1505 1510 1515
Phe Thr Asp Leu Ala Ser Val Asp Asn Ser Glu Phe Gln Gln Leu
1520 1525 1530
Leu Asn Gln Gly Ile Pro Val Ala Pro His Thr Thr Glu Pro Met
1535 1540 1545
Leu Met Glu Tyr Pro Glu Ala Ile Thr Arg Leu Val Thr Gly Ala
1550 1555 1560
Gln Arg Pro Pro Asp Pro Ala Pro Ala Pro Leu Gly Ala Pro Gly
1565 1570 1575
Leu Pro Asn Gly Leu Leu Ser Gly Asp Glu Asp Phe Ser Ser Ile
1580 1585 1590
Ala Asp Met Asp Phe Ser Ala Leu Leu Gly Ser Gly Ser Gly Ser
1595 1600 1605
Arg Asp Ser Arg Glu Gly Met Phe Leu Pro Lys Pro Glu Ala Gly
1610 1615 1620
Ser Ala Ile Ser Asp Val Phe Glu Gly Arg Glu Val Cys Gln Pro
1625 1630 1635
Lys Arg Ile Arg Pro Phe His Pro Pro Gly Ser Pro Trp Ala Asn
1640 1645 1650
Arg Pro Leu Pro Ala Ser Leu Ala Pro Thr Pro Thr Gly Pro Val
1655 1660 1665
His Glu Pro Val Gly Ser Leu Thr Pro Ala Pro Val Pro Gln Pro
1670 1675 1680
Leu Asp Pro Ala Pro Ala Val Thr Pro Glu Ala Ser His Leu Leu
1685 1690 1695
Glu Asp Pro Asp Glu Glu Thr Ser Gln Ala Val Lys Ala Leu Arg
1700 1705 1710
Glu Met Ala Asp Thr Val Ile Pro Gln Lys Glu Glu Ala Ala Ile
1715 1720 1725
Cys Gly Gln Met Asp Leu Ser His Pro Pro Pro Arg Gly His Leu
1730 1735 1740
Asp Glu Leu Thr Thr Thr Leu Glu Ser Met Thr Glu Asp Leu Asn
1745 1750 1755
Leu Asp Ser Pro Leu Thr Pro Glu Leu Asn Glu Ile Leu Asp Thr
1760 1765 1770
Phe Leu Asn Asp Glu Cys Leu Leu His Ala Met His Ile Ser Thr
1775 1780 1785
Gly Leu Ser Ile Phe Asp Thr Ser Leu Phe
1790 1795
<210> 11
<211> 1597
<212> PRT
<213> artificial sequence
<220>
<223> synthetic constructs produced by rational design
<400> 11
Met Pro Lys Lys Lys Arg Lys Val Gly Gly Gly Ser Pro Gly Lys Arg
1 5 10 15
Asn Tyr Ile Leu Gly Leu Ala Ile Gly Ile Thr Ser Val Gly Tyr Gly
20 25 30
Ile Ile Asp Tyr Glu Thr Arg Asp Val Ile Asp Ala Gly Val Arg Leu
35 40 45
Phe Lys Glu Ala Asn Val Glu Asn Asn Glu Gly Arg Arg Ser Lys Arg
50 55 60
Gly Ala Arg Arg Leu Lys Arg Arg Arg Arg His Arg Ile Gln Arg Val
65 70 75 80
Lys Lys Leu Leu Phe Asp Tyr Asn Leu Leu Thr Asp His Ser Glu Leu
85 90 95
Ser Gly Ile Asn Pro Tyr Glu Ala Arg Val Lys Gly Leu Ser Gln Lys
100 105 110
Leu Ser Glu Glu Glu Phe Ser Ala Ala Leu Leu His Leu Ala Lys Arg
115 120 125
Arg Gly Val His Asn Val Asn Glu Val Glu Glu Asp Thr Gly Asn Glu
130 135 140
Leu Ser Thr Lys Glu Gln Ile Ser Arg Asn Ser Lys Ala Leu Glu Glu
145 150 155 160
Lys Tyr Val Ala Glu Leu Gln Leu Glu Arg Leu Lys Lys Asp Gly Glu
165 170 175
Val Arg Gly Ser Ile Asn Arg Phe Lys Thr Ser Asp Tyr Val Lys Glu
180 185 190
Ala Lys Gln Leu Leu Lys Val Gln Lys Ala Tyr His Gln Leu Asp Gln
195 200 205
Ser Phe Ile Asp Thr Tyr Ile Asp Leu Leu Glu Thr Arg Arg Thr Tyr
210 215 220
Tyr Glu Gly Pro Gly Glu Gly Ser Pro Phe Gly Trp Lys Asp Ile Lys
225 230 235 240
Glu Trp Tyr Glu Met Leu Met Gly His Cys Thr Tyr Phe Pro Glu Glu
245 250 255
Leu Arg Ser Val Lys Tyr Ala Tyr Asn Ala Asp Leu Tyr Asn Ala Leu
260 265 270
Asn Asp Leu Asn Asn Leu Val Ile Thr Arg Asp Glu Asn Glu Lys Leu
275 280 285
Glu Tyr Tyr Glu Lys Phe Gln Ile Ile Glu Asn Val Phe Lys Gln Lys
290 295 300
Lys Lys Pro Thr Leu Lys Gln Ile Ala Lys Glu Ile Leu Val Asn Glu
305 310 315 320
Glu Asp Ile Lys Gly Tyr Arg Val Thr Ser Thr Gly Lys Pro Glu Phe
325 330 335
Thr Asn Leu Lys Val Tyr His Asp Ile Lys Asp Ile Thr Ala Arg Lys
340 345 350
Glu Ile Ile Glu Asn Ala Glu Leu Leu Asp Gln Ile Ala Lys Ile Leu
355 360 365
Thr Ile Tyr Gln Ser Ser Glu Asp Ile Gln Glu Glu Leu Thr Asn Leu
370 375 380
Asn Ser Glu Leu Thr Gln Glu Glu Ile Glu Gln Ile Ser Asn Leu Lys
385 390 395 400
Gly Tyr Thr Gly Thr His Asn Leu Ser Leu Lys Ala Ile Asn Leu Ile
405 410 415
Leu Asp Glu Leu Trp His Thr Asn Asp Asn Gln Ile Ala Ile Phe Asn
420 425 430
Arg Leu Lys Leu Val Pro Lys Lys Val Asp Leu Ser Gln Gln Lys Glu
435 440 445
Ile Pro Thr Thr Leu Val Asp Asp Phe Ile Leu Ser Pro Val Val Lys
450 455 460
Arg Ser Phe Ile Gln Ser Ile Lys Val Ile Asn Ala Ile Ile Lys Lys
465 470 475 480
Tyr Gly Leu Pro Asn Asp Ile Ile Ile Glu Leu Ala Arg Glu Lys Asn
485 490 495
Ser Lys Asp Ala Gln Lys Met Ile Asn Glu Met Gln Lys Arg Asn Arg
500 505 510
Gln Thr Asn Glu Arg Ile Glu Glu Ile Ile Arg Thr Thr Gly Lys Glu
515 520 525
Asn Ala Lys Tyr Leu Ile Glu Lys Ile Lys Leu His Asp Met Gln Glu
530 535 540
Gly Lys Cys Leu Tyr Ser Leu Glu Ala Ile Pro Leu Glu Asp Leu Leu
545 550 555 560
Asn Asn Pro Phe Asn Tyr Glu Val Asp His Ile Ile Pro Arg Ser Val
565 570 575
Ser Phe Asp Asn Ser Phe Asn Asn Lys Val Leu Val Lys Gln Glu Glu
580 585 590
Ala Ser Lys Lys Gly Asn Arg Thr Pro Phe Gln Tyr Leu Ser Ser Ser
595 600 605
Asp Ser Lys Ile Ser Tyr Glu Thr Phe Lys Lys His Ile Leu Asn Leu
610 615 620
Ala Lys Gly Lys Gly Arg Ile Ser Lys Thr Lys Lys Glu Tyr Leu Leu
625 630 635 640
Glu Glu Arg Asp Ile Asn Arg Phe Ser Val Gln Lys Asp Phe Ile Asn
645 650 655
Arg Asn Leu Val Asp Thr Arg Tyr Ala Thr Arg Gly Leu Met Asn Leu
660 665 670
Leu Arg Ser Tyr Phe Arg Val Asn Asn Leu Asp Val Lys Val Lys Ser
675 680 685
Ile Asn Gly Gly Phe Thr Ser Phe Leu Arg Arg Lys Trp Lys Phe Lys
690 695 700
Lys Glu Arg Asn Lys Gly Tyr Lys His His Ala Glu Asp Ala Leu Ile
705 710 715 720
Ile Ala Asn Ala Asp Phe Ile Phe Lys Glu Trp Lys Lys Leu Asp Lys
725 730 735
Ala Lys Lys Val Met Glu Asn Gln Met Phe Glu Glu Lys Gln Ala Glu
740 745 750
Ser Met Pro Glu Ile Glu Thr Glu Gln Glu Tyr Lys Glu Ile Phe Ile
755 760 765
Thr Pro His Gln Ile Lys His Ile Lys Asp Phe Lys Asp Tyr Lys Tyr
770 775 780
Ser His Arg Val Asp Lys Lys Pro Asn Arg Lys Leu Ile Asn Asp Thr
785 790 795 800
Leu Tyr Ser Thr Arg Lys Asp Asp Lys Gly Asn Thr Leu Ile Val Asn
805 810 815
Asn Leu Asn Gly Leu Tyr Asp Lys Asp Asn Asp Lys Leu Lys Lys Leu
820 825 830
Ile Asn Lys Ser Pro Glu Lys Leu Leu Met Tyr His His Asp Pro Gln
835 840 845
Thr Tyr Gln Lys Leu Lys Leu Ile Met Glu Gln Tyr Gly Asp Glu Lys
850 855 860
Asn Pro Leu Tyr Lys Tyr Tyr Glu Glu Thr Gly Asn Tyr Leu Thr Lys
865 870 875 880
Tyr Ser Lys Lys Asp Asn Gly Pro Val Ile Lys Lys Ile Lys Tyr Tyr
885 890 895
Gly Asn Lys Leu Asn Ala His Leu Asp Ile Thr Asp Asp Tyr Pro Asn
900 905 910
Ser Arg Asn Lys Val Val Lys Leu Ser Leu Lys Pro Tyr Arg Phe Asp
915 920 925
Val Tyr Leu Asp Asn Gly Val Tyr Lys Phe Val Thr Val Lys Asn Leu
930 935 940
Asp Val Ile Lys Lys Glu Asn Tyr Tyr Glu Val Asn Ser Lys Cys Tyr
945 950 955 960
Glu Glu Ala Lys Lys Leu Lys Lys Ile Ser Asn Gln Ala Glu Phe Ile
965 970 975
Ala Ser Phe Tyr Lys Asn Asp Leu Ile Lys Ile Asn Gly Glu Leu Tyr
980 985 990
Arg Val Ile Gly Val Asn Asn Asp Leu Leu Asn Arg Ile Glu Val Asn
995 1000 1005
Met Ile Asp Ile Thr Tyr Arg Glu Tyr Leu Glu Asn Met Asn Asp
1010 1015 1020
Lys Arg Pro Pro His Ile Ile Lys Thr Ile Ala Ser Lys Thr Gln
1025 1030 1035
Ser Ile Lys Lys Tyr Ser Thr Asp Ile Leu Gly Asn Leu Tyr Glu
1040 1045 1050
Val Lys Ser Lys Lys His Pro Gln Ile Ile Lys Lys Gly Thr Ser
1055 1060 1065
Arg Ala Asp Pro Lys Lys Lys Arg Lys Val Glu Ala Ser Gly Ser
1070 1075 1080
Gly Arg Ala Asp Ala Leu Asp Asp Phe Asp Leu Asp Met Leu Gly
1085 1090 1095
Ser Asp Ala Leu Asp Asp Phe Asp Leu Asp Met Leu Gly Ser Asp
1100 1105 1110
Ala Leu Asp Asp Phe Asp Leu Asp Met Leu Gly Ser Asp Ala Leu
1115 1120 1125
Asp Asp Phe Asp Leu Asp Met Leu Ile Asn Ser Arg Ser Gln Tyr
1130 1135 1140
Leu Pro Asp Thr Asp Asp Arg His Arg Ile Glu Glu Lys Arg Lys
1145 1150 1155
Arg Thr Tyr Glu Thr Phe Lys Ser Ile Met Lys Lys Ser Pro Phe
1160 1165 1170
Ser Gly Pro Thr Asp Pro Arg Pro Pro Pro Arg Arg Ile Ala Val
1175 1180 1185
Pro Ser Arg Ser Ser Ala Ser Val Pro Lys Pro Ala Pro Gln Pro
1190 1195 1200
Tyr Pro Phe Thr Ser Ser Leu Ser Thr Ile Asn Tyr Asp Glu Phe
1205 1210 1215
Pro Thr Met Val Phe Pro Ser Gly Gln Ile Ser Gln Ala Ser Ala
1220 1225 1230
Leu Ala Pro Ala Pro Pro Gln Val Leu Pro Gln Ala Pro Ala Pro
1235 1240 1245
Ala Pro Ala Pro Ala Met Val Ser Ala Leu Ala Gln Ala Pro Ala
1250 1255 1260
Pro Val Pro Val Leu Ala Pro Gly Pro Pro Gln Ala Val Ala Pro
1265 1270 1275
Pro Ala Pro Lys Pro Thr Gln Ala Gly Glu Gly Thr Leu Ser Glu
1280 1285 1290
Ala Leu Leu Gln Leu Gln Phe Asp Asp Glu Asp Leu Gly Ala Leu
1295 1300 1305
Leu Gly Asn Ser Thr Asp Pro Ala Val Phe Thr Asp Leu Ala Ser
1310 1315 1320
Val Asp Asn Ser Glu Phe Gln Gln Leu Leu Asn Gln Gly Ile Pro
1325 1330 1335
Val Ala Pro His Thr Thr Glu Pro Met Leu Met Glu Tyr Pro Glu
1340 1345 1350
Ala Ile Thr Arg Leu Val Thr Gly Ala Gln Arg Pro Pro Asp Pro
1355 1360 1365
Ala Pro Ala Pro Leu Gly Ala Pro Gly Leu Pro Asn Gly Leu Leu
1370 1375 1380
Ser Gly Asp Glu Asp Phe Ser Ser Ile Ala Asp Met Asp Phe Ser
1385 1390 1395
Ala Leu Leu Gly Ser Gly Ser Gly Ser Arg Asp Ser Arg Glu Gly
1400 1405 1410
Met Phe Leu Pro Lys Pro Glu Ala Gly Ser Ala Ile Ser Asp Val
1415 1420 1425
Phe Glu Gly Arg Glu Val Cys Gln Pro Lys Arg Ile Arg Pro Phe
1430 1435 1440
His Pro Pro Gly Ser Pro Trp Ala Asn Arg Pro Leu Pro Ala Ser
1445 1450 1455
Leu Ala Pro Thr Pro Thr Gly Pro Val His Glu Pro Val Gly Ser
1460 1465 1470
Leu Thr Pro Ala Pro Val Pro Gln Pro Leu Asp Pro Ala Pro Ala
1475 1480 1485
Val Thr Pro Glu Ala Ser His Leu Leu Glu Asp Pro Asp Glu Glu
1490 1495 1500
Thr Ser Gln Ala Val Lys Ala Leu Arg Glu Met Ala Asp Thr Val
1505 1510 1515
Ile Pro Gln Lys Glu Glu Ala Ala Ile Cys Gly Gln Met Asp Leu
1520 1525 1530
Ser His Pro Pro Pro Arg Gly His Leu Asp Glu Leu Thr Thr Thr
1535 1540 1545
Leu Glu Ser Met Thr Glu Asp Leu Asn Leu Asp Ser Pro Leu Thr
1550 1555 1560
Pro Glu Leu Asn Glu Ile Leu Asp Thr Phe Leu Asn Asp Glu Cys
1565 1570 1575
Leu Leu His Ala Met His Ile Ser Thr Gly Leu Ser Ile Phe Asp
1580 1585 1590
Thr Ser Leu Phe
1595
<210> 12
<211> 1393
<212> PRT
<213> artificial sequence
<220>
<223> synthetic constructs produced by rational design
<400> 12
Met Pro Lys Lys Lys Arg Lys Val Gly Gly Gly Ser Pro Gly Lys Arg
1 5 10 15
Asn Tyr Ile Leu Gly Leu Asp Ile Gly Ile Thr Ser Val Gly Tyr Gly
20 25 30
Ile Ile Asp Tyr Glu Thr Arg Asp Val Ile Asp Ala Gly Val Arg Leu
35 40 45
Phe Lys Glu Ala Asn Val Glu Asn Asn Glu Gly Arg Arg Ser Lys Arg
50 55 60
Gly Ala Arg Arg Leu Lys Arg Arg Arg Arg His Arg Ile Gln Arg Val
65 70 75 80
Lys Lys Leu Leu Phe Asp Tyr Asn Leu Leu Thr Asp His Ser Glu Leu
85 90 95
Ser Gly Ile Asn Pro Tyr Glu Ala Arg Val Lys Gly Leu Ser Gln Lys
100 105 110
Leu Ser Glu Glu Glu Phe Ser Ala Ala Leu Leu His Leu Ala Lys Arg
115 120 125
Arg Gly Val His Asn Val Asn Glu Val Glu Glu Asp Thr Gly Asn Glu
130 135 140
Leu Ser Thr Lys Glu Gln Ile Ser Arg Asn Ser Lys Ala Leu Glu Glu
145 150 155 160
Lys Tyr Val Ala Glu Leu Gln Leu Glu Arg Leu Lys Lys Asp Gly Glu
165 170 175
Val Arg Gly Ser Ile Asn Arg Phe Lys Thr Ser Asp Tyr Val Lys Glu
180 185 190
Ala Lys Gln Leu Leu Lys Val Gln Lys Ala Tyr His Gln Leu Asp Gln
195 200 205
Ser Phe Ile Asp Thr Tyr Ile Asp Leu Leu Glu Thr Arg Arg Thr Tyr
210 215 220
Tyr Glu Gly Pro Gly Glu Gly Ser Pro Phe Gly Trp Lys Asp Ile Lys
225 230 235 240
Glu Trp Tyr Glu Met Leu Gly Gly Gly Ser Gly Gly Gly Ile Leu Ser
245 250 255
Pro Val Val Lys Arg Ser Phe Ile Gln Ser Ile Lys Val Ile Asn Ala
260 265 270
Ile Ile Lys Lys Tyr Gly Leu Pro Asn Asp Ile Ile Ile Glu Leu Ala
275 280 285
Arg Glu Lys Asn Ser Lys Asp Ala Gln Lys Met Ile Asn Glu Met Gln
290 295 300
Lys Arg Asn Arg Gln Thr Asn Glu Arg Ile Glu Glu Ile Ile Arg Thr
305 310 315 320
Thr Gly Lys Glu Asn Ala Lys Tyr Leu Ile Glu Lys Ile Lys Leu His
325 330 335
Asp Met Gln Glu Gly Lys Cys Leu Tyr Ser Leu Glu Ala Ile Pro Leu
340 345 350
Glu Asp Leu Leu Asn Asn Pro Phe Asn Tyr Glu Val Asp His Ile Ile
355 360 365
Pro Arg Ser Val Ser Phe Asp Asn Ser Phe Asn Asn Lys Val Leu Val
370 375 380
Lys Gln Glu Glu Asn Ser Lys Lys Gly Asn Arg Thr Pro Phe Gln Tyr
385 390 395 400
Leu Ser Ser Ser Asp Ser Lys Ile Ser Tyr Glu Thr Phe Lys Lys His
405 410 415
Ile Leu Asn Leu Ala Lys Gly Lys Gly Arg Ile Ser Lys Thr Lys Lys
420 425 430
Glu Tyr Leu Leu Glu Glu Arg Asp Ile Asn Arg Phe Ser Val Gln Lys
435 440 445
Asp Phe Ile Asn Arg Asn Leu Val Asp Thr Arg Tyr Ala Thr Arg Gly
450 455 460
Leu Met Asn Leu Leu Arg Ser Tyr Phe Arg Val Asn Asn Leu Asp Val
465 470 475 480
Lys Val Lys Ser Ile Asn Gly Gly Phe Thr Ser Phe Leu Arg Arg Lys
485 490 495
Trp Lys Phe Lys Lys Glu Arg Asn Lys Gly Tyr Lys His His Ala Glu
500 505 510
Asp Ala Leu Ile Ile Ala Asn Ala Asp Phe Ile Phe Lys Glu Trp Lys
515 520 525
Lys Leu Asp Lys Ala Lys Lys Val Met Glu Asn Gln Met Phe Glu Glu
530 535 540
Lys Gln Ala Glu Ser Met Pro Glu Ile Glu Thr Glu Gln Glu Tyr Lys
545 550 555 560
Glu Ile Phe Ile Thr Pro His Gln Ile Lys His Ile Lys Asp Phe Lys
565 570 575
Asp Tyr Lys Tyr Ser His Arg Val Asp Lys Lys Pro Asn Arg Lys Leu
580 585 590
Ile Asn Asp Thr Leu Tyr Ser Thr Arg Lys Asp Asp Lys Gly Asn Thr
595 600 605
Leu Ile Val Asn Asn Leu Asn Gly Leu Tyr Asp Lys Asp Asn Asp Lys
610 615 620
Leu Lys Lys Leu Ile Asn Lys Ser Pro Glu Lys Leu Leu Met Tyr His
625 630 635 640
His Asp Pro Gln Thr Tyr Gln Lys Leu Lys Leu Ile Met Glu Gln Tyr
645 650 655
Gly Asp Glu Lys Asn Pro Leu Tyr Lys Tyr Tyr Glu Glu Thr Gly Asn
660 665 670
Tyr Leu Thr Lys Tyr Ser Lys Lys Asp Asn Gly Pro Val Ile Lys Lys
675 680 685
Ile Lys Tyr Tyr Gly Asn Lys Leu Asn Ala His Leu Asp Ile Thr Asp
690 695 700
Asp Tyr Pro Asn Ser Arg Asn Lys Val Val Lys Leu Ser Leu Lys Pro
705 710 715 720
Tyr Arg Phe Asp Val Tyr Leu Asp Asn Gly Val Tyr Lys Phe Val Thr
725 730 735
Val Lys Asn Leu Asp Val Ile Lys Lys Glu Asn Tyr Tyr Glu Val Asn
740 745 750
Ser Lys Cys Tyr Glu Glu Ala Lys Lys Leu Lys Lys Ile Ser Asn Gln
755 760 765
Ala Glu Phe Ile Ala Ser Phe Tyr Lys Asn Asp Leu Ile Lys Ile Asn
770 775 780
Gly Glu Leu Tyr Arg Val Ile Gly Val Asn Asn Asp Leu Leu Asn Arg
785 790 795 800
Ile Glu Val Asn Met Ile Asp Ile Thr Tyr Arg Glu Tyr Leu Glu Asn
805 810 815
Met Asn Asp Lys Arg Pro Pro His Ile Ile Lys Thr Ile Ala Ser Lys
820 825 830
Thr Gln Ser Ile Lys Lys Tyr Ser Thr Asp Ile Leu Gly Asn Leu Tyr
835 840 845
Glu Val Lys Ser Lys Lys His Pro Gln Ile Ile Lys Lys Gly Thr Ser
850 855 860
Arg Ala Asp Pro Lys Lys Lys Arg Lys Val Glu Ala Ser Gly Ser Gly
865 870 875 880
Arg Ala Asp Ala Leu Asp Asp Phe Asp Leu Asp Met Leu Gly Ser Asp
885 890 895
Ala Leu Asp Asp Phe Asp Leu Asp Met Leu Gly Ser Asp Ala Leu Asp
900 905 910
Asp Phe Asp Leu Asp Met Leu Gly Ser Asp Ala Leu Asp Asp Phe Asp
915 920 925
Leu Asp Met Leu Ile Asn Ser Arg Ser Gln Tyr Leu Pro Asp Thr Asp
930 935 940
Asp Arg His Arg Ile Glu Glu Lys Arg Lys Arg Thr Tyr Glu Thr Phe
945 950 955 960
Lys Ser Ile Met Lys Lys Ser Pro Phe Ser Gly Pro Thr Asp Pro Arg
965 970 975
Pro Pro Pro Arg Arg Ile Ala Val Pro Ser Arg Ser Ser Ala Ser Val
980 985 990
Pro Lys Pro Ala Pro Gln Pro Tyr Pro Phe Thr Ser Ser Leu Ser Thr
995 1000 1005
Ile Asn Tyr Asp Glu Phe Pro Thr Met Val Phe Pro Ser Gly Gln
1010 1015 1020
Ile Ser Gln Ala Ser Ala Leu Ala Pro Ala Pro Pro Gln Val Leu
1025 1030 1035
Pro Gln Ala Pro Ala Pro Ala Pro Ala Pro Ala Met Val Ser Ala
1040 1045 1050
Leu Ala Gln Ala Pro Ala Pro Val Pro Val Leu Ala Pro Gly Pro
1055 1060 1065
Pro Gln Ala Val Ala Pro Pro Ala Pro Lys Pro Thr Gln Ala Gly
1070 1075 1080
Glu Gly Thr Leu Ser Glu Ala Leu Leu Gln Leu Gln Phe Asp Asp
1085 1090 1095
Glu Asp Leu Gly Ala Leu Leu Gly Asn Ser Thr Asp Pro Ala Val
1100 1105 1110
Phe Thr Asp Leu Ala Ser Val Asp Asn Ser Glu Phe Gln Gln Leu
1115 1120 1125
Leu Asn Gln Gly Ile Pro Val Ala Pro His Thr Thr Glu Pro Met
1130 1135 1140
Leu Met Glu Tyr Pro Glu Ala Ile Thr Arg Leu Val Thr Gly Ala
1145 1150 1155
Gln Arg Pro Pro Asp Pro Ala Pro Ala Pro Leu Gly Ala Pro Gly
1160 1165 1170
Leu Pro Asn Gly Leu Leu Ser Gly Asp Glu Asp Phe Ser Ser Ile
1175 1180 1185
Ala Asp Met Asp Phe Ser Ala Leu Leu Gly Ser Gly Ser Gly Ser
1190 1195 1200
Arg Asp Ser Arg Glu Gly Met Phe Leu Pro Lys Pro Glu Ala Gly
1205 1210 1215
Ser Ala Ile Ser Asp Val Phe Glu Gly Arg Glu Val Cys Gln Pro
1220 1225 1230
Lys Arg Ile Arg Pro Phe His Pro Pro Gly Ser Pro Trp Ala Asn
1235 1240 1245
Arg Pro Leu Pro Ala Ser Leu Ala Pro Thr Pro Thr Gly Pro Val
1250 1255 1260
His Glu Pro Val Gly Ser Leu Thr Pro Ala Pro Val Pro Gln Pro
1265 1270 1275
Leu Asp Pro Ala Pro Ala Val Thr Pro Glu Ala Ser His Leu Leu
1280 1285 1290
Glu Asp Pro Asp Glu Glu Thr Ser Gln Ala Val Lys Ala Leu Arg
1295 1300 1305
Glu Met Ala Asp Thr Val Ile Pro Gln Lys Glu Glu Ala Ala Ile
1310 1315 1320
Cys Gly Gln Met Asp Leu Ser His Pro Pro Pro Arg Gly His Leu
1325 1330 1335
Asp Glu Leu Thr Thr Thr Leu Glu Ser Met Thr Glu Asp Leu Asn
1340 1345 1350
Leu Asp Ser Pro Leu Thr Pro Glu Leu Asn Glu Ile Leu Asp Thr
1355 1360 1365
Phe Leu Asn Asp Glu Cys Leu Leu His Ala Met His Ile Ser Thr
1370 1375 1380
Gly Leu Ser Ile Phe Asp Thr Ser Leu Phe
1385 1390
<210> 13
<211> 1393
<212> PRT
<213> artificial sequence
<220>
<223> synthetic constructs produced by rational design
<400> 13
Met Pro Lys Lys Lys Arg Lys Val Gly Gly Gly Ser Pro Gly Lys Arg
1 5 10 15
Asn Tyr Ile Leu Gly Leu Asp Ile Gly Ile Thr Ser Val Gly Tyr Gly
20 25 30
Ile Ile Asp Tyr Glu Thr Arg Asp Val Ile Asp Ala Gly Val Arg Leu
35 40 45
Phe Lys Glu Ala Asn Val Glu Asn Asn Glu Gly Arg Arg Ser Lys Arg
50 55 60
Gly Ala Arg Arg Leu Lys Arg Arg Arg Arg His Arg Ile Gln Arg Val
65 70 75 80
Lys Lys Leu Leu Phe Asp Tyr Asn Leu Leu Thr Asp His Ser Glu Leu
85 90 95
Ser Gly Ile Asn Pro Tyr Glu Ala Arg Val Lys Gly Leu Ser Gln Lys
100 105 110
Leu Ser Glu Glu Glu Phe Ser Ala Ala Leu Leu His Leu Ala Lys Arg
115 120 125
Arg Gly Val His Asn Val Asn Glu Val Glu Glu Asp Thr Gly Asn Glu
130 135 140
Leu Ser Thr Lys Glu Gln Ile Ser Arg Asn Ser Lys Ala Leu Glu Glu
145 150 155 160
Lys Tyr Val Ala Glu Leu Gln Leu Glu Arg Leu Lys Lys Asp Gly Glu
165 170 175
Val Arg Gly Ser Ile Asn Arg Phe Lys Thr Ser Asp Tyr Val Lys Glu
180 185 190
Ala Lys Gln Leu Leu Lys Val Gln Lys Ala Tyr His Gln Leu Asp Gln
195 200 205
Ser Phe Ile Asp Thr Tyr Ile Asp Leu Leu Glu Thr Arg Arg Thr Tyr
210 215 220
Tyr Glu Gly Pro Gly Glu Gly Ser Pro Phe Gly Trp Lys Asp Ile Lys
225 230 235 240
Glu Trp Tyr Glu Met Leu Lys Arg Arg Arg Arg His Arg Ile Leu Ser
245 250 255
Pro Val Val Lys Arg Ser Phe Ile Gln Ser Ile Lys Val Ile Asn Ala
260 265 270
Ile Ile Lys Lys Tyr Gly Leu Pro Asn Asp Ile Ile Ile Glu Leu Ala
275 280 285
Arg Glu Lys Asn Ser Lys Asp Ala Gln Lys Met Ile Asn Glu Met Gln
290 295 300
Lys Arg Asn Arg Gln Thr Asn Glu Arg Ile Glu Glu Ile Ile Arg Thr
305 310 315 320
Thr Gly Lys Glu Asn Ala Lys Tyr Leu Ile Glu Lys Ile Lys Leu His
325 330 335
Asp Met Gln Glu Gly Lys Cys Leu Tyr Ser Leu Glu Ala Ile Pro Leu
340 345 350
Glu Asp Leu Leu Asn Asn Pro Phe Asn Tyr Glu Val Asp His Ile Ile
355 360 365
Pro Arg Ser Val Ser Phe Asp Asn Ser Phe Asn Asn Lys Val Leu Val
370 375 380
Lys Gln Glu Glu Asn Ser Lys Lys Gly Asn Arg Thr Pro Phe Gln Tyr
385 390 395 400
Leu Ser Ser Ser Asp Ser Lys Ile Ser Tyr Glu Thr Phe Lys Lys His
405 410 415
Ile Leu Asn Leu Ala Lys Gly Lys Gly Arg Ile Ser Lys Thr Lys Lys
420 425 430
Glu Tyr Leu Leu Glu Glu Arg Asp Ile Asn Arg Phe Ser Val Gln Lys
435 440 445
Asp Phe Ile Asn Arg Asn Leu Val Asp Thr Arg Tyr Ala Thr Arg Gly
450 455 460
Leu Met Asn Leu Leu Arg Ser Tyr Phe Arg Val Asn Asn Leu Asp Val
465 470 475 480
Lys Val Lys Ser Ile Asn Gly Gly Phe Thr Ser Phe Leu Arg Arg Lys
485 490 495
Trp Lys Phe Lys Lys Glu Arg Asn Lys Gly Tyr Lys His His Ala Glu
500 505 510
Asp Ala Leu Ile Ile Ala Asn Ala Asp Phe Ile Phe Lys Glu Trp Lys
515 520 525
Lys Leu Asp Lys Ala Lys Lys Val Met Glu Asn Gln Met Phe Glu Glu
530 535 540
Lys Gln Ala Glu Ser Met Pro Glu Ile Glu Thr Glu Gln Glu Tyr Lys
545 550 555 560
Glu Ile Phe Ile Thr Pro His Gln Ile Lys His Ile Lys Asp Phe Lys
565 570 575
Asp Tyr Lys Tyr Ser His Arg Val Asp Lys Lys Pro Asn Arg Lys Leu
580 585 590
Ile Asn Asp Thr Leu Tyr Ser Thr Arg Lys Asp Asp Lys Gly Asn Thr
595 600 605
Leu Ile Val Asn Asn Leu Asn Gly Leu Tyr Asp Lys Asp Asn Asp Lys
610 615 620
Leu Lys Lys Leu Ile Asn Lys Ser Pro Glu Lys Leu Leu Met Tyr His
625 630 635 640
His Asp Pro Gln Thr Tyr Gln Lys Leu Lys Leu Ile Met Glu Gln Tyr
645 650 655
Gly Asp Glu Lys Asn Pro Leu Tyr Lys Tyr Tyr Glu Glu Thr Gly Asn
660 665 670
Tyr Leu Thr Lys Tyr Ser Lys Lys Asp Asn Gly Pro Val Ile Lys Lys
675 680 685
Ile Lys Tyr Tyr Gly Asn Lys Leu Asn Ala His Leu Asp Ile Thr Asp
690 695 700
Asp Tyr Pro Asn Ser Arg Asn Lys Val Val Lys Leu Ser Leu Lys Pro
705 710 715 720
Tyr Arg Phe Asp Val Tyr Leu Asp Asn Gly Val Tyr Lys Phe Val Thr
725 730 735
Val Lys Asn Leu Asp Val Ile Lys Lys Glu Asn Tyr Tyr Glu Val Asn
740 745 750
Ser Lys Cys Tyr Glu Glu Ala Lys Lys Leu Lys Lys Ile Ser Asn Gln
755 760 765
Ala Glu Phe Ile Ala Ser Phe Tyr Lys Asn Asp Leu Ile Lys Ile Asn
770 775 780
Gly Glu Leu Tyr Arg Val Ile Gly Val Asn Asn Asp Leu Leu Asn Arg
785 790 795 800
Ile Glu Val Asn Met Ile Asp Ile Thr Tyr Arg Glu Tyr Leu Glu Asn
805 810 815
Met Asn Asp Lys Arg Pro Pro His Ile Ile Lys Thr Ile Ala Ser Lys
820 825 830
Thr Gln Ser Ile Lys Lys Tyr Ser Thr Asp Ile Leu Gly Asn Leu Tyr
835 840 845
Glu Val Lys Ser Lys Lys His Pro Gln Ile Ile Lys Lys Gly Thr Ser
850 855 860
Arg Ala Asp Pro Lys Lys Lys Arg Lys Val Glu Ala Ser Gly Ser Gly
865 870 875 880
Arg Ala Asp Ala Leu Asp Asp Phe Asp Leu Asp Met Leu Gly Ser Asp
885 890 895
Ala Leu Asp Asp Phe Asp Leu Asp Met Leu Gly Ser Asp Ala Leu Asp
900 905 910
Asp Phe Asp Leu Asp Met Leu Gly Ser Asp Ala Leu Asp Asp Phe Asp
915 920 925
Leu Asp Met Leu Ile Asn Ser Arg Ser Gln Tyr Leu Pro Asp Thr Asp
930 935 940
Asp Arg His Arg Ile Glu Glu Lys Arg Lys Arg Thr Tyr Glu Thr Phe
945 950 955 960
Lys Ser Ile Met Lys Lys Ser Pro Phe Ser Gly Pro Thr Asp Pro Arg
965 970 975
Pro Pro Pro Arg Arg Ile Ala Val Pro Ser Arg Ser Ser Ala Ser Val
980 985 990
Pro Lys Pro Ala Pro Gln Pro Tyr Pro Phe Thr Ser Ser Leu Ser Thr
995 1000 1005
Ile Asn Tyr Asp Glu Phe Pro Thr Met Val Phe Pro Ser Gly Gln
1010 1015 1020
Ile Ser Gln Ala Ser Ala Leu Ala Pro Ala Pro Pro Gln Val Leu
1025 1030 1035
Pro Gln Ala Pro Ala Pro Ala Pro Ala Pro Ala Met Val Ser Ala
1040 1045 1050
Leu Ala Gln Ala Pro Ala Pro Val Pro Val Leu Ala Pro Gly Pro
1055 1060 1065
Pro Gln Ala Val Ala Pro Pro Ala Pro Lys Pro Thr Gln Ala Gly
1070 1075 1080
Glu Gly Thr Leu Ser Glu Ala Leu Leu Gln Leu Gln Phe Asp Asp
1085 1090 1095
Glu Asp Leu Gly Ala Leu Leu Gly Asn Ser Thr Asp Pro Ala Val
1100 1105 1110
Phe Thr Asp Leu Ala Ser Val Asp Asn Ser Glu Phe Gln Gln Leu
1115 1120 1125
Leu Asn Gln Gly Ile Pro Val Ala Pro His Thr Thr Glu Pro Met
1130 1135 1140
Leu Met Glu Tyr Pro Glu Ala Ile Thr Arg Leu Val Thr Gly Ala
1145 1150 1155
Gln Arg Pro Pro Asp Pro Ala Pro Ala Pro Leu Gly Ala Pro Gly
1160 1165 1170
Leu Pro Asn Gly Leu Leu Ser Gly Asp Glu Asp Phe Ser Ser Ile
1175 1180 1185
Ala Asp Met Asp Phe Ser Ala Leu Leu Gly Ser Gly Ser Gly Ser
1190 1195 1200
Arg Asp Ser Arg Glu Gly Met Phe Leu Pro Lys Pro Glu Ala Gly
1205 1210 1215
Ser Ala Ile Ser Asp Val Phe Glu Gly Arg Glu Val Cys Gln Pro
1220 1225 1230
Lys Arg Ile Arg Pro Phe His Pro Pro Gly Ser Pro Trp Ala Asn
1235 1240 1245
Arg Pro Leu Pro Ala Ser Leu Ala Pro Thr Pro Thr Gly Pro Val
1250 1255 1260
His Glu Pro Val Gly Ser Leu Thr Pro Ala Pro Val Pro Gln Pro
1265 1270 1275
Leu Asp Pro Ala Pro Ala Val Thr Pro Glu Ala Ser His Leu Leu
1280 1285 1290
Glu Asp Pro Asp Glu Glu Thr Ser Gln Ala Val Lys Ala Leu Arg
1295 1300 1305
Glu Met Ala Asp Thr Val Ile Pro Gln Lys Glu Glu Ala Ala Ile
1310 1315 1320
Cys Gly Gln Met Asp Leu Ser His Pro Pro Pro Arg Gly His Leu
1325 1330 1335
Asp Glu Leu Thr Thr Thr Leu Glu Ser Met Thr Glu Asp Leu Asn
1340 1345 1350
Leu Asp Ser Pro Leu Thr Pro Glu Leu Asn Glu Ile Leu Asp Thr
1355 1360 1365
Phe Leu Asn Asp Glu Cys Leu Leu His Ala Met His Ile Ser Thr
1370 1375 1380
Gly Leu Ser Ile Phe Asp Thr Ser Leu Phe
1385 1390
<210> 14
<211> 1225
<212> PRT
<213> artificial sequence
<220>
<223> synthetic constructs produced by rational design
<400> 14
Met Pro Lys Lys Lys Arg Lys Val Gly Gly Gly Ser Pro Gly Lys Arg
1 5 10 15
Asn Tyr Ile Leu Gly Leu Asp Ile Gly Ile Thr Ser Val Gly Tyr Gly
20 25 30
Ile Ile Asp Tyr Glu Thr Arg Asp Val Ile Asp Ala Gly Val Arg Leu
35 40 45
Phe Lys Glu Ala Asn Val Glu Asn Asn Glu Gly Arg Arg Ser Lys Arg
50 55 60
Gly Ala Arg Arg Leu Lys Arg Arg Arg Arg His Arg Ile Gln Arg Val
65 70 75 80
Lys Lys Leu Leu Phe Asp Tyr Asn Leu Leu Thr Asp His Ser Glu Leu
85 90 95
Ser Gly Ile Asn Pro Tyr Glu Ala Arg Val Lys Gly Leu Ser Gln Lys
100 105 110
Leu Ser Glu Glu Glu Phe Ser Ala Ala Leu Leu His Leu Ala Lys Arg
115 120 125
Arg Gly Val His Asn Val Asn Glu Val Glu Glu Asp Thr Gly Asn Glu
130 135 140
Leu Ser Thr Lys Glu Gln Ile Ser Arg Asn Ser Lys Ala Leu Glu Glu
145 150 155 160
Lys Tyr Val Ala Glu Leu Gln Leu Glu Arg Leu Lys Lys Asp Gly Glu
165 170 175
Val Arg Gly Ser Ile Asn Arg Phe Lys Thr Ser Asp Tyr Val Lys Glu
180 185 190
Ala Lys Gln Leu Leu Lys Val Gln Lys Ala Tyr His Gln Leu Asp Gln
195 200 205
Ser Phe Ile Asp Thr Tyr Ile Asp Leu Leu Glu Thr Arg Arg Thr Tyr
210 215 220
Tyr Glu Gly Pro Gly Glu Gly Ser Pro Phe Gly Trp Lys Asp Ile Lys
225 230 235 240
Glu Trp Tyr Glu Met Leu Lys Arg Arg Arg Arg His Arg Ile Leu Ser
245 250 255
Pro Val Val Lys Arg Ser Phe Ile Gln Ser Ile Lys Val Ile Asn Ala
260 265 270
Ile Ile Lys Lys Tyr Gly Leu Pro Asn Asp Ile Ile Ile Glu Leu Gly
275 280 285
Ser Lys Arg Tyr Ala Thr Arg Gly Leu Met Asn Leu Leu Arg Ser Tyr
290 295 300
Phe Arg Val Asn Asn Leu Asp Val Lys Val Lys Ser Ile Asn Gly Gly
305 310 315 320
Phe Thr Ser Phe Leu Arg Arg Lys Trp Lys Phe Lys Lys Glu Arg Asn
325 330 335
Lys Gly Tyr Lys His His Ala Glu Asp Ala Leu Ile Ile Ala Asn Ala
340 345 350
Asp Phe Ile Phe Lys Glu Trp Lys Lys Leu Asp Lys Ala Lys Lys Val
355 360 365
Met Glu Asn Gln Met Phe Glu Glu Lys Gln Ala Glu Ser Met Pro Glu
370 375 380
Ile Glu Thr Glu Gln Glu Tyr Lys Glu Ile Phe Ile Thr Pro His Gln
385 390 395 400
Ile Lys His Ile Lys Asp Phe Lys Asp Tyr Lys Tyr Ser His Arg Val
405 410 415
Asp Lys Lys Pro Asn Arg Lys Leu Ile Asn Asp Thr Leu Tyr Ser Thr
420 425 430
Arg Lys Asp Asp Lys Gly Asn Thr Leu Ile Val Asn Asn Leu Asn Gly
435 440 445
Leu Tyr Asp Lys Asp Asn Asp Lys Leu Lys Lys Leu Ile Asn Lys Ser
450 455 460
Pro Glu Lys Leu Leu Met Tyr His His Asp Pro Gln Thr Tyr Gln Lys
465 470 475 480
Leu Lys Leu Ile Met Glu Gln Tyr Gly Asp Glu Lys Asn Pro Leu Tyr
485 490 495
Lys Tyr Tyr Glu Glu Thr Gly Asn Tyr Leu Thr Lys Tyr Ser Lys Lys
500 505 510
Asp Asn Gly Pro Val Ile Lys Lys Ile Lys Tyr Tyr Gly Asn Lys Leu
515 520 525
Asn Ala His Leu Asp Ile Thr Asp Asp Tyr Pro Asn Ser Arg Asn Lys
530 535 540
Val Val Lys Leu Ser Leu Lys Pro Tyr Arg Phe Asp Val Tyr Leu Asp
545 550 555 560
Asn Gly Val Tyr Lys Phe Val Thr Val Lys Asn Leu Asp Val Ile Lys
565 570 575
Lys Glu Asn Tyr Tyr Glu Val Asn Ser Lys Cys Tyr Glu Glu Ala Lys
580 585 590
Lys Leu Lys Lys Ile Ser Asn Gln Ala Glu Phe Ile Ala Ser Phe Tyr
595 600 605
Lys Asn Asp Leu Ile Lys Ile Asn Gly Glu Leu Tyr Arg Val Ile Gly
610 615 620
Val Asn Asn Asp Leu Leu Asn Arg Ile Glu Val Asn Met Ile Asp Ile
625 630 635 640
Thr Tyr Arg Glu Tyr Leu Glu Asn Met Asn Asp Lys Arg Pro Pro His
645 650 655
Ile Ile Lys Thr Ile Ala Ser Lys Thr Gln Ser Ile Lys Lys Tyr Ser
660 665 670
Thr Asp Ile Leu Gly Asn Leu Tyr Glu Val Lys Ser Lys Lys His Pro
675 680 685
Gln Ile Ile Lys Lys Gly Thr Ser Arg Ala Asp Pro Lys Lys Lys Arg
690 695 700
Lys Val Glu Ala Ser Gly Ser Gly Arg Ala Asp Ala Leu Asp Asp Phe
705 710 715 720
Asp Leu Asp Met Leu Gly Ser Asp Ala Leu Asp Asp Phe Asp Leu Asp
725 730 735
Met Leu Gly Ser Asp Ala Leu Asp Asp Phe Asp Leu Asp Met Leu Gly
740 745 750
Ser Asp Ala Leu Asp Asp Phe Asp Leu Asp Met Leu Ile Asn Ser Arg
755 760 765
Ser Gln Tyr Leu Pro Asp Thr Asp Asp Arg His Arg Ile Glu Glu Lys
770 775 780
Arg Lys Arg Thr Tyr Glu Thr Phe Lys Ser Ile Met Lys Lys Ser Pro
785 790 795 800
Phe Ser Gly Pro Thr Asp Pro Arg Pro Pro Pro Arg Arg Ile Ala Val
805 810 815
Pro Ser Arg Ser Ser Ala Ser Val Pro Lys Pro Ala Pro Gln Pro Tyr
820 825 830
Pro Phe Thr Ser Ser Leu Ser Thr Ile Asn Tyr Asp Glu Phe Pro Thr
835 840 845
Met Val Phe Pro Ser Gly Gln Ile Ser Gln Ala Ser Ala Leu Ala Pro
850 855 860
Ala Pro Pro Gln Val Leu Pro Gln Ala Pro Ala Pro Ala Pro Ala Pro
865 870 875 880
Ala Met Val Ser Ala Leu Ala Gln Ala Pro Ala Pro Val Pro Val Leu
885 890 895
Ala Pro Gly Pro Pro Gln Ala Val Ala Pro Pro Ala Pro Lys Pro Thr
900 905 910
Gln Ala Gly Glu Gly Thr Leu Ser Glu Ala Leu Leu Gln Leu Gln Phe
915 920 925
Asp Asp Glu Asp Leu Gly Ala Leu Leu Gly Asn Ser Thr Asp Pro Ala
930 935 940
Val Phe Thr Asp Leu Ala Ser Val Asp Asn Ser Glu Phe Gln Gln Leu
945 950 955 960
Leu Asn Gln Gly Ile Pro Val Ala Pro His Thr Thr Glu Pro Met Leu
965 970 975
Met Glu Tyr Pro Glu Ala Ile Thr Arg Leu Val Thr Gly Ala Gln Arg
980 985 990
Pro Pro Asp Pro Ala Pro Ala Pro Leu Gly Ala Pro Gly Leu Pro Asn
995 1000 1005
Gly Leu Leu Ser Gly Asp Glu Asp Phe Ser Ser Ile Ala Asp Met
1010 1015 1020
Asp Phe Ser Ala Leu Leu Gly Ser Gly Ser Gly Ser Arg Asp Ser
1025 1030 1035
Arg Glu Gly Met Phe Leu Pro Lys Pro Glu Ala Gly Ser Ala Ile
1040 1045 1050
Ser Asp Val Phe Glu Gly Arg Glu Val Cys Gln Pro Lys Arg Ile
1055 1060 1065
Arg Pro Phe His Pro Pro Gly Ser Pro Trp Ala Asn Arg Pro Leu
1070 1075 1080
Pro Ala Ser Leu Ala Pro Thr Pro Thr Gly Pro Val His Glu Pro
1085 1090 1095
Val Gly Ser Leu Thr Pro Ala Pro Val Pro Gln Pro Leu Asp Pro
1100 1105 1110
Ala Pro Ala Val Thr Pro Glu Ala Ser His Leu Leu Glu Asp Pro
1115 1120 1125
Asp Glu Glu Thr Ser Gln Ala Val Lys Ala Leu Arg Glu Met Ala
1130 1135 1140
Asp Thr Val Ile Pro Gln Lys Glu Glu Ala Ala Ile Cys Gly Gln
1145 1150 1155
Met Asp Leu Ser His Pro Pro Pro Arg Gly His Leu Asp Glu Leu
1160 1165 1170
Thr Thr Thr Leu Glu Ser Met Thr Glu Asp Leu Asn Leu Asp Ser
1175 1180 1185
Pro Leu Thr Pro Glu Leu Asn Glu Ile Leu Asp Thr Phe Leu Asn
1190 1195 1200
Asp Glu Cys Leu Leu His Ala Met His Ile Ser Thr Gly Leu Ser
1205 1210 1215
Ile Phe Asp Thr Ser Leu Phe
1220 1225
<210> 15
<211> 1852
<212> PRT
<213> artificial sequence
<220>
<223> synthetic constructs produced by rational design
<400> 15
Met Pro Lys Lys Lys Arg Lys Val Gly Gly Gly Ser Pro Gly Met Thr
1 5 10 15
Gln Phe Glu Gly Phe Thr Asn Leu Tyr Gln Val Ser Lys Thr Leu Arg
20 25 30
Phe Glu Leu Ile Pro Gln Gly Lys Thr Leu Lys His Ile Gln Glu Gln
35 40 45
Gly Phe Ile Glu Glu Asp Lys Ala Arg Asn Asp His Tyr Lys Glu Leu
50 55 60
Lys Pro Ile Ile Asp Arg Ile Tyr Lys Thr Tyr Ala Asp Gln Cys Leu
65 70 75 80
Gln Leu Val Gln Leu Asp Trp Glu Asn Leu Ser Ala Ala Ile Asp Ser
85 90 95
Tyr Arg Lys Glu Lys Thr Glu Glu Thr Arg Asn Ala Leu Ile Glu Glu
100 105 110
Gln Ala Thr Tyr Arg Asn Ala Ile His Asp Tyr Phe Ile Gly Arg Thr
115 120 125
Asp Asn Leu Thr Asp Ala Ile Asn Lys Arg His Ala Glu Ile Tyr Lys
130 135 140
Gly Leu Phe Lys Ala Glu Leu Phe Asn Gly Lys Val Leu Lys Gln Leu
145 150 155 160
Gly Thr Val Thr Thr Thr Glu His Glu Asn Ala Leu Leu Arg Ser Phe
165 170 175
Asp Lys Phe Thr Thr Tyr Phe Ser Gly Phe Tyr Glu Asn Arg Lys Asn
180 185 190
Val Phe Ser Ala Glu Asp Ile Ser Thr Ala Ile Pro His Arg Ile Val
195 200 205
Gln Asp Asn Phe Pro Lys Phe Lys Glu Asn Cys His Ile Phe Thr Arg
210 215 220
Leu Ile Thr Ala Val Pro Ser Leu Arg Glu His Phe Glu Asn Val Lys
225 230 235 240
Lys Ala Ile Gly Ile Phe Val Ser Thr Ser Ile Glu Glu Val Phe Ser
245 250 255
Phe Pro Phe Tyr Asn Gln Leu Leu Thr Gln Thr Gln Ile Asp Leu Tyr
260 265 270
Asn Gln Leu Leu Gly Gly Ile Ser Arg Glu Ala Gly Thr Glu Lys Ile
275 280 285
Lys Gly Leu Asn Glu Val Leu Asn Leu Ala Ile Gln Lys Asn Asp Glu
290 295 300
Thr Ala His Ile Ile Ala Ser Leu Pro His Arg Phe Ile Pro Leu Phe
305 310 315 320
Lys Gln Ile Leu Ser Asp Arg Asn Thr Leu Ser Phe Ile Leu Glu Glu
325 330 335
Phe Lys Ser Asp Glu Glu Val Ile Gln Ser Phe Cys Lys Tyr Lys Thr
340 345 350
Leu Leu Arg Asn Glu Asn Val Leu Glu Thr Ala Glu Ala Leu Phe Asn
355 360 365
Glu Leu Asn Ser Ile Asp Leu Thr His Ile Phe Ile Ser His Lys Lys
370 375 380
Leu Glu Thr Ile Ser Ser Ala Leu Cys Asp His Trp Asp Thr Leu Arg
385 390 395 400
Asn Ala Leu Tyr Glu Arg Arg Ile Ser Glu Leu Thr Gly Lys Ile Thr
405 410 415
Lys Ser Ala Lys Glu Lys Val Gln Arg Ser Leu Lys His Glu Asp Ile
420 425 430
Asn Leu Gln Glu Ile Ile Ser Ala Ala Gly Lys Glu Leu Ser Glu Ala
435 440 445
Phe Lys Gln Lys Thr Ser Glu Ile Leu Ser His Ala His Ala Ala Leu
450 455 460
Asp Gln Pro Leu Pro Thr Thr Leu Lys Lys Gln Glu Glu Lys Glu Ile
465 470 475 480
Leu Lys Ser Gln Leu Asp Ser Leu Leu Gly Leu Tyr His Leu Leu Asp
485 490 495
Trp Phe Ala Val Asp Glu Ser Asn Glu Val Asp Pro Glu Phe Ser Ala
500 505 510
Arg Leu Thr Gly Ile Lys Leu Glu Met Glu Pro Ser Leu Ser Phe Tyr
515 520 525
Asn Lys Ala Arg Asn Tyr Ala Thr Lys Lys Pro Tyr Ser Val Glu Lys
530 535 540
Phe Lys Leu Asn Phe Gln Met Pro Thr Leu Ala Ser Gly Trp Asp Val
545 550 555 560
Asn Lys Glu Lys Asn Asn Gly Ala Ile Leu Phe Val Lys Asn Gly Leu
565 570 575
Tyr Tyr Leu Gly Ile Met Pro Lys Gln Lys Gly Arg Tyr Lys Ala Leu
580 585 590
Ser Phe Glu Pro Thr Glu Lys Thr Ser Glu Gly Phe Asp Lys Met Tyr
595 600 605
Tyr Asp Tyr Phe Pro Asp Ala Ala Lys Met Ile Pro Lys Cys Ser Thr
610 615 620
Gln Leu Lys Ala Val Thr Ala His Phe Gln Thr His Thr Thr Pro Ile
625 630 635 640
Leu Leu Ser Asn Asn Phe Ile Glu Pro Leu Glu Ile Thr Lys Glu Ile
645 650 655
Tyr Asp Leu Asn Asn Pro Glu Lys Glu Pro Lys Lys Phe Gln Thr Ala
660 665 670
Tyr Ala Lys Lys Thr Gly Asp Gln Lys Gly Tyr Arg Glu Ala Leu Cys
675 680 685
Lys Trp Ile Asp Phe Thr Arg Asp Phe Leu Ser Lys Tyr Thr Lys Thr
690 695 700
Thr Ser Ile Asp Leu Ser Ser Leu Arg Pro Ser Ser Gln Tyr Lys Asp
705 710 715 720
Leu Gly Glu Tyr Tyr Ala Glu Leu Asn Pro Leu Leu Tyr His Ile Ser
725 730 735
Phe Gln Arg Ile Ala Glu Lys Glu Ile Met Asp Ala Val Glu Thr Gly
740 745 750
Lys Leu Tyr Leu Phe Gln Ile Tyr Asn Lys Asp Phe Ala Lys Gly His
755 760 765
His Gly Lys Pro Asn Leu His Thr Leu Tyr Trp Thr Gly Leu Phe Ser
770 775 780
Pro Glu Asn Leu Ala Lys Thr Ser Ile Lys Leu Asn Gly Gln Ala Glu
785 790 795 800
Leu Phe Tyr Arg Pro Lys Ser Arg Met Lys Arg Met Ala His Arg Leu
805 810 815
Gly Glu Lys Met Leu Asn Lys Lys Leu Lys Asp Gln Lys Thr Pro Ile
820 825 830
Pro Asp Thr Leu Tyr Gln Glu Leu Tyr Asp Tyr Val Asn His Arg Leu
835 840 845
Ser His Asp Leu Ser Asp Glu Ala Arg Ala Leu Leu Pro Asn Val Ile
850 855 860
Thr Lys Glu Val Ser His Glu Ile Ile Lys Asp Arg Arg Phe Thr Ser
865 870 875 880
Asp Lys Phe Phe Phe His Val Pro Ile Thr Leu Asn Tyr Gln Ala Ala
885 890 895
Asn Ser Pro Ser Lys Phe Asn Gln Arg Val Asn Ala Tyr Leu Lys Glu
900 905 910
His Pro Glu Thr Pro Ile Ile Gly Ile Ala Arg Gly Glu Arg Asn Leu
915 920 925
Ile Tyr Ile Thr Val Ile Asp Ser Thr Gly Lys Ile Leu Glu Gln Arg
930 935 940
Ser Leu Asn Thr Ile Gln Gln Phe Asp Tyr Gln Lys Lys Leu Asp Asn
945 950 955 960
Arg Glu Lys Glu Arg Val Ala Ala Arg Gln Ala Trp Ser Val Val Gly
965 970 975
Thr Ile Lys Asp Leu Lys Gln Gly Tyr Leu Ser Gln Val Ile His Glu
980 985 990
Ile Val Asp Leu Met Ile His Tyr Gln Ala Val Val Val Leu Glu Asn
995 1000 1005
Leu Asn Phe Gly Phe Lys Ser Lys Arg Thr Gly Ile Ala Glu Lys
1010 1015 1020
Ala Val Tyr Gln Gln Phe Glu Lys Met Leu Ile Asp Lys Leu Asn
1025 1030 1035
Cys Leu Val Leu Lys Asp Tyr Pro Ala Glu Lys Val Gly Gly Val
1040 1045 1050
Leu Asn Pro Tyr Gln Leu Thr Asp Gln Phe Thr Ser Phe Ala Lys
1055 1060 1065
Met Gly Thr Gln Ser Gly Phe Leu Phe Tyr Val Pro Ala Pro Tyr
1070 1075 1080
Thr Ser Lys Ile Asp Pro Leu Thr Gly Phe Val Asp Pro Phe Val
1085 1090 1095
Trp Lys Thr Ile Lys Asn His Glu Ser Arg Lys His Phe Leu Glu
1100 1105 1110
Gly Phe Asp Phe Leu His Tyr Asp Val Lys Thr Gly Asp Phe Ile
1115 1120 1125
Leu His Phe Lys Met Asn Arg Asn Leu Ser Phe Gln Arg Gly Leu
1130 1135 1140
Pro Gly Phe Met Pro Ala Trp Asp Ile Val Phe Glu Lys Asn Glu
1145 1150 1155
Thr Gln Phe Asp Ala Lys Gly Thr Pro Phe Ile Ala Gly Lys Arg
1160 1165 1170
Ile Val Pro Val Ile Glu Asn His Arg Phe Thr Gly Arg Tyr Arg
1175 1180 1185
Asp Leu Tyr Pro Ala Asn Glu Leu Ile Ala Leu Leu Glu Glu Lys
1190 1195 1200
Gly Ile Val Phe Arg Asp Gly Ser Asn Ile Leu Pro Lys Leu Leu
1205 1210 1215
Glu Asn Asp Asp Ser His Ala Ile Asp Thr Met Val Ala Leu Ile
1220 1225 1230
Arg Ser Val Leu Gln Met Arg Asn Ser Asn Ala Ala Thr Gly Glu
1235 1240 1245
Asp Tyr Ile Asn Ser Pro Val Arg Asp Leu Asn Gly Val Cys Phe
1250 1255 1260
Asp Ser Arg Phe Gln Asn Pro Glu Trp Pro Met Asp Ala Asp Ala
1265 1270 1275
Asn Gly Ala Tyr His Ile Ala Leu Lys Gly Gln Leu Leu Leu Asn
1280 1285 1290
His Leu Lys Glu Ser Lys Asp Leu Lys Leu Gln Asn Gly Ile Ser
1295 1300 1305
Asn Gln Asp Trp Leu Ala Tyr Ile Gln Glu Leu Arg Asn Thr Ser
1310 1315 1320
Arg Ala Asp Pro Lys Lys Lys Arg Lys Val Glu Ala Ser Gly Ser
1325 1330 1335
Gly Arg Ala Asp Ala Leu Asp Asp Phe Asp Leu Asp Met Leu Gly
1340 1345 1350
Ser Asp Ala Leu Asp Asp Phe Asp Leu Asp Met Leu Gly Ser Asp
1355 1360 1365
Ala Leu Asp Asp Phe Asp Leu Asp Met Leu Gly Ser Asp Ala Leu
1370 1375 1380
Asp Asp Phe Asp Leu Asp Met Leu Ile Asn Ser Arg Ser Gln Tyr
1385 1390 1395
Leu Pro Asp Thr Asp Asp Arg His Arg Ile Glu Glu Lys Arg Lys
1400 1405 1410
Arg Thr Tyr Glu Thr Phe Lys Ser Ile Met Lys Lys Ser Pro Phe
1415 1420 1425
Ser Gly Pro Thr Asp Pro Arg Pro Pro Pro Arg Arg Ile Ala Val
1430 1435 1440
Pro Ser Arg Ser Ser Ala Ser Val Pro Lys Pro Ala Pro Gln Pro
1445 1450 1455
Tyr Pro Phe Thr Ser Ser Leu Ser Thr Ile Asn Tyr Asp Glu Phe
1460 1465 1470
Pro Thr Met Val Phe Pro Ser Gly Gln Ile Ser Gln Ala Ser Ala
1475 1480 1485
Leu Ala Pro Ala Pro Pro Gln Val Leu Pro Gln Ala Pro Ala Pro
1490 1495 1500
Ala Pro Ala Pro Ala Met Val Ser Ala Leu Ala Gln Ala Pro Ala
1505 1510 1515
Pro Val Pro Val Leu Ala Pro Gly Pro Pro Gln Ala Val Ala Pro
1520 1525 1530
Pro Ala Pro Lys Pro Thr Gln Ala Gly Glu Gly Thr Leu Ser Glu
1535 1540 1545
Ala Leu Leu Gln Leu Gln Phe Asp Asp Glu Asp Leu Gly Ala Leu
1550 1555 1560
Leu Gly Asn Ser Thr Asp Pro Ala Val Phe Thr Asp Leu Ala Ser
1565 1570 1575
Val Asp Asn Ser Glu Phe Gln Gln Leu Leu Asn Gln Gly Ile Pro
1580 1585 1590
Val Ala Pro His Thr Thr Glu Pro Met Leu Met Glu Tyr Pro Glu
1595 1600 1605
Ala Ile Thr Arg Leu Val Thr Gly Ala Gln Arg Pro Pro Asp Pro
1610 1615 1620
Ala Pro Ala Pro Leu Gly Ala Pro Gly Leu Pro Asn Gly Leu Leu
1625 1630 1635
Ser Gly Asp Glu Asp Phe Ser Ser Ile Ala Asp Met Asp Phe Ser
1640 1645 1650
Ala Leu Leu Gly Ser Gly Ser Gly Ser Arg Asp Ser Arg Glu Gly
1655 1660 1665
Met Phe Leu Pro Lys Pro Glu Ala Gly Ser Ala Ile Ser Asp Val
1670 1675 1680
Phe Glu Gly Arg Glu Val Cys Gln Pro Lys Arg Ile Arg Pro Phe
1685 1690 1695
His Pro Pro Gly Ser Pro Trp Ala Asn Arg Pro Leu Pro Ala Ser
1700 1705 1710
Leu Ala Pro Thr Pro Thr Gly Pro Val His Glu Pro Val Gly Ser
1715 1720 1725
Leu Thr Pro Ala Pro Val Pro Gln Pro Leu Asp Pro Ala Pro Ala
1730 1735 1740
Val Thr Pro Glu Ala Ser His Leu Leu Glu Asp Pro Asp Glu Glu
1745 1750 1755
Thr Ser Gln Ala Val Lys Ala Leu Arg Glu Met Ala Asp Thr Val
1760 1765 1770
Ile Pro Gln Lys Glu Glu Ala Ala Ile Cys Gly Gln Met Asp Leu
1775 1780 1785
Ser His Pro Pro Pro Arg Gly His Leu Asp Glu Leu Thr Thr Thr
1790 1795 1800
Leu Glu Ser Met Thr Glu Asp Leu Asn Leu Asp Ser Pro Leu Thr
1805 1810 1815
Pro Glu Leu Asn Glu Ile Leu Asp Thr Phe Leu Asn Asp Glu Cys
1820 1825 1830
Leu Leu His Ala Met His Ile Ser Thr Gly Leu Ser Ile Phe Asp
1835 1840 1845
Thr Ser Leu Phe
1850
<210> 16
<211> 1654
<212> PRT
<213> artificial sequence
<220>
<223> synthetic constructs produced by rational design
<400> 16
Met Pro Lys Lys Lys Arg Lys Val Gly Gly Gly Ser Pro Gly Met Thr
1 5 10 15
Gln Phe Glu Gly Phe Thr Asn Leu Tyr Gln Val Ser Lys Thr Leu Arg
20 25 30
Phe Glu Leu Ile Pro Gln Gly Lys Thr Leu Lys His Ile Gln Glu Gln
35 40 45
Gly Phe Ile Glu Glu Asp Lys Ala Arg Asn Asp His Tyr Lys Glu Leu
50 55 60
Lys Pro Ile Ile Asp Arg Ile Tyr Lys Thr Tyr Ala Asp Gln Cys Leu
65 70 75 80
Gln Leu Val Gln Leu Asp Trp Glu Asn Leu Ser Ala Ala Ile Asp Ser
85 90 95
Tyr Arg Lys Glu Lys Thr Glu Glu Thr Arg Asn Ala Leu Ile Glu Glu
100 105 110
Gln Ala Thr Tyr Arg Asn Ala Ile His Asp Tyr Phe Ile Gly Arg Thr
115 120 125
Asp Asn Leu Thr Asp Ala Ile Asn Lys Arg His Ala Glu Ile Tyr Lys
130 135 140
Gly Leu Phe Lys Ala Glu Leu Phe Asn Gly Lys Val Leu Lys Gln Leu
145 150 155 160
Gly Thr Val Thr Thr Thr Glu His Glu Asn Ala Leu Leu Arg Ser Phe
165 170 175
Asp Lys Phe Thr Thr Tyr Phe Ser Gly Phe Tyr Glu Asn Arg Lys Asn
180 185 190
Val Phe Ser Ala Glu Asp Ile Ser Thr Ala Ile Pro His Arg Ile Val
195 200 205
Gln Asp Asn Phe Pro Lys Phe Lys Glu Asn Cys His Ile Phe Thr Arg
210 215 220
Leu Ile Thr Ala Val Pro Ser Leu Arg Glu His Phe Glu Asn Val Lys
225 230 235 240
Lys Ala Ile Gly Ile Phe Val Ser Thr Ser Ile Glu Glu Val Phe Ser
245 250 255
Phe Pro Phe Tyr Asn Gln Leu Leu Thr Gln Thr Gln Ile Asp Leu Tyr
260 265 270
Asn Gln Leu Leu Gly Gly Ile Ser Arg Glu Ala Gly Thr Glu Lys Ile
275 280 285
Lys Gly Leu Asn Glu Val Leu Asn Leu Ala Ile Gln Lys Asn Asp Glu
290 295 300
Thr Ala His Ile Ile Ala Ser Leu Pro His Arg Phe Ile Pro Leu Phe
305 310 315 320
Lys Gln Ile Leu Ser Asp Arg Asn Thr Leu Ser Phe Ile Leu Glu Glu
325 330 335
Phe Gly Gly Gly Ser Tyr Ser Val Glu Lys Phe Lys Leu Asn Phe Gln
340 345 350
Met Pro Thr Leu Ala Ser Gly Trp Asp Val Asn Lys Glu Lys Asn Asn
355 360 365
Gly Ala Ile Leu Phe Val Lys Asn Gly Leu Tyr Tyr Leu Gly Ile Met
370 375 380
Pro Lys Gln Lys Gly Arg Tyr Lys Ala Leu Ser Phe Glu Pro Thr Glu
385 390 395 400
Lys Thr Ser Glu Gly Phe Asp Lys Met Tyr Tyr Asp Tyr Phe Pro Asp
405 410 415
Ala Ala Lys Met Ile Pro Lys Cys Ser Thr Gln Leu Lys Ala Val Thr
420 425 430
Ala His Phe Gln Thr His Thr Thr Pro Ile Leu Leu Ser Asn Asn Phe
435 440 445
Ile Glu Pro Leu Glu Ile Thr Lys Glu Ile Tyr Asp Leu Asn Asn Pro
450 455 460
Glu Lys Glu Pro Lys Lys Phe Gln Thr Ala Tyr Ala Lys Lys Thr Gly
465 470 475 480
Asp Gln Lys Gly Tyr Arg Glu Ala Leu Cys Lys Trp Ile Asp Phe Thr
485 490 495
Arg Asp Phe Leu Ser Lys Tyr Thr Lys Thr Thr Ser Ile Asp Leu Ser
500 505 510
Ser Leu Arg Pro Ser Ser Gln Tyr Lys Asp Leu Gly Glu Tyr Tyr Ala
515 520 525
Glu Leu Asn Pro Leu Leu Tyr His Ile Ser Phe Gln Arg Ile Ala Glu
530 535 540
Lys Glu Ile Met Asp Ala Val Glu Thr Gly Lys Leu Tyr Leu Phe Gln
545 550 555 560
Ile Tyr Asn Lys Asp Phe Ala Lys Gly His His Gly Lys Pro Asn Leu
565 570 575
His Thr Leu Tyr Trp Thr Gly Leu Phe Ser Pro Glu Asn Leu Ala Lys
580 585 590
Thr Ser Ile Lys Leu Asn Gly Gln Ala Glu Leu Phe Tyr Arg Pro Lys
595 600 605
Ser Arg Met Lys Arg Met Ala His Arg Leu Gly Glu Lys Met Leu Asn
610 615 620
Lys Lys Leu Lys Asp Gln Lys Thr Pro Ile Pro Asp Thr Leu Tyr Gln
625 630 635 640
Glu Leu Tyr Asp Tyr Val Asn His Arg Leu Ser His Asp Leu Ser Asp
645 650 655
Glu Ala Arg Ala Leu Leu Pro Asn Val Ile Thr Lys Glu Val Ser His
660 665 670
Glu Ile Ile Lys Asp Arg Arg Phe Thr Ser Asp Lys Phe Phe Phe His
675 680 685
Val Pro Ile Thr Leu Asn Tyr Gln Ala Ala Asn Ser Pro Ser Lys Phe
690 695 700
Asn Gln Arg Val Asn Ala Tyr Leu Lys Glu His Pro Glu Thr Pro Ile
705 710 715 720
Ile Gly Ile Asp Arg Gly Glu Arg Asn Leu Ile Tyr Ile Thr Val Ile
725 730 735
Asp Ser Thr Gly Lys Ile Leu Glu Gln Arg Ser Leu Asn Thr Ile Gln
740 745 750
Gln Phe Asp Tyr Gln Lys Lys Leu Asp Asn Arg Glu Lys Glu Arg Val
755 760 765
Ala Ala Arg Gln Ala Trp Ser Val Val Gly Thr Ile Lys Asp Leu Lys
770 775 780
Gln Gly Tyr Leu Ser Gln Val Ile His Glu Ile Val Asp Leu Met Ile
785 790 795 800
His Tyr Gln Ala Val Val Val Leu Glu Asn Leu Asn Phe Gly Phe Lys
805 810 815
Ser Lys Arg Thr Gly Ile Ala Glu Lys Ala Val Tyr Gln Gln Phe Glu
820 825 830
Lys Met Leu Ile Asp Lys Leu Asn Cys Leu Val Leu Lys Asp Tyr Pro
835 840 845
Ala Glu Lys Val Gly Gly Val Leu Asn Pro Tyr Gln Leu Thr Asp Gln
850 855 860
Phe Thr Ser Phe Ala Lys Met Gly Thr Gln Ser Gly Phe Leu Phe Tyr
865 870 875 880
Val Pro Ala Pro Tyr Thr Ser Lys Ile Asp Pro Leu Thr Gly Phe Val
885 890 895
Asp Pro Phe Val Trp Lys Thr Ile Lys Asn His Glu Ser Arg Lys His
900 905 910
Phe Leu Glu Gly Phe Asp Phe Leu His Tyr Asp Val Lys Thr Gly Asp
915 920 925
Phe Ile Leu His Phe Lys Met Asn Arg Asn Leu Ser Phe Gln Arg Gly
930 935 940
Leu Pro Gly Phe Met Pro Ala Trp Asp Ile Val Phe Glu Lys Asn Glu
945 950 955 960
Thr Gln Phe Asp Ala Lys Gly Thr Pro Phe Ile Ala Gly Lys Arg Ile
965 970 975
Val Pro Val Ile Glu Asn His Arg Phe Thr Gly Arg Tyr Arg Asp Leu
980 985 990
Tyr Pro Ala Asn Glu Leu Ile Ala Leu Leu Glu Glu Lys Gly Ile Val
995 1000 1005
Phe Arg Asp Gly Ser Asn Ile Leu Pro Lys Leu Leu Glu Asn Asp
1010 1015 1020
Asp Ser His Ala Ile Asp Thr Met Val Ala Leu Ile Arg Ser Val
1025 1030 1035
Leu Gln Met Arg Asn Ser Asn Ala Ala Thr Gly Glu Asp Tyr Ile
1040 1045 1050
Asn Ser Pro Val Arg Asp Leu Asn Gly Val Cys Phe Asp Ser Arg
1055 1060 1065
Phe Gln Asn Pro Glu Trp Pro Met Asp Ala Asp Ala Asn Gly Ala
1070 1075 1080
Tyr His Ile Ala Leu Lys Gly Gln Leu Leu Leu Asn His Leu Lys
1085 1090 1095
Glu Ser Lys Asp Leu Lys Leu Gln Asn Gly Ile Ser Asn Gln Asp
1100 1105 1110
Trp Leu Ala Tyr Ile Gln Glu Leu Arg Asn Thr Ser Arg Ala Asp
1115 1120 1125
Pro Lys Lys Lys Arg Lys Val Glu Ala Ser Gly Ser Gly Arg Ala
1130 1135 1140
Asp Ala Leu Asp Asp Phe Asp Leu Asp Met Leu Gly Ser Asp Ala
1145 1150 1155
Leu Asp Asp Phe Asp Leu Asp Met Leu Gly Ser Asp Ala Leu Asp
1160 1165 1170
Asp Phe Asp Leu Asp Met Leu Gly Ser Asp Ala Leu Asp Asp Phe
1175 1180 1185
Asp Leu Asp Met Leu Ile Asn Ser Arg Ser Gln Tyr Leu Pro Asp
1190 1195 1200
Thr Asp Asp Arg His Arg Ile Glu Glu Lys Arg Lys Arg Thr Tyr
1205 1210 1215
Glu Thr Phe Lys Ser Ile Met Lys Lys Ser Pro Phe Ser Gly Pro
1220 1225 1230
Thr Asp Pro Arg Pro Pro Pro Arg Arg Ile Ala Val Pro Ser Arg
1235 1240 1245
Ser Ser Ala Ser Val Pro Lys Pro Ala Pro Gln Pro Tyr Pro Phe
1250 1255 1260
Thr Ser Ser Leu Ser Thr Ile Asn Tyr Asp Glu Phe Pro Thr Met
1265 1270 1275
Val Phe Pro Ser Gly Gln Ile Ser Gln Ala Ser Ala Leu Ala Pro
1280 1285 1290
Ala Pro Pro Gln Val Leu Pro Gln Ala Pro Ala Pro Ala Pro Ala
1295 1300 1305
Pro Ala Met Val Ser Ala Leu Ala Gln Ala Pro Ala Pro Val Pro
1310 1315 1320
Val Leu Ala Pro Gly Pro Pro Gln Ala Val Ala Pro Pro Ala Pro
1325 1330 1335
Lys Pro Thr Gln Ala Gly Glu Gly Thr Leu Ser Glu Ala Leu Leu
1340 1345 1350
Gln Leu Gln Phe Asp Asp Glu Asp Leu Gly Ala Leu Leu Gly Asn
1355 1360 1365
Ser Thr Asp Pro Ala Val Phe Thr Asp Leu Ala Ser Val Asp Asn
1370 1375 1380
Ser Glu Phe Gln Gln Leu Leu Asn Gln Gly Ile Pro Val Ala Pro
1385 1390 1395
His Thr Thr Glu Pro Met Leu Met Glu Tyr Pro Glu Ala Ile Thr
1400 1405 1410
Arg Leu Val Thr Gly Ala Gln Arg Pro Pro Asp Pro Ala Pro Ala
1415 1420 1425
Pro Leu Gly Ala Pro Gly Leu Pro Asn Gly Leu Leu Ser Gly Asp
1430 1435 1440
Glu Asp Phe Ser Ser Ile Ala Asp Met Asp Phe Ser Ala Leu Leu
1445 1450 1455
Gly Ser Gly Ser Gly Ser Arg Asp Ser Arg Glu Gly Met Phe Leu
1460 1465 1470
Pro Lys Pro Glu Ala Gly Ser Ala Ile Ser Asp Val Phe Glu Gly
1475 1480 1485
Arg Glu Val Cys Gln Pro Lys Arg Ile Arg Pro Phe His Pro Pro
1490 1495 1500
Gly Ser Pro Trp Ala Asn Arg Pro Leu Pro Ala Ser Leu Ala Pro
1505 1510 1515
Thr Pro Thr Gly Pro Val His Glu Pro Val Gly Ser Leu Thr Pro
1520 1525 1530
Ala Pro Val Pro Gln Pro Leu Asp Pro Ala Pro Ala Val Thr Pro
1535 1540 1545
Glu Ala Ser His Leu Leu Glu Asp Pro Asp Glu Glu Thr Ser Gln
1550 1555 1560
Ala Val Lys Ala Leu Arg Glu Met Ala Asp Thr Val Ile Pro Gln
1565 1570 1575
Lys Glu Glu Ala Ala Ile Cys Gly Gln Met Asp Leu Ser His Pro
1580 1585 1590
Pro Pro Arg Gly His Leu Asp Glu Leu Thr Thr Thr Leu Glu Ser
1595 1600 1605
Met Thr Glu Asp Leu Asn Leu Asp Ser Pro Leu Thr Pro Glu Leu
1610 1615 1620
Asn Glu Ile Leu Asp Thr Phe Leu Asn Asp Glu Cys Leu Leu His
1625 1630 1635
Ala Met His Ile Ser Thr Gly Leu Ser Ile Phe Asp Thr Ser Leu
1640 1645 1650
Phe
<210> 17
<211> 977
<212> PRT
<213> artificial sequence
<220>
<223> synthetic constructs produced by rational design
<400> 17
Met Pro Lys Lys Lys Arg Lys Val Gly Gly Gly Ser Pro Gly Lys Arg
1 5 10 15
Asn Tyr Ile Leu Gly Leu Ala Ile Gly Ile Thr Ser Val Gly Tyr Gly
20 25 30
Ile Ile Asp Tyr Glu Thr Arg Asp Val Ile Asp Ala Gly Val Arg Leu
35 40 45
Phe Lys Glu Ala Asn Val Glu Asn Asn Glu Gly Arg Arg Ser Lys Arg
50 55 60
Gly Ala Arg Arg Leu Lys Arg Arg Arg Arg His Arg Ile Gln Arg Val
65 70 75 80
Lys Lys Leu Leu Phe Asp Tyr Asn Leu Leu Thr Asp His Ser Glu Leu
85 90 95
Ser Gly Ile Asn Pro Tyr Glu Ala Arg Val Lys Gly Leu Ser Gln Lys
100 105 110
Leu Ser Glu Glu Glu Phe Ser Ala Ala Leu Leu His Leu Ala Lys Arg
115 120 125
Arg Gly Val His Asn Val Asn Glu Val Glu Glu Asp Thr Gly Asn Glu
130 135 140
Leu Ser Thr Lys Glu Gln Ile Ser Arg Asn Ser Lys Ala Leu Glu Glu
145 150 155 160
Lys Tyr Val Ala Glu Leu Gln Leu Glu Arg Leu Lys Lys Asp Gly Glu
165 170 175
Val Arg Gly Ser Ile Asn Arg Phe Lys Thr Ser Asp Tyr Val Lys Glu
180 185 190
Ala Lys Gln Leu Leu Lys Val Gln Lys Ala Tyr His Gln Leu Asp Gln
195 200 205
Ser Phe Ile Asp Thr Tyr Ile Asp Leu Leu Glu Thr Arg Arg Thr Tyr
210 215 220
Tyr Glu Gly Pro Gly Glu Gly Ser Pro Phe Gly Trp Lys Asp Ile Lys
225 230 235 240
Glu Trp Tyr Glu Met Leu Met Gly His Cys Thr Tyr Phe Pro Glu Glu
245 250 255
Leu Arg Ser Val Lys Tyr Ala Tyr Asn Ala Asp Leu Tyr Asn Ala Leu
260 265 270
Asn Asp Leu Asn Asn Leu Val Ile Thr Arg Asp Glu Asn Glu Lys Leu
275 280 285
Glu Tyr Tyr Glu Lys Phe Gln Ile Ile Glu Asn Val Phe Lys Gln Lys
290 295 300
Lys Lys Pro Thr Leu Lys Gln Ile Ala Lys Glu Ile Leu Val Asn Glu
305 310 315 320
Glu Asp Ile Lys Gly Tyr Arg Val Thr Ser Thr Gly Lys Pro Glu Phe
325 330 335
Thr Asn Leu Lys Val Tyr His Asp Ile Lys Asp Ile Thr Ala Arg Lys
340 345 350
Glu Ile Ile Glu Asn Ala Glu Leu Leu Asp Gln Ile Ala Lys Ile Leu
355 360 365
Thr Ile Tyr Gln Ser Ser Glu Asp Ile Gln Glu Glu Leu Thr Asn Leu
370 375 380
Asn Ser Glu Leu Thr Gln Glu Glu Ile Glu Gln Ile Ser Asn Leu Lys
385 390 395 400
Gly Tyr Thr Gly Thr His Asn Leu Ser Leu Lys Ala Ile Asn Leu Ile
405 410 415
Leu Asp Glu Leu Trp His Thr Asn Asp Asn Gln Ile Ala Ile Phe Asn
420 425 430
Arg Leu Lys Leu Val Pro Lys Lys Val Asp Leu Ser Gln Gln Lys Glu
435 440 445
Ile Pro Thr Thr Leu Val Asp Asp Phe Ile Leu Ser Pro Val Val Lys
450 455 460
Arg Ser Phe Ile Gln Ser Ile Lys Val Ile Asn Ala Ile Ile Lys Lys
465 470 475 480
Tyr Gly Leu Pro Asn Asp Ile Ile Ile Glu Leu Ala Arg Glu Lys Asn
485 490 495
Ser Lys Asp Ala Gln Lys Met Ile Asn Glu Met Gln Lys Arg Asn Arg
500 505 510
Gln Thr Asn Glu Arg Ile Glu Glu Ile Ile Arg Thr Thr Gly Lys Glu
515 520 525
Asn Ala Lys Tyr Leu Ile Glu Lys Ile Lys Leu His Asp Met Gln Glu
530 535 540
Gly Lys Cys Leu Tyr Ser Leu Glu Ala Ile Pro Leu Glu Asp Leu Leu
545 550 555 560
Asn Asn Pro Phe Asn Tyr Glu Val Asp His Ile Ile Pro Arg Ser Val
565 570 575
Ser Phe Asp Asn Ser Phe Asn Asn Lys Val Leu Val Lys Gln Glu Glu
580 585 590
Ala Ser Lys Lys Gly Asn Arg Thr Pro Phe Gln Tyr Leu Ser Ser Ser
595 600 605
Asp Ser Lys Ile Ser Tyr Glu Thr Phe Lys Lys His Ile Leu Asn Leu
610 615 620
Ala Lys Gly Lys Gly Arg Ile Ser Lys Thr Lys Lys Glu Tyr Leu Leu
625 630 635 640
Glu Glu Arg Asp Ile Asn Arg Phe Ser Val Gln Lys Asp Phe Ile Asn
645 650 655
Arg Asn Leu Val Asp Thr Arg Tyr Ala Thr Arg Gly Leu Met Asn Leu
660 665 670
Leu Arg Ser Tyr Phe Arg Val Asn Asn Leu Asp Val Lys Val Lys Ser
675 680 685
Ile Asn Gly Gly Phe Thr Ser Phe Leu Arg Arg Lys Trp Lys Phe Lys
690 695 700
Lys Glu Arg Asn Lys Gly Tyr Lys His His Ala Glu Asp Ala Leu Ile
705 710 715 720
Ile Ala Asn Ala Asp Phe Ile Phe Lys Glu Trp Lys Lys Leu Asp Lys
725 730 735
Ala Lys Lys Val Met Glu Asn Gln Met Phe Gly Gly Gly Ser Gly Gly
740 745 750
Gly Ser Gly Gly Gly Ser Leu Val Lys Ser Glu Leu Glu Glu Lys Lys
755 760 765
Ser Glu Leu Arg His Lys Leu Lys Tyr Val Pro His Glu Tyr Ile Glu
770 775 780
Leu Ile Glu Ile Ala Arg Asn Ser Thr Gln Asp Arg Ile Leu Glu Met
785 790 795 800
Lys Val Met Glu Phe Phe Met Lys Val Tyr Gly Tyr Arg Gly Lys His
805 810 815
Leu Gly Gly Ser Arg Lys Pro Asp Gly Ala Ile Tyr Thr Val Gly Ser
820 825 830
Pro Ile Asp Tyr Gly Val Ile Val Asp Thr Lys Ala Tyr Ser Gly Gly
835 840 845
Tyr Asn Leu Pro Ile Gly Gln Ala Asp Glu Met Gln Arg Tyr Val Glu
850 855 860
Glu Asn Gln Thr Arg Asn Lys His Ile Asn Pro Asn Glu Trp Trp Lys
865 870 875 880
Val Tyr Pro Ser Ser Val Thr Glu Phe Lys Phe Leu Phe Val Ser Gly
885 890 895
His Phe Lys Gly Asn Tyr Lys Ala Gln Leu Thr Arg Leu Asn His Ile
900 905 910
Thr Asn Cys Asn Gly Ala Val Leu Ser Val Glu Glu Leu Leu Ile Gly
915 920 925
Gly Glu Met Ile Lys Ala Gly Thr Leu Thr Leu Glu Glu Val Arg Arg
930 935 940
Lys Phe Asn Asn Gly Glu Ile Asn Phe Thr Ser Gly Gly Gly Ser Lys
945 950 955 960
Arg Pro Ala Ala Thr Lys Lys Ala Gly Gln Ala Lys Lys Lys Lys Ser
965 970 975
Arg
<210> 18
<211> 354
<212> PRT
<213> artificial sequence
<220>
<223> synthetic constructs produced by rational design
<400> 18
Met Pro Lys Lys Lys Arg Lys Val Gly Gly Gly Ser Pro Gly Ala Glu
1 5 10 15
Ser Met Pro Glu Ile Glu Thr Glu Gln Glu Tyr Lys Glu Ile Phe Ile
20 25 30
Thr Pro His Gln Ile Lys His Ile Lys Asp Phe Lys Asp Tyr Lys Tyr
35 40 45
Ser His Arg Val Asp Lys Lys Pro Asn Arg Lys Leu Ile Asn Asp Thr
50 55 60
Leu Tyr Ser Thr Arg Lys Asp Asp Lys Gly Asn Thr Leu Ile Val Asn
65 70 75 80
Asn Leu Asn Gly Leu Tyr Asp Lys Asp Asn Asp Lys Leu Lys Lys Leu
85 90 95
Ile Asn Lys Ser Pro Glu Lys Leu Leu Met Tyr His His Asp Pro Gln
100 105 110
Thr Tyr Gln Lys Leu Lys Leu Ile Met Glu Gln Tyr Gly Asp Glu Lys
115 120 125
Asn Pro Leu Tyr Lys Tyr Tyr Glu Glu Thr Gly Asn Tyr Leu Thr Lys
130 135 140
Tyr Ser Lys Lys Asp Asn Gly Pro Val Ile Lys Lys Ile Lys Tyr Tyr
145 150 155 160
Gly Asn Lys Leu Asn Ala His Leu Asp Ile Thr Asp Asp Tyr Pro Asn
165 170 175
Ser Arg Asn Lys Val Val Lys Leu Ser Leu Lys Pro Tyr Arg Phe Asp
180 185 190
Val Tyr Leu Asp Asn Gly Val Tyr Lys Phe Val Thr Val Lys Asn Leu
195 200 205
Asp Val Ile Lys Lys Glu Asn Tyr Tyr Glu Val Asn Ser Lys Cys Tyr
210 215 220
Glu Glu Ala Lys Lys Leu Lys Lys Ile Ser Asn Gln Ala Glu Phe Ile
225 230 235 240
Ala Ser Phe Tyr Lys Asn Asp Leu Ile Lys Ile Asn Gly Glu Leu Tyr
245 250 255
Arg Val Ile Gly Val Asn Asn Asp Leu Leu Asn Arg Ile Glu Val Asn
260 265 270
Met Ile Asp Ile Thr Tyr Arg Glu Tyr Leu Glu Asn Met Asn Asp Lys
275 280 285
Arg Pro Pro His Ile Ile Lys Thr Ile Ala Ser Lys Thr Gln Ser Ile
290 295 300
Lys Lys Tyr Ser Thr Asp Ile Leu Gly Asn Leu Tyr Glu Val Lys Ser
305 310 315 320
Lys Lys His Pro Gln Ile Ile Lys Lys Gly Thr Ser Gly Gly Gly Ser
325 330 335
Lys Arg Pro Ala Ala Thr Lys Lys Ala Gly Gln Ala Lys Lys Lys Lys
340 345 350
Ser Arg
<210> 19
<211> 809
<212> PRT
<213> artificial sequence
<220>
<223> synthetic constructs produced by rational design
<400> 19
Met Pro Lys Lys Lys Arg Lys Val Gly Gly Gly Ser Pro Gly Lys Arg
1 5 10 15
Asn Tyr Ile Leu Gly Leu Ala Ile Gly Ile Thr Ser Val Gly Tyr Gly
20 25 30
Ile Ile Asp Tyr Glu Thr Arg Asp Val Ile Asp Ala Gly Val Arg Leu
35 40 45
Phe Lys Glu Ala Asn Val Glu Asn Asn Glu Gly Arg Arg Ser Lys Arg
50 55 60
Gly Ala Arg Arg Leu Lys Arg Arg Arg Arg His Arg Ile Gln Arg Val
65 70 75 80
Lys Lys Leu Leu Phe Asp Tyr Asn Leu Leu Thr Asp His Ser Glu Leu
85 90 95
Ser Gly Ile Asn Pro Tyr Glu Ala Arg Val Lys Gly Leu Ser Gln Lys
100 105 110
Leu Ser Glu Glu Glu Phe Ser Ala Ala Leu Leu His Leu Ala Lys Arg
115 120 125
Arg Gly Val His Asn Val Asn Glu Val Glu Glu Asp Thr Gly Asn Glu
130 135 140
Leu Ser Thr Lys Glu Gln Ile Ser Arg Asn Ser Lys Ala Leu Glu Glu
145 150 155 160
Lys Tyr Val Ala Glu Leu Gln Leu Glu Arg Leu Lys Lys Asp Gly Glu
165 170 175
Val Arg Gly Ser Ile Asn Arg Phe Lys Thr Ser Asp Tyr Val Lys Glu
180 185 190
Ala Lys Gln Leu Leu Lys Val Gln Lys Ala Tyr His Gln Leu Asp Gln
195 200 205
Ser Phe Ile Asp Thr Tyr Ile Asp Leu Leu Glu Thr Arg Arg Thr Tyr
210 215 220
Tyr Glu Gly Pro Gly Glu Gly Ser Pro Phe Gly Trp Lys Asp Ile Lys
225 230 235 240
Glu Trp Tyr Glu Met Leu Met Gly His Cys Thr Tyr Phe Pro Glu Glu
245 250 255
Leu Arg Ser Val Lys Tyr Ala Tyr Asn Ala Asp Leu Tyr Asn Ala Leu
260 265 270
Asn Asp Leu Asn Asn Leu Val Ile Thr Arg Asp Glu Asn Glu Lys Leu
275 280 285
Glu Tyr Tyr Glu Lys Phe Gln Ile Ile Glu Asn Val Phe Lys Gln Lys
290 295 300
Lys Lys Pro Thr Leu Lys Gln Ile Ala Lys Glu Ile Leu Val Asn Glu
305 310 315 320
Glu Asp Ile Lys Gly Tyr Arg Val Thr Ser Thr Gly Lys Pro Glu Phe
325 330 335
Thr Asn Leu Lys Val Tyr His Asp Ile Lys Asp Ile Thr Ala Arg Lys
340 345 350
Glu Ile Ile Glu Asn Ala Glu Leu Leu Asp Gln Ile Ala Lys Ile Leu
355 360 365
Thr Ile Tyr Gln Ser Ser Glu Asp Ile Gln Glu Glu Leu Thr Asn Leu
370 375 380
Asn Ser Glu Leu Thr Gln Glu Glu Ile Glu Gln Ile Ser Asn Leu Lys
385 390 395 400
Gly Tyr Thr Gly Thr His Asn Leu Ser Leu Lys Ala Ile Asn Leu Ile
405 410 415
Leu Asp Glu Leu Trp His Thr Asn Asp Asn Gln Ile Ala Ile Phe Asn
420 425 430
Arg Leu Lys Leu Val Pro Lys Lys Val Asp Leu Ser Gln Gln Lys Glu
435 440 445
Ile Pro Thr Thr Leu Val Asp Asp Phe Ile Leu Ser Pro Val Val Lys
450 455 460
Arg Ser Phe Ile Gln Ser Ile Lys Val Ile Asn Ala Ile Ile Lys Lys
465 470 475 480
Tyr Gly Leu Pro Asn Asp Ile Ile Ile Glu Leu Gly Ser Lys Arg Tyr
485 490 495
Ala Thr Arg Gly Leu Met Asn Leu Leu Arg Ser Tyr Phe Arg Val Asn
500 505 510
Asn Leu Asp Val Lys Val Lys Ser Ile Asn Gly Gly Phe Thr Ser Phe
515 520 525
Leu Arg Arg Lys Trp Lys Phe Lys Lys Glu Arg Asn Lys Gly Tyr Lys
530 535 540
His His Ala Glu Asp Ala Leu Ile Ile Ala Asn Ala Asp Phe Ile Phe
545 550 555 560
Lys Glu Trp Lys Lys Leu Asp Lys Ala Lys Lys Val Met Glu Asn Gln
565 570 575
Met Phe Gly Gly Gly Ser Gly Gly Gly Ser Gly Gly Gly Ser Leu Val
580 585 590
Lys Ser Glu Leu Glu Glu Lys Lys Ser Glu Leu Arg His Lys Leu Lys
595 600 605
Tyr Val Pro His Glu Tyr Ile Glu Leu Ile Glu Ile Ala Arg Asn Ser
610 615 620
Thr Gln Asp Arg Ile Leu Glu Met Lys Val Met Glu Phe Phe Met Lys
625 630 635 640
Val Tyr Gly Tyr Arg Gly Lys His Leu Gly Gly Ser Arg Lys Pro Asp
645 650 655
Gly Ala Ile Tyr Thr Val Gly Ser Pro Ile Asp Tyr Gly Val Ile Val
660 665 670
Asp Thr Lys Ala Tyr Ser Gly Gly Tyr Asn Leu Pro Ile Gly Gln Ala
675 680 685
Asp Glu Met Gln Arg Tyr Val Glu Glu Asn Gln Thr Arg Asn Lys His
690 695 700
Ile Asn Pro Asn Glu Trp Trp Lys Val Tyr Pro Ser Ser Val Thr Glu
705 710 715 720
Phe Lys Phe Leu Phe Val Ser Gly His Phe Lys Gly Asn Tyr Lys Ala
725 730 735
Gln Leu Thr Arg Leu Asn His Ile Thr Asn Cys Asn Gly Ala Val Leu
740 745 750
Ser Val Glu Glu Leu Leu Ile Gly Gly Glu Met Ile Lys Ala Gly Thr
755 760 765
Leu Thr Leu Glu Glu Val Arg Arg Lys Phe Asn Asn Gly Glu Ile Asn
770 775 780
Phe Thr Ser Gly Gly Gly Ser Lys Arg Pro Ala Ala Thr Lys Lys Ala
785 790 795 800
Gly Gln Ala Lys Lys Lys Lys Ser Arg
805
<210> 20
<211> 1409
<212> PRT
<213> artificial sequence
<220>
<223> synthetic constructs produced by rational design
<400> 20
Met Pro Lys Lys Lys Arg Lys Val Gly Gly Gly Ser Pro Gly Met Ser
1 5 10 15
Ser Glu Thr Gly Pro Val Ala Val Asp Pro Thr Leu Arg Arg Arg Ile
20 25 30
Glu Pro His Glu Phe Glu Val Phe Phe Asp Pro Arg Glu Leu Arg Lys
35 40 45
Glu Thr Cys Leu Leu Tyr Glu Ile Asn Trp Gly Gly Arg His Ser Ile
50 55 60
Trp Arg His Thr Ser Gln Asn Thr Asn Lys His Val Glu Val Asn Phe
65 70 75 80
Ile Glu Lys Phe Thr Thr Glu Arg Tyr Phe Cys Pro Asn Thr Arg Cys
85 90 95
Ser Ile Thr Trp Phe Leu Ser Trp Ser Pro Cys Gly Glu Cys Ser Arg
100 105 110
Ala Ile Thr Glu Phe Leu Ser Arg Tyr Pro His Val Thr Leu Phe Ile
115 120 125
Tyr Ile Ala Arg Leu Tyr His His Ala Asp Pro Arg Asn Arg Gln Gly
130 135 140
Leu Arg Asp Leu Ile Ser Ser Gly Val Thr Ile Gln Ile Met Thr Glu
145 150 155 160
Gln Glu Ser Gly Tyr Cys Trp Arg Asn Phe Val Asn Tyr Ser Pro Ser
165 170 175
Asn Glu Ala His Trp Pro Arg Tyr Pro His Leu Trp Val Arg Leu Tyr
180 185 190
Val Leu Glu Leu Tyr Cys Ile Ile Leu Gly Leu Pro Pro Cys Leu Asn
195 200 205
Ile Leu Arg Arg Lys Gln Pro Gln Leu Thr Phe Phe Thr Ile Ala Leu
210 215 220
Gln Ser Cys His Tyr Gln Arg Leu Pro Pro His Ile Leu Trp Ala Thr
225 230 235 240
Gly Leu Lys Ser Gly Ser Glu Thr Pro Gly Thr Ser Glu Ser Ala Thr
245 250 255
Pro Glu Lys Arg Asn Tyr Ile Leu Gly Leu Ala Ile Gly Ile Thr Ser
260 265 270
Val Gly Tyr Gly Ile Ile Asp Tyr Glu Thr Arg Asp Val Ile Asp Ala
275 280 285
Gly Val Arg Leu Phe Lys Glu Ala Asn Val Glu Asn Asn Glu Gly Arg
290 295 300
Arg Ser Lys Arg Gly Ala Arg Arg Leu Lys Arg Arg Arg Arg His Arg
305 310 315 320
Ile Gln Arg Val Lys Lys Leu Leu Phe Asp Tyr Asn Leu Leu Thr Asp
325 330 335
His Ser Glu Leu Ser Gly Ile Asn Pro Tyr Glu Ala Arg Val Lys Gly
340 345 350
Leu Ser Gln Lys Leu Ser Glu Glu Glu Phe Ser Ala Ala Leu Leu His
355 360 365
Leu Ala Lys Arg Arg Gly Val His Asn Val Asn Glu Val Glu Glu Asp
370 375 380
Thr Gly Asn Glu Leu Ser Thr Lys Glu Gln Ile Ser Arg Asn Ser Lys
385 390 395 400
Ala Leu Glu Glu Lys Tyr Val Ala Glu Leu Gln Leu Glu Arg Leu Lys
405 410 415
Lys Asp Gly Glu Val Arg Gly Ser Ile Asn Arg Phe Lys Thr Ser Asp
420 425 430
Tyr Val Lys Glu Ala Lys Gln Leu Leu Lys Val Gln Lys Ala Tyr His
435 440 445
Gln Leu Asp Gln Ser Phe Ile Asp Thr Tyr Ile Asp Leu Leu Glu Thr
450 455 460
Arg Arg Thr Tyr Tyr Glu Gly Pro Gly Glu Gly Ser Pro Phe Gly Trp
465 470 475 480
Lys Asp Ile Lys Glu Trp Tyr Glu Met Leu Met Gly His Cys Thr Tyr
485 490 495
Phe Pro Glu Glu Leu Arg Ser Val Lys Tyr Ala Tyr Asn Ala Asp Leu
500 505 510
Tyr Asn Ala Leu Asn Asp Leu Asn Asn Leu Val Ile Thr Arg Asp Glu
515 520 525
Asn Glu Lys Leu Glu Tyr Tyr Glu Lys Phe Gln Ile Ile Glu Asn Val
530 535 540
Phe Lys Gln Lys Lys Lys Pro Thr Leu Lys Gln Ile Ala Lys Glu Ile
545 550 555 560
Leu Val Asn Glu Glu Asp Ile Lys Gly Tyr Arg Val Thr Ser Thr Gly
565 570 575
Lys Pro Glu Phe Thr Asn Leu Lys Val Tyr His Asp Ile Lys Asp Ile
580 585 590
Thr Ala Arg Lys Glu Ile Ile Glu Asn Ala Glu Leu Leu Asp Gln Ile
595 600 605
Ala Lys Ile Leu Thr Ile Tyr Gln Ser Ser Glu Asp Ile Gln Glu Glu
610 615 620
Leu Thr Asn Leu Asn Ser Glu Leu Thr Gln Glu Glu Ile Glu Gln Ile
625 630 635 640
Ser Asn Leu Lys Gly Tyr Thr Gly Thr His Asn Leu Ser Leu Lys Ala
645 650 655
Ile Asn Leu Ile Leu Asp Glu Leu Trp His Thr Asn Asp Asn Gln Ile
660 665 670
Ala Ile Phe Asn Arg Leu Lys Leu Val Pro Lys Lys Val Asp Leu Ser
675 680 685
Gln Gln Lys Glu Ile Pro Thr Thr Leu Val Asp Asp Phe Ile Leu Ser
690 695 700
Pro Val Val Lys Arg Ser Phe Ile Gln Ser Ile Lys Val Ile Asn Ala
705 710 715 720
Ile Ile Lys Lys Tyr Gly Leu Pro Asn Asp Ile Ile Ile Glu Leu Ala
725 730 735
Arg Glu Lys Asn Ser Lys Asp Ala Gln Lys Met Ile Asn Glu Met Gln
740 745 750
Lys Arg Asn Arg Gln Thr Asn Glu Arg Ile Glu Glu Ile Ile Arg Thr
755 760 765
Thr Gly Lys Glu Asn Ala Lys Tyr Leu Ile Glu Lys Ile Lys Leu His
770 775 780
Asp Met Gln Glu Gly Lys Cys Leu Tyr Ser Leu Glu Ala Ile Pro Leu
785 790 795 800
Glu Asp Leu Leu Asn Asn Pro Phe Asn Tyr Glu Val Asp His Ile Ile
805 810 815
Pro Arg Ser Val Ser Phe Asp Asn Ser Phe Asn Asn Lys Val Leu Val
820 825 830
Lys Gln Glu Glu Ala Ser Lys Lys Gly Asn Arg Thr Pro Phe Gln Tyr
835 840 845
Leu Ser Ser Ser Asp Ser Lys Ile Ser Tyr Glu Thr Phe Lys Lys His
850 855 860
Ile Leu Asn Leu Ala Lys Gly Lys Gly Arg Ile Ser Lys Thr Lys Lys
865 870 875 880
Glu Tyr Leu Leu Glu Glu Arg Asp Ile Asn Arg Phe Ser Val Gln Lys
885 890 895
Asp Phe Ile Asn Arg Asn Leu Val Asp Thr Arg Tyr Ala Thr Arg Gly
900 905 910
Leu Met Asn Leu Leu Arg Ser Tyr Phe Arg Val Asn Asn Leu Asp Val
915 920 925
Lys Val Lys Ser Ile Asn Gly Gly Phe Thr Ser Phe Leu Arg Arg Lys
930 935 940
Trp Lys Phe Lys Lys Glu Arg Asn Lys Gly Tyr Lys His His Ala Glu
945 950 955 960
Asp Ala Leu Ile Ile Ala Asn Ala Asp Phe Ile Phe Lys Glu Trp Lys
965 970 975
Lys Leu Asp Lys Ala Lys Lys Val Met Glu Asn Gln Met Phe Glu Glu
980 985 990
Lys Gln Ala Glu Ser Met Pro Glu Ile Glu Thr Glu Gln Glu Tyr Lys
995 1000 1005
Glu Ile Phe Ile Thr Pro His Gln Ile Lys His Ile Lys Asp Phe
1010 1015 1020
Lys Asp Tyr Lys Tyr Ser His Arg Val Asp Lys Lys Pro Asn Arg
1025 1030 1035
Lys Leu Ile Asn Asp Thr Leu Tyr Ser Thr Arg Lys Asp Asp Lys
1040 1045 1050
Gly Asn Thr Leu Ile Val Asn Asn Leu Asn Gly Leu Tyr Asp Lys
1055 1060 1065
Asp Asn Asp Lys Leu Lys Lys Leu Ile Asn Lys Ser Pro Glu Lys
1070 1075 1080
Leu Leu Met Tyr His His Asp Pro Gln Thr Tyr Gln Lys Leu Lys
1085 1090 1095
Leu Ile Met Glu Gln Tyr Gly Asp Glu Lys Asn Pro Leu Tyr Lys
1100 1105 1110
Tyr Tyr Glu Glu Thr Gly Asn Tyr Leu Thr Lys Tyr Ser Lys Lys
1115 1120 1125
Asp Asn Gly Pro Val Ile Lys Lys Ile Lys Tyr Tyr Gly Asn Lys
1130 1135 1140
Leu Asn Ala His Leu Asp Ile Thr Asp Asp Tyr Pro Asn Ser Arg
1145 1150 1155
Asn Lys Val Val Lys Leu Ser Leu Lys Pro Tyr Arg Phe Asp Val
1160 1165 1170
Tyr Leu Asp Asn Gly Val Tyr Lys Phe Val Thr Val Lys Asn Leu
1175 1180 1185
Asp Val Ile Lys Lys Glu Asn Tyr Tyr Glu Val Asn Ser Lys Cys
1190 1195 1200
Tyr Glu Glu Ala Lys Lys Leu Lys Lys Ile Ser Asn Gln Ala Glu
1205 1210 1215
Phe Ile Ala Ser Phe Tyr Lys Asn Asp Leu Ile Lys Ile Asn Gly
1220 1225 1230
Glu Leu Tyr Arg Val Ile Gly Val Asn Asn Asp Leu Leu Asn Arg
1235 1240 1245
Ile Glu Val Asn Met Ile Asp Ile Thr Tyr Arg Glu Tyr Leu Glu
1250 1255 1260
Asn Met Asn Asp Lys Arg Pro Pro His Ile Ile Lys Thr Ile Ala
1265 1270 1275
Ser Lys Thr Gln Ser Ile Lys Lys Tyr Ser Thr Asp Ile Leu Gly
1280 1285 1290
Asn Leu Tyr Glu Val Lys Ser Lys Lys His Pro Gln Ile Ile Lys
1295 1300 1305
Lys Gly Ser Gly Gly Ser Thr Asn Leu Ser Asp Ile Ile Glu Lys
1310 1315 1320
Glu Thr Gly Lys Gln Leu Val Ile Gln Glu Ser Ile Leu Met Leu
1325 1330 1335
Pro Glu Glu Val Glu Glu Val Ile Gly Asn Lys Pro Glu Ser Asp
1340 1345 1350
Ile Leu Val His Thr Ala Tyr Asp Glu Ser Thr Asp Glu Asn Val
1355 1360 1365
Met Leu Leu Thr Ser Asp Ala Pro Glu Tyr Lys Pro Trp Ala Leu
1370 1375 1380
Val Ile Gln Asp Ser Asn Gly Glu Asn Lys Ile Lys Met Leu Ser
1385 1390 1395
Gly Gly Ser Pro Pro Lys Lys Lys Arg Lys Val
1400 1405
<210> 21
<211> 1241
<212> PRT
<213> artificial sequence
<220>
<223> synthetic constructs produced by rational design
<400> 21
Met Pro Lys Lys Lys Arg Lys Val Gly Gly Gly Ser Pro Gly Met Ser
1 5 10 15
Ser Glu Thr Gly Pro Val Ala Val Asp Pro Thr Leu Arg Arg Arg Ile
20 25 30
Glu Pro His Glu Phe Glu Val Phe Phe Asp Pro Arg Glu Leu Arg Lys
35 40 45
Glu Thr Cys Leu Leu Tyr Glu Ile Asn Trp Gly Gly Arg His Ser Ile
50 55 60
Trp Arg His Thr Ser Gln Asn Thr Asn Lys His Val Glu Val Asn Phe
65 70 75 80
Ile Glu Lys Phe Thr Thr Glu Arg Tyr Phe Cys Pro Asn Thr Arg Cys
85 90 95
Ser Ile Thr Trp Phe Leu Ser Trp Ser Pro Cys Gly Glu Cys Ser Arg
100 105 110
Ala Ile Thr Glu Phe Leu Ser Arg Tyr Pro His Val Thr Leu Phe Ile
115 120 125
Tyr Ile Ala Arg Leu Tyr His His Ala Asp Pro Arg Asn Arg Gln Gly
130 135 140
Leu Arg Asp Leu Ile Ser Ser Gly Val Thr Ile Gln Ile Met Thr Glu
145 150 155 160
Gln Glu Ser Gly Tyr Cys Trp Arg Asn Phe Val Asn Tyr Ser Pro Ser
165 170 175
Asn Glu Ala His Trp Pro Arg Tyr Pro His Leu Trp Val Arg Leu Tyr
180 185 190
Val Leu Glu Leu Tyr Cys Ile Ile Leu Gly Leu Pro Pro Cys Leu Asn
195 200 205
Ile Leu Arg Arg Lys Gln Pro Gln Leu Thr Phe Phe Thr Ile Ala Leu
210 215 220
Gln Ser Cys His Tyr Gln Arg Leu Pro Pro His Ile Leu Trp Ala Thr
225 230 235 240
Gly Leu Lys Ser Gly Ser Glu Thr Pro Gly Thr Ser Glu Ser Ala Thr
245 250 255
Pro Glu Lys Arg Asn Tyr Ile Leu Gly Leu Ala Ile Gly Ile Thr Ser
260 265 270
Val Gly Tyr Gly Ile Ile Asp Tyr Glu Thr Arg Asp Val Ile Asp Ala
275 280 285
Gly Val Arg Leu Phe Lys Glu Ala Asn Val Glu Asn Asn Glu Gly Arg
290 295 300
Arg Ser Lys Arg Gly Ala Arg Arg Leu Lys Arg Arg Arg Arg His Arg
305 310 315 320
Ile Gln Arg Val Lys Lys Leu Leu Phe Asp Tyr Asn Leu Leu Thr Asp
325 330 335
His Ser Glu Leu Ser Gly Ile Asn Pro Tyr Glu Ala Arg Val Lys Gly
340 345 350
Leu Ser Gln Lys Leu Ser Glu Glu Glu Phe Ser Ala Ala Leu Leu His
355 360 365
Leu Ala Lys Arg Arg Gly Val His Asn Val Asn Glu Val Glu Glu Asp
370 375 380
Thr Gly Asn Glu Leu Ser Thr Lys Glu Gln Ile Ser Arg Asn Ser Lys
385 390 395 400
Ala Leu Glu Glu Lys Tyr Val Ala Glu Leu Gln Leu Glu Arg Leu Lys
405 410 415
Lys Asp Gly Glu Val Arg Gly Ser Ile Asn Arg Phe Lys Thr Ser Asp
420 425 430
Tyr Val Lys Glu Ala Lys Gln Leu Leu Lys Val Gln Lys Ala Tyr His
435 440 445
Gln Leu Asp Gln Ser Phe Ile Asp Thr Tyr Ile Asp Leu Leu Glu Thr
450 455 460
Arg Arg Thr Tyr Tyr Glu Gly Pro Gly Glu Gly Ser Pro Phe Gly Trp
465 470 475 480
Lys Asp Ile Lys Glu Trp Tyr Glu Met Leu Met Gly His Cys Thr Tyr
485 490 495
Phe Pro Glu Glu Leu Arg Ser Val Lys Tyr Ala Tyr Asn Ala Asp Leu
500 505 510
Tyr Asn Ala Leu Asn Asp Leu Asn Asn Leu Val Ile Thr Arg Asp Glu
515 520 525
Asn Glu Lys Leu Glu Tyr Tyr Glu Lys Phe Gln Ile Ile Glu Asn Val
530 535 540
Phe Lys Gln Lys Lys Lys Pro Thr Leu Lys Gln Ile Ala Lys Glu Ile
545 550 555 560
Leu Val Asn Glu Glu Asp Ile Lys Gly Tyr Arg Val Thr Ser Thr Gly
565 570 575
Lys Pro Glu Phe Thr Asn Leu Lys Val Tyr His Asp Ile Lys Asp Ile
580 585 590
Thr Ala Arg Lys Glu Ile Ile Glu Asn Ala Glu Leu Leu Asp Gln Ile
595 600 605
Ala Lys Ile Leu Thr Ile Tyr Gln Ser Ser Glu Asp Ile Gln Glu Glu
610 615 620
Leu Thr Asn Leu Asn Ser Glu Leu Thr Gln Glu Glu Ile Glu Gln Ile
625 630 635 640
Ser Asn Leu Lys Gly Tyr Thr Gly Thr His Asn Leu Ser Leu Lys Ala
645 650 655
Ile Asn Leu Ile Leu Asp Glu Leu Trp His Thr Asn Asp Asn Gln Ile
660 665 670
Ala Ile Phe Asn Arg Leu Lys Leu Val Pro Lys Lys Val Asp Leu Ser
675 680 685
Gln Gln Lys Glu Ile Pro Thr Thr Leu Val Asp Asp Phe Ile Leu Ser
690 695 700
Pro Val Val Lys Arg Ser Phe Ile Gln Ser Ile Lys Val Ile Asn Ala
705 710 715 720
Ile Ile Lys Lys Tyr Gly Leu Pro Asn Asp Ile Ile Ile Glu Leu Gly
725 730 735
Ser Lys Arg Tyr Ala Thr Arg Gly Leu Met Asn Leu Leu Arg Ser Tyr
740 745 750
Phe Arg Val Asn Asn Leu Asp Val Lys Val Lys Ser Ile Asn Gly Gly
755 760 765
Phe Thr Ser Phe Leu Arg Arg Lys Trp Lys Phe Lys Lys Glu Arg Asn
770 775 780
Lys Gly Tyr Lys His His Ala Glu Asp Ala Leu Ile Ile Ala Asn Ala
785 790 795 800
Asp Phe Ile Phe Lys Glu Trp Lys Lys Leu Asp Lys Ala Lys Lys Val
805 810 815
Met Glu Asn Gln Met Phe Glu Glu Lys Gln Ala Glu Ser Met Pro Glu
820 825 830
Ile Glu Thr Glu Gln Glu Tyr Lys Glu Ile Phe Ile Thr Pro His Gln
835 840 845
Ile Lys His Ile Lys Asp Phe Lys Asp Tyr Lys Tyr Ser His Arg Val
850 855 860
Asp Lys Lys Pro Asn Arg Lys Leu Ile Asn Asp Thr Leu Tyr Ser Thr
865 870 875 880
Arg Lys Asp Asp Lys Gly Asn Thr Leu Ile Val Asn Asn Leu Asn Gly
885 890 895
Leu Tyr Asp Lys Asp Asn Asp Lys Leu Lys Lys Leu Ile Asn Lys Ser
900 905 910
Pro Glu Lys Leu Leu Met Tyr His His Asp Pro Gln Thr Tyr Gln Lys
915 920 925
Leu Lys Leu Ile Met Glu Gln Tyr Gly Asp Glu Lys Asn Pro Leu Tyr
930 935 940
Lys Tyr Tyr Glu Glu Thr Gly Asn Tyr Leu Thr Lys Tyr Ser Lys Lys
945 950 955 960
Asp Asn Gly Pro Val Ile Lys Lys Ile Lys Tyr Tyr Gly Asn Lys Leu
965 970 975
Asn Ala His Leu Asp Ile Thr Asp Asp Tyr Pro Asn Ser Arg Asn Lys
980 985 990
Val Val Lys Leu Ser Leu Lys Pro Tyr Arg Phe Asp Val Tyr Leu Asp
995 1000 1005
Asn Gly Val Tyr Lys Phe Val Thr Val Lys Asn Leu Asp Val Ile
1010 1015 1020
Lys Lys Glu Asn Tyr Tyr Glu Val Asn Ser Lys Cys Tyr Glu Glu
1025 1030 1035
Ala Lys Lys Leu Lys Lys Ile Ser Asn Gln Ala Glu Phe Ile Ala
1040 1045 1050
Ser Phe Tyr Lys Asn Asp Leu Ile Lys Ile Asn Gly Glu Leu Tyr
1055 1060 1065
Arg Val Ile Gly Val Asn Asn Asp Leu Leu Asn Arg Ile Glu Val
1070 1075 1080
Asn Met Ile Asp Ile Thr Tyr Arg Glu Tyr Leu Glu Asn Met Asn
1085 1090 1095
Asp Lys Arg Pro Pro His Ile Ile Lys Thr Ile Ala Ser Lys Thr
1100 1105 1110
Gln Ser Ile Lys Lys Tyr Ser Thr Asp Ile Leu Gly Asn Leu Tyr
1115 1120 1125
Glu Val Lys Ser Lys Lys His Pro Gln Ile Ile Lys Lys Gly Ser
1130 1135 1140
Gly Gly Ser Thr Asn Leu Ser Asp Ile Ile Glu Lys Glu Thr Gly
1145 1150 1155
Lys Gln Leu Val Ile Gln Glu Ser Ile Leu Met Leu Pro Glu Glu
1160 1165 1170
Val Glu Glu Val Ile Gly Asn Lys Pro Glu Ser Asp Ile Leu Val
1175 1180 1185
His Thr Ala Tyr Asp Glu Ser Thr Asp Glu Asn Val Met Leu Leu
1190 1195 1200
Thr Ser Asp Ala Pro Glu Tyr Lys Pro Trp Ala Leu Val Ile Gln
1205 1210 1215
Asp Ser Asn Gly Glu Asn Lys Ile Lys Met Leu Ser Gly Gly Ser
1220 1225 1230
Pro Pro Lys Lys Lys Arg Lys Val
1235 1240
<210> 22
<211> 771
<212> PRT
<213> artificial sequence
<220>
<223> synthetic constructs produced by rational design
<400> 22
Met Pro Lys Lys Lys Arg Lys Val Gly Gly Gly Ser Pro Gly Lys Arg
1 5 10 15
Asn Tyr Ile Leu Gly Leu Ala Ile Gly Ile Thr Ser Val Gly Tyr Gly
20 25 30
Ile Ile Asp Tyr Glu Thr Arg Asp Val Ile Asp Ala Gly Val Arg Leu
35 40 45
Phe Lys Glu Ala Asn Val Glu Asn Asn Glu Gly Arg Arg Ser Lys Arg
50 55 60
Gly Ala Arg Arg Leu Lys Arg Arg Arg Arg His Arg Ile Gln Arg Val
65 70 75 80
Lys Lys Leu Leu Phe Asp Tyr Asn Leu Leu Thr Asp His Ser Glu Leu
85 90 95
Ser Gly Ile Asn Pro Tyr Glu Ala Arg Val Lys Gly Leu Ser Gln Lys
100 105 110
Leu Ser Glu Glu Glu Phe Ser Ala Ala Leu Leu His Leu Ala Lys Arg
115 120 125
Arg Gly Val His Asn Val Asn Glu Val Glu Glu Asp Thr Gly Asn Glu
130 135 140
Leu Ser Thr Lys Glu Gln Ile Ser Arg Asn Ser Lys Ala Leu Glu Glu
145 150 155 160
Lys Tyr Val Ala Glu Leu Gln Leu Glu Arg Leu Lys Lys Asp Gly Glu
165 170 175
Val Arg Gly Ser Ile Asn Arg Phe Lys Thr Ser Asp Tyr Val Lys Glu
180 185 190
Ala Lys Gln Leu Leu Lys Val Gln Lys Ala Tyr His Gln Leu Asp Gln
195 200 205
Ser Phe Ile Asp Thr Tyr Ile Asp Leu Leu Glu Thr Arg Arg Thr Tyr
210 215 220
Tyr Glu Gly Pro Gly Glu Gly Ser Pro Phe Gly Trp Lys Asp Ile Lys
225 230 235 240
Glu Trp Tyr Glu Met Leu Met Gly His Cys Thr Tyr Phe Pro Glu Glu
245 250 255
Leu Arg Ser Val Lys Tyr Ala Tyr Asn Ala Asp Leu Tyr Asn Ala Leu
260 265 270
Asn Asp Leu Asn Asn Leu Val Ile Thr Arg Asp Glu Asn Glu Lys Leu
275 280 285
Glu Tyr Tyr Glu Lys Phe Gln Ile Ile Glu Asn Val Phe Lys Gln Lys
290 295 300
Lys Lys Pro Thr Leu Lys Gln Ile Ala Lys Glu Ile Leu Val Asn Glu
305 310 315 320
Glu Asp Ile Lys Gly Tyr Arg Val Thr Ser Thr Gly Lys Pro Glu Phe
325 330 335
Thr Asn Leu Lys Val Tyr His Asp Ile Lys Asp Ile Thr Ala Arg Lys
340 345 350
Glu Ile Ile Glu Asn Ala Glu Leu Leu Asp Gln Ile Ala Lys Ile Leu
355 360 365
Thr Ile Tyr Gln Ser Ser Glu Asp Ile Gln Glu Glu Leu Thr Asn Leu
370 375 380
Asn Ser Glu Leu Thr Gln Glu Glu Ile Glu Gln Ile Ser Asn Leu Lys
385 390 395 400
Gly Tyr Thr Gly Thr His Asn Leu Ser Leu Lys Ala Ile Asn Leu Ile
405 410 415
Leu Asp Glu Leu Trp His Thr Asn Asp Asn Gln Ile Ala Ile Phe Asn
420 425 430
Arg Leu Lys Leu Val Pro Lys Lys Val Asp Leu Ser Gln Gln Lys Glu
435 440 445
Ile Pro Thr Thr Leu Val Asp Asp Phe Ile Leu Ser Pro Val Val Lys
450 455 460
Arg Ser Phe Ile Gln Ser Ile Lys Val Ile Asn Ala Ile Ile Lys Lys
465 470 475 480
Tyr Gly Leu Pro Asn Asp Ile Ile Ile Glu Leu Ala Arg Glu Lys Gly
485 490 495
Gly Gly Ser Gly Gly Gly Ser Met Ser Ser Glu Thr Gly Pro Val Ala
500 505 510
Val Asp Pro Thr Leu Arg Arg Arg Ile Glu Pro His Glu Phe Glu Val
515 520 525
Phe Phe Asp Pro Arg Glu Leu Arg Lys Glu Thr Cys Leu Leu Tyr Glu
530 535 540
Ile Asn Trp Gly Gly Arg His Ser Ile Trp Arg His Thr Ser Gln Asn
545 550 555 560
Thr Asn Lys His Val Glu Val Asn Phe Ile Glu Lys Phe Thr Thr Glu
565 570 575
Arg Tyr Phe Cys Pro Asn Thr Arg Cys Ser Ile Thr Trp Phe Leu Ser
580 585 590
Trp Ser Pro Cys Gly Glu Cys Ser Arg Ala Ile Thr Glu Phe Leu Ser
595 600 605
Arg Tyr Pro His Val Thr Leu Phe Ile Tyr Ile Ala Arg Leu Tyr His
610 615 620
His Ala Asp Pro Arg Asn Arg Gln Gly Leu Arg Asp Leu Ile Ser Ser
625 630 635 640
Gly Val Thr Ile Gln Ile Met Thr Glu Gln Glu Ser Gly Tyr Cys Trp
645 650 655
Arg Asn Phe Val Asn Tyr Ser Pro Ser Asn Glu Ala His Trp Pro Arg
660 665 670
Tyr Pro His Leu Trp Val Arg Leu Tyr Val Leu Glu Leu Tyr Cys Ile
675 680 685
Ile Leu Gly Leu Pro Pro Cys Leu Asn Ile Leu Arg Arg Lys Gln Pro
690 695 700
Gln Leu Thr Phe Phe Thr Ile Ala Leu Gln Ser Cys His Tyr Gln Arg
705 710 715 720
Leu Pro Pro His Ile Leu Trp Ala Thr Gly Leu Lys Ser Gly Ser Glu
725 730 735
Thr Pro Gly Thr Ser Glu Ser Ala Thr Pro Glu Thr Ser Gly Gly Gly
740 745 750
Ser Lys Arg Pro Ala Ala Thr Lys Lys Ala Gly Gln Ala Lys Lys Lys
755 760 765
Lys Ser Arg
770
<210> 23
<211> 518
<212> PRT
<213> artificial sequence
<220>
<223> synthetic constructs produced by rational design
<400> 23
Met Pro Lys Lys Lys Arg Lys Val Gly Gly Gly Ser Pro Gly Thr Arg
1 5 10 15
Tyr Ala Thr Arg Gly Leu Met Asn Leu Leu Arg Ser Tyr Phe Arg Val
20 25 30
Asn Asn Leu Asp Val Lys Val Lys Ser Ile Asn Gly Gly Phe Thr Ser
35 40 45
Phe Leu Arg Arg Lys Trp Lys Phe Lys Lys Glu Arg Asn Lys Gly Tyr
50 55 60
Lys His His Ala Glu Asp Ala Leu Ile Ile Ala Asn Ala Asp Phe Ile
65 70 75 80
Phe Lys Glu Trp Lys Lys Leu Asp Lys Ala Lys Lys Val Met Glu Asn
85 90 95
Gln Met Phe Glu Glu Lys Gln Ala Glu Ser Met Pro Glu Ile Glu Thr
100 105 110
Glu Gln Glu Tyr Lys Glu Ile Phe Ile Thr Pro His Gln Ile Lys His
115 120 125
Ile Lys Asp Phe Lys Asp Tyr Lys Tyr Ser His Arg Val Asp Lys Lys
130 135 140
Pro Asn Arg Lys Leu Ile Asn Asp Thr Leu Tyr Ser Thr Arg Lys Asp
145 150 155 160
Asp Lys Gly Asn Thr Leu Ile Val Asn Asn Leu Asn Gly Leu Tyr Asp
165 170 175
Lys Asp Asn Asp Lys Leu Lys Lys Leu Ile Asn Lys Ser Pro Glu Lys
180 185 190
Leu Leu Met Tyr His His Asp Pro Gln Thr Tyr Gln Lys Leu Lys Leu
195 200 205
Ile Met Glu Gln Tyr Gly Asp Glu Lys Asn Pro Leu Tyr Lys Tyr Tyr
210 215 220
Glu Glu Thr Gly Asn Tyr Leu Thr Lys Tyr Ser Lys Lys Asp Asn Gly
225 230 235 240
Pro Val Ile Lys Lys Ile Lys Tyr Tyr Gly Asn Lys Leu Asn Ala His
245 250 255
Leu Asp Ile Thr Asp Asp Tyr Pro Asn Ser Arg Asn Lys Val Val Lys
260 265 270
Leu Ser Leu Lys Pro Tyr Arg Phe Asp Val Tyr Leu Asp Asn Gly Val
275 280 285
Tyr Lys Phe Val Thr Val Lys Asn Leu Asp Val Ile Lys Lys Glu Asn
290 295 300
Tyr Tyr Glu Val Asn Ser Lys Cys Tyr Glu Glu Ala Lys Lys Leu Lys
305 310 315 320
Lys Ile Ser Asn Gln Ala Glu Phe Ile Ala Ser Phe Tyr Lys Asn Asp
325 330 335
Leu Ile Lys Ile Asn Gly Glu Leu Tyr Arg Val Ile Gly Val Asn Asn
340 345 350
Asp Leu Leu Asn Arg Ile Glu Val Asn Met Ile Asp Ile Thr Tyr Arg
355 360 365
Glu Tyr Leu Glu Asn Met Asn Asp Lys Arg Pro Pro His Ile Ile Lys
370 375 380
Thr Ile Ala Ser Lys Thr Gln Ser Ile Lys Lys Tyr Ser Thr Asp Ile
385 390 395 400
Leu Gly Asn Leu Tyr Glu Val Lys Ser Lys Lys His Pro Gln Ile Ile
405 410 415
Lys Lys Gly Ser Gly Gly Ser Thr Asn Leu Ser Asp Ile Ile Glu Lys
420 425 430
Glu Thr Gly Lys Gln Leu Val Ile Gln Glu Ser Ile Leu Met Leu Pro
435 440 445
Glu Glu Val Glu Glu Val Ile Gly Asn Lys Pro Glu Ser Asp Ile Leu
450 455 460
Val His Thr Ala Tyr Asp Glu Ser Thr Asp Glu Asn Val Met Leu Leu
465 470 475 480
Thr Ser Asp Ala Pro Glu Tyr Lys Pro Trp Ala Leu Val Ile Gln Asp
485 490 495
Ser Asn Gly Glu Asn Lys Ile Lys Met Leu Ser Gly Gly Ser Pro Pro
500 505 510
Lys Lys Lys Arg Lys Val
515
<210> 24
<211> 391
<212> PRT
<213> artificial sequence
<220>
<223> synthetic constructs produced by rational design
<400> 24
Gly Ser Gly Arg Ala Asp Ala Leu Asp Asp Phe Asp Leu Asp Met Leu
1 5 10 15
Gly Ser Asp Ala Leu Asp Asp Phe Asp Leu Asp Met Leu Gly Ser Asp
20 25 30
Ala Leu Asp Asp Phe Asp Leu Asp Met Leu Gly Ser Asp Ala Leu Asp
35 40 45
Asp Phe Asp Leu Asp Met Leu Ile Asn Ser Ala Leu Leu Gln Leu Gln
50 55 60
Phe Asp Asp Glu Asp Leu Gly Ala Leu Leu Gly Asn Ser Thr Asp Pro
65 70 75 80
Ala Val Phe Thr Asp Leu Ala Ser Val Asp Asn Ser Glu Phe Gln Gln
85 90 95
Leu Leu Asn Gln Gly Ile Pro Val Ala Pro His Thr Thr Glu Pro Met
100 105 110
Leu Met Glu Tyr Pro Glu Ala Ile Thr Arg Leu Val Thr Gly Ala Gln
115 120 125
Arg Pro Pro Asp Pro Ala Pro Ala Pro Leu Gly Ala Pro Gly Leu Pro
130 135 140
Asn Gly Leu Leu Ser Gly Asp Glu Asp Phe Ser Ser Ile Ala Asp Met
145 150 155 160
Asp Phe Ser Ala Leu Leu Ser Gln Ile Ser Ser Gly Ser Gly Ser Gly
165 170 175
Ser Arg Asp Ser Arg Glu Gly Met Phe Leu Pro Lys Pro Glu Ala Gly
180 185 190
Ser Ala Ile Ser Asp Val Phe Glu Gly Arg Glu Val Cys Gln Pro Lys
195 200 205
Arg Ile Arg Pro Phe His Pro Pro Gly Ser Pro Trp Ala Asn Arg Pro
210 215 220
Leu Pro Ala Ser Leu Ala Pro Thr Pro Thr Gly Pro Val His Glu Pro
225 230 235 240
Val Gly Ser Leu Thr Pro Ala Pro Val Pro Gln Pro Leu Asp Pro Ala
245 250 255
Pro Ala Val Thr Pro Glu Ala Ser His Leu Leu Glu Asp Pro Asp Glu
260 265 270
Glu Thr Ser Gln Ala Val Lys Ala Leu Arg Glu Met Ala Asp Thr Val
275 280 285
Ile Pro Gln Lys Glu Glu Ala Ala Ile Cys Gly Gln Met Asp Leu Ser
290 295 300
His Pro Pro Pro Arg Gly His Leu Asp Glu Leu Thr Thr Thr Leu Glu
305 310 315 320
Ser Met Thr Glu Asp Leu Asn Leu Asp Ser Pro Leu Thr Pro Glu Leu
325 330 335
Asn Glu Ile Leu Asp Thr Phe Leu Asn Asp Glu Cys Leu Leu His Ala
340 345 350
Met His Ile Ser Thr Gly Leu Ser Ile Phe Asp Thr Ser Leu Phe Thr
355 360 365
Ser Gly Gly Gly Ser Lys Arg Pro Ala Ala Thr Lys Lys Ala Gly Gln
370 375 380
Ala Lys Lys Lys Lys Ser Arg
385 390
<210> 25
<211> 382
<212> PRT
<213> artificial sequence
<220>
<223> synthetic constructs produced by rational design
<400> 25
Gly Ser Gly Arg Ala Asp Ala Leu Asp Asp Phe Asp Leu Asp Met Leu
1 5 10 15
Gly Ser Asp Ala Leu Asp Asp Phe Asp Leu Asp Met Leu Gly Ser Asp
20 25 30
Ala Leu Asp Asp Phe Asp Leu Asp Met Leu Gly Ser Asp Ala Leu Asp
35 40 45
Asp Phe Asp Leu Asp Met Leu Ile Asn Ser Arg Leu Val Thr Gly Ala
50 55 60
Gln Arg Pro Pro Asp Pro Ala Pro Ala Pro Leu Gly Ala Pro Gly Leu
65 70 75 80
Pro Asn Gly Leu Leu Ser Gly Asp Glu Asp Phe Ser Ser Ile Ala Asp
85 90 95
Met Asp Phe Ser Ala Leu Leu Ser Gln Ile Ser Ser Gly Gly Gly Ser
100 105 110
Arg Leu Val Thr Gly Ala Gln Arg Pro Pro Asp Pro Ala Pro Ala Pro
115 120 125
Leu Gly Ala Pro Gly Leu Pro Asn Gly Leu Leu Ser Gly Asp Glu Asp
130 135 140
Phe Ser Ser Ile Ala Asp Met Asp Phe Ser Ala Leu Leu Ser Gln Ile
145 150 155 160
Ser Ser Gly Ser Gly Ser Gly Ser Arg Asp Ser Arg Glu Gly Met Phe
165 170 175
Leu Pro Lys Pro Glu Ala Gly Ser Ala Ile Ser Asp Val Phe Glu Gly
180 185 190
Arg Glu Val Cys Gln Pro Lys Arg Ile Arg Pro Phe His Pro Pro Gly
195 200 205
Ser Pro Trp Ala Asn Arg Pro Leu Pro Ala Ser Leu Ala Pro Thr Pro
210 215 220
Thr Gly Pro Val His Glu Pro Val Gly Ser Leu Thr Pro Ala Pro Val
225 230 235 240
Pro Gln Pro Leu Asp Pro Ala Pro Ala Val Thr Pro Glu Ala Ser His
245 250 255
Leu Leu Glu Asp Pro Asp Glu Glu Thr Ser Gln Ala Val Lys Ala Leu
260 265 270
Arg Glu Met Ala Asp Thr Val Ile Pro Gln Lys Glu Glu Ala Ala Ile
275 280 285
Cys Gly Gln Met Asp Leu Ser His Pro Pro Pro Arg Gly His Leu Asp
290 295 300
Glu Leu Thr Thr Thr Leu Glu Ser Met Thr Glu Asp Leu Asn Leu Asp
305 310 315 320
Ser Pro Leu Thr Pro Glu Leu Asn Glu Ile Leu Asp Thr Phe Leu Asn
325 330 335
Asp Glu Cys Leu Leu His Ala Met His Ile Ser Thr Gly Leu Ser Ile
340 345 350
Phe Asp Thr Ser Leu Phe Thr Ser Gly Gly Gly Ser Lys Arg Pro Ala
355 360 365
Ala Thr Lys Lys Ala Gly Gln Ala Lys Lys Lys Lys Ser Arg
370 375 380
<210> 26
<211> 663
<212> PRT
<213> artificial sequence
<220>
<223> synthetic constructs produced by rational design
<400> 26
Met Pro Lys Lys Lys Arg Lys Val Gly Gly Gly Ser Pro Gly Gly Ala
1 5 10 15
Met Val Asp Thr Leu Ser Gly Leu Ser Ser Glu Gln Gly Gln Ser Gly
20 25 30
Asp Met Thr Ile Glu Glu Asp Ser Ala Thr His Ile Lys Phe Ser Lys
35 40 45
Arg Asp Glu Asp Gly Lys Glu Leu Ala Gly Ala Thr Met Glu Leu Arg
50 55 60
Asp Ser Ser Gly Lys Thr Ile Ser Thr Trp Ile Ser Asp Gly Gln Val
65 70 75 80
Lys Asp Phe Tyr Leu Tyr Pro Gly Lys Tyr Thr Phe Val Glu Thr Ala
85 90 95
Ala Pro Asp Gly Tyr Glu Val Ala Thr Ala Ile Thr Phe Thr Val Asn
100 105 110
Glu Gln Gly Gln Val Thr Val Asn Gly Lys Ala Thr Lys Gly Asp Ala
115 120 125
His Ile Thr Gly Thr Ser Arg Ala Asp Pro Lys Lys Lys Arg Lys Val
130 135 140
Glu Ala Ser Gly Ser Gly Arg Ala Asp Ala Leu Asp Asp Phe Asp Leu
145 150 155 160
Asp Met Leu Gly Ser Asp Ala Leu Asp Asp Phe Asp Leu Asp Met Leu
165 170 175
Gly Ser Asp Ala Leu Asp Asp Phe Asp Leu Asp Met Leu Gly Ser Asp
180 185 190
Ala Leu Asp Asp Phe Asp Leu Asp Met Leu Ile Asn Ser Arg Ser Gln
195 200 205
Tyr Leu Pro Asp Thr Asp Asp Arg His Arg Ile Glu Glu Lys Arg Lys
210 215 220
Arg Thr Tyr Glu Thr Phe Lys Ser Ile Met Lys Lys Ser Pro Phe Ser
225 230 235 240
Gly Pro Thr Asp Pro Arg Pro Pro Pro Arg Arg Ile Ala Val Pro Ser
245 250 255
Arg Ser Ser Ala Ser Val Pro Lys Pro Ala Pro Gln Pro Tyr Pro Phe
260 265 270
Thr Ser Ser Leu Ser Thr Ile Asn Tyr Asp Glu Phe Pro Thr Met Val
275 280 285
Phe Pro Ser Gly Gln Ile Ser Gln Ala Ser Ala Leu Ala Pro Ala Pro
290 295 300
Pro Gln Val Leu Pro Gln Ala Pro Ala Pro Ala Pro Ala Pro Ala Met
305 310 315 320
Val Ser Ala Leu Ala Gln Ala Pro Ala Pro Val Pro Val Leu Ala Pro
325 330 335
Gly Pro Pro Gln Ala Val Ala Pro Pro Ala Pro Lys Pro Thr Gln Ala
340 345 350
Gly Glu Gly Thr Leu Ser Glu Ala Leu Leu Gln Leu Gln Phe Asp Asp
355 360 365
Glu Asp Leu Gly Ala Leu Leu Gly Asn Ser Thr Asp Pro Ala Val Phe
370 375 380
Thr Asp Leu Ala Ser Val Asp Asn Ser Glu Phe Gln Gln Leu Leu Asn
385 390 395 400
Gln Gly Ile Pro Val Ala Pro His Thr Thr Glu Pro Met Leu Met Glu
405 410 415
Tyr Pro Glu Ala Ile Thr Arg Leu Val Thr Gly Ala Gln Arg Pro Pro
420 425 430
Asp Pro Ala Pro Ala Pro Leu Gly Ala Pro Gly Leu Pro Asn Gly Leu
435 440 445
Leu Ser Gly Asp Glu Asp Phe Ser Ser Ile Ala Asp Met Asp Phe Ser
450 455 460
Ala Leu Leu Gly Ser Gly Ser Gly Ser Arg Asp Ser Arg Glu Gly Met
465 470 475 480
Phe Leu Pro Lys Pro Glu Ala Gly Ser Ala Ile Ser Asp Val Phe Glu
485 490 495
Gly Arg Glu Val Cys Gln Pro Lys Arg Ile Arg Pro Phe His Pro Pro
500 505 510
Gly Ser Pro Trp Ala Asn Arg Pro Leu Pro Ala Ser Leu Ala Pro Thr
515 520 525
Pro Thr Gly Pro Val His Glu Pro Val Gly Ser Leu Thr Pro Ala Pro
530 535 540
Val Pro Gln Pro Leu Asp Pro Ala Pro Ala Val Thr Pro Glu Ala Ser
545 550 555 560
His Leu Leu Glu Asp Pro Asp Glu Glu Thr Ser Gln Ala Val Lys Ala
565 570 575
Leu Arg Glu Met Ala Asp Thr Val Ile Pro Gln Lys Glu Glu Ala Ala
580 585 590
Ile Cys Gly Gln Met Asp Leu Ser His Pro Pro Pro Arg Gly His Leu
595 600 605
Asp Glu Leu Thr Thr Thr Leu Glu Ser Met Thr Glu Asp Leu Asn Leu
610 615 620
Asp Ser Pro Leu Thr Pro Glu Leu Asn Glu Ile Leu Asp Thr Phe Leu
625 630 635 640
Asn Asp Glu Cys Leu Leu His Ala Met His Ile Ser Thr Gly Leu Ser
645 650 655
Ile Phe Asp Thr Ser Leu Phe
660
<210> 27
<211> 88
<212> PRT
<213> artificial sequence
<220>
<223> synthetic constructs produced by rational design
<400> 27
Gly Gly Gly Ser Gly Gly Ala His Ile Val Met Val Asp Ala Tyr Lys
1 5 10 15
Pro Thr Lys Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Ala His Ile
20 25 30
Val Met Val Asp Ala Tyr Lys Pro Thr Lys Gly Gly Ser Gly Gly Gly
35 40 45
Gly Ser Gly Gly Ala His Ile Val Met Val Asp Ala Tyr Lys Pro Thr
50 55 60
Lys Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Ala His Ile Val Met
65 70 75 80
Val Asp Ala Tyr Lys Pro Thr Lys
85
<210> 28
<211> 643
<212> PRT
<213> artificial sequence
<220>
<223> synthetic constructs produced by rational design
<400> 28
Met Pro Lys Lys Lys Arg Lys Val Gly Gly Gly Ser Pro Gly Asn His
1 5 10 15
Val Ile Glu Thr Glu Gln Asn Leu Pro Asn Glu Asp Gly Gln Ser Gly
20 25 30
Asn Ile Ile Glu Gln Glu Asp Ser Lys Thr Leu Val Lys Phe Ser Lys
35 40 45
Arg Asp Ile Lys Gly Asn Glu Leu Ala Gly Ala Thr Ile Glu Leu Arg
50 55 60
Asp Leu Ser Gly Lys Ser Ile Gln Ser Trp Val Ser Asp Gly Lys Ala
65 70 75 80
Lys Asp Phe Tyr Leu Leu Pro Gly Ser Tyr Glu Phe Val Glu Thr Ala
85 90 95
Ala Pro Glu Gly Tyr Gln Ile Ala Thr Lys Ile Met Phe Thr Ile Ser
100 105 110
Thr Asp Gly Arg Ile Thr Val Asp Gly Gln Leu Val Thr Gly Thr Gly
115 120 125
Ser Gly Arg Ala Asp Ala Leu Asp Asp Phe Asp Leu Asp Met Leu Gly
130 135 140
Ser Asp Ala Leu Asp Asp Phe Asp Leu Asp Met Leu Gly Ser Asp Ala
145 150 155 160
Leu Asp Asp Phe Asp Leu Asp Met Leu Gly Ser Asp Ala Leu Asp Asp
165 170 175
Phe Asp Leu Asp Met Leu Ile Asn Ser Arg Ser Gln Tyr Leu Pro Asp
180 185 190
Thr Asp Asp Arg His Arg Ile Glu Glu Lys Arg Lys Arg Thr Tyr Glu
195 200 205
Thr Phe Lys Ser Ile Met Lys Lys Ser Pro Phe Ser Gly Pro Thr Asp
210 215 220
Pro Arg Pro Pro Pro Arg Arg Ile Ala Val Pro Ser Arg Ser Ser Ala
225 230 235 240
Ser Val Pro Lys Pro Ala Pro Gln Pro Tyr Pro Phe Thr Ser Ser Leu
245 250 255
Ser Thr Ile Asn Tyr Asp Glu Phe Pro Thr Met Val Phe Pro Ser Gly
260 265 270
Gln Ile Ser Gln Ala Ser Ala Leu Ala Pro Ala Pro Pro Gln Val Leu
275 280 285
Pro Gln Ala Pro Ala Pro Ala Pro Ala Pro Ala Met Val Ser Ala Leu
290 295 300
Ala Gln Ala Pro Ala Pro Val Pro Val Leu Ala Pro Gly Pro Pro Gln
305 310 315 320
Ala Val Ala Pro Pro Ala Pro Lys Pro Thr Gln Ala Gly Glu Gly Thr
325 330 335
Leu Ser Glu Ala Leu Leu Gln Leu Gln Phe Asp Asp Glu Asp Leu Gly
340 345 350
Ala Leu Leu Gly Asn Ser Thr Asp Pro Ala Val Phe Thr Asp Leu Ala
355 360 365
Ser Val Asp Asn Ser Glu Phe Gln Gln Leu Leu Asn Gln Gly Ile Pro
370 375 380
Val Ala Pro His Thr Thr Glu Pro Met Leu Met Glu Tyr Pro Glu Ala
385 390 395 400
Ile Thr Arg Leu Val Thr Gly Ala Gln Arg Pro Pro Asp Pro Ala Pro
405 410 415
Ala Pro Leu Gly Ala Pro Gly Leu Pro Asn Gly Leu Leu Ser Gly Asp
420 425 430
Glu Asp Phe Ser Ser Ile Ala Asp Met Asp Phe Ser Ala Leu Leu Gly
435 440 445
Ser Gly Ser Gly Ser Arg Asp Ser Arg Glu Gly Met Phe Leu Pro Lys
450 455 460
Pro Glu Ala Gly Ser Ala Ile Ser Asp Val Phe Glu Gly Arg Glu Val
465 470 475 480
Cys Gln Pro Lys Arg Ile Arg Pro Phe His Pro Pro Gly Ser Pro Trp
485 490 495
Ala Asn Arg Pro Leu Pro Ala Ser Leu Ala Pro Thr Pro Thr Gly Pro
500 505 510
Val His Glu Pro Val Gly Ser Leu Thr Pro Ala Pro Val Pro Gln Pro
515 520 525
Leu Asp Pro Ala Pro Ala Val Thr Pro Glu Ala Ser His Leu Leu Glu
530 535 540
Asp Pro Asp Glu Glu Thr Ser Gln Ala Val Lys Ala Leu Arg Glu Met
545 550 555 560
Ala Asp Thr Val Ile Pro Gln Lys Glu Glu Ala Ala Ile Cys Gly Gln
565 570 575
Met Asp Leu Ser His Pro Pro Pro Arg Gly His Leu Asp Glu Leu Thr
580 585 590
Thr Thr Leu Glu Ser Met Thr Glu Asp Leu Asn Leu Asp Ser Pro Leu
595 600 605
Thr Pro Glu Leu Asn Glu Ile Leu Asp Thr Phe Leu Asn Asp Glu Cys
610 615 620
Leu Leu His Ala Met His Ile Ser Thr Gly Leu Ser Ile Phe Asp Thr
625 630 635 640
Ser Leu Phe
<210> 29
<211> 78
<212> PRT
<213> artificial sequence
<220>
<223> synthetic constructs produced by rational design
<400> 29
Gly Gly Gly Ser Gly Gly Ala His Ile Val Met Val Asp Asn Tyr Lys
1 5 10 15
Pro Ile Val Gly Gly Ser Gly Gly Ala His Ile Val Met Val Asp Asn
20 25 30
Tyr Lys Pro Ile Val Gly Gly Ser Gly Gly Ala His Ile Val Met Val
35 40 45
Asp Asn Tyr Lys Pro Ile Val Gly Gly Ser Gly Gly Gly Gly Ser Gly
50 55 60
Gly Ala His Ile Val Met Val Asp Asn Tyr Lys Pro Ile Val
65 70 75
<210> 30
<211> 285
<212> PRT
<213> artificial sequence
<220>
<223> synthetic constructs produced by rational design
<400> 30
Gly Ser Gly Arg Ala Asp Ala Leu Asp Asp Phe Asp Leu Asp Met Leu
1 5 10 15
Gly Ser Asp Ala Leu Asp Asp Phe Asp Leu Asp Met Leu Gly Ser Asp
20 25 30
Ala Leu Asp Asp Phe Asp Leu Asp Met Leu Gly Ser Asp Ala Leu Asp
35 40 45
Asp Phe Asp Leu Asp Met Leu Ile Asn Ser Arg Leu Val Thr Gly Ala
50 55 60
Gln Arg Pro Pro Asp Pro Ala Pro Ala Pro Leu Gly Ala Pro Gly Leu
65 70 75 80
Pro Asn Gly Leu Leu Ser Gly Asp Glu Asp Phe Ser Ser Ile Ala Asp
85 90 95
Met Asp Phe Ser Ala Leu Leu Ser Gln Ile Ser Ser Gly Gly Gly Ser
100 105 110
Arg Leu Val Thr Gly Ala Gln Arg Pro Pro Asp Pro Ala Pro Ala Pro
115 120 125
Leu Gly Ala Pro Gly Leu Pro Asn Gly Leu Leu Ser Gly Asp Glu Asp
130 135 140
Phe Ser Ser Ile Ala Asp Met Asp Phe Ser Ala Leu Leu Ser Gln Ile
145 150 155 160
Ser Ser Gly Ser Gly Ser Gln Pro Leu Asp Pro Ala Pro Ala Val Thr
165 170 175
Pro Glu Ala Ser His Leu Leu Glu Asp Pro Asp Glu Glu Thr Ser Gln
180 185 190
Ala Val Lys Ala Leu Arg Glu Met Ala Asp Thr Val Ile Pro Gln Lys
195 200 205
Glu Glu Ala Ala Ile Cys Gly Gln Met Asp Leu Ser His Pro Pro Pro
210 215 220
Arg Gly His Leu Asp Glu Leu Thr Thr Thr Leu Glu Ser Met Thr Glu
225 230 235 240
Asp Leu Asn Leu Asp Ser Pro Leu Thr Pro Glu Leu Asn Glu Ile Leu
245 250 255
Asp Thr Phe Leu Asn Asp Glu Cys Leu Leu His Ala Met His Ile Ser
260 265 270
Thr Gly Leu Ser Ile Phe Asp Thr Ser Leu Phe Thr Ser
275 280 285
<210> 31
<211> 20
<212> DNA
<213> artificial sequence
<220>
<223> synthetic constructs produced by rational design
<400> 31
gctgtgtttg cgtctctccc 20
<210> 32
<211> 20
<212> DNA
<213> artificial sequence
<220>
<223> synthetic constructs produced by rational design
<400> 32
ggggtggtga caagtgtgat 20
<210> 33
<211> 1433
<212> PRT
<213> artificial sequence
<220>
<223> synthetic constructs produced by rational design
<400> 33
Met Pro Lys Lys Lys Arg Lys Val Gly Gly Gly Ser Pro Gly Gly Gly
1 5 10 15
Gly Ser Lys Arg Asn Tyr Ile Leu Gly Leu Asp Ile Gly Ile Thr Ser
20 25 30
Val Gly Tyr Gly Ile Ile Asp Tyr Glu Thr Arg Asp Val Ile Asp Ala
35 40 45
Gly Val Arg Leu Phe Lys Glu Ala Asn Val Glu Asn Asn Glu Gly Arg
50 55 60
Arg Ser Lys Arg Gly Ala Arg Arg Leu Lys Arg Arg Arg Arg His Arg
65 70 75 80
Ile Gln Arg Val Lys Lys Leu Leu Phe Asp Tyr Asn Leu Leu Thr Asp
85 90 95
His Ser Glu Leu Ser Gly Ile Asn Pro Tyr Glu Ala Arg Val Lys Gly
100 105 110
Leu Ser Gln Lys Leu Ser Glu Glu Glu Phe Ser Ala Ala Leu Leu His
115 120 125
Leu Ala Lys Arg Arg Gly Val His Asn Val Asn Glu Val Glu Glu Asp
130 135 140
Thr Gly Asn Glu Leu Ser Thr Lys Glu Gln Ile Ser Arg Asn Ser Lys
145 150 155 160
Ala Leu Glu Glu Lys Tyr Val Ala Glu Leu Gln Leu Glu Arg Leu Lys
165 170 175
Lys Asp Gly Glu Val Arg Gly Ser Ile Asn Arg Phe Lys Thr Ser Asp
180 185 190
Tyr Val Lys Glu Ala Lys Gln Leu Leu Lys Val Gln Lys Ala Tyr His
195 200 205
Gln Leu Asp Gln Ser Phe Ile Asp Thr Tyr Ile Asp Leu Leu Glu Thr
210 215 220
Arg Arg Thr Tyr Tyr Glu Gly Pro Gly Glu Gly Ser Pro Phe Gly Trp
225 230 235 240
Lys Asp Ile Lys Glu Trp Tyr Glu Met Leu Met Gly His Cys Thr Tyr
245 250 255
Phe Pro Glu Glu Leu Arg Ser Val Lys Tyr Ala Tyr Asn Ala Asp Leu
260 265 270
Tyr Asn Ala Leu Asn Asp Leu Asn Asn Leu Val Ile Thr Arg Asp Glu
275 280 285
Asn Glu Lys Leu Glu Tyr Tyr Glu Lys Phe Gln Ile Ile Glu Asn Val
290 295 300
Phe Lys Gln Lys Lys Lys Pro Thr Leu Lys Gln Ile Ala Lys Glu Ile
305 310 315 320
Leu Val Asn Glu Glu Asp Ile Lys Gly Tyr Arg Val Thr Ser Thr Gly
325 330 335
Lys Pro Glu Phe Thr Asn Leu Lys Val Tyr His Asp Ile Lys Asp Ile
340 345 350
Thr Ala Arg Lys Glu Ile Ile Glu Asn Ala Glu Leu Leu Asp Gln Ile
355 360 365
Ala Lys Ile Leu Thr Ile Tyr Gln Ser Ser Glu Asp Ile Gln Glu Glu
370 375 380
Leu Thr Asn Leu Asn Ser Glu Leu Thr Gln Glu Glu Ile Glu Gln Ile
385 390 395 400
Ser Asn Leu Lys Gly Tyr Thr Gly Thr His Asn Leu Ser Leu Lys Ala
405 410 415
Ile Asn Leu Ile Leu Asp Glu Leu Trp His Thr Asn Asp Asn Gln Ile
420 425 430
Ala Ile Phe Asn Arg Leu Lys Leu Val Pro Lys Lys Val Asp Leu Ser
435 440 445
Gln Gln Lys Glu Ile Pro Thr Thr Leu Val Asp Asp Phe Ile Leu Ser
450 455 460
Pro Val Val Lys Arg Ser Phe Ile Gln Ser Ile Lys Val Ile Asn Ala
465 470 475 480
Ile Ile Lys Lys Tyr Gly Leu Pro Asn Asp Ile Ile Ile Glu Leu Gly
485 490 495
Ser Lys Arg Tyr Ala Thr Arg Gly Leu Met Asn Leu Leu Arg Ser Tyr
500 505 510
Phe Arg Val Asn Asn Leu Asp Val Lys Val Lys Ser Ile Asn Gly Gly
515 520 525
Phe Thr Ser Phe Leu Arg Arg Lys Trp Lys Phe Lys Lys Glu Arg Asn
530 535 540
Lys Gly Tyr Lys His His Ala Glu Asp Ala Leu Ile Ile Ala Asn Ala
545 550 555 560
Asp Phe Ile Phe Lys Glu Trp Lys Lys Leu Asp Lys Ala Lys Lys Val
565 570 575
Met Glu Asn Gln Met Phe Glu Glu Lys Gln Ala Glu Ser Met Pro Glu
580 585 590
Ile Glu Thr Glu Gln Glu Tyr Lys Glu Ile Phe Ile Thr Pro His Gln
595 600 605
Ile Lys His Ile Lys Asp Phe Lys Asp Tyr Lys Tyr Ser His Arg Val
610 615 620
Asp Lys Lys Pro Asn Arg Lys Leu Ile Asn Asp Thr Leu Tyr Ser Thr
625 630 635 640
Arg Lys Asp Asp Lys Gly Asn Thr Leu Ile Val Asn Asn Leu Asn Gly
645 650 655
Leu Tyr Asp Lys Asp Asn Asp Lys Leu Lys Lys Leu Ile Asn Lys Ser
660 665 670
Pro Glu Lys Leu Leu Met Tyr His His Asp Pro Gln Thr Tyr Gln Lys
675 680 685
Leu Lys Leu Ile Met Glu Gln Tyr Gly Asp Glu Lys Asn Pro Leu Tyr
690 695 700
Lys Tyr Tyr Glu Glu Thr Gly Asn Tyr Leu Thr Lys Tyr Ser Lys Lys
705 710 715 720
Asp Asn Gly Pro Val Ile Lys Lys Ile Lys Tyr Tyr Gly Asn Lys Leu
725 730 735
Asn Ala His Leu Asp Ile Thr Asp Asp Tyr Pro Asn Ser Arg Asn Lys
740 745 750
Val Val Lys Leu Ser Leu Lys Pro Tyr Arg Phe Asp Val Tyr Leu Asp
755 760 765
Asn Gly Val Tyr Lys Phe Val Thr Val Lys Asn Leu Asp Val Ile Lys
770 775 780
Lys Glu Asn Tyr Tyr Glu Val Asn Ser Lys Cys Tyr Glu Glu Ala Lys
785 790 795 800
Lys Leu Lys Lys Ile Ser Asn Gln Ala Glu Phe Ile Ala Ser Phe Tyr
805 810 815
Lys Asn Asp Leu Ile Lys Ile Asn Gly Glu Leu Tyr Arg Val Ile Gly
820 825 830
Val Asn Asn Asp Leu Leu Asn Arg Ile Glu Val Asn Met Ile Asp Ile
835 840 845
Thr Tyr Arg Glu Tyr Leu Glu Asn Met Asn Asp Lys Arg Pro Pro His
850 855 860
Ile Ile Lys Thr Ile Ala Ser Lys Thr Gln Ser Ile Lys Lys Tyr Ser
865 870 875 880
Thr Asp Ile Leu Gly Asn Leu Tyr Glu Val Lys Ser Lys Lys His Pro
885 890 895
Gln Ile Ile Lys Lys Gly Thr Ser Arg Ala Asp Pro Lys Lys Lys Arg
900 905 910
Lys Val Glu Ala Ser Gly Ser Gly Arg Ala Asp Ala Leu Asp Asp Phe
915 920 925
Asp Leu Asp Met Leu Gly Ser Asp Ala Leu Asp Asp Phe Asp Leu Asp
930 935 940
Met Leu Gly Ser Asp Ala Leu Asp Asp Phe Asp Leu Asp Met Leu Gly
945 950 955 960
Ser Asp Ala Leu Asp Asp Phe Asp Leu Asp Met Leu Ile Asn Ser Arg
965 970 975
Ser Gln Tyr Leu Pro Asp Thr Asp Asp Arg His Arg Ile Glu Glu Lys
980 985 990
Arg Lys Arg Thr Tyr Glu Thr Phe Lys Ser Ile Met Lys Lys Ser Pro
995 1000 1005
Phe Ser Gly Pro Thr Asp Pro Arg Pro Pro Pro Arg Arg Ile Ala
1010 1015 1020
Val Pro Ser Arg Ser Ser Ala Ser Val Pro Lys Pro Ala Pro Gln
1025 1030 1035
Pro Tyr Pro Phe Thr Ser Ser Leu Ser Thr Ile Asn Tyr Asp Glu
1040 1045 1050
Phe Pro Thr Met Val Phe Pro Ser Gly Gln Ile Ser Gln Ala Ser
1055 1060 1065
Ala Leu Ala Pro Ala Pro Pro Gln Val Leu Pro Gln Ala Pro Ala
1070 1075 1080
Pro Ala Pro Ala Pro Ala Met Val Ser Ala Leu Ala Gln Ala Pro
1085 1090 1095
Ala Pro Val Pro Val Leu Ala Pro Gly Pro Pro Gln Ala Val Ala
1100 1105 1110
Pro Pro Ala Pro Lys Pro Thr Gln Ala Gly Glu Gly Thr Leu Ser
1115 1120 1125
Glu Ala Leu Leu Gln Leu Gln Phe Asp Asp Glu Asp Leu Gly Ala
1130 1135 1140
Leu Leu Gly Asn Ser Thr Asp Pro Ala Val Phe Thr Asp Leu Ala
1145 1150 1155
Ser Val Asp Asn Ser Glu Phe Gln Gln Leu Leu Asn Gln Gly Ile
1160 1165 1170
Pro Val Ala Pro His Thr Thr Glu Pro Met Leu Met Glu Tyr Pro
1175 1180 1185
Glu Ala Ile Thr Arg Leu Val Thr Gly Ala Gln Arg Pro Pro Asp
1190 1195 1200
Pro Ala Pro Ala Pro Leu Gly Ala Pro Gly Leu Pro Asn Gly Leu
1205 1210 1215
Leu Ser Gly Asp Glu Asp Phe Ser Ser Ile Ala Asp Met Asp Phe
1220 1225 1230
Ser Ala Leu Leu Gly Ser Gly Ser Gly Ser Arg Asp Ser Arg Glu
1235 1240 1245
Gly Met Phe Leu Pro Lys Pro Glu Ala Gly Ser Ala Ile Ser Asp
1250 1255 1260
Val Phe Glu Gly Arg Glu Val Cys Gln Pro Lys Arg Ile Arg Pro
1265 1270 1275
Phe His Pro Pro Gly Ser Pro Trp Ala Asn Arg Pro Leu Pro Ala
1280 1285 1290
Ser Leu Ala Pro Thr Pro Thr Gly Pro Val His Glu Pro Val Gly
1295 1300 1305
Ser Leu Thr Pro Ala Pro Val Pro Gln Pro Leu Asp Pro Ala Pro
1310 1315 1320
Ala Val Thr Pro Glu Ala Ser His Leu Leu Glu Asp Pro Asp Glu
1325 1330 1335
Glu Thr Ser Gln Ala Val Lys Ala Leu Arg Glu Met Ala Asp Thr
1340 1345 1350
Val Ile Pro Gln Lys Glu Glu Ala Ala Ile Cys Gly Gln Met Asp
1355 1360 1365
Leu Ser His Pro Pro Pro Arg Gly His Leu Asp Glu Leu Thr Thr
1370 1375 1380
Thr Leu Glu Ser Met Thr Glu Asp Leu Asn Leu Asp Ser Pro Leu
1385 1390 1395
Thr Pro Glu Leu Asn Glu Ile Leu Asp Thr Phe Leu Asn Asp Glu
1400 1405 1410
Cys Leu Leu His Ala Met His Ile Ser Thr Gly Leu Ser Ile Phe
1415 1420 1425
Asp Thr Ser Leu Phe
1430
<210> 34
<211> 1414
<212> PRT
<213> artificial sequence
<220>
<223> synthetic constructs produced by rational design
<400> 34
Met Pro Lys Lys Lys Arg Lys Val Gly Gly Gly Ser Pro Gly Gly Gly
1 5 10 15
Gly Ser Lys Arg Asn Tyr Ile Leu Gly Leu Asp Ile Gly Ile Thr Ser
20 25 30
Val Gly Tyr Gly Ile Ile Asp Tyr Glu Thr Arg Asp Val Ile Asp Ala
35 40 45
Gly Val Arg Leu Phe Lys Glu Ala Asn Val Glu Asn Asn Glu Gly Arg
50 55 60
Arg Ser Lys Arg Gly Ala Arg Arg Leu Lys Arg Arg Arg Arg His Arg
65 70 75 80
Ile Gln Arg Val Lys Lys Leu Leu Phe Asp Tyr Asn Leu Leu Thr Asp
85 90 95
His Ser Glu Leu Ser Gly Ile Asn Pro Tyr Glu Ala Arg Val Lys Gly
100 105 110
Leu Ser Gln Lys Leu Ser Glu Glu Glu Phe Ser Ala Ala Leu Leu His
115 120 125
Leu Ala Lys Arg Arg Gly Val His Asn Val Asn Glu Val Glu Glu Asp
130 135 140
Thr Gly Asn Glu Leu Ser Thr Lys Glu Gln Ile Ser Arg Asn Ser Lys
145 150 155 160
Ala Leu Glu Glu Lys Tyr Val Ala Glu Leu Gln Leu Glu Arg Leu Lys
165 170 175
Lys Asp Gly Glu Val Arg Gly Ser Ile Asn Arg Phe Lys Thr Ser Asp
180 185 190
Tyr Val Lys Glu Ala Lys Gln Leu Leu Lys Val Gln Lys Ala Tyr His
195 200 205
Gln Leu Asp Gln Ser Phe Ile Asp Thr Tyr Ile Asp Leu Leu Glu Thr
210 215 220
Arg Arg Thr Tyr Tyr Glu Gly Pro Gly Glu Gly Ser Pro Phe Gly Trp
225 230 235 240
Lys Asp Ile Lys Glu Trp Tyr Glu Met Leu Met Gly His Cys Thr Lys
245 250 255
Lys Val Asp Leu Ser Gln Gln Lys Glu Ile Pro Thr Thr Leu Val Asp
260 265 270
Asp Phe Ile Leu Ser Pro Val Val Lys Arg Ser Phe Ile Gln Ser Ile
275 280 285
Lys Val Ile Asn Ala Ile Ile Lys Lys Tyr Gly Leu Pro Asn Asp Ile
290 295 300
Ile Ile Glu Leu Ala Arg Glu Lys Asn Ser Lys Asp Ala Gln Lys Met
305 310 315 320
Ile Asn Glu Met Gln Lys Arg Asn Arg Gln Thr Asn Glu Arg Ile Glu
325 330 335
Glu Ile Ile Arg Thr Thr Gly Lys Glu Asn Ala Lys Tyr Leu Ile Glu
340 345 350
Lys Ile Lys Leu His Asp Met Gln Glu Gly Lys Cys Leu Tyr Ser Leu
355 360 365
Glu Ala Ile Pro Leu Glu Asp Leu Leu Asn Asn Pro Phe Asn Tyr Glu
370 375 380
Val Asp His Ile Ile Pro Arg Ser Val Ser Phe Asp Asn Ser Phe Asn
385 390 395 400
Asn Lys Val Leu Val Lys Gln Glu Glu Asn Ser Lys Lys Gly Asn Arg
405 410 415
Thr Pro Phe Gln Tyr Leu Ser Ser Ser Asp Ser Lys Ile Ser Tyr Glu
420 425 430
Thr Phe Lys Lys His Ile Leu Asn Leu Ala Lys Gly Lys Gly Arg Ile
435 440 445
Ser Lys Thr Lys Lys Glu Tyr Leu Leu Glu Glu Arg Asp Ile Asn Arg
450 455 460
Phe Ser Val Gln Lys Asp Phe Ile Asn Arg Asn Leu Val Asp Thr Arg
465 470 475 480
Tyr Ala Thr Arg Gly Leu Met Asn Leu Leu Arg Ser Tyr Phe Arg Val
485 490 495
Asn Asn Leu Asp Val Lys Val Lys Ser Ile Asn Gly Gly Phe Thr Ser
500 505 510
Phe Leu Arg Arg Lys Trp Lys Phe Lys Lys Glu Arg Asn Lys Gly Tyr
515 520 525
Lys His His Ala Glu Asp Ala Leu Ile Ile Ala Asn Ala Asp Phe Ile
530 535 540
Phe Lys Glu Trp Lys Lys Leu Asp Lys Ala Lys Lys Val Met Glu Asn
545 550 555 560
Gln Met Phe Glu Glu Lys Gln Ala Glu Ser Met Pro Glu Ile Glu Thr
565 570 575
Glu Gln Glu Tyr Lys Glu Ile Phe Ile Thr Pro His Gln Ile Lys His
580 585 590
Ile Lys Asp Phe Lys Asp Tyr Lys Tyr Ser His Arg Val Asp Lys Lys
595 600 605
Pro Asn Arg Lys Leu Ile Asn Asp Thr Leu Tyr Ser Thr Arg Lys Asp
610 615 620
Asp Lys Gly Asn Thr Leu Ile Val Asn Asn Leu Asn Gly Leu Tyr Asp
625 630 635 640
Lys Asp Asn Asp Lys Leu Lys Lys Leu Ile Asn Lys Ser Pro Glu Lys
645 650 655
Leu Leu Met Tyr His His Asp Pro Gln Thr Tyr Gln Lys Leu Lys Leu
660 665 670
Ile Met Glu Gln Tyr Gly Asp Glu Lys Asn Pro Leu Tyr Lys Tyr Tyr
675 680 685
Glu Glu Thr Gly Asn Tyr Leu Thr Lys Tyr Ser Lys Lys Asp Asn Gly
690 695 700
Pro Val Ile Lys Lys Ile Lys Tyr Tyr Gly Asn Lys Leu Asn Ala His
705 710 715 720
Leu Asp Ile Thr Asp Asp Tyr Pro Asn Ser Arg Asn Lys Val Val Lys
725 730 735
Leu Ser Leu Lys Pro Tyr Arg Phe Asp Val Tyr Leu Asp Asn Gly Val
740 745 750
Tyr Lys Phe Val Thr Val Lys Asn Leu Asp Val Ile Lys Lys Glu Asn
755 760 765
Tyr Tyr Glu Val Asn Ser Lys Cys Tyr Glu Glu Ala Lys Lys Leu Lys
770 775 780
Lys Ile Ser Asn Gln Ala Glu Phe Ile Ala Ser Phe Tyr Lys Asn Asp
785 790 795 800
Leu Ile Lys Ile Asn Gly Glu Leu Tyr Arg Val Ile Gly Val Asn Asn
805 810 815
Asp Leu Leu Asn Arg Ile Glu Val Asn Met Ile Asp Ile Thr Tyr Arg
820 825 830
Glu Tyr Leu Glu Asn Met Asn Asp Lys Arg Pro Pro His Ile Ile Lys
835 840 845
Thr Ile Ala Ser Lys Thr Gln Ser Ile Lys Lys Tyr Ser Thr Asp Ile
850 855 860
Leu Gly Asn Leu Tyr Glu Val Lys Ser Lys Lys His Pro Gln Ile Ile
865 870 875 880
Lys Lys Gly Thr Ser Arg Ala Asp Pro Lys Lys Lys Arg Lys Val Glu
885 890 895
Ala Ser Gly Ser Gly Arg Ala Asp Ala Leu Asp Asp Phe Asp Leu Asp
900 905 910
Met Leu Gly Ser Asp Ala Leu Asp Asp Phe Asp Leu Asp Met Leu Gly
915 920 925
Ser Asp Ala Leu Asp Asp Phe Asp Leu Asp Met Leu Gly Ser Asp Ala
930 935 940
Leu Asp Asp Phe Asp Leu Asp Met Leu Ile Asn Ser Arg Ser Gln Tyr
945 950 955 960
Leu Pro Asp Thr Asp Asp Arg His Arg Ile Glu Glu Lys Arg Lys Arg
965 970 975
Thr Tyr Glu Thr Phe Lys Ser Ile Met Lys Lys Ser Pro Phe Ser Gly
980 985 990
Pro Thr Asp Pro Arg Pro Pro Pro Arg Arg Ile Ala Val Pro Ser Arg
995 1000 1005
Ser Ser Ala Ser Val Pro Lys Pro Ala Pro Gln Pro Tyr Pro Phe
1010 1015 1020
Thr Ser Ser Leu Ser Thr Ile Asn Tyr Asp Glu Phe Pro Thr Met
1025 1030 1035
Val Phe Pro Ser Gly Gln Ile Ser Gln Ala Ser Ala Leu Ala Pro
1040 1045 1050
Ala Pro Pro Gln Val Leu Pro Gln Ala Pro Ala Pro Ala Pro Ala
1055 1060 1065
Pro Ala Met Val Ser Ala Leu Ala Gln Ala Pro Ala Pro Val Pro
1070 1075 1080
Val Leu Ala Pro Gly Pro Pro Gln Ala Val Ala Pro Pro Ala Pro
1085 1090 1095
Lys Pro Thr Gln Ala Gly Glu Gly Thr Leu Ser Glu Ala Leu Leu
1100 1105 1110
Gln Leu Gln Phe Asp Asp Glu Asp Leu Gly Ala Leu Leu Gly Asn
1115 1120 1125
Ser Thr Asp Pro Ala Val Phe Thr Asp Leu Ala Ser Val Asp Asn
1130 1135 1140
Ser Glu Phe Gln Gln Leu Leu Asn Gln Gly Ile Pro Val Ala Pro
1145 1150 1155
His Thr Thr Glu Pro Met Leu Met Glu Tyr Pro Glu Ala Ile Thr
1160 1165 1170
Arg Leu Val Thr Gly Ala Gln Arg Pro Pro Asp Pro Ala Pro Ala
1175 1180 1185
Pro Leu Gly Ala Pro Gly Leu Pro Asn Gly Leu Leu Ser Gly Asp
1190 1195 1200
Glu Asp Phe Ser Ser Ile Ala Asp Met Asp Phe Ser Ala Leu Leu
1205 1210 1215
Gly Ser Gly Ser Gly Ser Arg Asp Ser Arg Glu Gly Met Phe Leu
1220 1225 1230
Pro Lys Pro Glu Ala Gly Ser Ala Ile Ser Asp Val Phe Glu Gly
1235 1240 1245
Arg Glu Val Cys Gln Pro Lys Arg Ile Arg Pro Phe His Pro Pro
1250 1255 1260
Gly Ser Pro Trp Ala Asn Arg Pro Leu Pro Ala Ser Leu Ala Pro
1265 1270 1275
Thr Pro Thr Gly Pro Val His Glu Pro Val Gly Ser Leu Thr Pro
1280 1285 1290
Ala Pro Val Pro Gln Pro Leu Asp Pro Ala Pro Ala Val Thr Pro
1295 1300 1305
Glu Ala Ser His Leu Leu Glu Asp Pro Asp Glu Glu Thr Ser Gln
1310 1315 1320
Ala Val Lys Ala Leu Arg Glu Met Ala Asp Thr Val Ile Pro Gln
1325 1330 1335
Lys Glu Glu Ala Ala Ile Cys Gly Gln Met Asp Leu Ser His Pro
1340 1345 1350
Pro Pro Arg Gly His Leu Asp Glu Leu Thr Thr Thr Leu Glu Ser
1355 1360 1365
Met Thr Glu Asp Leu Asn Leu Asp Ser Pro Leu Thr Pro Glu Leu
1370 1375 1380
Asn Glu Ile Leu Asp Thr Phe Leu Asn Asp Glu Cys Leu Leu His
1385 1390 1395
Ala Met His Ile Ser Thr Gly Leu Ser Ile Phe Asp Thr Ser Leu
1400 1405 1410
Phe

Claims (12)

1. A non-naturally occurring rationally designed reduced Cas9 system consisting of a reduced size staphylococcus aureus mini-SaCas9 that retains DNA binding activity by deleting a conserved functional domain and an effector domain, wherein the conserved functional domain of mini-SaCas9 is removed at amino acid positions 479-649 between RuvC2 and RuvC3 regions and connected by a GSK linker, the effector domain being a VPR domain or FOK i domain, wherein the reduced Cas9 system has one of the following sequences:
a mini-SaCas9:VPR sequence as shown in SEQ ID NO. 33;
the mini-SaCas9:FOK I sequence is shown as SEQ ID NO. 19.
2. The non-naturally occurring reduced form Cas9 system of claim 1, wherein the mini-SaCas9 is mini-dscas 9 and further comprises a plurality of gene regulatory elements to form a fokl dimer configuration upon formation of a complex.
3. The non-naturally occurring reduced form Cas9 system according to claim 2, wherein the fokl dimer configuration creates a 4-40 base pair spacing between dimerized mini-dscas 9.
4. The non-naturally occurring rationally designed reduced Cas9 according to claim 2, wherein the fokl dimer configuration produces a 14 base pair spacing between dimerized mini-dscas 9.
5. The non-naturally occurring reduced form Cas9 system according to claim 2, wherein the fokl dimer configuration creates a 12-24 base pair spacing between dimerized mini-dscas 9.
6. The non-naturally occurring reduced form Cas9 system according to any one of claims 2 to 5, wherein the fokl dimer configuration restores nuclease activity when in the dimer configuration.
7. The non-naturally occurring reduced form Cas9 system of claim 1, wherein the mini-SaCas9 is linked to merge effector domains, gRNA expression cassettes in a single AAV vector for transcriptional activation, DNA cleavage, and base editing.
8. The non-naturally occurring reduced form Cas9 system according to claim 7, wherein the gRNA expression cassette comprises an optimized guide RNA that is a putative RNA Pol III terminator sequence in the first stem loop that introduces a-U inversion and U-G conversion to disrupt the gRNA scaffold, wherein four repeat U-a regions in the putative RNA Pol III terminator sequence are optimized by a-U inversion by U-a at the third position and G-C substitution by U-a at the fourth position.
9. The non-naturally occurring reduced form Cas9 system according to claim 8, wherein the gRNA expression cassette further comprises a Glu tRNA coupled to the optimized guide RNA.
10. A CRISPR/Cas9 system comprising a guide RNA and the non-naturally occurring reduced Cas9 system of any one of claims 1 to 9.
11. The CRISPR/Cas9 system according to claim 10, wherein the guide RNA is an optimized guide RNA that is a putative RNA Pol III terminator sequence in the first stem loop that introduces a-U inversion and U-G conversion to disrupt a gRNA scaffold, wherein four repeat U-a regions in the putative RNA Pol III terminator sequence are the optimized by a-U inversion at the third position and G-C substitution at the fourth position.
12. The CRISPR/Cas9 system according to claim 11, wherein the guide RNA further comprises a Glu tRNA coupled to the optimized guide RNA.
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