| 质粒名称: | pyEvolvR (enCas9-PolI5M) |
|---|---|
| 出品公司: | Addgene |
| 目录编号: | 158073 |
| 存储实验室: | John Dueber |
|
载体骨架基本信息 |
|
| 载体名称: | custom |
| 载体出品公司: | -- |
| 空载体大小: | -- |
| 完整质粒大小: | -- |
| 载体修饰: | -- |
| 载体类型: | 细菌表达，酵母表达 |
| 筛选标记: | LEU2 |
| 载体抗性: | 卡那霉素50 μg/mL |
| 生长温度: | 37 ℃ |
| 宿主菌: | DH5a |
| 拷贝数: | 低拷贝 |
|
插入基因信息 |
|
| 插入基因名称: | enCas9-PolI5M |
| 物种来源: | -- |
| 插入大小: | -- |
| 基因突变: | -- |
| GenBank编号 | -- |
| 启动子: | -- |
| 标签/融合蛋白: | -- |
| 克隆方法: | -- |
| 5’克隆位点: | -- |
| 3’克隆位点: | -- |
| 5’测序引物 | -- |
| 3’测序引物 | -- |
|
文献引用方法 |
|
| 材料和方法部分: | pyEvolvR (enCas9-PolI5M) was a gift from John Dueber (Addgene plasmid # 158073 ; http://n2t.net/addgene:158073 ; RRID:Addgene_158073) |
| 参考文献部分: | Targeted Diversification in the S. cerevisiae Genome with CRISPR-Guided DNA Polymerase I. Tou CJ, Schaffer DV, Dueber JE. ACS Synth Biol. 2020 Jun 16. doi: 10.1021/acssynbio.0c00149. 10.1021/acssynbio.0c00149 PubMed 32485105 |
买家导航
载体基本信息
载体图谱质粒图谱和多克隆位点信息
质粒图谱.png)
载体简介
载体序列
LOCUS pyEvolvR (enCas9-PolI5M) 14085 bp ds-DNA circular SYN 12-MAY-2021
DEFINITION Expresses enCas9 fused to PolI5M in S. cerevisiae; contains a
BsmbI-flanked GFP cassette for sgRNA cloning..
ACCESSION .
VERSION .
KEYWORDS pyEvolvR (enCas9-PolI5M)
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 14085)
AUTHORS Tou CJ, Schaffer DV, Dueber JE
TITLE Targeted Diversification in the S. cerevisiae Genome with
CRISPR-Guided DNA Polymerase I.
JOURNAL ACS Synth Biol. 2020 Jun 16. doi: 10.1021/acssynbio.0c00149.
PUBMED 32485105
REFERENCE 2 (bases 1 to 14085)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58
http://www.snapgene.com
FEATURES Location/Qualifiers
source 1..14085
/organism="synthetic DNA construct"
/mol_type="other DNA"
RBS 675..686
/note="strong bacterial ribosome binding site (Elowitz and
Leibler, 2000)"
CDS 693..1406
/codon_start=1
/product="GFP variant that folds robustly even when fused
to poorly folded proteins (Pedelacq et al., 2006)"
/label=superfolder GFP
/translation="MRKGEELFTGVVPILVELDGDVNGHKFSVRGEGEGDATNGKLTLK
FICTTGKLPVPWPTLVTTLTYGVQCFARYPDHMKQHDFFKSAMPEGYVQERTISFKDDG
TYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNFNSHNVYITADKQKNGIKA
NFKIRHNVEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSVLSKDPNEKRDHMVLLE
FVTAAGITHGMDELYK"
terminator 1418..1489
/label=rrnB T1 terminator
/note="transcription terminator T1 from the E. coli rrnB
gene"
terminator 1505..1532
/label=T7Te terminator
/note="phage T7 early transcription terminator"
misc_RNA 1546..1621
/label=gRNA scaffold
/note="guide RNA scaffold for the Streptococcus pyogenes
CRISPR/Cas9 system"
promoter 1989..2396
/gene="S. cerevisiae LEU2"
/label=LEU2 promoter
CDS 2397..3491
/codon_start=1
/gene="S. cerevisiae LEU2"
/product="3-isopropylmalate dehydrogenase, required for
leucine biosynthesis"
/label=LEU2
/note="yeast auxotrophic marker"
/translation="MSAPKKIVVLPGDHVGQEITAEAIKVLKAISDVRSNVKFDFENHL
IGGAAIDATGVPLPDEALEASKKADAVLLGAVGGPKWGTGSVRPEQGLLKIRKELQLYA
NLRPCNFASDSLLDLSPIKPQFAKGTDFVVVRELVGGIYFGKRKEDDGDGVAWDSEQYT
VPEVQRITRMAAFMALQHEPPLPIWSLDKANVLASSRLWRKTVEETIKNEFPTLKVQHQ
LIDSAAMILVKNPTHLNGIIITSNMFGDIISDEASVIPGSLGLLPSASLASLPDKNTAF
GLYEPCHGSAPDLPKNKVNPIATILSAAMMLKLSLNLPEEGKAIEDAVKKVLDAGIRTG
DLGGSNSTTEVGDAVAEEVKKILA"
misc_feature 3973..4476
/label=CEN/ARS
/note="S. cerevisiae CEN6 centromere fused to an
autonomously replicating sequence"
CDS complement(4520..5335)
/codon_start=1
/gene="aph(3')-Ia"
/product="aminoglycoside phosphotransferase"
/label=KanR
/note="confers resistance to kanamycin in bacteria or G418
(Geneticin(R)) in eukaryotes"
/translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
primer_bind 5243..5262
/label=Kan-R
/note="Kanamycin resistance gene, reverse primer"
primer_bind 5327..5346
/label=pENTR-R
/note="pENTR vectors, reverse primer"
rep_origin 5560..6148
/direction=RIGHT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind 6049..6068
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
promoter 6583..6990
/gene="S. cerevisiae TEF1"
/label=TEF1 promoter
/note="promoter for EF-1-alpha"
CDS 7011..7031
/codon_start=1
/product="nuclear localization signal of SV40 (simian virus
40) large T antigen"
/label=SV40 NLS
/translation="PKKKRKV"
CDS 7059..8774
/codon_start=1
/product="N-terminal portion of Streptococcus pyogenes Cas9
(Zetsche et al., 2015)"
/label=Cas9(N)
/translation="DKKYSIGLAIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKKN
LIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEESF
LVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIKF
RGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRLE
NLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQI
GDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVRQ
QLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLLR
KQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARGN
SRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEYF
TVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIE"
CDS 11181..11201
/codon_start=1
/product="nuclear localization signal of SV40 (simian virus
40) large T antigen"
/label=SV40 NLS
/translation="PKKKRKV"
CDS 11208..11228
/codon_start=1
/product="nuclear localization signal of SV40 (simian virus
40) large T antigen"
/label=SV40 NLS
/translation="PKKKRKV"
RBS 12008..12016
/label=Shine-Dalgarno sequence
/note="full consensus sequence for ribosome-binding sites
upstream of start codons in E. coli; complementary to a
region in the 3' end of the 16S rRNA (Chen et al., 1994)"
ORIGIN
1 agcttttgat taagccttct agtccaaaaa acacgttttt ttgtcattta tttcattttc
61 ttagaatagt ttagtttatt cattttatag tcacgaatgt tttatgattc tatatagggt
121 tgcaaacaag catttttcat tttatgttaa aacaatttca ggtttacctt ttattctgct
181 tgtggtgacg cgtgtatccg cccgctcttt tggtcaccca tgtatgctgc caaaccagat
241 gtcaacacag ctacaacgat gtgcttcagt attacatttt ttgccttcaa cgccttgatt
301 gttctatttt tgctaataat aaatctattt catcggacta aaagtccatt agttgtaagc
361 ggatttagct cagttgggag agcgccagac tgaagaaaaa cttcggtcaa gtcatctgga
421 ggtcctgtgt tcgatccaca gaattcgcag atggccggca tggtcccagc ctcctcgctg
481 gcgccggctg ggcaacacct tcgggtggcg aatgggactt gagacggaaa gtgaaacgtg
541 atttcatgcg tcattttgaa cattttgtaa atcttattta ataatgtgtg cggcaattca
601 catttaattt atgaatgttt tcttaacatc gcggcaactc aagaaacggc aggttcggat
661 cttagctact agagaaagag gagaaatact agatgcgtaa aggcgaagag ctgttcactg
721 gtgtcgtccc tattctggtg gaactggatg gtgatgtcaa cggtcataag ttttccgtgc
781 gtggcgaggg tgaaggtgac gcaactaatg gtaaactgac gctgaagttc atctgtacta
841 ctggtaaact gccggttcct tggccgactc tggtaacgac gctgacttat ggtgttcagt
901 gctttgctcg ttatccggac catatgaagc agcatgactt cttcaagtcc gccatgccgg
961 aaggctatgt gcaggaacgc acgatttcct ttaaggatga cggcacgtac aaaacgcgtg
1021 cggaagtgaa atttgaaggc gataccctgg taaaccgcat tgagctgaaa ggcattgact
1081 ttaaagagga cggcaatatc ctgggccata agctggaata caattttaac agccacaatg
1141 tttacatcac cgccgataaa caaaaaaatg gcattaaagc gaattttaaa attcgccaca
1201 acgtggagga tggcagcgtg cagctggctg atcactacca gcaaaacact ccaatcggtg
1261 atggtcctgt tctgctgcca gacaatcact atctgagcac gcaaagcgtt ctgtctaaag
1321 atccgaacga gaaacgcgat catatggttc tgctggagtt cgtaaccgca gcgggcatca
1381 cgcatggtat ggatgaactg tacaaatgac caggcatcaa ataaaacgaa aggctcagtc
1441 gaaagactgg gcctttcgtt ttatctgttg tttgtcggtg aacgctctct actagagtca
1501 cactggctca ccttcgggtg ggcctttctg cgtttatacg tctcagtttt agagctagaa
1561 atagcaagtt aaaataaggc tagtccgtta tcaacttgaa aaagtggcac cgagtcggtg
1621 ctttttttat tttttgtcac tattgttatg taaaatgcca cctctgacag tatggaacgc
1681 aaacttctgt ctagtggata gctgagcagt tacagagatg ttacgaacca ctagtgcact
1741 gcagtacatc gaggagaact tctagtatat ctacatacct aatattattg ccttattaaa
1801 aatggaatcc caacaattac atcaaaatcc acattctctt caaaatcaat tgtcctgtac
1861 ttccttgttc atgtgtgttc aaaaacgtta tatttatagg ataattatac tctatttctc
1921 aacaagtaat tggttgtttg gccgagcggt ctaaggcgcc tgattcaaga aatatcttga
1981 ccgcagttaa ctgtgggaat actcaggtat cgtaagatgc aagagttcga atctcttagc
2041 aaccattatt tttttcctca acataacgag aacacacagg ggcgctatcg cacagaatca
2101 aattcgatga ctggaaattt tttgttaatt tcagaggtcg cctgacgcat ataccttttt
2161 caactgaaaa attgggagaa aaaggaaagg tgagagcgcc ggaaccggct tttcatatag
2221 aatagagaag cgttcatgac taaatgcttg catcacaata cttgaagttg acaatattat
2281 ttaaggacct attgtttttt ccaataggtg gttagcaatc gtcttacttt ctaacttttc
2341 ttacctttta catttcagca atatatatat atatatttca aggatatacc attctaatgt
2401 ctgcccctaa gaagatcgtc gttttgccag gtgaccacgt tggtcaagaa atcacagccg
2461 aagccattaa ggttcttaaa gctatttctg atgttcgttc caatgtcaag ttcgatttcg
2521 aaaatcattt aattggtggt gctgctatcg atgctacagg tgttccactt ccagatgagg
2581 cgctggaagc ctccaagaag gctgatgccg ttttgttagg tgctgtgggt ggtcctaaat
2641 ggggaaccgg tagtgttaga cctgaacaag gtttactaaa aatccgtaaa gaacttcaat
2701 tgtacgccaa cttaagacca tgtaactttg catccgactc tcttttagac ttatctccaa
2761 tcaagccaca atttgctaaa ggtactgact tcgttgttgt cagagaatta gtgggaggta
2821 tttactttgg taagagaaag gaagacgatg gtgatggtgt cgcttgggat agtgaacaat
2881 acaccgttcc agaagtgcaa agaatcacaa gaatggccgc tttcatggcc ctacaacatg
2941 agccaccatt gcctatttgg tccttggata aagctaatgt tttggcctct tcaagattat
3001 ggagaaaaac tgtggaggaa accatcaaga acgaatttcc tacattgaag gttcaacatc
3061 aattgattga ttctgccgcc atgatcctag ttaagaaccc aacccaccta aatggtatta
3121 taatcaccag caacatgttt ggtgatatca tctccgatga agcctccgtt atcccaggtt
3181 ccttgggttt gttgccatct gcgtccttgg cctctttgcc agacaagaac accgcatttg
3241 gtttgtacga accatgccac ggttctgctc cagatttgcc aaagaataag gtcaacccta
3301 tcgccactat cttgtctgct gcaatgatgt tgaaattgtc attgaacttg cctgaagaag
3361 gtaaggccat tgaagatgca gttaaaaagg ttttggatgc aggtatcaga actggtgatt
3421 taggtggttc caacagtacc accgaagtcg gtgatgctgt cgccgaagaa gttaagaaaa
3481 tccttgctta aaaagattct ctttttttat gatatttgta cataaacttt ataaatgaaa
3541 ttcataatag aaacgacacg aaattacaaa atggaatatg ttcatagggt agacgaaact
3601 atatacgcaa tctacataca tttatcaaga aggagaaaaa ggaggatgta aaggaataca
3661 ggtaagcaaa ttgatactaa tggctcaacg tgataaggaa aaagaattgc actttaacat
3721 taatattgac aaggaggagg gcaccacaca aaaagttagg tgtaacagaa aatcatgaaa
3781 ctatgattcc taatttatat attggaggat tttctctaaa aaaaaaaaaa tacaacaaat
3841 aaaaaacact caatgacctg accatttgat ggagtttaag tcaatacctt cttgaaccat
3901 ttcccataat ggtgaaagtt ccctcaagaa ttttactctg tcagaaacgg ccttaacgac
3961 gtagtcgaga gtatcacgtg ctataaaaat aattataatt taaatttttt aatataaata
4021 tataaattaa aaatagaaag taaaaaaaga aattaaagaa aaaatagttt ttgttttccg
4081 aagatgtaaa agactctagg gggatcgcca acaaatacta ccttttatct tgctcttcct
4141 gctctcaggt attaatgccg aattgtttca tcttgtctgt gtagaaaacc acacacgaaa
4201 atcctgtgat tttacatttt acttatcgtt aatcgaatgt atatctattt aatctgcttt
4261 tcttgtctaa taaatatata tgtaaagtac gctttttgtt gaaatttttt aaacctttgt
4321 ttattttttt ttcttcattc cgtaactctt ctaccttctt tatttacttt ctaaaatcca
4381 aatacaaaac ataaaaataa ataaacacag agtaaattcc caaattattc catcattaaa
4441 agatacgagg cgcgtgtaag ttacaggcaa gcgatccgtc ccgagcggcc gcgtgttaca
4501 accaattaac caattctgat tagaaaaact catcgagcat caaatgaaac tgcaatttat
4561 tcatatcagg attatcaata ccatattttt gaaaaagccg tttctgtaat gaaggagaaa
4621 actcaccgag gcagttccat aggatggcaa gatcctggta tcggtctgcg attccgactc
4681 gtccaacatc aatacaacct attaatttcc cctcgtcaaa aataaggtta tcaagtgaga
4741 aatcaccatg agtgacgact gaatccggtg agaatggcaa aagcttatgc atttctttcc
4801 agacttgttc aacaggccag ccattacgct cgtcatcaaa atcactcgca tcaaccaaac
4861 cgttattcat tcgtgattgc gcctgagcga ggcgaaatac gcgatcgctg ttaaaaggac
4921 aattacaaac aggaatcgaa tgcaaccggc gcaggaacac tgccagcgca tcaacaatat
4981 tttcacctga atcaggatat tcttctaata cctggaatgc tgttttcccg gggatcgcag
5041 tggtgagtaa ccatgcatca tcaggagtac ggataaaatg cttgatggtc ggaagaggca
5101 taaattccgt cagccagttt agtctgacca tctcatctgt aacatcattg gcaacgctac
5161 ctttgccatg tttcagaaac aactctggcg catcgggctt cccatacaat cgatagattg
5221 tcgcacctga ttgcccgaca ttatcgcgag cccatttata cccatataaa tcagcatcca
5281 tgttggaatt taatcgcggc ctggagcaag acgtttcccg ttgaatatgg ctcataacac
5341 cccttgtatt actgtttatg taagcagaca gttttattgt tcatgatgat atatttttat
5401 cttgtgcaat gtaacatcag agattttgag acacaacgtg gctttgttga ataaatcgaa
5461 cttttgctga gttgaaggat cagtcatgac caaaatccct taacgtgagt tttcgttcca
5521 ctgagcgtca gaccccgtag aaaagatcaa aggatcttct tgagatcctt tttttctgcg
5581 cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca gcggtggttt gtttgccgga
5641 tcaagagcta ccaactcttt ttccgaaggt aactggcttc agcagagcgc agataccaaa
5701 tactgttctt ctagtgtagc cgtagttagg ccaccacttc aagaactctg tagcaccgcc
5761 tacatacctc gctctgctaa tcctgttacc agtggctgct gccagtggcg ataagtcgtg
5821 tcttaccggg ttggactcaa gacgatagtt accggataag gcgcagcggt cgggctgaac
5881 ggggggttcg tgcacacagc ccagcttgga gcgaacgacc tacaccgaac tgagatacct
5941 acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc
6001 ggtaagcggc agggtcggaa caggagagcg cacgagggag cttccagggg gaaacgcctg
6061 gtatctttat agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg
6121 ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac gcggcctttt tacggttcct
6181 ggccttttgc tggccttttg ctcacatgtt ctttcctgcg ttatcccctg attctgtgga
6241 taaccgtgcg gccgcccctg aattcgcatc tagactgaac tggccgataa ttgcagacga
6301 acgccttgcc aacagggagt tcttcagaga catggaggct caaaacgaaa ttattgacag
6361 cctagacatc aatagtcata caacagaaag cgaccaccca actttggctg ataatagcgt
6421 ataaacaatg catactttgt acgttcaaaa tacaatgcag tagatatatt tatgcatatt
6481 acatataata catatcacat aggaagcaac aggcgcgttg gacttttaat tttcgaggac
6541 cgcgaatcct tacatcacac ccaatccccc acaagtgatc ccccacacac catagcttca
6601 aaatgtttct actccttttt tactcttcca gattttctcg gactccgcgc atcgccgtac
6661 cacttcaaaa cacccaagca cagcatacta aatttcccct ctttcttcct ctagggtgtc
6721 gttaattacc cgtactaaag gtttggaaaa gaaaaaagac accgcctcgt ttctttttct
6781 tcgtcgaaaa aggcaataaa aatttttatc acgtttcttt ttcttgaaaa tttttttttt
6841 tgattttttt ctctttcgat gacctcccat tgatatttaa gttaataaac ggtcatcaat
6901 ttctcaagtt tcagtttcat ttttcttgtt ctattacaac tttttttact tcttgctcat
6961 tagaaagaaa gcatagcaat ctaatctaag ttttaattac aaatatggcc ccaaagaaga
7021 agcggaaggt cggtatccac ggagtcccag gatccggaga caagaagtac agcatcggcc
7081 tggccatcgg caccaactct gtgggctggg ccgtgatcac cgacgagtac aaggtgccca
7141 gcaagaaatt caaggtgctg ggcaacaccg accggcacag catcaagaag aacctgatcg
7201 gcgccctgct gttcgacagc ggagaaacag ccgaggccac ccggctgaag agaaccgcca
7261 gaagaagata caccagacgg aagaaccgga tctgctatct gcaagagatc ttcagcaacg
7321 agatggccaa ggtggacgac agcttcttcc acagactgga agagtccttc ctggtggaag
7381 aggataagaa gcacgagcgg caccccatct tcggcaacat cgtggacgag gtggcctacc
7441 acgagaagta ccccaccatc taccacctga gaaagaaact ggtggacagc accgacaagg
7501 ccgacctgcg gctgatctat ctggccctgg cccacatgat caagttccgg ggccacttcc
7561 tgatcgaggg cgacctgaac cccgacaaca gcgacgtgga caagctgttc atccagctgg
7621 tgcagaccta caaccagctg ttcgaggaaa accccatcaa cgccagcggc gtggacgcca
7681 aggccatcct gtctgccaga ctgagcaaga gcagacggct ggaaaatctg atcgcccagc
7741 tgcccggcga gaagaagaat ggcctgttcg gcaacctgat tgccctgagc ctgggcctga
7801 cccccaactt caagagcaac ttcgacctgg ccgaggatgc caaactgcag ctgagcaagg
7861 acacctacga cgacgacctg gacaacctgc tggcccagat cggcgaccag tacgccgacc
7921 tgtttctggc cgccaagaac ctgtccgacg ccatcctgct gagcgacatc ctgagagtga
7981 acaccgagat caccaaggcc cccctgagcg cctctatgat caagagatac gacgagcacc
8041 accaggacct gaccctgctg aaagctctcg tgcggcagca gctgcctgag aagtacaaag
8101 agattttctt cgaccagagc aagaacggct acgccggcta catcgatggc ggagccagcc
8161 aggaagagtt ctacaagttc atcaagccca tcctggaaaa gatggacggc accgaggaac
8221 tgctcgtgaa gctgaacaga gaggacctgc tgcggaagca gcggaccttc gacaacggca
8281 gcatccccca ccagatccac ctgggagagc tgcacgccat tctgcggcgg caggaagatt
8341 tttacccatt cctgaaggac aaccgggaaa agatcgagaa gatcctgacc ttccgcatcc
8401 cctactacgt gggccctctg gccaggggaa acagcagatt cgcctggatg accagaaaga
8461 gcgaggaaac catcaccccc tggaacttcg aggaagtggt ggacaagggc gccagcgccc
8521 agagcttcat cgagcggatg accaacttcg ataagaacct gcccaacgag aaggtgctgc
8581 ccaagcacag cctgctgtac gagtacttca ccgtgtacaa cgagctgacc aaagtgaaat
8641 acgtgaccga gggaatgaga aagcccgcct tcctgagcgg cgagcagaaa aaagccatcg
8701 tggacctgct gttcaagacc aaccggaaag tgaccgtgaa gcagctgaaa gaggactact
8761 tcaagaaaat cgagtgcttc gactccgtgg aaatctccgg cgtggaagat cggttcaacg
8821 cctccctggg cacataccac gatctgctga aaattatcaa ggacaaggac ttcctggaca
8881 atgaggaaaa cgaggacatt ctggaagata tcgtgctgac cctgacactg tttgaggaca
8941 gagagatgat cgaggaacgg ctgaaaacct atgcccacct gttcgacgac aaagtgatga
9001 agcagctgaa gcggcggaga tacaccggct ggggcaggct gagccggaag ctgatcaacg
9061 gcatccggga caagcagtcc ggcaagacaa tcctggattt cctgaagtcc gacggcttcg
9121 ccaacagaaa cttcatgcag ctgatccacg acgacagcct gacctttaaa gaggacatcc
9181 agaaagccca ggtgtccggc cagggcgata gcctgcacga gcacattgcc aatctggccg
9241 gcagccccgc cattaagaag ggcatcctgc agacagtgaa ggtggtggac gagctcgtga
9301 aagtgatggg ccggcacaag cccgagaaca tcgtgatcga aatggccaga gagaaccaga
9361 ccacccagaa gggacagaag aacagccgcg agagaatgaa gcggatcgaa gagggcatca
9421 aagagctggg cagccagatc ctgaaagaac accccgtgga aaacacccag ctgcagaacg
9481 agaagctgta cctgtactac ctgcagaatg ggcgggatat gtacgtggac caggaactgg
9541 acatcaaccg gctgtccgac tacgatgtgg accatatcgt gcctcagagc tttctggcag
9601 acgactccat cgataacaaa gtgctgactc ggagcgacaa gaaccggggc aagagcgaca
9661 acgtgccctc cgaagaggtc gtgaagaaga tgaagaacta ctggcgccag ctgctgaatg
9721 ccaagctgat tacccagagg aagttcgaca atctgaccaa ggccgagaga ggcggcctga
9781 gcgaactgga taaggccggc ttcatcaaga gacagctggt ggaaacccgg cagatcacaa
9841 agcacgtggc acagatcctg gactcccgga tgaacactaa gtacgacgag aacgacaaac
9901 tgatccggga agtgaaagtg atcaccctga agtccaagct ggtgtccgat ttccggaagg
9961 atttccagtt ttacaaagtg cgcgagatca acaactacca ccacgcccac gacgcctacc
10021 tgaacgccgt cgtgggaacc gccctgatca aaaagtaccc tgcgctggaa agcgagttcg
10081 tgtacggcga ctacaaggtg tacgacgtgc ggaagatgat cgccaagagc gagcaggaaa
10141 tcggcaaggc taccgccaag tacttcttct acagcaacat catgaacttt ttcaagaccg
10201 agattaccct ggccaacggc gagatccgga aggcgcctct gatcgagaca aacggcgaaa
10261 caggcgagat cgtgtgggat aagggccggg actttgccac cgtgcggaaa gtgctgtcta
10321 tgccccaagt gaatatcgtg aaaaagaccg aggtgcagac aggcggcttc agcaaagagt
10381 ctatcctgcc caagaggaac agcgacaagc tgatcgccag aaagaaggac tgggacccta
10441 agaagtacgg cggcttcgac agccccaccg tggcctattc tgtgctggtg gtggccaaag
10501 tggaaaaggg caagtccaag aaactgaaga gtgtgaaaga gctgctgggg atcaccatca
10561 tggaaagaag cagcttcgag aagaatccca tcgactttct ggaagccaag ggctacaaag
10621 aagtgaaaaa ggacctgatc atcaagctgc ctaagtactc cctgttcgag ctggaaaacg
10681 gccggaagag aatgctggcc tctgccggcg aactgcagaa gggaaacgaa ctggccctgc
10741 cctccaaata tgtgaacttc ctgtacctgg ccagccacta tgagaagctg aagggctccc
10801 ccgaggataa tgagcagaaa cagctgtttg tggaacagca caaacactac ctggacgaga
10861 tcatcgagca gatcagcgag ttctccaaga gagtgatcct ggccgacgct aatctggaca
10921 aggtgctgag cgcctacaac aagcacagag acaagcctat cagagagcag gccgagaata
10981 tcatccacct gtttaccctg accaatctgg gagcccctgc cgccttcaag tactttgaca
11041 ccaccatcga ccggaagagg tacaccagca ccaaagaggt gctggacgcc accctgatcc
11101 accagagcat caccggcctg tacgagacac ggatcgacct gtctcagctg ggaggcgacg
11161 ccagcctggg cagcggctcc cccaagaaaa aacgcaaggt ggaagatcct aagaaaaagc
11221 ggaaagtgga cagtgaaacc ccgggaacaa gtgagtcggc cacccctgaa ggtggatcag
11281 ggggtagcgg atccgttcag atcccgcaga acccgctgat tctggttgac ggatctagtt
11341 acctgtaccg tgcttaccat gctttcccgc ctttgaccaa ttctgctggt gaacctacgg
11401 gagctatgta cggagttctg aatatgttgc gttctttaat tatgcagtac aagcctaccc
11461 acgctgctgt tgttttcgat gctaaaggta agacgttccg cgacgagtta ttcgagcact
11521 ataagtctca ccgtcctccg atgcctgatg acttacgcgc tcagattgag ccgctgcatg
11581 ctatggtgaa ggctatgggt ttacctcttt tggctgtcag cggtgttgag gctgatgatg
11641 tcattggcac cttagctcgt gaggctgaga aggctggtcg ccctgttttg atttctaccg
11701 gtgacaagga catggctcaa ttggttaccc cgaacatcac cctgatcaac accatgacca
11761 acacgattct gggtcctgag gaagttgtta acaaatatgg tgttcctccg gagttgatta
11821 ttgactttct tgctcttatg ggcgattctt cagacaatat cccgggtgtt ccaggtgttg
11881 gagagaagac tgctcaagct ctgcttcagg gtctgggtgg tttggacacc ctttacgctg
11941 aaccggagaa gatcgccggt ctgtcttttc gcggtgctaa gaccatggct gctaaactgg
12001 aacagaataa ggaggtcgca tacctgtctt atcaattggc taccatcaag acggatgtgg
12061 agttagaact tacgtgcgag cagcttgagg ttcaacagcc tgctgctgag gaactgctgg
12121 gtctttttaa gaaatacgaa tttaagcgtt ggaccgccga cgttgaggct ggtaagtggc
12181 tgcaagctaa gggtgctaag ccggctgcta aaccgcaaga aacgagtgtc gctgatgagg
12241 ctccggaggt taccgctacc gttatctctt acgataatta tgttacgatt ctggacgagg
12301 aaaccttaaa ggcttggatc gctaaattag agaaggctcc tgttttcgct ttcgacacgg
12361 aaacggattc tctggacaat attagtgcga atcttgttgg tctgagtttc gcaattgaac
12421 cgggtgttgc tgcttacatc cctgtggcac acgactacct ggacgctccg gaccagattt
12481 cacgtgaacg cgctctggaa ctgctgaagc ctttattaga ggacgagaaa gctttgaaag
12541 ttggtcagaa tttgaagtat gctcgtggaa tcttagctaa ttatggtatc gagttgcgcg
12601 gtatcgcttt cgacacgatg ttggaatctt atatcctgaa ctctgtcgct ggtcgccatg
12661 acatggactc tctggctgag cgctggctga aacataagac gattaccttc gaggaaatcg
12721 caggaaaggg taagaaccag ctcacgttca atcaaatcgc tctggaggaa gctggtcgct
12781 atgctgctga ggacgctgac gttactctgc aactgcactt gaagatgtgg cctgacttgc
12841 agaagcataa gggtccactg aatgtttttg aaaacattga gatgcctttg gttccagttc
12901 tgtctcgtat cgagcgcaat ggcgttaaaa ttgacccaaa ggttttacat aaccactcag
12961 aggaactgac gctgcgctta gccgaattgg agaaaaaggc tcacgagatc gctggcgaag
13021 agttcaatct gtcatctacg aaacaactgc agactatcct gttcgagaag caaggtatca
13081 agccattaaa aaagacccct ggcggtgctc cgtctacctc tgaggaagtt ttggaggagt
13141 tagctttgga ttaccctctg ccgaaggtta tcttggaata ccgcggtttg gctaaattga
13201 agtctactta tacggataaa cttcctttga tgattaatcc aaagacgggt cgcgttcaca
13261 cgtcgtacca tcaagctgtt accgctaccg gtcgcctgtc ttctacggat ccgaatttac
13321 agaatattcc tgtgcgcaat gaggagggcc gccgcattcg tcaagctttt atcgctccgg
13381 aagactacgt tatcgtttct gctgattatt ctcaaaatga attacgtatc atggctcacc
13441 tgtctcgcga taagggtctg ttgacggcct ttgctgaggg taaggacatt catcgtgcta
13501 ccgctgctga ggtttacggc ctgccgttgg aaacggttac gtctgaacag cgtcgctctg
13561 ctaagcgtat taatttcggc ttaatctacg gtatgtctgc gtttggctta gctcgtcagc
13621 tgaatatccc gcgcaaggaa gctcaaaaat atatggatct gtattttgag cgttaccacg
13681 gtgttttgga atacatggag cgtacgcgcg cgcaagctaa ggaacaaggt tatgtggaaa
13741 ccttggatgg tcgtcgcttg tacttgcctg acattaagtc ttctaacggc gcccgccgcg
13801 ctgctgccga gcgcgctgct atcaatgctc cgatgcaagg tactgctgct gatattatta
13861 agcgtgctat gatcgctgtg gacgcttggc tgcaagctga acagcctcgc gttcgcatga
13921 ttatgcaagt tcatgacgag ttggttttcg aggtgcataa ggacgacgtg gacgctgttg
13981 ctaaacaaat ccaccagttg atggagaatt gcacgcgctt agacgttccg ctgctggttg
14041 aagttggttc tggtgaaaac tgggaccagg ctcactaata atggc
//
