| 质粒名称: | pCRISPomyces-1 |
|---|---|
| 出品公司: | 爱迪基因 |
| 目录编号: | 61736 |
| 存储实验室: | Huimin Zhao |
|
载体骨架基本信息 |
|
| 载体名称: | pKC1139 |
| 载体出品公司: | -- |
| 载体抗性: | 氨苄青霉素 |
| 空载体大小: | -- |
| 完整质粒大小: | 11474 bp |
| 载体修饰: | 突变G9173C从pSG5 rep中删除BbsI位点 |
| 载体类型: | 细菌表达，CRISPR |
| 筛选标记: | -- |
| 高拷贝/低拷贝: | 高拷贝 |
| 生长条件: | 37 ℃ |
|
插入基因信息 |
|
| 插入基因名称: | sSpCas9,Streptomyces codon-optimized Cas9 |
| 物种来源: | Synthetic |
| 插入大小: | 4107 bp |
| 基因突变: | -- |
| GenBank编号 | -- |
| 启动子 | rpsLp(XC)-BbsI |
| 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: | pCRISPomyces-1 was a gift from Huimin Zhao (Addgene plasmid # 61736 ; http://n2t.net/addgene:61736 ; RRID:Addgene_61736) |
| 参考文献部分: | High-Efficiency Multiplex Genome Editing of Streptomyces Species Using an Engineered CRISPR/Cas System. Cobb RE, Wang Y, Zhao H. ACS Synth Biol. 2014 Dec 8. 10.1021/sb500351f PubMed 25458909 |
买家导航
pCRISPomyces1载体基本信息
pCRISPomyces1载体图谱质粒图谱和多克隆位点信息
pCRISPomyces1载体简介
pCRISPomyces1载体序列
LOCUS pCRISPomyces-1 11473 bp ds-DNA circular SYN 17-SEP-2019
DEFINITION Streptomyces expression of codon-optimized Cas9, tracrRNA, and
custom crRNA.
ACCESSION .
VERSION .
KEYWORDS pCRISPomyces-1
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 11473)
AUTHORS Cobb RE, Wang Y, Zhao H
TITLE High-Efficiency Multiplex Genome Editing of Streptomyces Species
Using an Engineered CRISPR/Cas System.
JOURNAL ACS Synth Biol. 2014 Dec 8.
PUBMED 25458909
REFERENCE 2 (bases 1 to 11473)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Sep 17, 2019 from SnapGene Server 1.1.58
http://www.snapgene.com
FEATURES Location/Qualifiers
source 1..11473
/organism="synthetic DNA construct"
/mol_type="other DNA"
promoter 79..380
/label=rpsL promoter
/note="promoter for the Xylanimonas cellulosilytica
ribosomal protein S12 gene"
CDS 381..4487
/codon_start=1
/product="Cas9 (Csn1) endonuclease from the Streptococcus
pyogenes Type II CRISPR/Cas system"
/label=Cas9
/note="generates RNA-guided double strand breaks in DNA"
/translation="MDKKYSIGLDIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKK
NLIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEES
FLVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIK
FRGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRL
ENLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQ
IGDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVR
QQLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLL
RKQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARG
NSRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEY
FTVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFD
SVEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEER
LKTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFM
QLIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGR
HKPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLY
LYYLQNGRDMYVDQELDINRLSDYDVDHIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVP
SEEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKH
VAQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYL
NAVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKT
EITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSK
ESILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGI
TIMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNE
LALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILAD
ANLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVL
DATLIHQSITGLYETRIDLSQLGGD"
promoter 4687..5047
/label=rpsL promoter
/note="promoter for the Cellulomonas flavigena ribosomal
protein S12 gene"
misc_RNA 5048..5126
/label=tracrRNA
/note="trans-activating CRISPR RNA for the Streptococcus
pyogenes CRISPR/Cas9 system"
terminator 5150..5183
/label=lambda t0 terminator
/note="minimal transcription terminator from phage lambda
(Scholtissek and Grosse, 1987)"
promoter 5184..5468
/label=gapdh promoter
/note="glyceraldehyde-3-phosphate dehydrogenase promoter
from Eggerthella lenta"
CDS complement(5512..5808)
/codon_start=1
/gene="lacZ fragment"
/product="LacZ-alpha fragment of beta-galactosidase"
/label=lacZ-alpha
/translation="MTMITPSAQLTLTKGNKSWRTSRGGPVPNSPYSESYYARSLAVVL
QRRDWENPGVTQLNRLAAHPPFASWRNSEEARTDRPSQQLRSLNGEWDAPCSG"
primer_bind 5658..5680
/label=M13/pUC Forward
/note="In lacZ gene"
primer_bind 5672..5689
/label=M13 Forward
/note="In lacZ gene. Also called M13-F20 or M13 (-21)
Forward"
primer_bind 5673..5689
/label=M13 fwd
/note="common sequencing primer, one of multiple similar
variants"
primer_bind 5699..5718
/label=T7
/note="T7 promoter, forward primer"
promoter 5699..5717
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
primer_bind complement(5765..5785)
/label=T3
/note="T3 promoter, forward primer"
promoter complement(5765..5783)
/label=T3 promoter
/note="promoter for bacteriophage T3 RNA polymerase"
primer_bind complement(5804..5820)
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(5804..5820)
/label=M13 Reverse
/note="In lacZ gene. Also called M13-rev"
primer_bind complement(5817..5839)
/label=M13/pUC Reverse
/note="In lacZ gene"
protein_bind 5828..5844
/label=lac operator
/bound_moiety="lac repressor encoded by lacI"
/note="The lac repressor binds to the lac operator to
inhibit transcription in E. coli. This inhibition can be
relieved by adding lactose or
isopropyl-beta-D-thiogalactopyranoside (IPTG)."
promoter complement(5852..5882)
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 5897..5918
/label=CAP binding site
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding activates transcription in the presence
of cAMP."
terminator 5991..6038
/label=T7 terminator
/note="transcription terminator for bacteriophage T7 RNA
polymerase"
rep_origin complement(6081..6669)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind complement(6161..6180)
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
CDS 6873..7673
/codon_start=1
/gene="aac(3)IV"
/product="aminoglycoside 3-N-acetyltransferase type IV
"
/label=ApmR
/note="confers resistance to apramycin (Paget and Davies,
1996)"
/translation="MSSAVECNVVQYEWRKAELIGQLLNLGVTPGGVLLVHSSFRSVRP
LEDGPLGLIEALRAALGPGGTLVMPSWSGLDDEPFDPATSPVTPDLGVVSDTFWRLPNV
KRSAHPFAFAAAGPQAEQIISDPLPLPPHSPASPVARVHELDGQVLLLGVGHDANTTLH
LAELMAKVPYGVPRHCTILQDGKLVRVDYLENDHCCERFALADRWLKEKSLQKEGPVGH
AFARLIRSRDIVATALGQLGRDPLIFLHPPEAGCEECDAARQSIG"
CDS complement(8612..10054)
/codon_start=1
/gene="rep"
/product="replication initiator protein from the
Streptomyces ghanaensis plasmid pSG5"
/label=pSG5 Rep
/translation="MWKPGEATWGNTCRCNNVHTCPWCMSRILAVRGSNVQLAADGLAD
AGYGLHLGTNTLRHFERMAFGTVRKGMRHGLVAVLHDGWKGAYGSSGRRWRTMRDDFGI
IGYERAFEDTFGWGSGWHLHWHTLWVTREVLGPDAQAAFRDALAGAWAAGVESAGGYTV
SETCDRPGCSCEGKGHGTDVRPLNGADAADGDAGKQARYLYKDGDKTKGGVAKIGLELA
GQNFKAGRGDDRMGPLDLGDAAAAELQRLRRPGPFVEKYREREFGVFQVRKHYRSQNLN
RLIKELGIQQDVRTEEEITDDTEGLVAIAVIPAYIWYRYIARVAGRRLDLIKVAETYGL
PGVRRLVESWGLVWGKDVLDPPAPEAPAAPGDLDADQMRFEVMSEEEAAFREARRKANE
ARTEELAASLDRVRQPKKEAIRPTISLRKRLKPKPVTVDVKTPPPGAASPVCRRCKGKL
APVLQPWGQHPGDCLRVDTAVA"
oriT 11221..11330
/note="incP origin of transfer"
ORIGIN
1 ctgtcggcct acctgctggc cgtcggccag ggctacaaaa tcacgggcgt cgtggactat
61 gagcacgtcc gcgagctggc cctgcaggcg gaagtcaggt agacacgact tccgctagtc
121 cttgcaaggt ctgctgacgt gaggcggggc ggtcgttttt gaccgccctg ccttcgtcat
181 gtaggctcgc tcgctgtgcc tggcgtgtca tcagacgccc aggtcccggt gccgtgaggc
241 ccgggccatc gagccggtgg tacgtggctg cggtcccctt gtgagggctg cgcgccgtgt
301 gctgtccggc gcgcacagcc ttgaatccac ccgcgggggc cggccggtct ccgtgagctc
361 gagtagacga cggagacgta atggacaaga agtacagcat cggcctggac atcggcacca
421 acagcgtggg ctgggcggtc atcaccgacg agtacaaggt cccctccaag aagttcaagg
481 tcctgggcaa caccgaccgg cactcgatca agaagaacct gatcggcgcc ctgctcttcg
541 acagcggcga aaccgccgag gcgacccgcc tgaagcggac cgcccgtcgc cgctacaccc
601 ggcgcaagaa ccgcatctgc tacctgcagg agatcttctc caacgagatg gccaaggtcg
661 acgactcgtt cttccaccgg ctcgaggaga gcttcctggt ggaggaggac aagaagcacg
721 agcgccaccc gatcttcggc aacatcgtcg acgaggtggc ctaccacgag aagtacccca
781 ccatctacca cctccgcaag aagctggtcg actcgaccga caaggcggac ctgcggctca
841 tctacctggc cctcgcgcac atgatcaagt tccgcggcca cttcctcatc gagggcgacc
901 tgaacccgga caactccgac gtcgacaagc tcttcatcca gctggtgcag acctacaacc
961 agctgttcga ggagaacccc atcaacgcca gcggcgtcga cgccaaggcg atcctctccg
1021 cgcgcctgag caagtcccgg cgcctggaga acctcatcgc ccagctgccg ggcgagaaga
1081 agaacggcct cttcggcaac ctgatcgcgc tgtcgctcgg cctgaccccc aacttcaaga
1141 gcaacttcga cctggccgag gacgcgaagc tccagctgtc caaggacacc tacgacgacg
1201 acctggacaa cctgctcgcc cagatcggcg accagtacgc ggacctcttc ctggccgcga
1261 agaacctctc ggacgccatc ctgctcagcg acatcctgcg ggtcaacacc gagatcacca
1321 aggccccgct gtcggcgagc atgatcaagc ggtacgacga gcaccaccag gacctgaccc
1381 tgctcaaggc cctcgtgcgc cagcagctgc ccgagaagta caaggagatc ttcttcgacc
1441 agtccaagaa cggctacgcc ggctacatcg acggcggcgc gtcgcaggag gagttctaca
1501 agttcatcaa gccgatcctg gagaagatgg acggcaccga ggagctgctc gtcaagctga
1561 accgcgagga cctgctccgc aagcagcgga ccttcgacaa cggctccatc ccgcaccaga
1621 tccacctggg cgagctccac gccatcctcc ggcgccagga ggacttctac cccttcctga
1681 aggacaaccg cgagaagatc gagaagatcc tgaccttccg gatcccgtac tacgtcggcc
1741 ccctggcccg cggcaactcc cggttcgcgt ggatgacccg gaagtcggag gaaaccatca
1801 ccccgtggaa cttcgaggag gtcgtggaca agggcgcctc cgcgcagtcg ttcatcgagc
1861 gcatgaccaa cttcgacaag aacctcccga acgagaaggt cctgcccaag cacagcctgc
1921 tctacgagta cttcaccgtg tacaacgagc tgaccaaggt caagtacgtg accgagggca
1981 tgcggaagcc ggccttcctg tccggcgagc agaagaaggc gatcgtcgac ctgctcttca
2041 agaccaaccg caaggtcacc gtgaagcagc tgaaggagga ctacttcaag aagatcgagt
2101 gcttcgactc cgtcgagatc tcgggcgtgg aggaccgctt caacgcctcc ctgggcacct
2161 accacgacct gctcaagatc atcaaggaca aggacttcct cgacaacgag gagaacgagg
2221 acatcctgga ggacatcgtc ctcaccctga ccctcttcga ggaccgcgag atgatcgagg
2281 agcggctcaa gacctacgcc cacctgttcg acgacaaggt gatgaagcag ctgaagcggc
2341 gccggtacac cggctggggc cgcctctccc ggaagctgat caacggcatc cgggacaagc
2401 agagcggcaa gaccatcctg gacttcctca agtccgacgg cttcgccaac cgcaacttca
2461 tgcagctcat ccacgacgac tcgctgacct tcaaggagga catccagaag gcccaggtgt
2521 ccggccaggg cgacagcctc cacgagcaca tcgccaacct ggcgggctcc ccggcgatca
2581 agaagggcat cctccagacc gtcaaggtcg tggacgagct ggtcaaggtg atgggccgcc
2641 acaagcccga gaacatcgtg atcgagatgg cccgggagaa ccagaccacc cagaagggcc
2701 agaagaactc ccgcgagcgg atgaagcgca tcgaggaggg catcaaggag ctcggctcgc
2761 agatcctgaa ggagcacccg gtcgagaaca cccagctcca gaacgagaag ctgtacctct
2821 actacctgca gaacggccgc gacatgtacg tggaccagga gctcgacatc aaccggctga
2881 gcgactacga cgtcgaccac atcgtgccgc agtccttcct gaaggacgac tcgatcgaca
2941 acaaggtcct gacccgctcc gacaagaacc ggggcaagtc cgacaacgtg ccctcggagg
3001 aggtcgtgaa gaagatgaag aactactggc gccagctgct caacgccaag ctcatcaccc
3061 agcgcaagtt cgacaacctg accaaggccg agcggggcgg cctgtcggag ctcgacaagg
3121 cgggcttcat caagcgccag ctcgtcgaaa cccggcagat caccaagcac gtggcccaga
3181 tcctggacag ccggatgaac accaagtacg acgagaacga caagctgatc cgcgaggtca
3241 aggtgatcac cctcaagagc aagctggtgt ccgacttccg caaggacttc cagttctaca
3301 aggtccggga gatcaacaac taccaccacg cccacgacgc gtacctgaac gccgtcgtgg
3361 gcaccgcgct gatcaagaag tacccgaagc tggagtccga gttcgtctac ggcgactaca
3421 aggtctacga cgtgcgcaag atgatcgcca agtcggagca ggagatcggc aaggccaccg
3481 cgaagtactt cttctacagc aacatcatga acttcttcaa gaccgagatc accctggcca
3541 acggcgagat ccgcaagcgg cccctgatcg aaaccaacgg cgaaaccggc gagatcgtct
3601 gggacaaggg ccgcgacttc gccaccgtcc ggaaggtgct gtccatgccg caggtcaaca
3661 tcgtcaagaa aaccgaggtg cagaccggcg gcttcagcaa ggagtccatc ctccccaagc
3721 gcaactcgga caagctgatc gcccggaaga aggactggga cccgaagaag tacggcggct
3781 tcgacagccc caccgtcgcc tactccgtgc tggtcgtggc gaaggtcgag aagggcaaga
3841 gcaagaagct gaagtccgtg aaggagctgc tcggcatcac catcatggag cgctcctcgt
3901 tcgagaagaa cccgatcgac ttcctggagg ccaagggcta caaggaggtc aagaaggacc
3961 tcatcatcaa gctgcccaag tactcgctgt tcgagctcga gaacggccgc aagcggatgc
4021 tcgccagcgc gggcgagctg cagaagggca acgagctggc cctcccgtcc aagtacgtca
4081 acttcctgta cctcgcgtcc cactacgaga agctgaaggg ctcgcccgag gacaacgagc
4141 agaagcagct cttcgtggag cagcacaagc actacctgga cgagatcatc gagcagatct
4201 cggagttcag caagcgggtc atcctggccg acgcgaacct cgacaaggtg ctgtccgcct
4261 acaacaagca ccgcgacaag ccgatccggg agcaggcgga gaacatcatc cacctgttca
4321 ccctcaccaa cctgggcgcc cccgccgcgt tcaagtactt cgacaccacc atcgaccgca
4381 agcggtacac cagcaccaag gaggtcctcg acgcgaccct gatccaccag tccatcaccg
4441 gcctgtacga aacccgcatc gacctctccc agctcggcgg cgactgagaa ttcagatcta
4501 cgcgttcccc gcaaaagcgg cctttgactc cctgcaagcc tcagcgaccg aatatatcgg
4561 ttatgcgtgg gcgatggttg ttgtcattgt cggcgcaact atcggtatca agctgtttaa
4621 gaaattcacc tcgaaagcaa gctgataaac cgatacaatt aaaggctcct tttggagcct
4681 ttttttccgc ccgcgggacg tgacgagcgg cacgactcga cgactccggg ctcctttgac
4741 gctgtccgtc gcgccgggta gcgtaggaca ccgtgcccgc gccgtcgggc cctcgcgcgt
4801 gcactcggtc gaccgctccc tgccggagtg ggtgcgggtg cacggggtgg ctccccacct
4861 cctctcggat cggtcctcgc ggactgccgc cgtgcggagg accggggcga cacgcccggg
4921 cgcgggggtc ggtgcgggac tccagacctc cggggtagtc gtgcgacggg cgacgatccg
4981 ggccgagccg gccgtcctgg gtgacgggtg ccggtcagac cagagaacac cgacagacgg
5041 agacgtagga accattcaaa acagcatagc aagttaaaat aaggctagtc cgttatcaac
5101 ttgaaaaagt ggcaccgagt cggtgctttt tttactccat ctggatttgt tcagaacgct
5161 cggttgccgc cgggcgtttt ttagctgctc cttcggtcgg acgtgcgtct acgggcacct
5221 taccgcagcc gtcggctgtg cgacacggac ggatcgggcg aactggccga tgctgggaga
5281 agcgcgctgc tgtacggcgc gcaccgggtg cggagcccct cggcgagcgg tgtgaaactt
5341 ctgtgaatgg cctgttcggt tgcttttttt atacggctgc cagataaggc ttgcagcatc
5401 tgggcggcta ccgctatgat cggggcgttc ctgcaattct tagtgcgagt atctgaaagg
5461 ggatacgcgg gttttagagc tatgctgttt tgaatggtcc caaaagtctt ctcagccgct
5521 acagggcgcg tcccattcgc cattcaggct gcgcaactgt tgggaagggc gatcggtgcg
5581 ggcctcttcg ctattacgcc agctggcgaa agggggatgt gctgcaaggc gattaagttg
5641 ggtaacgcca gggttttccc agtcacgacg ttgtaaaacg acggccagtg agcgcgcgta
5701 atacgactca ctatagggcg aattgggtac cgggcccccc ctcgaggtcc tccagctttt
5761 gttcccttta gtgagggtta attgcgcgct tggcgtaatc atggtcatag ctgtttcctg
5821 tgtgaaattg ttatccgctc acaattccac acaacatacg agccggaagc ataaagtgta
5881 aagcctgggg tgcctaatga gtgagctaac tcacattaat tgcgttgcgc tcactgcccg
5941 ctttccaccg gtgaagacat agagctatgc tgttttgaat ggtcccaaaa ctagcataac
6001 cccttggggc ctctaaacgg gtcttgaggg gttttttgtc tagaggccag gaaccgtaaa
6061 aaggccgcgt tgctggcgtt tttccatagg ctccgccccc ctgacgagca tcacaaaaat
6121 cgacgctcaa gtcagaggtg gcgaaacccg acaggactat aaagatacca ggcgtttccc
6181 cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc
6241 gcctttctcc cttcgggaag cgtggcgctt tctcatagct cacgctgtag gtatctcagt
6301 tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg acccccccgt tcagcccgac
6361 cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggtaagaca cgacttatcg
6421 ccactggcag cagccactgg taacaggatt agcagagcga ggtatgtagg cggtgctaca
6481 gagttcttga agtggtggcc taactacggc tacactagaa gaacagtatt tggtatctgc
6541 gctctgctga agccagttac cttcggaaaa agagttggta gctcttgatc cggcaaacaa
6601 accaccgctg gtagcggtgg tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa
6661 ggatctcaag aagatccttt gatcttttct acggggtctg acgctcagtg gaacgaaaac
6721 tcacgttaag ggattttggt catgagatta tcaaaaagga tcttcaccta gatccttttg
6781 gttcatgtgc agctccatca gcaaaagggg atgataagtt tatcaccacc gactatttgc
6841 aacagtgccg ttgatcgtgc tatgatcgac tgatgtcatc agcggtggag tgcaatgtcg
6901 tgcaatacga atggcgaaaa gccgagctca tcggtcagct tctcaacctt ggggttaccc
6961 ccggcggtgt gctgctggtc cacagctcct tccgtagcgt ccggcccctc gaagatgggc
7021 cacttggact gatcgaggcc ctgcgtgctg cgctgggtcc gggagggacg ctcgtcatgc
7081 cctcgtggtc aggtctggac gacgagccgt tcgatcctgc cacgtcgccc gttacaccgg
7141 accttggagt tgtctctgac acattctggc gcctgccaaa tgtaaagcgc agcgcccatc
7201 catttgcctt tgcggcagcg gggccacagg cagagcagat catctctgat ccattgcccc
7261 tgccacctca ctcgcctgca agcccggtcg cccgtgtcca tgaactcgat gggcaggtac
7321 ttctcctcgg cgtgggacac gatgccaaca cgacgctgca tcttgccgag ttgatggcaa
7381 aggttcccta tggggtgccg agacactgca ccattcttca ggatggcaag ttggtacgcg
7441 tcgattatct cgagaatgac cactgctgtg agcgctttgc cttggcggac aggtggctca
7501 aggagaagag ccttcagaag gaaggtccag tcggtcatgc ctttgctcgg ttgatccgct
7561 cccgcgacat tgtggcgaca gccctgggtc aactgggccg agatccgttg atcttcctgc
7621 atccgccaga ggcgggatgc gaagaatgcg atgccgctcg ccagtcgatt ggctgagctc
7681 atgagcggag aacgagatga cgttggaggg gcaaggtcgc gctgattgct ggggcaacac
7741 gtggagcgga tcggggattg tctttcttca gctcgctgat gatatgctga cgctcaatgc
7801 cgtttggcct ccgactaacg aaaatcccgc atttggacgg ctgatccgat tggcacggcg
7861 gacggcgaat ggcggagcag acgctcgtcc gggggcaatg agatatgaaa aagcctgaac
7921 tcaccgcgac gtatcgatgt cgaggttcct caggggagcc accccagaga agccctcgga
7981 gctgagcgga gctatttcca aagccatgcc agctagagac agtgcacacc gccaagcatc
8041 tgcaaaaccc tcgcctggag ggaaagtgca atgtacgtac gctggtttcc ctccagaaag
8101 ggattctgcg gcttctcaac tttggaagaa gaggcggaac gaactgctgt ttcagcctac
8161 tcatgtgagg aggctggtcc tttaccctga tgcccgaggg catcagggta aaggaccagc
8221 ctcgccaact aggagcgcta ccaaggcgaa cacaacccag gacttggtag aacccttcgc
8281 cggagaagtt cagactcatc ggcatgagca cccctcgccg cgcactcggc accggtccgt
8341 cgaccaccac caggacgtcg ttgtcgacgt cggccccgcg gctcctgccc gccgaacgcg
8401 tcgtcgtcga cggcctggtg ctcatcgacg agcacccgga gccaggtgaa aagcgccggc
8461 ggacgctcgg actgggcgcg ggattccagc agtaacccag gtccgccggc cacctcacgg
8521 caggcagacc ctcggctttc gcgccgggac cgccatgaga ccgccacccg gatgtccggg
8581 gtggcggtct catggcggtc cctcagcggc cctacgcgac cgctgtgtcg acgcggaggc
8641 agtctccggg gtgctggccc cagggctgga gtaccggggc gagcttgccc ttgcagcggc
8701 ggcacacggg cgacgcggcg ccgggcgggg gagtcttgac gtcgacggtg accggcttcg
8761 gcttcaggcg cttccgaagg ctgatcgtcg ggcggatcgc ctccttcttc ggctgacgca
8821 ctcggtcgag cgaagcggcc agctcctccg tgcgggcctc gttggccttc cggcgcgcct
8881 cgcggaaagc agcttcctcc tcggacatga cctcgaacct catctggtca gcgtcaaggt
8941 cgcccggcgc ggccggcgct tccggggcgg gcgggtccag gacgtccttg ccccacacca
9001 ggccccagga ctcgacgagc cgccggacgc ccggtaggcc gtacgtctcg gcgaccttga
9061 tgaggtcgag gcgacgtccg gcgacgcggg cgatgtatcg gtaccagatg taggccggga
9121 tgaccgcgat ggcgaccagg ccctcggtgt cgtcggtgat ctcctcctcg gtgcggacgt
9181 cctgctggat gccgagttcc ttgatcagcc ggttcaggtt ctgcgaccgg tagtgcttgc
9241 ggacctggaa cacgccgaac tcgcgctcgc ggtacttctc gacgaacggg ccgggccgac
9301 gaagccgctg cagctcggcg gccgccgcgt cgcccaggtc gagcggtccc atgcggtcgt
9361 cgccgcgacc ggccttgaag ttctgtccgg ccagctccag gccgatcttg gcgacgccgc
9421 ccttggtctt gtcgccgtcc ttgtagaggt agcgggcctg cttgcccgca tcgccgtcag
9481 cggcgtccgc gccgttgagt gggcgcacgt cggtgccgtg gcccttgccc tcacaggagc
9541 aaccgggccg gtcgcacgtc tcgctgacgg tgtagccgcc cgcggattcg accccggcgg
9601 cccaggctcc ggcgagtgcg tcgcggaacg cggcctgggc gtccgggccg agcacctcgc
9661 gggtgaccca gagcgtgtgc cagtgcaggt gccagccgga gccccagccg aaggtgtcct
9721 cgaaggcccg ctcgtagccg atgatcccga agtcgtcgcg catcgtgcgc cagcggcggc
9781 cggacgagcc gtacgcgccc ttccagccgt cgtgcaagac cgcgaccagg ccgtgccgca
9841 ttcccttgcg gacggtgccg aacgccatgc gctcgaagtg gcgcaacgtg ttcgtgccaa
9901 ggtgcagccc gtacccggcg tccgcgagac cgtcggcggc gagctgcacg ttcgagcccc
9961 gtacggccag gatgcggctc atgcaccacg ggcaggtgtg gacgttgttg cagcggcacg
10021 tgttgcccca cgtcgcctcg cccggcttcc acatcagctc ggccgtcccg gcagtgagcc
10081 gggtcccgca gcccttgaac gcctcgttca gcgacaccgt ctggtgccgg tcccgccggg
10141 cgaaccgctc gtcgcgcggg tcctcccgcc cggctgtcgc ggcaccctcg tttggggtag
10201 aacccgttcc agttacagcg ctctgacctg cagtggacgg agattttccc ttactactaa
10261 agcccgcgtc cggattaccc gctgtagtcg tgcttgctac gctgcgtgac tggtccgcaa
10321 tgagacgctt tgcgcgcttt cggcaggcgt ccgagcagta gattttgggg cgcttcccgg
10381 ggatgtggac gatcggggtg ccgcagtggc acttcggtcc ggcggggcgc ggtggtgtcg
10441 acgcgctgtt ctctcgtacg ctcgtcacag agcaaacgtc ctcactcggc atgctgcgcc
10501 ggttcggggg cggcgagccc gggaggccaa tcccgggctc gtgccatttc tgggtcctgt
10561 tgatcctggc attggtgtgg ccgttcattg cccctgctcg ctcctgacgc gccgatagac
10621 gtccgatacg cccggtgctg gtgggatttg ataggtcgga agaagccccg cccgggcctg
10681 ggcggggctt cctgtgcgtc aggacctcct cgtcgtgagc ctcttcggcc tatggacgga
10741 gtgacctcgt gatccgttac agccgcgcgc gctcgcgtag agcggtctca tcagttccac
10801 gaacggtcct cttcgcagat cagggcgttg gggcggagtc tcaccaagga ctacgtctgc
10861 tggcgatttc cgttacaccc cgggcggtgg ccggcgcaca cgcgcgcccg cgttgggcag
10921 tgcagaaagt gcagaaacct aggcgctgat ggtccaggtc cacggttcgt cgtcggcggc
10981 ggcgcgggcg gcggcgtcgg ccagggcgcg ggcgagaccg gctacggcgg gcttgatgcg
11041 ccggttgcgg gcgaccttga gcagctagta tgcaggtcga cggatctttt ccgctgcata
11101 accctgcttc ggggtcatta tagcgatttt ttcggtatat ccatcctttt tcgcacgata
11161 tacaggattt tgccaaaggg ttcgtgtaga ctttccttgg tgtatccaac ggcgtcagcc
11221 gggcaggata ggtgaagtag gcccacccgc gagcgggtgt tccttcttca ctgtccctta
11281 ttcgcacctg gcggtgctca acgggaatcc tgctctgcga ggctggccgg ctaccgccgg
11341 cgtaacagat gagggcaagc ggatggctga tgaaaccaag ccaaccagga agggcagccc
11401 acctatcaag gtgtactgcc ttccagacga acgaagagcg attgaggaaa aggcggcggc
11461 ggccggcatg agc
//
