| 质粒名称: | pCRISPRyl |
|---|---|
| 出品公司: | 爱迪基因 |
| 目录编号: | 70007 |
| 存储实验室: | Ian Wheeldon |
|
载体骨架基本信息 |
|
| 载体名称: | pUC19 |
| 载体出品公司: | -- |
| 载体抗性: | 氨苄青霉素 |
| 空载体大小: | 2623 bp |
| 完整质粒大小: | 11804 bp |
| 载体修饰: | -- |
| 载体类型: | 酵母表达，CRISPR，合成生物学 |
| 筛选标记: | LEU2 |
| 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: | Codon optimized Cas9 from S. pyogenes |
| 物种来源: | Synthetic |
| 插入大小: | 4140 bp |
| 基因突变: | -- |
| GenBank编号 | -- |
| 启动子 | UAS1B8-TEF(136) |
| 标签/融合蛋白: | -- |
| 克隆方法: | 限制性内切酶 |
| 5' 克隆位点: | BssHII(未破坏) |
| 3' 克隆位点: | NheI(未破坏) |
| 5' 测序引物 | GTAAAACGACGGCCAGT |
| 3' 测序引物 | CAGGAAACAGCTATGAC |
|
文献引用方法 |
|
| 材料和方法部分: | pCRISPRyl was a gift from Ian Wheeldon (Addgene plasmid # 70007 ; http://n2t.net/addgene:70007 ; RRID:Addgene_70007) |
| 参考文献部分: | Synthetic RNA polymerase III promoters facilitate high efficiency CRISPR-Cas9 mediated genome editing in Yarrowia lipolytica. Schwartz CM, Hussain MS, Blenner M, Wheeldon I. ACS Synth Biol. 2015 Dec 29. 10.1021/acssynbio.5b00162 PubMed 26714206 |
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
载体简介
载体序列
LOCUS pCRISPRyl 11805 bp ds-DNA circular SYN 06-AUG-2018
DEFINITION CRISPR/Cas9 vector for Yarrowia lipolytica, with AvrII site for
sgRNA insertion.
ACCESSION .
VERSION .
KEYWORDS pCRISPRyl
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 11805)
AUTHORS Schwartz CM, Hussain MS, Blenner M, Wheeldon I
TITLE Synthetic RNA polymerase III promoters facilitate high efficiency
CRISPR-Cas9 mediated genome editing in Yarrowia lipolytica.
JOURNAL ACS Synth Biol. 2015 Dec 29.
PUBMED 26714206
REFERENCE 2 (bases 1 to 11805)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Aug 6, 2018 from SnapGene Server 1.1.58
http://www.snapgene.com
FEATURES Location/Qualifiers
source 1..11805
/organism="synthetic DNA construct"
/mol_type="other DNA"
primer_bind complement(2908..2926)
/label=pBRforEco
/note="pBR322 vectors, upsteam of EcoRI site, forward
primer"
misc_RNA complement(2976..3051)
/label=gRNA scaffold
/note="guide RNA scaffold for the Streptococcus pyogenes
CRISPR/Cas9 system"
promoter 3528..3631
/gene="bla"
/label=AmpR promoter
CDS 3632..4492
/codon_start=1
/gene="bla"
/product="beta-lactamase"
/label=AmpR
/note="confers resistance to ampicillin, carbenicillin, and
related antibiotics"
/translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
LIKHW"
primer_bind complement(3850..3869)
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
rep_origin 4663..5251
/direction=RIGHT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind 5152..5171
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
primer_bind 5405..5422
/label=L4440
/note="L4440 vector, forward primer"
protein_bind 5539..5560
/label=CAP binding site
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding activates transcription in the presence
of cAMP."
promoter 5575..5605
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 5613..5629
/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)."
primer_bind 5618..5640
/label=M13/pUC Reverse
/note="In lacZ gene"
primer_bind 5637..5653
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
primer_bind 5637..5653
/label=M13 Reverse
/note="In lacZ gene. Also called M13-rev"
terminator 5672..5919
/gene="S. cerevisiae CYC1"
/label=CYC1 terminator
/note="transcription terminator for CYC1"
primer_bind 5885..5903
/label=CYC1
/note="CYC1 transcription termination signal, reverse
primer"
CDS complement(5930..5950)
/codon_start=1
/product="nuclear localization signal of SV40 (simian virus
40) large T antigen"
/label=SV40 NLS
/translation="PKKKRKV"
CDS complement(5963..10066)
/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"
primer_bind complement(11411..11428)
/label=M13 Forward
/note="In lacZ gene. Also called M13-F20 or M13 (-21)
Forward"
primer_bind complement(11411..11427)
/label=M13 fwd
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(11420..11442)
/label=M13/pUC Forward
/note="In lacZ gene"
primer_bind complement(11636..11655)
/label=pRS-marker
/note="pRS vectors, use to sequence yeast selectable
marker"
primer_bind 11755..11777
/label=pGEX 3'
/note="pGEX vectors, reverse primer"
ORIGIN
1 gacgaaaggg ccagatctgt tcggaaatca acggatgctc aaccgatttc gacagtaata
61 atttgaatcg aatcggagcc taaaatgaac ccgagtatat ctcataaaat tctcggtgag
121 aggtctgtga ctgtcagtac aaggtgcctt cattatgccc tcaaccttac catacctcac
181 tggatgtagt gtacctctaa aaatgaaata cagtgccaaa agccatggca ctgagctcgt
241 ctaacggact tgatatacaa ccaattaaaa caaatgaaaa gaaatacagt tctttgtatc
301 atttgtaaca attaccctgt acaaactaag gtattgaaat cccacaatat tcccaaagtc
361 cacccctttc caaattgtca tgcctacaac tcatatacca gacactaacc taccaaacac
421 cactaaaaac cccacaaaat atatcttacc gaatatacag taacaagcta ccaccacact
481 cgttgggtgc agtcgccagc tttaaagata tctatccaca tcagccacaa ctcccttcct
541 ttaataaacc gactacaccc ttggctattg aggttatgag tgaatatact gtagacaaga
601 cactttcaag aagactgttt ccaaaacgta ccactgtcct ccactacaaa cacacccaat
661 ctgcttcttc tagtcaaggt tgctacaccg gtaaattata aatcatcatt tcattagcag
721 ggctgggccc tttttataga gtcttataca ctagcggacc ctgccggtag accaacccgc
781 aggcgcgtca gtttgctcct tccatcaatg cgtcgtagaa acgacttact ccttcttgag
841 cagctccttg accttgttgg caacaaagtc tccgacctcg gaggtggagg aagagcctcc
901 gatatcggcg gtagtgatac cagcctcgac ggactccttg acggcagcct caacagcgtc
961 accggcgggc ttcatgttaa gagagaactt gagcatcatg gcggcagaca gaatggtggc
1021 aatggggttg accttctgct tgccgagatc gggggcagat ccgtgacagg gctcgtacag
1081 accgaacgcc tcgttggtgt cgggcagaga agccagagag gcggagggca gcagacccag
1141 agaaccgggg atgacggagg cctcgtcgga gatgatatcg ccaaacatgt tggtggtgat
1201 gatgatacca ttcatcttgg agggctgctt gatgaggatc atggcggccg agtcgatcag
1261 ctggtggttg agctcgagct gggggaattc gtccttgagg actcgagtga cagtctttcg
1321 ccaaagtcga gaggaggcca gcacgttggc cttgtcaaga gaccacacgg gaagaggggg
1381 gttgtgctga agggccagga aggcggccat tcgggcaatt cgctcaacct caggaacgga
1441 gtaggtctcg gtgtcggaag cgacgccaga tccgtcatcc tcctttcgct ctccaaagta
1501 gatacctccg acgagctctc ggacaatgat gaagtcggtg ccctcaacgt ttcggatggg
1561 ggagagatcg gcgagcttgg gcgacagcag ctggcagggt cgcaggttgg cgtacaggtt
1621 caggtccttt cgcagcttga ggagaccctg ctcgggtcgc acgtcggttc gtccgtcggg
1681 agtggtccat acggtgttgg cagcgcctcc gacagcaccg agcataatag agtcagcctt
1741 tcggcagatg tcgagagtag cgtcggtgat gggctcgccc tccttctcaa tggcagctcc
1801 tccaatgagt cggtcctcaa acacaaactc ggtgccggag gcctcagcaa cagacttgag
1861 caccttgacg gcctcggcaa tcacctcggg gccacagaag tcgccgccga gaagaacaat
1921 cttcttggag tcagtcttgg tcttcttagt ttcgggttcc attgtggatg tgtgtggttg
1981 tatgtgtgat gtggtgtgtg gagtgaaaat ctgtggctgg caaacgctct tgtatatata
2041 cgcacttttg cccgtgctat gtggaagact aaacctccga agattgtgac tcaggtagtg
2101 cggtatcggc tagggaccca aaccttgtcg atgccgatag cgctatcgaa cgtaccccag
2161 ccggccggga gtatgtcgga ggggacatac gagatcgtca agggtttgtg gccaactggt
2221 aaataaatga tgactcaggc gacgacggaa ttcgacagca actactcctt tcaccaacca
2281 tgtgcatttt agctcgaata acattcacag gcttggtgat ctacatccat ggtgtctggc
2341 cgattaccgt ggtgttttgg cagtaacgag aatattgagt gaactcttcc catcaccaat
2401 aaagactcat actacaatca cgagcgcttc agctgccact atagtgttgg tgacacaata
2461 cccctcgatg ctgggcatta ctgtagcaag agatattatt tcatggcgca ttttccagtc
2521 tacctgactt tttagtgtga tttcttctcc acattttatg ctcagtgtga aaagttggag
2581 tgcacactta attatcgccg gttttcggaa agtactatgt gctcaaggtt gcaccccacg
2641 ttacgtatgc agcacattga gcagcctttg gaccgtggag ataacggtgt ggagatagca
2701 acgggtagtc ttcgtattaa ttcaatgcat tgttagtttt atatgatatg gtgtcgaagc
2761 ggccgcatac tactgtatat tcaagcaagt atatccgtgg gtgcgggtga tttggatcta
2821 aggttcgtac tcaacactca cgagcagctt gcctatgtta catcctttta tcagacatag
2881 cggccgcttc gactctagag gatctgggcc tcgtgatacg cctattttta taggttaatg
2941 tcatgataat aatggtttct tagacgtaaa aaaaagcacc gactcggtgc cactttttca
3001 agttgataac ggactagcct tattttaact tgctatttct agctctaaaa cctaggtcaa
3061 cctgcgccga cccggaatcg aaccgggggc ccatcgatgg caacgatgga ttttaccact
3121 aaaccatcgg cgcattagag gtatttttca aagtagccca gtgcagagtc cagaactcga
3181 acgcacaaca agaaggcgat ttaccactcc gacaaaatgc cattacagct agtaaaaaac
3241 tttgtttaca ccactatagc tgataataca tatcacgtga cggagccaaa aaatggagtc
3301 tcatacacga agaagtgtcc gatttgagat tttgaaaaag acaatatatt aaaaaggcat
3361 tagagcgtta ttttaaccaa cttttaaagc cctagcacat caattaagac caatattata
3421 taatcacacc ctaacccttg ttatgattct atatttggaa gcagatctag agttgtgtca
3481 acttttgcaa ctggggacgt caggtggcac ttttcgggga aatgtgtgcg gaacccctat
3541 ttgtttattt ttctaaatac attcaaatat gtatccgctc atgagacaat aaccctgata
3601 aatgcttcaa taatattgaa aaaggaagag tatgagtatt caacatttcc gtgtcgccct
3661 tattcccttt tttgcggcat tttgccttcc tgtttttgct cacccagaaa cgctggtgaa
3721 agtaaaagat gctgaagatc agttgggtgc acgagtgggt tacatcgaac tggatctcaa
3781 cagcggtaag atccttgaga gttttcgccc cgaagaacgt tttccaatga tgagcacttt
3841 taaagttctg ctatgtggcg cggtattatc ccgtattgac gccgggcaag agcaactcgg
3901 tcgccgcata cactattctc agaatgactt ggttgagtac tcaccagtca cagaaaagca
3961 tcttacggat ggcatgacag taagagaatt atgcagtgct gccataacca tgagtgataa
4021 cactgcggcc aacttacttc tgacaacgat cggaggaccg aaggagctaa ccgctttttt
4081 gcacaacatg ggggatcatg taactcgcct tgatcgttgg gaaccggagc tgaatgaagc
4141 cataccaaac gacgagcgtg acaccacgat gcctgtagca atggcaacaa cgttgcgcaa
4201 actattaact ggcgaactac ttactctagc ttcccggcaa caattaatag actggatgga
4261 ggcggataaa gttgcaggac cacttctgcg ctcggccctt ccggctggct ggtttattgc
4321 tgataaatct ggagccggtg agcgtgggtc tcgcggtatc attgcagcac tggggccaga
4381 tggtaagccc tcccgtatcg tagttatcta cacgacgggg agtcaggcaa ctatggatga
4441 acgaaataga cagatcgctg agataggtgc ctcactgatt aagcattggt aactgtcaga
4501 ccaagtttac tcatatatac tttagattga tttaaaactt catttttaat ttaaaaggat
4561 ctaggtgaag atcctttttg ataatctcat gaccaaaatc ccttaacgtg agttttcgtt
4621 ccactgagcg tcagaccccg tagaaaagat caaaggatct tcttgagatc ctttttttct
4681 gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc
4741 ggatcaagag ctaccaactc tttttccgaa ggtaactggc ttcagcagag cgcagatacc
4801 aaatactgtc cttctagtgt agccgtagtt aggccaccac ttcaagaact ctgtagcacc
4861 gcctacatac ctcgctctgc taatcctgtt accagtggct gctgccagtg gcgataagtc
4921 gtgtcttacc gggttggact caagacgata gttaccggat aaggcgcagc ggtcgggctg
4981 aacggggggt tcgtgcacac agcccagctt ggagcgaacg acctacaccg aactgagata
5041 cctacagcgt gagcattgag aaagcgccac gcttcccgaa gggagaaagg cggacaggta
5101 tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg gagcttccag ggggaaacgc
5161 ctggtatctt tatagtcctg tcgggtttcg ccacctctga cttgagcgtc gatttttgtg
5221 atgctcgtca ggggggcgga gcctatggaa aaacgccagc aacgcggcct ttttacggtt
5281 cctggccttt tgctggcctt ttgctcacat gttctttcct gcgttatccc ctgattctgt
5341 ggataaccgt attaccgcct ttgagtgagc tgataccgct cgccgcagcc gaacgaccga
5401 gcgcagcgag tcagtgagcg aggaagcgga agagcgccca atacgcaaac cgcctctccc
5461 cgcgcgttgg ccgattcatt aatgcagctg gcacgacagg tttcccgact ggaaagcggg
5521 cagtgagcgc aacgcaatta atgtgagtta gctcactcat taggcacccc aggctttaca
5581 ctttatgctt ccggctcgta tgttgtgtgg aattgtgagc ggataacaat ttcacacagg
5641 aaacagctat gaccatgatt acgccaagct tgcaaattaa agccttcgag cgtcccaaaa
5701 ccttctcaag caaggttttc agtataatgt tacatgcgta cacgcgtctg tacagaaaaa
5761 aaagaaaaat ttgaaatata aataacgttc ttaatactaa cataactata aaaaaataaa
5821 tagggaccta gacttcaggt tgtctaactc cttccttttc ggttagagcg gatgtggagg
5881 gagggcgtga atgtaagcgt gacataacta attacatgag gctagcttac acctttcgct
5941 tcttcttggg gtcggctcga gagtcgccgc ccagctgaga caggtcgatt cgggtctcgt
6001 acaggccggt gatagactgg tggatcaggg tggcgtccag cacctccttg gtagaggtgt
6061 atcgctttcg gtcgatggtg gtgtcgaagt acttgaaggc agcgggggct cccaggttgg
6121 tcagggtgaa caggtgaatg atgttctcag cctgctctcg aatgggcttg tctcggtgct
6181 tgttgtaagc agacagcacc ttatccaggt tggcgtcagc caggatcact cgcttagaga
6241 actcagagat ctgctcgatg atctcgtcca ggtagtgctt atgctgctcc acgaagagct
6301 gcttctgctc gttatcctcg ggagagccct tcagcttctc gtaatgggag gcgaggtaca
6361 ggaagttgac gtacttggag ggcagggcca gctcgtttcc cttctgcagc tcgccggcag
6421 aggccagcat tcgctttcgt ccgttctcca gctcgaacag agagtacttg gggagcttga
6481 taatcagatc tttcttcacc tccttgtatc ccttggcctc caggaagtca atggggttct
6541 tctcgaaaga agatcgctcc atgatggtaa tgcccagcag ctccttcaca gacttcagct
6601 tcttagactt gcccttctcc accttggcca ccaccagcac agagtaagcc acggtgggag
6661 agtcgaaacc tccgtacttc ttggggtccc agtccttctt tcgggcgatc agcttgtcag
6721 agtttcgctt gggcaggatg gactccttag agaaacctcc ggtctgcacc tcggtcttct
6781 tcacgatgtt cacctggggc atagacagca cctttcgcac ggtggcgaag tctcgtccct
6841 tgtcccacac gatctcaccg gtctctccgt tggtctcaat gaggggtcgc tttcgaatct
6901 cgccgttagc cagggtaatc tcggtcttga agaagttcat aatgttggag tagaagaagt
6961 acttggcggt agccttgcca atctcctgct cggacttggc aatcattttt cgcacatcgt
7021 agaccttgta gtcgccgtag acgaactcgg actccagctt gggatacttc ttgatgaggg
7081 cggtgcccac gacagcgttc aggtaggcat cgtgggcgtg gtggtagttg ttaatctctc
7141 gcaccttgta gaactggaag tcctttcgga agtcagacac cagcttagac ttcagggtga
7201 tcaccttcac ctctcggatc agcttgtcgt tctcgtcgta cttggtgttc attcgagagt
7261 ccaggatctg ggccacgtgc ttggtgatct gtcgggtctc caccagctgt cgcttgatga
7321 agccggcctt gtccagctca gacaggccgc ctcgctcggc cttggtcagg ttgtcgaact
7381 ttcgctgggt aatcagcttg gcgttcagca gctgtcgcca gtagttcttc atcttcttga
7441 ccacctcctc ggagggcacg ttgtcagact tgcctcggtt cttgtcggat cgggtcagca
7501 ccttgttatc gatagaatcg tccttcagga aggactgggg cacaatgtgg tccacatcgt
7561 agtcggagag tcggttgata tccagctcct ggtcgacgta catgtctcga ccgttctgca
7621 gatagtacag gtacagcttc tcgttctgga gctgagtgtt ctccacggga tgctccttga
7681 gaatctggga tcccagctcc ttaatacctt cctcaattcg cttcattcgc tctcgggagt
7741 ttttctgacc cttctgagta gtctggttct ctcgagccat ctcaatgaca atgttctcgg
7801 gcttgtgtcg gcccatcacc ttgacgagct catcgaccac cttgacggtc tgcaggatac
7861 ctttcttaat ggcgggagag ccggccaggt tggcgatgtg ctcgtgcaga gagtcgccct
7921 ggccagacac ctgggccttc tggatgtcct ccttgaaggt cagagagtcg tcgtggatca
7981 gctgcatgaa gtttcggttg gcgaagccgt cagacttcag gaagtccagg atggtcttgc
8041 cagactgctt gtctcggatg ccgttgatca gctttcgaga cagtcggccc cagccggtgt
8101 atcgtcgtcg cttcagctgc ttcatcacct tgtcgtcgaa caggtgagcg taggtcttga
8161 gtcgctcctc aatcatctct cgatcctcga acagggtgag ggtcaggaca atgtcctcga
8221 gaatatcctc gttctcctcg ttatccagga aatccttgtc cttgataatc ttcagcaggt
8281 cgtggtaggt acccagagag gcgttgaatc ggtcctcgac gccggagatc tcgacagaat
8341 cgaaacactc aatcttcttg aagtaatcct ccttcagctg cttgacggtg acctttcggt
8401 tagtcttgaa gagcagatcc acaatagcct tcttctgctc tccagacagg aaagcgggct
8461 ttcgcattcc ctcggtcacg tacttgacct tggtgagctc gttgtagacg gtgaagtact
8521 cgtagagcag agagtgcttg ggcaggacct tctcgttggg gaggttcttg tcgaagttgg
8581 tcattcgctc gatgaaagac tgagcagagg cgcccttatc caccacctcc tcgaagttcc
8641 agggggtaat ggtctcctca gactttcggg tcatccaagc gaatcgagag tttcctcggg
8701 ccaggggtcc cacgtagtag gggattcgga aggtcaggat cttctcgatc ttctctcggt
8761 tgtccttcag gaaggggtag aagtcctcct gtcgtcgcag gatggcgtgc agctcgccca
8821 ggtggatctg gtgggggata gagccgttgt cgaaggttcg ctgctttcgc aggaggtcct
8881 ctcggttcag cttcacgagc agttcctcgg ttccgtccat cttctccagg atgggcttaa
8941 tgaacttgta gaactcctcc tgagaggcgc cgccgtcgat gtagccggcg tagccgttct
9001 tagactggtc gaagaaaatc tccttgtact tctcgggcag ctgctgtcgg accagagcct
9061 tgaggagggt cagatcctgg tggtgctcat cgtatcgctt aatcatggag gcagacaggg
9121 gggccttggt gatctcggtg ttcactcgca ggatgtcaga cagcaggatg gcgtcagaca
9181 ggttcttggc ggccaggaac aggtcggcgt actggtcgcc aatctgggcg agcaggttat
9241 ccaggtcatc gtcgtaggtg tccttagaca gctgcagctt agcgtcctca gccagatcga
9301 agttggactt gaagttaggg gtgagaccca gggacagagc aatcaggttt ccgaacagac
9361 cgttcttctt ctctccgggg agctgagcaa tcaggttctc gagtcgtcga gacttagaca
9421 gtcgggcaga caggatggcc ttagcatcca cgccagaagc gttaatgggg ttctcttcga
9481 agagctggtt gtaggtctgc accagctgga tgaacagctt gtccacgtca gagttgtcgg
9541 ggttcaggtc gccctcgatc aggaagtggc ctcggaactt gatcatgtgg gccagggcca
9601 ggtagatcag tcgcaggtcg gccttgtcgg tagagtccac cagcttcttt cgcaggtggt
9661 agatggtggg gtacttctcg tggtaggcca cctcgtccac gatgttgccg aagatggggt
9721 gtcgctcgtg cttcttgtcc tcctccacca ggaaagactc ctccagtcgg tggaagaaag
9781 agtcgtccac cttggccatc tcgttagaga agatctcctg caggtaacag attcggttct
9841 ttcttcgggt gtatcgtctt cgagcggttc gcttcagtcg ggtggcctca gcggtctcgc
9901 cagagtcgaa gagcagagcg ccgatcaggt tcttcttgat ggagtgtcgg tcggtgttgc
9961 ccaggacctt gaatttctta gagggcacct tgtactcatc ggtgatcacg gcccaaccca
10021 cggagttggt tccgatgtcc aggccaatgg agtatttctt atccattgtg gcgcgccttt
10081 gaatgattct tatactcaga aggaaatgct taacgatttc gggtgtgagt tgacaaggag
10141 agagagaaaa gaagaggaaa ggtaattcgg ggacggtggt cttttatacc cttggctaaa
10201 gtcccaacca caaaagcttc tgcaggcatg cccggatcga ggtgggcggg aacaacggtc
10261 tcgataatgt acgtacacta cacacaaaga gaagcaagtg ggggcgggac tgcacggcac
10321 ttgtgagaca cctcaggtcg acccggatcg aggtgggcgg gaacaacggt ctcgataatg
10381 tacgtacact acacacaaag agaagcaagt gggggcggga ctgcacggca cttgtgagac
10441 acctcaggga tccccggatc gaggtgggcg ggaacaacgg tctcgataat gtacgtacac
10501 tacacacaaa gagaagcaag tgggggcggg actgcacggc acttgtgaga cacctcagtc
10561 tagaccggat cgaggtgggc gggaacaacg gtctcgataa tgtacgtaca ctacacacaa
10621 agagaagcaa gtgggggcgg gactgcacgg cacttgtgag acacctcagg agctcccgga
10681 tcgaggtggg cgggaacaac ggtctcgata atgtacgtac actacacaca aagagaagca
10741 agtgggggcg ggactgcacg gcacttgtga gacacctcag gtcgacccgg atcgaggtgg
10801 gcgggaacaa cggtctcgat aatgtacgta cactacacac aaagagaagc aagtgggggc
10861 gggactgcac ggcacttgtg agacacctca gggatccccg gatcgaggtg ggcgggaaca
10921 acggtctcga taatgtacgt acactacaca caaagagaag caagtggggg cgggactgca
10981 cggcacttgt gagacacctc agtctagacc ggatcgaggt gggcgggaac aacggtctcg
11041 ataatgtacg tacactacac acaaagagaa gcaagtgggg gcgggactgc acggcacttg
11101 tgagacacct caggaattcg ggtaccgcat gcttccttgg taccttcgaa cccgggggat
11161 ccccctttca tcaaatttag ggatgccatc aactttcagt tcataattaa tatcttacca
11221 aattaggtaa tctgcaaaag ttcagactgt gaaatgtaac attttatata tcaagctcta
11281 tttaacgcct cacagtagtt aacataaaga gatacagaat tgtcgtgtca gtgtatacta
11341 tccatgtgta tactctggat atccatttgt attccattat ctacgaaaag cgggtaccga
11401 gctcgaattc actggccgtc gttttacaac gtcgtgactg ggaaaaccct ggcgttaccc
11461 aacttaatcg ccttgcagca catccccctt tcgccagctg gcgtaatagc gaagaggccc
11521 gcaccgatcg cccttcccaa cagttgcgca gcctgaatgg cgaatggcgc ctgatgcggt
11581 attttctcct tacgcatctg tgcggtattt cacaccgcat atggtgcact ctcagtacaa
11641 tctgctctga tgccgcatag ttaagccagc cccgacaccc gccaacaccc gctgacgcgc
11701 cctgacgggc ttgtctgctc ccggcatccg cttacagaca agctgtgacc gtctccggga
11761 gctgcatgtg tcagaggttt tcaccgtcat caccgaaacg cgcga
//
