| 质粒名称: | pHCas9-Nours |
|---|---|
| 出品公司: | Addgene |
| 目录编号: | 107733 |
| 存储实验室: | Sheng Yang |
|
载体骨架基本信息 |
|
| 载体名称: | pSH47 |
| 空载体大小: | 6979bp |
| 完整质粒大小: | 10749 bp |
| 载体类型: | Yeast Expression, CRISPR |
| 筛选标记: | URA3 |
| 载体抗性: | 氨苄霉素 |
| 生长温度: | 37 ℃ |
| 宿主菌: | DH5a |
| 拷贝数: | 高拷贝 |
|
插入基因信息 |
|
| 插入基因名称: | Cas9 |
| 物种来源: | Human Optimized Streptococcus pyogenes Cas9 |
| 插入大小: | 4140 bp |
| 基因突变: | Human optimized |
| 启动子: | TEF1 |
| 标签/融合蛋白: |
SV40 NLS (N terminal on insert)
SV40 NLS (C terminal on insert)
|
| 克隆方法: | 未知 |
| 5’测序引物 | NA |
|
文献引用方法 |
|
| 材料和方法部分: | pHCas9-Nours was a gift from Sheng Yang (Addgene plasmid # 107733 ; http://n2t.net/addgene:107733 ; RRID:Addgene_107733) |
| 参考文献部分: | Unraveling the genetic basis of fast l-arabinose consumption on top of recombinant xylose-fermenting Saccharomyces cerevisiae. Wang X, Yang J, Yang S, Jiang Y. Biotechnol Bioeng. 2018 Sep 10. doi: 10.1002/bit.26827. 10.1002/bit.26827 PubMed 30199094 |
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 pHCas9-Nours 10749 bp ds-DNA circular SYN 13-MAY-2021
DEFINITION Escherichia coli- S. cerevisiae shuttle plasmid harbors a Cas9 gene,
a natMX6 and URA3 selection markers used in S. cerevisiae.
ACCESSION .
VERSION .
KEYWORDS pHCas9-Nours
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 10749)
AUTHORS Wang X, Yang J, Yang S, Jiang Y
TITLE Unraveling the genetic basis of fast l-arabinose consumption on top
of recombinant xylose-fermenting Saccharomyces cerevisiae.
JOURNAL Biotechnol Bioeng. 2018 Sep 10. doi: 10.1002/bit.26827.
PUBMED 30199094
REFERENCE 2 (bases 1 to 10749)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported May 13, 2021 from SnapGene Server 1.1.58
http://www.biofeng.com
FEATURES Location/Qualifiers
source 1..10749
/organism="synthetic DNA construct"
/mol_type="other DNA"
promoter 1..400
/gene="S. cerevisiae TEF1"
/label=TEF1 promoter
/note="promoter for EF-1-alpha"
CDS 420..440
/codon_start=1
/product="nuclear localization signal of SV40 (simian virus
40) large T antigen"
/label=SV40 NLS
/translation="PKKKRKV"
CDS 441..4541
/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="DKKYSIGLDIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKKN
LIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEESF
LVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIKF
RGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRLE
NLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQI
GDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVRQ
QLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLLR
KQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARGN
SRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEYF
TVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFDS
VEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEERL
KTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFMQ
LIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGRH
KPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLYL
YYLQNGRDMYVDQELDINRLSDYDVDHIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVPS
EEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKHV
AQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYLN
AVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKTE
ITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSKE
SILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGIT
IMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNEL
ALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILADA
NLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVLD
ATLIHQSITGLYETRIDLSQLGGD"
CDS 4554..4574
/codon_start=1
/product="nuclear localization signal of SV40 (simian virus
40) large T antigen"
/label=SV40 NLS
/translation="PKKKRKV"
terminator 4590..4837
/gene="S. cerevisiae CYC1"
/label=CYC1 terminator
/note="transcription terminator for CYC1"
primer_bind complement(4606..4624)
/label=CYC1
/note="CYC1 transcription termination signal, reverse
primer"
primer_bind complement(4969..4988)
/label=F1ori-R
/note="F1 origin, reverse primer"
promoter 5059..5402
/label=TEF promoter
/note="Ashbya gossypii TEF promoter"
promoter 5403..5450
/label=EM7 promoter
/note="synthetic bacterial promoter "
CDS 5469..6041
/codon_start=1
/gene="Streptomyces noursei nat1"
/product="nourseothricin acetyltransferase"
/label=NrsR
/note="confers resistance to nourseothricin"
/translation="MGTTLDDTAYRYRTSVPGDAEAIEALDGSFTTDTVFRVTATGDGF
TLREVPVDPPLTKVFPDDESDDESDDGEDGDPDSRTFVAYGDDGDLAGFVVVSYSGWNR
RLTVEDIEVAPEHRGHGVGRALMGLATEFARERGAGHLWLEVTNVNAPAIHAYRRMGFT
LCGLDTALYDGTASDGEQALYMSMPCP"
terminator 6047..6244
/label=TEF terminator
/note="Ashbya gossypii TEF terminator"
primer_bind 6456..6477
/label=F1ori-F
/note="F1 origin, forward primer"
CDS complement(6745..7548)
/codon_start=1
/gene="S. cerevisiae URA3"
/product="orotidine-5'-phosphate decarboxylase, required
for uracil biosynthesis"
/label=URA3
/note="yeast auxotrophic marker, counterselectable with
5-fluoroorotic acid (5-FOA)"
/translation="MSKATYKERAATHPSPVAAKLFNIMHEKQTNLCASLDVRTTKELL
ELVEALGPKICLLKTHVDILTDFSMEGTVKPLKALSAKYNFLLFEDRKFADIGNTVKLQ
YSAGVYRIAEWADITNAHGVVGPGIVSGLKQAAEEVTKEPRGLLMLAELSCKGSLSTGE
YTKGTVDIAKSDKDFVIGFIAQRDMGGRDEGYDWLIMTPGVGLDDKGDALGQQYRTVDD
VVSTGSDIIIVGRGLFAKGRDAKVEGERYRKAGWEAYLRRCGQQN"
promoter complement(7549..7764)
/gene="S. cerevisiae URA3"
/label=URA3 promoter
primer_bind complement(7788..7807)
/label=pRS-marker
/note="pRS vectors, use to sequence yeast selectable
marker"
primer_bind 7907..7929
/label=pGEX 3'
/note="pGEX vectors, reverse primer"
primer_bind complement(7967..7985)
/label=pBRforEco
/note="pBR322 vectors, upsteam of EcoRI site, forward
primer"
misc_feature 8026..8529
/label=CEN/ARS
/note="S. cerevisiae CEN6 centromere fused to an
autonomously replicating sequence"
promoter 8566..8670
/gene="bla"
/label=AmpR promoter
CDS 8671..9531
/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(8889..8908)
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
rep_origin 9702..10290
/direction=RIGHT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind 10191..10210
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
primer_bind 10444..10461
/label=L4440
/note="L4440 vector, forward primer"
protein_bind 10578..10599
/label=CAP binding site
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding activates transcription in the presence
of cAMP."
promoter 10614..10644
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 10652..10668
/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 10657..10679
/label=M13/pUC Reverse
/note="In lacZ gene"
primer_bind 10676..10692
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
primer_bind 10676..10692
/label=M13 Reverse
/note="In lacZ gene. Also called M13-rev"
primer_bind 10711..10731
/label=T3
/note="T3 promoter, forward primer"
promoter 10713..10731
/label=T3 promoter
/note="promoter for bacteriophage T3 RNA polymerase"
ORIGIN
1 catagcttca aaatgtttct actccttttt tactcttcca gattttctcg gactccgcgc
61 atcgccgtac cacttcaaaa cacccaagca cagcatacta aatttcccct ctttcttcct
121 ctagggtgtc gttaattacc cgtactaaag gtttggaaaa gaaaaaagag accgcctcgt
181 ttctttttct tcgtcgaaaa aggcaataaa aatttttatc acgtttcttt ttcttgaaaa
241 tttttttttt tgattttttt ctctttcgat gacctcccat tgatatttaa gttaataaac
301 ggtcttcaat ttctcaagtt tcagtttcat ttttcttgtt ctattacaac tttttttact
361 tcttgctcat tagaaagaaa gcatagcaat ctaatctaag ttttaattac aaaatgcctc
421 caaaaaagaa gagaaaggtc gacaagaagt actccattgg gctcgatatc ggcacaaaca
481 gcgtcggttg ggccgtcatt acggacgagt acaaggtgcc gagcaaaaaa ttcaaagttc
541 tgggcaatac cgatcgccac agcataaaga agaacctcat tggcgccctc ctgttcgact
601 ccggggagac ggccgaagcc acgcggctca aaagaacagc acggcgcaga tatacccgca
661 gaaagaatcg gatctgctac ctgcaggaga tctttagtaa tgagatggct aaggtggatg
721 actctttctt ccataggctg gaggagtcct ttttggtgga ggaggataaa aagcacgagc
781 gccacccaat ctttggcaat atcgtggacg aggtggcgta ccatgaaaag tacccaacca
841 tatatcatct gaggaagaag cttgtagaca gtactgataa ggctgacttg cggttgatct
901 atctcgcgct ggcgcatatg atcaaatttc ggggacactt cctcatcgag ggggacctga
961 acccagacaa cagcgatgtc gacaaactct ttatccaact ggttcagact tacaatcagc
1021 ttttcgaaga gaacccgatc aacgcatccg gagttgacgc caaagcaatc ctgagcgcta
1081 ggctgtccaa atcccggcgg ctcgaaaacc tcatcgcaca gctccctggg gagaagaaga
1141 acggcctgtt tggtaatctt atcgccctgt cactcgggct gacccccaac tttaaatcta
1201 acttcgacct ggccgaagat gccaagcttc aactgagcaa agacacctac gatgatgatc
1261 tcgacaatct gctggcccag atcggcgacc agtacgcaga cctttttttg gcggcaaaga
1321 acctgtcaga cgccattctg ctgagtgata ttctgcgagt gaacacggag atcaccaaag
1381 ctccgctgag cgctagtatg atcaagcgct atgatgagca ccaccaagac ttgactttgc
1441 tgaaggccct tgtcagacag caactgcctg agaagtacaa ggaaattttc ttcgatcagt
1501 ctaaaaatgg ctacgccgga tacattgacg gcggagcaag ccaggaggaa ttttacaaat
1561 ttattaagcc catcttggaa aaaatggacg gcaccgagga gctgctggta aagcttaaca
1621 gagaagatct gttgcgcaaa cagcgcactt tcgacaatgg aagcatcccc caccagattc
1681 acctgggcga actgcacgct atcctcaggc ggcaagagga tttctacccc tttttgaaag
1741 ataacaggga aaagattgag aaaatcctca catttcggat accctactat gtaggccccc
1801 tcgcccgggg aaattccaga ttcgcgtgga tgactcgcaa atcagaagag accatcactc
1861 cctggaactt cgaggaagtc gtggataagg gggcctctgc ccagtccttc atcgaaagga
1921 tgactaactt tgataaaaat ctgcctaacg aaaaggtgct tcctaaacac tctctgctgt
1981 acgagtactt cacagtttat aacgagctca ccaaggtcaa atacgtcaca gaagggatga
2041 gaaagccagc attcctgtct ggagagcaga agaaagctat cgtggacctc ctcttcaaga
2101 cgaaccggaa agttaccgtg aaacagctca aagaagacta tttcaaaaag attgaatgtt
2161 tcgactctgt tgaaatcagc ggagtggagg atcgcttcaa cgcatccctg ggaacgtatc
2221 acgatctcct gaaaatcatt aaagacaagg acttcctgga caatgaggag aacgaggaca
2281 ttcttgagga cattgtcctc acccttacgt tgtttgaaga tagggagatg attgaagaac
2341 gcttgaaaac ttacgctcat ctcttcgacg acaaagtcat gaaacagctc aagaggcgcc
2401 gatatacagg atgggggcgg ctgtcaagaa aactgatcaa tgggatccga gacaagcaga
2461 gtggaaagac aatcctggat tttcttaagt ccgatggatt tgccaaccgg aacttcatgc
2521 agttgatcca tgatgactct ctcaccttta aggaggacat ccagaaagca caagtttctg
2581 gccaggggga cagtcttcac gagcacatcg ctaatcttgc aggtagccca gctatcaaaa
2641 agggaatact gcagaccgtt aaggtcgtgg atgaactcgt caaagtaatg ggaaggcata
2701 agcccgagaa tatcgttatc gagatggccc gagagaacca aactacccag aagggacaga
2761 agaacagtag ggaaaggatg aagaggattg aagagggtat aaaagaactg gggtcccaaa
2821 tccttaagga acacccagtt gaaaacaccc agcttcagaa tgagaagctc tacctgtact
2881 acctgcagaa cggcagggac atgtacgtgg atcaggaact ggacatcaat cggctctccg
2941 actacgacgt ggatcatatc gtgccccagt cttttctcaa agatgattct attgataata
3001 aagtgttgac aagatccgat aaaaatagag ggaagagtga taacgtcccc tcagaagaag
3061 ttgtcaagaa aatgaaaaat tattggcggc agctgctgaa cgccaaactg atcacacaac
3121 ggaagttcga taatctgact aaggctgaac gaggtggcct gtctgagttg gataaagccg
3181 gcttcatcaa aaggcagctt gttgagacac gccagatcac caagcacgtg gcccaaattc
3241 tcgattcacg catgaacacc aagtacgatg aaaatgacaa actgattcga gaggtgaaag
3301 ttattactct gaagtctaag ctggtctcag atttcagaaa ggactttcag ttttataagg
3361 tgagagagat caacaattac caccatgcgc atgatgccta cctgaatgca gtggtaggca
3421 ctgcacttat caaaaaatat cccaagcttg aatctgaatt tgtttacgga gactataaag
3481 tgtacgatgt taggaaaatg atcgcaaagt ctgagcagga aataggcaag gccaccgcta
3541 agtacttctt ttacagcaat attatgaatt ttttcaagac cgagattaca ctggccaatg
3601 gagagattcg gaagcgacca cttatcgaaa caaacggaga aacaggagaa atcgtgtggg
3661 acaagggtag ggatttcgcg acagtccgga aggtcctgtc catgccgcag gtgaacatcg
3721 ttaaaaagac cgaagtacag accggaggct tctccaagga aagtatcctc ccgaaaagga
3781 acagcgacaa gctgatcgca cgcaaaaaag attgggaccc caagaaatac ggcggattcg
3841 attctcctac agtcgcttac agtgtactgg ttgtggccaa agtggagaaa gggaagtcta
3901 aaaaactcaa aagcgtcaag gaactgctgg gcatcacaat catggagcga tcaagcttcg
3961 aaaaaaaccc catcgacttt ctcgaggcga aaggatataa agaggtcaaa aaagacctca
4021 tcattaagct tcccaagtac tctctctttg agcttgaaaa cggccggaaa cgaatgctcg
4081 ctagtgcggg cgagctgcag aaaggtaacg agctggcact gccctctaaa tacgttaatt
4141 tcttgtatct ggccagccac tatgaaaagc tcaaagggtc tcccgaagat aatgagcaga
4201 agcagctgtt cgtggaacaa cacaaacact accttgatga gatcatcgag caaataagcg
4261 aattctccaa aagagtgatc ctcgccgacg ctaacctcga taaggtgctt tctgcttaca
4321 ataagcacag ggataagccc atcagggagc aggcagaaaa cattatccac ttgtttactc
4381 tgaccaactt gggcgcgcct gcagccttca agtacttcga caccaccata gacagaaagc
4441 ggtacacctc tacaaaggag gtcctggacg ccacactgat tcatcagtca attacggggc
4501 tctatgaaac aagaatcgac ctctctcagc tcggtggaga cagcagggct gaccccaaga
4561 agaagaggaa ggtgtgatct cttctcgagt catgtaatta gttatgtcac gcttacattc
4621 acgccctccc cccacatccg ctctaaccga aaaggaagga gttagacaac ctgaagtcta
4681 ggtccctatt tattttttta tagttatgtt agtattaaga acgttattta tatttcaaat
4741 ttttcttttt tttctgtaca gacgcgtgta cgcatgtaac attatactga aaaccttgct
4801 tgagaaggtt ttgggacgct cgaaggcttt aatttgcggc cggtaccttc ccaacagttg
4861 cgcagcctga atggcgaatg gacgcgccct gtagcggcgc attaagcgcg gcgggtgtgg
4921 tggttacgcg cagcgtgacc gctacacttg ccagcgccct agcgcccgct cctttcgctt
4981 tcttcccttc ctttctcgcc acgttcgccc tagagatctg tttagcttgc ctcgtccccg
5041 ccgggtcacc cggccagcga catggaggcc cagaataccc tccttgacag tcttgacgtg
5101 cgcagctcag gggcatgatg tgactgtcgc ccgtacattt agcccataca tccccatgta
5161 taatcatttg catccataca ttttgatggc cgcacggcgc gaagcaaaaa ttacggctcc
5221 tcgctgcaga cctgcgagca gggaaacgct cccctcacag acgcgttgaa ttgtccccac
5281 gccgcgcccc tgtagagaaa tataaaaggt taggatttgc cactgaggtt cttctttcat
5341 atacttcctt ttaaaatctt gctaggatac agttctcaca tcacatccga acataaacaa
5401 ccgttgacaa ttaatcatcg gcatagtata tcggcatagt ataatacgac aaggtgagga
5461 actaaaccat gggtaccact cttgacgaca cggcttaccg gtaccgcacc agtgtcccgg
5521 gggacgccga ggccatcgag gcactggatg ggtccttcac caccgacacc gtcttccgcg
5581 tcaccgccac cggggacggc ttcaccctgc gggaggtgcc ggtggacccg cccctgacca
5641 aggtgttccc cgacgacgaa tcggacgacg aatcggacga cggggaggac ggcgacccgg
5701 actcccggac gttcgtcgcg tacggggacg acggcgacct ggcgggcttc gtggtcgtct
5761 cgtactccgg ctggaaccgc cggctgaccg tcgaggacat cgaggtcgcc ccggagcacc
5821 gggggcacgg ggtcgggcgc gcgttgatgg ggctcgcgac ggagttcgcc cgcgagcggg
5881 gcgccgggca cctctggctg gaggtcacca acgtcaacgc accggcgatc cacgcgtacc
5941 ggcggatggg gttcaccctc tgcggcctgg acaccgccct gtacgacggc accgcctcgg
6001 acggcgagca ggcgctctac atgagcatgc cctgccccta atcagtactg acaataaaaa
6061 gattcttgtt ttcaagaact tgtcatttgt atagtttttt tatattgtag ttgttctatt
6121 ttaatcaaat gttagcgtga tttatatttt ttttcgcctc gacatcatct gcccagatgc
6181 gaagttaagt gcgcagaaag taatatcatg cgtcaatcgt atgtgaatgc tggtcgctat
6241 actgctgtcg attcgatact aacgccgcca tccagtgtcg aaaacgggct ttccccgtca
6301 agctctaaat cgggggctcc ctttagggtt ccgatttagt gctttacggc acctcgaccc
6361 caaaaaactt gattagggtg atggttcacg tagtgggcca tcgccctgat agacggtttt
6421 tcgccctttg acgttggagt ccacgttctt taatagtgga ctcttgttcc aaactggaac
6481 aacactcaac cctatctcgg tctattcttt tgatttataa gggattttgc cgatttcggc
6541 ctattggtta aaaaatgagc tgatttaaca aaaatttaac gcgaatttta acaaaatatt
6601 aacgcttaca atttcctgat gcggtatttt ctccttacgc atctgtgcgg tatttcacac
6661 cgcatagggt aataactgat ataattaaat tgaagctcta atttgtgagt ttagtataca
6721 tgcatttact tataatacag ttttttagtt ttgctggccg catcttctca aatatgcttc
6781 ccagcctgct tttctgtaac gttcaccctc taccttagca tcccttccct ttgcaaatag
6841 tcctcttcca acaataataa tgtcagatcc tgtagagacc acatcatcca cggttctata
6901 ctgttgaccc aatgcgtctc ccttgtcatc taaacccaca ccgggtgtca taatcaacca
6961 atcgtaacct tcatctcttc cacccatgtc tctttgagca ataaagccga taacaaaatc
7021 tttgtcgctc ttcgcaatgt caacagtacc cttagtatat tctccagtag atagggagcc
7081 cttgcatgac aattctgcta acatcaaaag gcctctaggt tcctttgtta cttcttctgc
7141 cgcctgcttc aaaccgctaa caatacctgg gcccaccaca ccgtgtgcat tcgtaatgtc
7201 tgcccattct gctattctgt atacacccgc agagtactgc aatttgactg tattaccaat
7261 gtcagcaaat tttctgtctt cgaagagtaa aaaattgtac ttggcggata atgcctttag
7321 cggcttaact gtgccctcca tggaaaaatc agtcaagata tccacatgtg tttttagtaa
7381 acaaattttg ggacctaatg cttcaactaa ctccagtaat tccttggtgg tacgaacatc
7441 caatgaagca cacaagtttg tttgcttttc gtgcatgata ttaaatagct tggcagcaac
7501 aggactagga tgagtagcag cacgttcctt atatgtagct ttcgacatga tttatcttcg
7561 tttcctgcag gtttttgttc tgtgcagttg ggttaagaat actgggcaat ttcatgtttc
7621 ttcaacacta catatgcgta tatataccaa tctaagtctg tgctccttcc ttcgttcttc
7681 cttctgttcg gagattaccg aatcaaaaaa atttcaagga aaccgaaatc aaaaaaaaga
7741 ataaaaaaaa aatgatgaat tgaaaaggtg gtatggtgca ctctcagtac aatctgctct
7801 gatgccgcat agttaagcca gccccgacac ccgccaacac ccgctgacgc gccctgacgg
7861 gcttgtctgc tcccggcatc cgcttacaga caagctgtga ccgtctccgg gagctgcatg
7921 tgtcagaggt tttcaccgtc atcaccgaaa cgcgcgagac gaaagggcct cgtgatacgc
7981 ctatttttat aggttaatgt catgataata atggtttctt agacggatcg cttgcctgta
8041 acttacacgc gcctcgtatc ttttaatgat ggaataattt gggaatttac tctgtgttta
8101 tttattttta tgttttgtat ttggatttta gaaagtaaat aaagaaggta gaagagttac
8161 ggaatgaaga aaaaaaaata aacaaaggtt taaaaaattt caacaaaaag cgtactttac
8221 atatatattt attagacaag aaaagcagat taaatagata tacattcgat taacgataag
8281 taaaatgtaa aatcacagga ttttcgtgtg tggtcttcta cacagacaag atgaaacaat
8341 tcggcattaa tacctgagag caggaagagc aagataaaag gtagtatttg ttggcgatcc
8401 ccctagagtc ttttacatct tcggaaaaca aaaactattt tttctttaat ttcttttttt
8461 actttctatt tttaatttat atatttatat taaaaaattt aaattataat tatttttata
8521 gcacgtgatg aaaaggaccc aggtggcact tttcggggaa atgtgcgcgg aacccctatt
8581 tgtttatttt tctaaataca ttcaaatatg tatccgctca tgagacaata accctgataa
8641 atgcttcaat aatattgaaa aaggaagagt atgagtattc aacatttccg tgtcgccctt
8701 attccctttt ttgcggcatt ttgccttcct gtttttgctc acccagaaac gctggtgaaa
8761 gtaaaagatg ctgaagatca gttgggtgca cgagtgggtt acatcgaact ggatctcaac
8821 agcggtaaga tccttgagag ttttcgcccc gaagaacgtt ttccaatgat gagcactttt
8881 aaagttctgc tatgtggcgc ggtattatcc cgtattgacg ccgggcaaga gcaactcggt
8941 cgccgcatac actattctca gaatgacttg gttgagtact caccagtcac agaaaagcat
9001 cttacggatg gcatgacagt aagagaatta tgcagtgctg ccataaccat gagtgataac
9061 actgcggcca acttacttct gacaacgatc ggaggaccga aggagctaac cgcttttttg
9121 cacaacatgg gggatcatgt aactcgcctt gatcgttggg aaccggagct gaatgaagcc
9181 ataccaaacg acgagcgtga caccacgatg cctgtagcaa tggcaacaac gttgcgcaaa
9241 ctattaactg gcgaactact tactctagct tcccggcaac aattaataga ctggatggag
9301 gcggataaag ttgcaggacc acttctgcgc tcggcccttc cggctggctg gtttattgct
9361 gataaatctg gagccggtga gcgtgggtct cgcggtatca ttgcagcact ggggccagat
9421 ggtaagccct cccgtatcgt agttatctac acgacgggga gtcaggcaac tatggatgaa
9481 cgaaatagac agatcgctga gataggtgcc tcactgatta agcattggta actgtcagac
9541 caagtttact catatatact ttagattgat ttaaaacttc atttttaatt taaaaggatc
9601 taggtgaaga tcctttttga taatctcatg accaaaatcc cttaacgtga gttttcgttc
9661 cactgagcgt cagaccccgt agaaaagatc aaaggatctt cttgagatcc tttttttctg
9721 cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg
9781 gatcaagagc taccaactct ttttccgaag gtaactggct tcagcagagc gcagatacca
9841 aatactgttc ttctagtgta gccgtagtta ggccaccact tcaagaactc tgtagcaccg
9901 cctacatacc tcgctctgct aatcctgtta ccagtggctg ctgccagtgg cgataagtcg
9961 tgtcttaccg ggttggactc aagacgatag ttaccggata aggcgcagcg gtcgggctga
10021 acggggggtt cgtgcacaca gcccagcttg gagcgaacga cctacaccga actgagatac
10081 ctacagcgtg agctatgaga aagcgccacg cttcccgaag ggagaaaggc ggacaggtat
10141 ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg agcttccagg gggaaacgcc
10201 tggtatcttt atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg atttttgtga
10261 tgctcgtcag gggggcggag cctatggaaa aacgccagca acgcggcctt tttacggttc
10321 ctggcctttt gctggccttt tgctcacatg ttctttcctg cgttatcccc tgattctgtg
10381 gataaccgta ttaccgcctt tgagtgagct gataccgctc gccgcagccg aacgaccgag
10441 cgcagcgagt cagtgagcga ggaagcggaa gagcgcccaa tacgcaaacc gcctctcccc
10501 gcgcgttggc cgattcatta atgcagctgg cacgacaggt ttcccgactg gaaagcgggc
10561 agtgagcgca acgcaattaa tgtgagttag ctcactcatt aggcacccca ggctttacac
10621 tttatgcttc cggctcgtat gttgtgtgga attgtgagcg gataacaatt tcacacagga
10681 aacagctatg accatgatta cgccaagcgc gcaattaacc ctcactaaag ggaacaaaag
10741 ctggagctc
//
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