| 质粒名称: | pRCC-N |
|---|---|
| 出品公司: | 爱迪基因 |
| 目录编号: | 81192 |
| 存储实验室: | Eckhard Boles |
|
载体骨架基本信息 |
|
| 载体名称: | pRS42N |
| 载体出品公司: | Euroscarf |
| 载体抗性: | 氨苄青霉素 |
| 空载体大小: | -- |
| 完整质粒大小: | -- |
| 载体修饰: | -- |
| 载体类型: | 酵母表达，CRISPR |
| 筛选标记: | Nourseothricin |
| 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: | Cas9 |
| 物种来源: | S. pyogenes |
| 插入大小: | -- |
| 基因突变: | WT - codon optimized |
| GenBank编号 | -- |
| 启动子 | ROX3 |
| 标签/融合蛋白: | NTS（主干上的C终端） |
| 克隆方法: | Gibson克隆 |
| 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: | pRCC-N was a gift from Eckhard Boles (Addgene plasmid # 81192 ; http://n2t.net/addgene:81192 ; RRID:Addgene_81192) |
| 参考文献部分: | Simplified CRISPR-Cas genome editing for Saccharomyces cerevisiae. Generoso WC, Gottardi M, Oreb M, Boles E. J Microbiol Methods. 2016 Aug;127:203-5. doi: 10.1016/j.mimet.2016.06.020. Epub 2016 Jun 17. 10.1016/j.mimet.2016.06.020 PubMed 27327211 |
买家导航
pRCCN载体基本信息
pRCCN载体图谱质粒图谱和多克隆位点信息
pRCCN载体简介
pRCCN载体序列
LOCUS pRCC-N 10018 bp ds-DNA circular SYN 06-AUG-2018
DEFINITION Expression of Cas9 and gRNA cassette in S. cerevisiae;
Nourseothricin Resistance.
ACCESSION .
VERSION .
KEYWORDS pRCC-N
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 10018)
AUTHORS Generoso WC, Gottardi M, Oreb M, Boles E
TITLE Simplified CRISPR-Cas genome editing for Saccharomyces cerevisiae.
JOURNAL J Microbiol Methods. 2016 Aug;127:203-5. doi:
10.1016/j.mimet.2016.06.020. Epub 2016 Jun 17.
PUBMED 27327211
REFERENCE 2 (bases 1 to 10018)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Aug 6, 2018 from SnapGene Server 1.1.58
http://www.snapgene.com
FEATURES Location/Qualifiers
source 1..10018
/organism="synthetic DNA construct"
/mol_type="other DNA"
primer_bind complement(10..29)
/label=pRS-marker
/note="pRS vectors, use to sequence yeast selectable
marker"
primer_bind 129..151
/label=pGEX 3'
/note="pGEX vectors, reverse primer"
primer_bind complement(189..207)
/label=pBRforEco
/note="pBR322 vectors, upsteam of EcoRI site, forward
primer"
rep_origin 248..1590
/label=2u ori
/note="yeast 2u plasmid origin of replication"
protein_bind 580..627
/label=FRT
/bound_moiety="FLP recombinase from the Saccharomyces
cerevisiae 2u plasmid"
/note="FLP-mediated recombination occurs in the 8-bp core
sequence TCTAGAAA (Turan and Bode, 2011)."
promoter 1617..1721
/gene="bla"
/label=AmpR promoter
CDS 1722..2582
/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(1940..1959)
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
rep_origin 2753..3341
/direction=RIGHT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind 3242..3261
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
promoter 3378..3646
/label=SNR52 promoter
/note="promoter for the S. cerevisiae small nucleolar RNA
gene SNR52"
misc_RNA 3670..3744
/label=gRNA scaffold
/note="guide RNA scaffold for the Streptococcus pyogenes
CRISPR/Cas9 system"
terminator 3749..3768
/label=SUP4 terminator
/note="transcription terminator for the S. cerevisiae SUP4
tRNA gene"
terminator 3789..4036
/gene="S. cerevisiae CYC1"
/label=CYC1 terminator
/note="transcription terminator for CYC1"
primer_bind 4002..4020
/label=CYC1
/note="CYC1 transcription termination signal, reverse
primer"
CDS complement(4052..4072)
/codon_start=1
/product="nuclear localization signal of SV40 (simian virus
40) large T antigen"
/label=SV40 NLS
/translation="PKKKRKV"
CDS complement(4085..8188)
/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"
gene complement(8877..9996)
/label=natMX6
/note="yeast selectable marker conferring nourseothricin
resistance"
terminator 8877..9074
/label=TEF terminator
/note="Ashbya gossypii TEF terminator"
CDS complement(9080..9652)
/codon_start=1
/gene="Streptomyces noursei nat1"
/product="nourseothricin acetyltransferase"
/label=NrsR
/note="confers resistance to nourseothricin"
/translation="MGTTLDDTAYRYRTSVPGDAEAIEALDGSFTTDTVFRVTATGDGF
TLREVPVDPPLTKVFPDDESDDESDDGEDGDPDSRTFVAYGDDGDLAGFVVVSYSGWNR
RLTVEDIEVAPEHRGHGVGRALMGLATEFARERGAGHLWLEVTNVNAPAIHAYRRMGFT
LCGLDTALYDGTASDGEQALYMSMPCP"
promoter complement(9653..9996)
/label=TEF promoter
/note="Ashbya gossypii TEF promoter"
ORIGIN
1 cactctcagt acaatctgct ctgatgccgc atagttaagc cagccccgac acccgccaac
61 acccgctgac gcgccctgac gggcttgtct gctcccggca tccgcttaca gacaagctgt
121 gaccgtctcc gggagctgca tgtgtcagag gttttcaccg tcatcaccga aacgcgcgag
181 acgaaagggc ctcgtgatac gcctattttt ataggttaat gtcatgataa taatggtttc
241 ttagtatgat ccaatatcaa aggaaatgat agcattgaag gatgagacta atccaattga
301 ggagtggcag catatagaac agctaaaggg tagtgctgaa ggaagcatac gataccccgc
361 atggaatggg ataatatcac aggaggtact agactacctt tcatcctaca taaatagacg
421 catataagta cgcatttaag cataaacacg cactatgccg ttcttctcat gtatatatat
481 atacaggcaa cacgcagata taggtgcgac gtgaacagtg agctgtatgt gcgcagctcg
541 cgttgcattt tcggaagcgc tcgttttcgg aaacgctttg aagttcctat tccgaagttc
601 ctattctcta gaaagtatag gaacttcaga gcgcttttga aaaccaaaag cgctctgaag
661 acgcactttc aaaaaaccaa aaacgcaccg gactgtaacg agctactaaa atattgcgaa
721 taccgcttcc acaaacattg ctcaaaagta tctctttgct atatatctct gtgctatatc
781 cctatataac ctacccatcc acctttcgct ccttgaactt gcatctaaac tcgacctcta
841 cattttttat gtttatctct agtattactc tttagacaaa aaaattgtag taagaactat
901 tcatagagtg aatcgaaaac aatacgaaaa tgtaaacatt tcctatacgt agtatataga
961 gacaaaatag aagaaaccgt tcataatttt ctgaccaatg aagaatcatc aacgctatca
1021 ctttctgttc acaaagtatg cgcaatccac atcggtatag aatataatcg gggatgcctt
1081 tatcttgaaa aaatgcaccc gcagcttcgc tagtaatcag taaacgcggg aagtggagtc
1141 aggctttttt tatggaagag aaaatagaca ccaaagtagc cttcttctaa ccttaacgga
1201 cctacagtgc aaaaagttat caagagactg cattatagag cgcacaaagg agaaaaaaag
1261 taatctaaga tgctttgtta gaaaaatagc gctctcggga tgcatttttg tagaacaaaa
1321 aagaagtata gattctttgt tggtaaaata gcgctctcgc gttgcatttc tgttctgtaa
1381 aaatgcagct cagattcttt gtttgaaaaa ttagcgctct cgcgttgcat ttttgtttta
1441 caaaaatgaa gcacagattc ttcgttggta aaatagcgct ttcgcgttgc atttctgttc
1501 tgtaaaaatg cagctcagat tctttgtttg aaaaattagc gctctcgcgt tgcatttttg
1561 ttctacaaaa tgaagcacag atgcttcgtt caggtggcac ttttcgggga aatgtgcgcg
1621 gaacccctat ttgtttattt ttctaaatac attcaaatat gtatccgctc atgagacaat
1681 aaccctgata aatgcttcaa taatattgaa aaaggaagag tatgagtatt caacatttcc
1741 gtgtcgccct tattcccttt tttgcggcat tttgccttcc tgtttttgct cacccagaaa
1801 cgctggtgaa agtaaaagat gctgaagatc agttgggtgc acgagtgggt tacatcgaac
1861 tggatctcaa cagcggtaag atccttgaga gttttcgccc cgaagaacgt tttccaatga
1921 tgagcacttt taaagttctg ctatgtggcg cggtattatc ccgtattgac gccgggcaag
1981 agcaactcgg tcgccgcata cactattctc agaatgactt ggttgagtac tcaccagtca
2041 cagaaaagca tcttacggat ggcatgacag taagagaatt atgcagtgct gccataacca
2101 tgagtgataa cactgcggcc aacttacttc tgacaacgat cggaggaccg aaggagctaa
2161 ccgctttttt gcacaacatg ggggatcatg taactcgcct tgatcgttgg gaaccggagc
2221 tgaatgaagc cataccaaac gacgagcgtg acaccacgat gcctgtagca atggcaacaa
2281 cgttgcgcaa actattaact ggcgaactac ttactctagc ttcccggcaa caattaatag
2341 actggatgga ggcggataaa gttgcaggac cacttctgcg ctcggccctt ccggctggct
2401 ggtttattgc tgataaatct ggagccggtg agcgtgggtc tcgcggtatc attgcagcac
2461 tggggccaga tggtaagccc tcccgtatcg tagttatcta cacgacgggg agtcaggcaa
2521 ctatggatga acgaaataga cagatcgctg agataggtgc ctcactgatt aagcattggt
2581 aactgtcaga ccaagtttac tcatatatac tttagattga tttaaaactt catttttaat
2641 ttaaaaggat ctaggtgaag atcctttttg ataatctcat gaccaaaatc ccttaacgtg
2701 agttttcgtt ccactgagcg tcagaccccg tagaaaagat caaaggatct tcttgagatc
2761 ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg
2821 tttgtttgcc ggatcaagag ctaccaactc tttttccgaa ggtaactggc ttcagcagag
2881 cgcagatacc aaatactgtc cttctagtgt agccgtagtt aggccaccac ttcaagaact
2941 ctgtagcacc gcctacatac ctcgctctgc taatcctgtt accagtggct gctgccagtg
3001 gcgataagtc gtgtcttacc gggttggact caagacgata gttaccggat aaggcgcagc
3061 ggtcgggctg aacggggggt tcgtgcacac agcccagctt ggagcgaacg acctacaccg
3121 aactgagata cctacagcgt gagctatgag aaagcgccac gcttcccgaa gggagaaagg
3181 cggacaggta tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg gagcttccag
3241 ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg ccacctctga cttgagcgtc
3301 gatttttgtg atgctcgtca ggggggcgga gcctatggaa aaacgccagc aacgcgggct
3361 agggataaca gggtaattct ttgaaaagat aatgtatgat tatgctttca ctcatattta
3421 tacagaaact tgatgttttc tttcgagtat atacaaggtg attacatgta cgtttgaagt
3481 acaactctag attttgtagt gccctcttgg gctagcggta aaggtgcgca ttttttcaca
3541 ccctacaatg ttctgttcaa aagattttgg tcaaacgctg tagaagtgaa agttggtgcg
3601 catgtttcgg cgttcgaaac ttctccgcag tgaaagataa atgatcggat ccacgttctc
3661 tgcggccgcg ttttagagct agaaatagca agttaaaata aggctagtcc gttatcaact
3721 tgaaaaagtg gcaccgagtc ggtggtgctt tttttgtttt ttatgtctcg gcgaattggg
3781 taccggccgc aaattaaagc cttcgagcgt cccaaaacct tctcaagcaa ggttttcagt
3841 ataatgttac atgcgtacac gcgtctgtac agaaaaaaaa gaaaaatttg aaatataaat
3901 aacgttctta atactaacat aactataaaa aaataaatag ggacctagac ttcaggttgt
3961 ctaactcctt ccttttcggt tagagcggat gtggggggag ggcgtgaatg taagcgtgac
4021 ataactaatt acatgactcg aggtcgactt aaacctttct cttcttcttt gggtcagctc
4081 tagagtcacc acccaattga gacaagtcaa ttctggtttc gtacaaaccg gtaatagatt
4141 ggtgaatcaa ggtagcgtcc aaaacttcct tggtagaggt gtatctcttt ctgtcaatgg
4201 tggtgtcgaa gtacttgaaa gcagctggag cacccaagtt ggtcaaggtg aacaagtgaa
4261 taatgttttc agcttgttct ctaattggct tgtctctgtg cttgttgtaa gcagacaaaa
4321 ccttgtccaa gttagcgtca gccaaaataa ctctcttaga gaattcagaa atttgttcaa
4381 taatttcgtc caagtagtgc ttgtgttgtt caacgaacaa ttgcttttgt tcgttgtctt
4441 ctggagaacc cttcaacttt tcgtagtgag aagccaagta caagaagtta acgtacttag
4501 atggcaaagc caattcgtta cccttttgca attcaccagc agaagccaac attctctttc
4561 taccgttttc caattcgaac aaagagtact ttggcaactt aataatcaag tccttcttaa
4621 cttccttgta acccttagct tccaagaagt caattgggtt cttttcgaaa gaagatcttt
4681 ccataatggt aatacccaac aattccttaa cagacttcaa cttcttagac ttaccctttt
4741 caaccttagc aacaaccaaa acagagtaag caacggttgg agagtcgaaa ccaccgtact
4801 tctttgggtc ccagtccttc tttctagcaa tcaacttgtc agagtttctc tttggcaaaa
4861 tagattcctt agagaaacca ccggtttgaa cttcggtctt cttaacaatg ttaacttgtg
4921 gcatagacaa aacctttcta acggtagcga agtctctacc cttgtcccaa acaatttcac
4981 cggtttcacc gttggtttca atcaatggtc tctttctaat ttcaccgtta gccaaggtaa
5041 tttcggtctt gaagaagttc ataatgttag agtagaagaa gtacttagcg gtagccttac
5101 caatttcttg ttcagactta gcaatcatct ttctaacgtc gtaaaccttg tagtcaccgt
5161 aaacgaattc agattccaac tttgggtact tcttaatcaa agcggtacca acaacagcgt
5221 tcaagtaagc gtcgtgagcg tggtggtagt tgttaatttc tctaaccttg tagaattgga
5281 agtcctttct gaagtcagaa accaacttag acttcaaggt aataacctta acttctctaa
5341 tcaacttgtc gttttcgtcg tacttggtgt tcattctaga gtccaaaatt tgagcaacgt
5401 gcttggtaat ttgtctggtt tcaaccaatt gtctcttaat gaaaccagcc ttgtccaatt
5461 cagacaaacc acctctttca gccttggtca agttgtcgaa ctttctttgg gtaatcaact
5521 tagcgttcaa caattgtctc cagtagttct tcatcttctt aacaacttct tcagatggaa
5581 cgttgtcaga cttacctctg ttcttgtcag atctggtcaa aaccttgttg tcaatagagt
5641 cgtccttcaa gaaagattgt ggaacaatgt ggtcaacgtc gtagtcagac aatctgttaa
5701 tgtccaattc ttggtcaacg tacatgtctc taccgttttg caagtagtac aagtacaact
5761 tttcgttttg caattgggtg ttttcaactg ggtgttcctt caaaatttga gaacccaatt
5821 ccttaatacc ttcttcaatt ctcttcattc tttctctaga gttcttttga cccttttggg
5881 tggtttggtt ttctctagcc atttcaataa caatgttttc tggcttgtgt ctacccataa
5941 ccttaaccaa ttcgtcaaca accttaacgg tttgcaaaat acccttctta atagctggag
6001 aaccagccaa gttagcaatg tgttcgtgca aagagtcacc ttgaccagaa acttgagcct
6061 tttgaatgtc ttccttgaag gtcaaagagt cgtcgtgaat caattgcatg aagtttctgt
6121 tagcgaaacc gtcagacttc aagaagtcca aaatggtctt accagattgc ttgtctctaa
6181 taccgttaat caactttcta gacaatctac cccaaccggt gtatcttctt ctcttcaatt
6241 gcttcataac cttgtcgtcg aacaagtgag cgtaggtctt caatctttct tcaatcattt
6301 ctctgtcttc gaacaaggtc aaggtcaaaa caatgtcttc caaaatgtct tcgttttctt
6361 cgttgtccaa gaagtccttg tccttaataa tcttcaacaa gtcgtggtag gtacccaaag
6421 aagcgttgaa tctgtcttca acaccagaaa tttcaacaga gtcgaaacat tcaatcttct
6481 tgaagtagtc ttccttcaat tgcttaacgg taacctttct gttggtcttg aacaacaagt
6541 caacaatagc cttcttttgt tcaccagaca agaaagctgg ctttctcata ccttcggtaa
6601 cgtacttaac cttggtcaat tcgttgtaaa cggtgaagta ttcgtacaac aaagagtgct
6661 ttggcaaaac cttttcgttt ggcaagttct tgtcgaagtt ggtcattctt tcaatgaaag
6721 attgagcaga agcacccttg tcaacaactt cttcgaagtt ccatggggta atggtttctt
6781 cagactttct ggtcatccaa gcgaatctag agttacctct agccaatgga ccaacgtagt
6841 atggaattct gaaggtcaaa atcttttcaa tcttttctct gttgtccttc aagaatgggt
6901 agaagtcttc ttgtcttctc aaaatagcgt gcaattcacc caagtgaatt tggtgtggaa
6961 tagaaccgtt gtcgaaggtt ctttgctttc tcaacaagtc ttctctgttc aacttaacca
7021 acaattcttc ggtaccgtcc atcttttcca aaattggctt aatgaacttg tagaattctt
7081 cttgagaagc accaccgtca atgtaaccag cgtaaccgtt cttagattgg tcgaagaaaa
7141 tttccttgta cttttctggc aattgttgtc taaccaaagc cttcaacaag gtcaagtctt
7201 ggtggtgttc gtcgtatctc ttaatcatag aagcagacaa tggagccttg gtaatttcgg
7261 tgttaactct caaaatgtca gacaacaaaa tagcgtcaga caagttctta gcagccaaga
7321 acaagtcagc gtattggtca ccaatttgag ccaacaagtt gtccaagtcg tcgtcgtagg
7381 tgtccttaga caattgcaac ttagcgtctt cagccaagtc gaagttagac ttgaagtttg
7441 gggtcaaacc caaagacaaa gcaatcaagt taccgaacaa accgttcttc ttttcacctg
7501 gcaattgagc aatcaagttt tccaatcttc tagacttaga caatctagca gacaaaatag
7561 ccttagcgtc aacaccagaa gcgttaattg ggttttcttc gaacaattgg ttgtaggttt
7621 gaaccaattg aatgaacaac ttgtcaacgt cagagttgtc tgggttcaag tcaccttcaa
7681 tcaagaagtg acctctgaac ttaatcatgt gagccaaagc caagtaaatc aatctcaagt
7741 cagccttgtc ggtagagtca accaacttct ttctcaagtg gtaaatggtt gggtactttt
7801 cgtggtaagc aacttcgtca acaatgttac cgaaaattgg gtgtctttcg tgcttcttgt
7861 cttcttcaac caagaaagat tcttccaatc tgtggaagaa agagtcgtca accttagcca
7921 tttcgttaga gaaaatttct tgcaagtaac aaattctgtt ctttcttctg gtgtatcttc
7981 ttctagcggt tctcttcaat ctggtagctt cagcggtttc accagagtcg aacaacaaag
8041 caccaatcaa gttcttctta atagagtgtc tgtcggtgtt acccaaaacc ttgaacttct
8101 tagatggaac cttgtattcg tcggtaataa cagcccaacc aacagagttg gtaccaatgt
8161 ccaaaccaat agagtacttc ttgtccatat gtattatcgt attccttttt cgttcactta
8221 ctattgctat tcgcactcac tacagatcta tattttatgt tccaccctgt tgtcagatca
8281 ggaaaggaat gaagaggctc tatttaaacc aaatacacct cttcgtttct caatattgtt
8341 gggctttcct tttttttttc cttccacgtc tatctattgt ctctatcaaa attaggaaaa
8401 agaaatgaag aacgatgagc caaacgaggc tgagacaatc atgcatccgt acattctatg
8461 tatgtttggg tgtatctttc cagattcggt tctttttttt tttcagtcca attgaaaaaa
8521 aaaaggaaca ggaacaacgg gtcgaaattt gccgtcacat tcagagcgaa gagcgacagg
8581 ggaattggtg ggctaaccac ctgttgagcc cagctagcag ctgtcggtaa accgtcgttg
8641 ggcggggcgc gacaatgcag ccagcggagg cattgtatgg tttccgtctg gagagtgggc
8701 ttagccggcc tggcactcct tctctaagca gagtgagaag ggaatctttt tactggagaa
8761 gagtgtttga atccagcaga aggtaatacg ctgcggtatt ttctccttac gcatctgtgc
8821 ggtatttcac accgcgtttt cgacactgga tggcggcgtt agtatcgaat cgacagcagt
8881 atagcgacca gcattcacat acgattgacg catgatatta ctttctgcgc acttaacttc
8941 gcatctgggc agatgatgtc gaggcgaaaa aaaatataaa tcacgctaac atttgattaa
9001 aatagaacaa ctacaatata aaaaaactat acaaatgaca agttcttgaa aacaagaatc
9061 tttttattgt cagtactgat taggggcagg gcatgctcat gtagagcgcc tgctcgccgt
9121 ccgaggcggt gccgtcgtac agggcggtgt ccaggccgca gagggtgaac cccatccgcc
9181 ggtacgcgtg gatcgccggt gcgttgacgt tggtgacctc cagccagagg tgcccggcgc
9241 cccgctcgcg ggcgaactcc gtcgcgagcc ccatcaacgc gcgcccgacc ccgtgccccc
9301 ggtgctccgg ggcgacctcg atgtcctcga cggtcagccg gcggttccag ccggagtacg
9361 agacgaccac gaagcccgcc aggtcgccgt cgtccccgta cgcgacgaac gtccgggagt
9421 ccgggtcgcc gtcctccccg tcgtccgatt cgtcgtccga ttcgtcgtcg gggaacacct
9481 tggtcagggg cgggtccacc ggcacctccc gcagggtgaa gccgtccccg gtggcggtga
9541 cgcggaagac ggtgtcggtg gtgaaggacc catccagtgc ctcgatggcc tcggcgtccc
9601 ccgggacact ggtgcggtac cggtaagccg tgtcgtcaag agtggtaccc atggttgttt
9661 atgttcggat gtgatgtgag aactgtatcc tagcaagatt ttaaaaggaa gtatatgaaa
9721 gaagaacctc agtggcaaat cctaaccttt tatatttctc tacaggggcg cggcgtgggg
9781 acaattcaac gcgtctgtga ggggagcgtt tccctgctcg caggtctgca gcgaggagcc
9841 gtaatttttg cttcgcgccg tgcggccatc aaaatgtatg gatgcaaatg attatacatg
9901 gggatgtatg ggctaaatgt acgggcgaca gtcacatcat gcccctgagc tgcgcacgtc
9961 aagactgtca aggagggtat tctgggcctc catgtcgctg attaccctgt tatcccta
//
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