| 质粒名称: | EC2_10_dCas9_VPR_HC_sgRNA |
|---|---|
| 出品公司: | Addgene |
| 目录编号: | 163712 |
| 存储实验室: | Yvonne Nygård |
|
载体骨架基本信息 |
|
| 载体名称: | MoClo-YTK |
| 载体出品公司: | -- |
| 空载体大小: | 11420 bp |
| 完整质粒大小: | -- |
| 载体修饰: | -- |
| 载体类型: | 酵母表达 |
| 筛选标记: | 新霉素（G418） |
| 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: | dCas9,Nuclease-deficient Cas9 |
| 物种来源: | Streptococcus pyogenes |
| 插入大小: | 4101 bp |
| 基因突变: | 点突变D10A和H840A |
| GenBank编号 | -- |
| 启动子: | ScTEF1 |
| 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 | Golden Gate Assembly |
| 3’测序引物 | -- |
|
文献引用方法 |
|
| 材料和方法部分: | EC2_10_dCas9_VPR_HC_sgRNA was a gift from Yvonne Nygård (Addgene plasmid # 163712 ; http://n2t.net/addgene:163712 ; RRID:Addgene_163712) |
| 参考文献部分: | A CRISPR activation and interference toolkit for industrial Saccharomyces cerevisiae strain KE6-12. Camara E, Lenitz I, Nygard Y. Sci Rep. 2020 Sep 3;10(1):14605. doi: 10.1038/s41598-020-71648-w. 10.1038/s41598-020-71648-w PubMed 32884066 |
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 EC2_10_dCas9_VPR_HC_sgRNA 11420 bp ds-DNA circular SYN 01-NOV-2021
DEFINITION Yeast high copy plasmid with dCas9, sgRNA expression cassette and
VPR activation domains.
ACCESSION .
VERSION .
KEYWORDS EC2_10_dCas9_VPR_HC_sgRNA
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 11420)
AUTHORS Camara E, Lenitz I, Nygard Y
TITLE A CRISPR activation and interference toolkit for industrial
Saccharomyces cerevisiae strain KE6-12.
JOURNAL Sci Rep. 2020 Sep 3;10(1):14605. doi: 10.1038/s41598-020-71648-w.
PUBMED 32884066
REFERENCE 2 (bases 1 to 11420)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Nov 1, 2021 from SnapGene Server 1.1.58
http://www.snapgene.com
FEATURES Location/Qualifiers
source 1..11420
/organism="synthetic DNA construct"
/mol_type="other DNA"
promoter 328..735
/gene="S. cerevisiae TEF1"
/label=TEF1 promoter
/note="promoter for EF-1-alpha"
CDS 758..778
/codon_start=1
/product="nuclear localization signal of SV40 (simian virus
40) large T antigen"
/label=SV40 NLS
/translation="PKKKRKV"
CDS 4886..4906
/codon_start=1
/product="nuclear localization signal of SV40 (simian virus
40) large T antigen"
/label=SV40 NLS
/translation="PKKKRKV"
CDS 4913..5062
/codon_start=1
/product="tetrameric repeat of the minimal activation
domain of herpes simplex virus VP16 (Beerli et al., 1998)"
/label=VP64
/translation="DALDDFDLDMLGSDALDDFDLDMLGSDALDDFDLDMLGSDALDDF
DLDML"
CDS 5087..5107
/codon_start=1
/product="nuclear localization signal of SV40 (simian virus
40) large T antigen"
/label=SV40 NLS
/translation="PKKKRKV"
CDS 5537..5893
/codon_start=1
/product="transcriptional activation domain of human RelA,
also known as p65 (O'Shea and Perkins, 2008)"
/label=RelA (p65) AD
/translation="PTQAGEGTLSEALLQLQFDDEDLGALLGNSTDPAVFTDLASVDNS
EFQQLLNQGIPVAPHTTEPMLMEYPEAITRLVTGAQRPPDPAPAPLGAPGLPNGLLSGD
EDFSSIADMDFSALL"
CDS 5912..6481
/codon_start=1
/product="transcriptional activation domain from the human
herpesvirus 4 (Epstein-Barr virus) replication and
transcription activator Rta/BRLF1 (Hardwick et al., 1992;
Chavez et al., 2015)"
/label=Rta AD
/translation="RDSREGMFLPKPEAGSAISDVFEGREVCQPKRIRPFHPPGSPWAN
RPLPASLAPTPTGPVHEPVGSLTPAPVPQPLDPAPAVTPEASHLLEDPDEETSQAVKAL
REMADTVIPQKEEAAICGQMDLSHPPPRGHLDELTTTLESMTEDLNLDSPLTPELNEIL
DTFLNDECLLHAMHISTGLSIFDTSLF"
misc_RNA 7043..7118
/label=gRNA scaffold
/note="guide RNA scaffold for the Streptococcus pyogenes
CRISPR/Cas9 system"
CDS 7629..8438
/codon_start=1
/gene="aph(3')-Ia"
/product="aminoglycoside phosphotransferase"
/label=KanR
/note="confers resistance to kanamycin"
/translation="MGKEKTHVSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYGKP
DAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGKTA
FQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDASD
FDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGIAD
RYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
terminator 8443..8640
/label=TEF terminator
/note="Ashbya gossypii TEF terminator"
rep_origin 8645..9627
/label=2u ori
/note="yeast 2u plasmid origin of replication"
CDS complement(9685..10500)
/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 10492..10511
/label=pENTR-R
/note="pENTR vectors, reverse primer"
rep_origin 10725..11313
/direction=RIGHT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind 11214..11233
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
ORIGIN
1 ccctgaattc gcatctagac tgaactggcc gataattgca gacgaacgcc ttgccaacag
61 ggagttcttc agagacatgg aggctcaaaa cgaaattatt gacagcctag acatcaatag
121 tcatacaaca gaaagcgacc acccaacttt ggctgataat agcgtataaa caatgcatac
181 tttgtacgtt caaaatacaa tgcagtagat atatttatgc atattacata taatacatat
241 cacataggaa gcaacaggcg cgttggactt ttaattttcg aggaccgcga atccttacat
301 cacacccaat cccccacaag tgatccccca cacaccatag cttcaaaatg tttctactcc
361 ttttttactc ttccagattt tctcggactc cgcgcatcgc cgtaccactt caaaacaccc
421 aagcacagca tactaaattt cccctctttc ttcctctagg gtgtcgttaa ttacccgtac
481 taaaggtttg gaaaagaaaa aagacaccgc ctcgtttctt tttcttcgtc gaaaaaggca
541 ataaaaattt ttatcacgtt tctttttctt gaaaattttt ttttttgatt tttttctctt
601 tcgatgacct cccattgata tttaagttaa taaacggtca tcaatttctc aagtttcagt
661 ttcatttttc ttgttctatt acaacttttt ttacttcttg ctcattagaa agaaagcata
721 gcaatctaat ctaagtttta attacaaaag atctatgcca aagaagaaga gaaaggtagg
781 cagcgacaag aagtattcta tcggactggc catcgggact aatagcgtcg ggtgggccgt
841 gatcactgac gagtacaagg tgccctctaa gaagttcaag gtgctcggga acaccgaccg
901 gcattccatc aagaaaaatc tgatcggagc tctcctcttt gattcagggg aaaccgctga
961 agcaacccgc ctcaagcgga ctgctagacg gcggtacacc aggaggaaga accggatttg
1021 ttaccttcaa gagatattct ccaacgaaat ggcaaaggtc gacgacagct tcttccatag
1081 gctggaagaa tcattcctcg tggaagagga taagaagcat gaacggcatc ccatcttcgg
1141 taatatcgtc gacgaggtgg cctatcacga gaaataccca accatctacc atcttcgcaa
1201 aaagctggtg gactcaaccg acaaggcaga cctccggctt atctacctgg ccctggccca
1261 catgatcaag ttcagaggcc acttcctgat cgagggcgac ctcaatcctg acaatagcga
1321 tgtggataaa ctgttcatcc agctggtgca gacttacaac cagctctttg aagagaaccc
1381 catcaatgca agcggagtcg atgccaaggc cattctgtca gcccggctgt caaagagccg
1441 cagacttgag aatcttatcg ctcagctgcc gggtgaaaag aaaaatggac tgttcgggaa
1501 cctgattgct ctttcacttg ggctgactcc caatttcaag tctaatttcg acctggcaga
1561 ggatgccaag ctgcaactgt ccaaggacac ctatgatgac gatctcgaca acctcctggc
1621 ccagatcggt gaccaatacg ccgacctttt ccttgctgct aagaatcttt ctgacgccat
1681 cctgctgtct gacattctcc gcgtgaacac tgaaatcacc aaggcccctc tttcagcttc
1741 aatgattaag cggtatgatg agcaccacca ggacctgacc ctgcttaagg cactcgtccg
1801 gcagcagctt ccggagaagt acaaggaaat cttctttgac cagtcaaaga atggatacgc
1861 cggctacatc gacggaggtg cctcccaaga ggaattttat aagtttatca aacctatcct
1921 tgagaagatg gacggcaccg aagagctcct cgtgaaactg aatcgggagg atctgctgcg
1981 gaagcagcgc actttcgaca atgggagcat tccccaccag atccatcttg gggagcttca
2041 cgccatcctt cggcgccaag aggacttcta cccctttctt aaggacaaca gggagaagat
2101 tgagaaaatt ctcactttcc gcatccccta ctacgtggga cccctcgcca gaggaaatag
2161 ccggtttgct tggatgacca gaaagtcaga agaaactatc actccctgga acttcgaaga
2221 ggtggtggac aagggagcca gcgctcagtc attcatcgaa cggatgacta acttcgataa
2281 gaacctcccc aatgagaagg tcctgccgaa acattccctg ctctacgagt actttaccgt
2341 gtacaacgag ctgaccaagg tgaaatatgt caccgaaggg atgaggaagc ccgcattcct
2401 gtcaggcgaa caaaagaagg caattgtgga ccttctgttc aagaccaata gaaaggtgac
2461 cgtgaagcag ctgaaggagg actatttcaa gaaaattgaa tgcttcgact ctgtggagat
2521 tagcggggtc gaagatcggt tcaacgcaag cctgggtacc taccatgatc tgcttaagat
2581 catcaaggac aaggattttc tggacaatga ggagaaagag gacatccttg aggacattgt
2641 cctgactctc actctgttcg aggaccggga aatgatcgag gagaggctta agacctacgc
2701 ccatctgttc gacgataaag tgatgaagca acttaaacgg agaagatata ccggatgggg
2761 acgccttagc cgcaaactca tcaacggaat ccgggacaaa cagagcggaa agaccattct
2821 tgatttcctt aagagcgacg gattcgctaa tcgcaacttc atgcaactta tccatgatga
2881 ttccctgacc tttaaggagg acatccagaa ggcccaagtg tctggacaag gtgactcact
2941 gcacgagcat atcgcaaatc tggctggttc acccgctatt aagaagggta ttctccagac
3001 cgtgaaagtc gtggacgagc tggtcaaggt gatgggtcgc cataaaccag agaacattgt
3061 catcgagatg gccagggaaa accagactac ccagaaggga cagaagaaca gcagggagcg
3121 gatgaaaaga attgaggaag ggattaagga gctcgggtca cagatcctta aagagcaccc
3181 ggtggaaaac acccagcttc agaatgagaa gctctatctg tactaccttc aaaatggacg
3241 cgatatgtat gtggaccaag agcttgatat caacaggctc tcagactacg acgtggacgc
3301 catcgtccct cagagcttcc tcaaagacga ctcaattgac aataaggtgc tgactcgctc
3361 agacaagaac cggggaaagt cagataacgt gccctcagag gaagtcgtga aaaagatgaa
3421 gaactattgg cgccagcttc tgaacgcaaa gctgatcact cagcggaagt tcgacaatct
3481 cactaaggct gagaggggcg gactgagcga actggacaaa gcaggattca ttaaacggca
3541 acttgtggag actcggcaga ttactaaaca tgtagcccaa atccttgact cacgcatgaa
3601 taccaagtac gacgaaaacg acaaacttat ccgcgaggtg aaggtgatta ccctgaagtc
3661 caagctggtc agcgatttca gaaaggactt tcaattctac aaagtgcggg agatcaataa
3721 ctatcatcat gctcatgacg catatctgaa tgccgtggtg ggaaccgccc tgatcaagaa
3781 gtacccaaag ctggaaagcg agttcgtgta cggagactac aaggtctacg acgtgcgcaa
3841 gatgattgcc aaatctgagc aggagatcgg aaaggccacc gcaaagtact tcttctacag
3901 caacatcatg aatttcttca agaccgaaat cacccttgca aacggtgaga tccggaagag
3961 gccgctcatc gagactaatg gggagactgg cgaaatcgtg tgggacaagg gcagagattt
4021 cgctaccgtg cgcaaagtgc tttctatgcc tcaagtgaac atcgtgaaga aaaccgaggt
4081 gcaaaccgga ggcttttcta aggaatcaat cctccccaag cgcaactccg acaagctcat
4141 tgcaaggaag aaggattggg accctaagaa gtacggcgga ttcgattcac caactgtggc
4201 ttattctgtc ctggtcgtgg ctaaggtgga aaaaggaaag tctaagaagc tcaagagcgt
4261 gaaggaactg ctgggtatca ccattatgga gcgcagctcc ttcgagaaga acccaattga
4321 ctttctcgaa gccaaaggtt acaaggaagt caagaaggac cttatcatca agctcccaaa
4381 gtatagcctg ttcgaactgg agaatgggcg gaagcggatg ctcgcctccg ctggcgaact
4441 tcagaagggt aatgagctgg ctctcccctc caagtacgtg aatttcctct accttgcaag
4501 ccattacgag aagctgaagg ggagccccga ggacaacgag caaaagcaac tgtttgtgga
4561 gcagcataag cattatctgg acgagatcat tgagcagatt tccgagtttt ctaaacgcgt
4621 cattctcgct gatgccaacc tcgataaagt ccttagcgca tacaataagc acagagacaa
4681 accaattcgg gagcaggctg agaatatcat ccacctgttc accctcacca atcttggtgc
4741 ccctgccgca ttcaagtact tcgacaccac catcgaccgg aaacgctata cctccaccaa
4801 agaagtgctg gacgccaccc tcatccacca gagcatcacc ggactttacg aaactcggat
4861 tgacctctca cagctcggag gggatcccaa gaaaaagcgc aaggtaggat ccgacgcatt
4921 ggacgatttt gatctggata tgctgggaag tgacgccctc gatgattttg accttgacat
4981 gcttggttcg gatgcccttg atgactttga cctcgacatg ctcggcagtg acgcccttga
5041 tgatttcgac ctggacatgc tgattaactc tagaagttcc ggatctccga aaaagaaacg
5101 caaagttggt agccagtacc tgcccgacac cgacgaccgg caccggatcg aggaaaagcg
5161 gaagcggacc tacgagacat tcaagagcat catgaagaag tcccccttca gcggccccac
5221 cgaccctaga cctccaccta gaagaatcgc cgtgcccagc agatccagcg ccagcgtgcc
5281 aaaacctgcc ccccagcctt accccttcac cagcagcctg agcaccatca actacgacga
5341 gttccctacc atggtgttcc ccagcggcca gatctctcag gcctctgctc tggctccagc
5401 ccctcctcag gtgctgcctc aggctcctgc tcctgcacca gctccagcca tggtgtctgc
5461 actggctcag gcaccagcac ccgtgcctgt gctggctcct ggacctccac aggctgtggc
5521 tccaccagcc cctaaaccta cacaggccgg cgagggcaca ctgtctgaag ctctgctgca
5581 gctgcagttc gacgacgagg atctgggagc cctgctggga aacagcaccg atcctgccgt
5641 gttcaccgac ctggccagcg tggacaacag cgagttccag cagctgctga accagggcat
5701 ccctgtggcc cctcacacca ccgagcccat gctgatggaa taccccgagg ccatcacccg
5761 gctcgtgaca ggcgctcaga ggcctcctga tccagctcct gcccctctgg gagcaccagg
5821 cctgcctaat ggactgctgt ctggcgacga ggacttcagc tctatcgccg acatggactt
5881 ctccgcactg ctgggtagcg gatcgggatc tcgggattcc agggaaggga tgtttttgcc
5941 gaagcctgag gccggctccg ctattagtga cgtgtttgag ggccgcgagg tgtgccagcc
6001 aaaacgaatc cggccatttc atcctccagg aagtccatgg gccaaccgcc cactccccgc
6061 cagcctcgca ccaacaccaa ccggtccagt acatgagcca gtcgggtcac tgaccccggc
6121 accagtccct cagccactgg atccagcgcc cgcagtgact cccgaggcca gtcacctgtt
6181 ggaggatccc gatgaagaaa cgagccaggc tgtcaaagcc cttcgggaga tggccgatac
6241 tgtgattccc cagaaggaag aggctgcaat ctgtggccaa atggaccttt cccatccgcc
6301 cccaaggggc catctggatg agctgacaac cacacttgag tccatgaccg aggatctgaa
6361 cctggactca cccctgaccc cggaattgaa cgagattctg gataccttcc tgaacgacga
6421 gtgcctcttg catgccatgc atatcagcac aggactgtcc atcttcgaca catctctgtt
6481 ttaactcgag tggcataaag caatcttgat gaggataatg attttttttt gaatatacat
6541 aaatactacc gtttttctgc tagattttgt gatgacgtaa ataagtacat attacttttt
6601 aagccaagac aagattaagc attaacttta cccttttctt tctaagtttc aatattagtt
6661 atcactgttt aaaagttatg gcgagaacgt cggcggttaa aatatattac cctgaacggc
6721 tgccaaacca gatgtcaaca cagctacaac gatgtgcttc agtattacat tttttgcctt
6781 caacgccttg attgttctat ttttgctaat aataaatcta tttcatcgga ctaaaagtcc
6841 attagttgta agcggattta gctcagttgg gagagcgcca gactgaagaa aaacttcggt
6901 caagtcatct ggaggtcctg tgttcgatcc acagaattcg cagatggccg gcatggtccc
6961 agcctcctcg ctggcgccgg ctgggcaaca ccttcgggtg gcgaatggga ctcgctatcg
7021 tcttctcacg aagacttact cagttttaga gctagaaata gcaagttaaa ataaggctag
7081 tccgttatca acttgaaaaa gtggcaccga gtcggtgctt tttttatttt ttgtcactat
7141 tgttatgtaa aatgccacct ctgacagtat ggaacgcaaa cttctgtcta gtggatagct
7201 gagcagttac agagatgtta cgaaccacta gtgcactgca gtacagttta gcttgcctcg
7261 tccccgccgg gtcacccggc cagcgacatg gaggcccaga ataccctcct tgacagtctt
7321 gacgtgcgca gctcaggggc atgatgtgac tgtcgcccgt acatttagcc catacatccc
7381 catgtataat catttgcatc catacatttt gatggccgca cggcgcgaag caaaaattac
7441 ggctcctcgc tccagacctg cgagcaggga aacgctcccc tcacagacgc gttgaattgt
7501 ccccacgccg cgcccctgta gagaaatata aaaggttagg atttgccact gaggttcttc
7561 tttcatatac ttccttttaa aatcttgcta ggatacagtt ctcacatcac atccgaacat
7621 aaacaaaaat gggtaaggaa aagactcacg tttcgaggcc gcgattaaat tccaacatgg
7681 atgctgattt atatgggtat aaatgggctc gcgataatgt cgggcaatca ggtgcgacaa
7741 tctatcgatt gtatgggaag cccgatgcgc cagagttgtt tctgaaacat ggcaaaggta
7801 gcgttgccaa tgatgttaca gatgagatgg tcagactaaa ctggctgacg gaatttatgc
7861 ctcttccgac catcaagcat tttatccgta ctcctgatga tgcatggtta ctcaccactg
7921 cgatccccgg caaaacagca ttccaggtat tagaagaata tcctgattca ggtgaaaata
7981 ttgttgatgc gctggcagtg ttcctgcgcc ggttgcattc gattcctgtt tgtaattgtc
8041 cttttaacag cgatcgcgta tttcgtctgg ctcaggcgca atcacgaatg aataacggtt
8101 tggttgatgc gagtgatttt gatgacgagc gtaatggctg gcctgttgaa caagtctgga
8161 aagaaatgca taagcttttg ccattctcac cggattcagt cgtcactcat ggtgatttct
8221 cacttgataa ccttattttt gacgagggga aattaatagg ttgtattgat gttggacgag
8281 tcggaatcgc agaccgatac caggatcttg ccatcctatg gaactgcctc ggtgagtttt
8341 ctccttcatt acagaaacgg ctttttcaaa aatatggtat tgataatcct gatatgaata
8401 aattgcagtt tcatttgatg ctcgatgagt ttttctaaag taactgacaa taaaaagatt
8461 cttgttttca agaacttgtc atttgtatag tttttttata ttgtagttgt tctattttaa
8521 tcaaatgtta gcgtgattta tatttttttt cgcctcgaca tcatctgccc agatgcgaag
8581 ttaagtgcgc agaaagtaat atcatgcgtc aatcgtatgt gaatgctggt cgctatactg
8641 gagtaacgaa gcatctgtgc ttcattttgt agaacaaaaa tgcaacgcga gagcgctaat
8701 ttttcaaaca aagaatctga gctgcatttt tacagaacag aaatgcaacg cgaaagcgct
8761 attttaccaa cgaagaatct gtgcttcatt tttgtaaaac aaaaatgcaa cgcgagagcg
8821 ctaatttttc aaacaaagaa tctgagctgc atttttacag aacagaaatg caacgcgaga
8881 gcgctatttt accaacaaag aatctatact tcttttttgt tctacaaaaa tgcatcccga
8941 gagcgctatt tttctaacaa agcatcttag attacttttt ttctcctttg tgcgctctat
9001 aatgcagtct cttgataact ttttgcactg taggtccgtt aaggttagaa gaaggctact
9061 ttggtgtcta ttttctcttc cataaaaaaa gcctgactcc acttcccgcg tttactgatt
9121 actagcgaag ctgcgggtgc attttttcaa gataaaggca tccccgatta tattctatac
9181 cgatgtggat tgcgcatact ttgtgaacag aaagtgatag cgttgatgat tcttcattgg
9241 tcagaaaatt atgaacggtt tcttctattt tgtctctata tactacgtat aggaaatgtt
9301 tacattttcg tattgttttc gattcactct atgaatagtt cttactacaa tttttttgtc
9361 taaagagtaa tactagagat aaacataaaa aatgtagagg tcgagtttag atgcaagttc
9421 aaggagcgaa aggtggatgg gtaggttata tagggatata gcacagagat atatagcaaa
9481 gagatacttt tgagcaatgt ttgtggaagc ggtattcgca atattttagt agctcgttac
9541 agtccggtgc gtttttggtt ttttgaaagt gcgtcatcag agcgcttttg gttttcaaaa
9601 gcgctctgaa gttcctatac tttctagcta gagaatagga acttcccgag cggccgcgtg
9661 ttacaaccaa ttaaccaatt ctgattagaa aaactcatcg agcatcaaat gaaactgcaa
9721 tttattcata tcaggattat caataccata tttttgaaaa agccgtttct gtaatgaagg
9781 agaaaactca ccgaggcagt tccataggat ggcaagatcc tggtatcggt ctgcgattcc
9841 gactcgtcca acatcaatac aacctattaa tttcccctcg tcaaaaataa ggttatcaag
9901 tgagaaatca ccatgagtga cgactgaatc cggtgagaat ggcaaaagct tatgcatttc
9961 tttccagact tgttcaacag gccagccatt acgctcgtca tcaaaatcac tcgcatcaac
10021 caaaccgtta ttcattcgtg attgcgcctg agcgaggcga aatacgcgat cgctgttaaa
10081 aggacaatta caaacaggaa tcgaatgcaa ccggcgcagg aacactgcca gcgcatcaac
10141 aatattttca cctgaatcag gatattcttc taatacctgg aatgctgttt tcccggggat
10201 cgcagtggtg agtaaccatg catcatcagg agtacggata aaatgcttga tggtcggaag
10261 aggcataaat tccgtcagcc agtttagtct gaccatctca tctgtaacat cattggcaac
10321 gctacctttg ccatgtttca gaaacaactc tggcgcatcg ggcttcccat acaatcgata
10381 gattgtcgca cctgattgcc cgacattatc gcgagcccat ttatacccat ataaatcagc
10441 atccatgttg gaatttaatc gcggcctgga gcaagacgtt tcccgttgaa tatggctcat
10501 aacacccctt gtattactgt ttatgtaagc agacagtttt attgttcatg atgatatatt
10561 tttatcttgt gcaatgtaac atcagagatt ttgagacaca acgtggcttt gttgaataaa
10621 tcgaactttt gctgagttga aggatcagtc atgaccaaaa tcccttaacg tgagttttcg
10681 ttccactgag cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga tccttttttt
10741 ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg
10801 ccggatcaag agctaccaac tctttttccg aaggtaactg gcttcagcag agcgcagata
10861 ccaaatactg ttcttctagt gtagccgtag ttaggccacc acttcaagaa ctctgtagca
10921 ccgcctacat acctcgctct gctaatcctg ttaccagtgg ctgctgccag tggcgataag
10981 tcgtgtctta ccgggttgga ctcaagacga tagttaccgg ataaggcgca gcggtcgggc
11041 tgaacggggg gttcgtgcac acagcccagc ttggagcgaa cgacctacac cgaactgaga
11101 tacctacagc gtgagctatg agaaagcgcc acgcttcccg aagggagaaa ggcggacagg
11161 tatccggtaa gcggcagggt cggaacagga gagcgcacga gggagcttcc agggggaaac
11221 gcctggtatc tttatagtcc tgtcgggttt cgccacctct gacttgagcg tcgatttttg
11281 tgatgctcgt caggggggcg gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg
11341 ttcctggcct tttgctggcc ttttgctcac atgttctttc ctgcgttatc ccctgattct
11401 gtggataacc gtgcggccgc
//
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