別稱: | Plasmid#82416;pLentiCRISPRV2-GFP |
---|---|
啟動(dòng)子: | U6 |
復(fù)制子: | pUC |
原核抗性: | Amp |
篩選標(biāo)記: | 博來霉素Zeo,綠色熒光GFP |
克隆菌株: | Stbl3 |
培養(yǎng)條件: | 37度 |
載體宿主: | 哺乳細(xì)胞,慢病毒 |
---|---|
載體用途: | 基因敲除 |
基因種屬: | 空載體 |
基因類型: | CRISPR,Cas9,sgRNA |
原核抗性: | Amp |
真核抗性: | |
熒光蛋白: | 綠色 |
LentiCRISPRV2GFP是一個(gè)哺乳細(xì)胞Cas9和gRNA表達(dá)質(zhì)粒。
LOCUS Exported 13131 bp ds-DNA circular SYN 07-JUN-2018
DEFINITION synthetic circular DNA
ACCESSION .
VERSION .
KEYWORDS LentiCRISPRv2GFP
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 13131)
AUTHORS Triple Threat
TITLE Direct Submission
FEATURES Location/Qualifiers
source 1..13131
/organism="synthetic DNA construct"
/mol_type="other DNA"
promoter 1..241
/note="U6 promoter"
/note="RNA polymerase III promoter for human U6 snRNA"
misc_RNA 266..341
/note="gRNA scaffold"
/note="guide RNA scaffold for the Streptococcus pyogenes
CRISPR/Cas9 system"
promoter 403..614
/note="EF-1-alpha core promoter"
/note="core promoter for human elongation factor
EF-1-alpha"
CDS 639..4742
/codon_start=1
/product="Cas9 (Csn1) endonuclease from the Streptococcus
pyogenes Type II CRISPR/Cas system"
/note="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"
CDS 4743..4790
/codon_start=1
/product="bipartite nuclear localization signal from
nucleoplasmin"
/note="nucleoplasmin NLS"
/translation="KRPAATKKAGQAKKKK"
CDS 4791..4814
/codon_start=1
/product="FLAG(R) epitope tag, followed by an enterokinase
cleavage site"
/note="FLAG"
/translation="DYKDDDDK"
CDS 4824..4880
/codon_start=1
/product="2A peptide from porcine teschovirus-1
polyprotein"
/note="P2A"
/note="Eukaryotic ribosomes fail to insert a peptide bond
between the Gly and Pro residues, yielding separate
polypeptides."
/translation="ATNFSLLKQAGDVEENPGP"
misc_feature 5616..6204
/note="WPRE"
/note="woodchuck hepatitis virus posttranscriptional
regulatory element"
LTR 6276..6509
/note="3' LTR (Delta-U3)"
/note="self-inactivating 3' long terminal repeat (LTR) from
HIV-1"
polyA_signal 6541..6765
/note="bGH poly(A) signal"
/note="bovine growth hormone polyadenylation signal"
rep_origin 6811..7239
/direction=RIGHT
/note="f1 ori"
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
promoter 7253..7582
/note="SV40 promoter"
/note="SV40 enhancer and early promoter"
rep_origin 7433..7568
/note="SV40 ori"
/note="SV40 origin of replication"
promoter 7630..7677
/note="EM7 promoter"
/note="synthetic bacterial promoter "
CDS 7696..8070
/codon_start=1
/gene="Sh ble from Streptoalloteichus hindustanus"
/product="antibiotic-binding protein"
/note="Zeo"
/note="confers resistance to bleomycin, phleomycin, and
Zeocin(TM)"
/translation="MAKLTSAVPVLTARDVAGAVEFWTDRLGFSRDFVEDDFAGVVRDD
VTLFISAVQDQVVPDNTLAWVWVRGLDELYAEWSEVVSTNFRDASGPAMTEIGEQPWGR
EFALRDPAGNCVHFVAEEQD"
polyA_signal 8200..8321
/note="SV40 poly(A) signal"
/note="SV40 polyadenylation signal"
rep_origin complement(8772..9360)
/direction=LEFT
/note="ori"
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
CDS complement(9531..10391)
/codon_start=1
/gene="bla"
/product="beta-lactamase"
/note="AmpR"
/note="confers resistance to ampicillin, carbenicillin, and
related antibiotics"
/translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
LIKHW"
promoter complement(10392..10496)
/gene="bla"
/note="AmpR promoter"
enhancer 10762..11141
/note="CMV enhancer"
/note="human cytomegalovirus immediate early enhancer"
promoter 11143..11341
/note="CMV promoter"
/note="human cytomegalovirus (CMV) immediate early
promoter"
LTR 11359..11539
/note="5' LTR (truncated)"
/note="truncated 5' long terminal repeat (LTR) from HIV-1"
misc_feature 11586..11711
/note="HIV-1 Psi"
/note="packaging signal of human immunodeficiency virus
type 1"
misc_feature 12204..12437
/note="RRE"
/note="The Rev response element (RRE) of HIV-1 allows for
Rev-dependent mRNA export from the nucleus to the
cytoplasm."
misc_feature 12964..13081
/note="cPPT/CTS"
/note="central polypurine tract and central termination
sequence of HIV-1"
ORIGIN
1 gagggcctat ttcccatgat tccttcatat ttgcatatac gatacaaggc tgttagagag
61 ataattagaa ttaatttgac tgtaaacaca aagatattag tacaaaatac gtgacgtaga
121 aagtaataat ttcttgggta gtttgcagtt ttaaaattat gttttaaaat ggactatcat
181 atgcttaccg taacttgaaa gtatttcgat ttcttggctt tatatatctt gtggaaagga
241 cgaaacaccg gagacggacg tctctgtttt agagctagaa atagcaagtt aaaataaggc
301 tagtccgtta tcaacttgaa aaagtggcac cgagtcggtg cttttttgaa ttcgctagct
361 aggtcttgaa aggagtggga attggctccg gtgcccgtca gtgggcagag cgcacatcgc
421 ccacagtccc cgagaagttg gggggagggg tcggcaattg atccggtgcc tagagaaggt
481 ggcgcggggt aaactgggaa agtgatgtcg tgtactggct ccgccttttt cccgagggtg
541 ggggagaacc gtatataagt gcagtagtcg ccgtgaacgt tctttttcgc aacgggtttg
601 ccgccagaac acaggaccgg ttctagagcg ctgccaccat ggacaagaag tacagcatcg
661 gcctggacat cggcaccaac tctgtgggct gggccgtgat caccgacgag tacaaggtgc
721 ccagcaagaa attcaaggtg ctgggcaaca ccgaccggca cagcatcaag aagaacctga
781 tcggagccct gctgttcgac agcggcgaaa cagccgaggc cacccggctg aagagaaccg
841 ccagaagaag atacaccaga cggaagaacc ggatctgcta tctgcaagag atcttcagca
901 acgagatggc caaggtggac gacagcttct tccacagact ggaagagtcc ttcctggtgg
961 aagaggataa gaagcacgag cggcacccca tcttcggcaa catcgtggac gaggtggcct
1021 accacgagaa gtaccccacc atctaccacc tgagaaagaa actggtggac agcaccgaca
1081 aggccgacct gcggctgatc tatctggccc tggcccacat gatcaagttc cggggccact
1141 tcctgatcga gggcgacctg aaccccgaca acagcgacgt ggacaagctg ttcatccagc
1201 tggtgcagac ctacaaccag ctgttcgagg aaaaccccat caacgccagc ggcgtggacg
1261 ccaaggccat cctgtctgcc agactgagca agagcagacg gctggaaaat ctgatcgccc
1321 agctgcccgg cgagaagaag aatggcctgt tcggaaacct gattgccctg agcctgggcc
1381 tgacccccaa cttcaagagc aacttcgacc tggccgagga tgccaaactg cagctgagca
1441 aggacaccta cgacgacgac ctggacaacc tgctggccca gatcggcgac cagtacgccg
1501 acctgtttct ggccgccaag aacctgtccg acgccatcct gctgagcgac atcctgagag
1561 tgaacaccga gatcaccaag gcccccctga gcgcctctat gatcaagaga tacgacgagc
1621 accaccagga cctgaccctg ctgaaagctc tcgtgcggca gcagctgcct gagaagtaca
1681 aagagatttt cttcgaccag agcaagaacg gctacgccgg ctacattgac ggcggagcca
1741 gccaggaaga gttctacaag ttcatcaagc ccatcctgga aaagatggac ggcaccgagg
1801 aactgctcgt gaagctgaac agagaggacc tgctgcggaa gcagcggacc ttcgacaacg
1861 gcagcatccc ccaccagatc cacctgggag agctgcacgc cattctgcgg cggcaggaag
1921 atttttaccc attcctgaag gacaaccggg aaaagatcga gaagatcctg accttccgca
1981 tcccctacta cgtgggccct ctggccaggg gaaacagcag attcgcctgg atgaccagaa
2041 agagcgagga aaccatcacc ccctggaact tcgaggaagt ggtggacaag ggcgcttccg
2101 cccagagctt catcgagcgg atgaccaact tcgataagaa cctgcccaac gagaaggtgc
2161 tgcccaagca cagcctgctg tacgagtact tcaccgtgta taacgagctg accaaagtga
2221 aatacgtgac cgagggaatg agaaagcccg ccttcctgag cggcgagcag aaaaaggcca
2281 tcgtggacct gctgttcaag accaaccgga aagtgaccgt gaagcagctg aaagaggact
2341 acttcaagaa aatcgagtgc ttcgactccg tggaaatctc cggcgtggaa gatcggttca
2401 acgcctccct gggcacatac cacgatctgc tgaaaattat caaggacaag gacttcctgg
2461 acaatgagga aaacgaggac attctggaag atatcgtgct gaccctgaca ctgtttgagg
2521 acagagagat gatcgaggaa cggctgaaaa cctatgccca cctgttcgac gacaaagtga
2581 tgaagcagct gaagcggcgg agatacaccg gctggggcag gctgagccgg aagctgatca
2641 acggcatccg ggacaagcag tccggcaaga caatcctgga tttcctgaag tccgacggct
2701 tcgccaacag aaacttcatg cagctgatcc acgacgacag cctgaccttt aaagaggaca
2761 tccagaaagc ccaggtgtcc ggccagggcg atagcctgca cgagcacatt gccaatctgg
2821 ccggcagccc cgccattaag aagggcatcc tgcagacagt gaaggtggtg gacgagctcg
2881 tgaaagtgat gggccggcac aagcccgaga acatcgtgat cgaaatggcc agagagaacc
2941 agaccaccca gaagggacag aagaacagcc gcgagagaat gaagcggatc gaagagggca
3001 tcaaagagct gggcagccag atcctgaaag aacaccccgt ggaaaacacc cagctgcaga
3061 acgagaagct gtacctgtac tacctgcaga atgggcggga tatgtacgtg gaccaggaac
3121 tggacatcaa ccggctgtcc gactacgatg tggaccatat cgtgcctcag agctttctga
3181 aggacgactc catcgacaac aaggtgctga ccagaagcga caagaaccgg ggcaagagcg
3241 acaacgtgcc ctccgaagag gtcgtgaaga agatgaagaa ctactggcgg cagctgctga
3301 acgccaagct gattacccag agaaagttcg acaatctgac caaggccgag agaggcggcc
3361 tgagcgaact ggataaggcc ggcttcatca agagacagct ggtggaaacc cggcagatca
3421 caaagcacgt ggcacagatc ctggactccc ggatgaacac taagtacgac gagaatgaca
3481 agctgatccg ggaagtgaaa gtgatcaccc tgaagtccaa gctggtgtcc gatttccgga
3541 aggatttcca gttttacaaa gtgcgcgaga tcaacaacta ccaccacgcc cacgacgcct
3601 acctgaacgc cgtcgtggga accgccctga tcaaaaagta ccctaagctg gaaagcgagt
3661 tcgtgtacgg cgactacaag gtgtacgacg tgcggaagat gatcgccaag agcgagcagg
3721 aaatcggcaa ggctaccgcc aagtacttct tctacagcaa catcatgaac tttttcaaga
3781 ccgagattac cctggccaac ggcgagatcc ggaagcggcc tctgatcgag acaaacggcg
3841 aaaccgggga gatcgtgtgg gataagggcc gggattttgc caccgtgcgg aaagtgctga
3901 gcatgcccca agtgaatatc gtgaaaaaga ccgaggtgca gacaggcggc ttcagcaaag
3961 agtctatcct gcccaagagg aacagcgata agctgatcgc cagaaagaag gactgggacc
4021 ctaagaagta cggcggcttc gacagcccca ccgtggccta ttctgtgctg gtggtggcca
4081 aagtggaaaa gggcaagtcc aagaaactga agagtgtgaa agagctgctg gggatcacca
4141 tcatggaaag aagcagcttc gagaagaatc ccatcgactt tctggaagcc aagggctaca
4201 aagaagtgaa aaaggacctg atcatcaagc tgcctaagta ctccctgttc gagctggaaa
4261 acggccggaa gagaatgctg gcctctgccg gcgaactgca gaagggaaac gaactggccc
4321 tgccctccaa atatgtgaac ttcctgtacc tggccagcca ctatgagaag ctgaagggct
4381 cccccgagga taatgagcag aaacagctgt ttgtggaaca gcacaagcac tacctggacg
4441 agatcatcga gcagatcagc gagttctcca agagagtgat cctggccgac gctaatctgg
4501 acaaagtgct gtccgcctac aacaagcacc gggataagcc catcagagag caggccgaga
4561 atatcatcca cctgtttacc ctgaccaatc tgggagcccc tgccgccttc aagtactttg
4621 acaccaccat cgaccggaag aggtacacca gcaccaaaga ggtgctggac gccaccctga
4681 tccaccagag catcaccggc ctgtacgaga cacggatcga cctgtctcag ctgggaggcg
4741 acaagcgacc tgccgccaca aagaaggctg gacaggctaa gaagaagaaa gattacaaag
4801 acgatgacga taagggatcc ggcgcaacaa acttctctct gctgaaacaa gccggagatg
4861 tcgaagagaa tcctggaccg atggtgtcta agggcgaaga gctgttcacc ggggtggtgc
4921 ccatcctggt cgagctggac ggcgacgtaa acggccacaa gttcagcgtg tccggcgagg
4981 gcgagggcga tgccacctac ggcaagctga ccctgaagtt catctgcacc accggcaagc
5041 tgcccgtgcc ctggcccacc ctcgtgacca ccctgaccta cggcgtgcag tgcttcagcc
5101 gctaccccga ccacatgaag cagcacgact tcttcaagtc cgccatgccc gaaggctacg
5161 tccaggagcg caccatcttc ttcaaggacg acggcaacta caagacccgc gccgaggtga
5221 agttcgaggg cgacaccctg gtgaaccgca tcgagctgaa gggcatcgac ttcaaggagg
5281 acggcaacat cctggggcac aagctggagt acaactacaa cagccacaac gtctatatca
5341 tggccgacaa gcagaagaac ggcatcaagg tgaacttcaa gatccgccac aacatcgagg
5401 acggcagcgt gcagctcgcc gaccactacc agcagaacac ccccatcggc gacggccccg
5461 tgctgctgcc cgacaaccac tacctgagca cccagtccgc cctgagcaaa gaccccaacg
5521 agaagcgcga tcacatggtc ctgctggagt tcgtgaccgc cgccgggatc actctcggca
5581 tggacgagct gtacaagtaa acgcgttaag tcgacaatca acctctggat tacaaaattt
5641 gtgaaagatt gactggtatt cttaactatg ttgctccttt tacgctatgt ggatacgctg
5701 ctttaatgcc tttgtatcat gctattgctt cccgtatggc tttcattttc tcctccttgt
5761 ataaatcctg gttgctgtct ctttatgagg agttgtggcc cgttgtcagg caacgtggcg
5821 tggtgtgcac tgtgtttgct gacgcaaccc ccactggttg gggcattgcc accacctgtc
5881 agctcctttc cgggactttc gctttccccc tccctattgc cacggcggaa ctcatcgccg
5941 cctgccttgc ccgctgctgg acaggggctc ggctgttggg cactgacaat tccgtggtgt
6001 tgtcggggaa atcatcgtcc tttccttggc tgctcgcctg tgttgccacc tggattctgc
6061 gcgggacgtc cttctgctac gtcccttcgg ccctcaatcc agcggacctt ccttcccgcg
6121 gcctgctgcc ggctctgcgg cctcttccgc gtcttcgcct tcgccctcag acgagtcgga
6181 tctccctttg ggccgcctcc ccgcgtcgac tttaagacca atgacttaca aggcagctgt
6241 agatcttagc cactttttaa aagaaaaggg gggactggaa gggctaattc actcccaacg
6301 aagacaagat ctgctttttg cttgtactgg gtctctctgg ttagaccaga tctgagcctg
6361 ggagctctct ggctaactag ggaacccact gcttaagcct caataaagct tgccttgagt
6421 gcttcaagta gtgtgtgccc gtctgttgtg tgactctggt aactagagat ccctcagacc
6481 cttttagtca gtgtggaaaa tctctagcag ggcccgttta aacccgctga tcagcctcga
6541 ctgtgccttc tagttgccag ccatctgttg tttgcccctc ccccgtgcct tccttgaccc
6601 tggaaggtgc cactcccact gtcctttcct aataaaatga ggaaattgca tcgcattgtc
6661 tgagtaggtg tcattctatt ctggggggtg gggtggggca ggacagcaag ggggaggatt
6721 gggaagacaa tagcaggcat gctggggatg cggtgggctc tatggcttct gaggcggaaa
6781 gaaccagctg gggctctagg gggtatcccc acgcgccctg tagcggcgca ttaagcgcgg
6841 cgggtgtggt ggttacgcgc agcgtgaccg ctacacttgc cagcgcccta gcgcccgctc
6901 ctttcgcttt cttcccttcc tttctcgcca cgttcgccgg ctttccccgt caagctctaa
6961 atcgggggct ccctttaggg ttccgattta gtgctttacg gcacctcgac cccaaaaaac
7021 ttgattaggg tgatggttca cgtagtgggc catcgccctg atagacggtt tttcgccctt
7081 tgacgttgga gtccacgttc tttaatagtg gactcttgtt ccaaactgga acaacactca
7141 accctatctc ggtctattct tttgatttat aagggatttt gccgatttcg gcctattggt
7201 taaaaaatga gctgatttaa caaaaattta acgcgaatta attctgtgga atgtgtgtca
7261 gttagggtgt ggaaagtccc caggctcccc agcaggcaga agtatgcaaa gcatgcatct
7321 caattagtca gcaaccaggt gtggaaagtc cccaggctcc ccagcaggca gaagtatgca
7381 aagcatgcat ctcaattagt cagcaaccat agtcccgccc ctaactccgc ccatcccgcc
7441 cctaactccg cccagttccg cccattctcc gccccatggc tgactaattt tttttattta
7501 tgcagaggcc gaggccgcct ctgcctctga gctattccag aagtagtgag gaggcttttt
7561 tggaggccta ggcttttgca aaaagctccc gggagcttgt atatccattt tcggatctga
7621 tcagcacgtg ttgacaatta atcatcggca tagtatatcg gcatagtata atacgacaag
7681 gtgaggaact aaaccatggc caagttgacc agtgccgttc cggtgctcac cgcgcgcgac
7741 gtcgccggag cggtcgagtt ctggaccgac cggctcgggt tctcccggga cttcgtggag
7801 gacgacttcg ccggtgtggt ccgggacgac gtgaccctgt tcatcagcgc ggtccaggac
7861 caggtggtgc cggacaacac cctggcctgg gtgtgggtgc gcggcctgga cgagctgtac
7921 gccgagtggt cggaggtcgt gtccacgaac ttccgggacg cctccgggcc ggccatgacc
7981 gagatcggcg agcagccgtg ggggcgggag ttcgccctgc gcgacccggc cggcaactgc
8041 gtgcacttcg tggccgagga gcaggactga cacgtgctac gagatttcga ttccaccgcc
8101 gccttctatg aaaggttggg cttcggaatc gttttccggg acgccggctg gatgatcctc
8161 cagcgcgggg atctcatgct ggagttcttc gcccacccca acttgtttat tgcagcttat
8221 aatggttaca aataaagcaa tagcatcaca aatttcacaa ataaagcatt tttttcactg
8281 cattctagtt gtggtttgtc caaactcatc aatgtatctt atcatgtctg tataccgtcg
8341 acctctagct agagcttggc gtaatcatgg tcatagctgt ttcctgtgtg aaattgttat
8401 ccgctcacaa ttccacacaa catacgagcc ggaagcataa agtgtaaagc ctggggtgcc
8461 taatgagtga gctaactcac attaattgcg ttgcgctcac tgcccgcttt ccagtcggga
8521 aacctgtcgt gccagctgca ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt
8581 attgggcgct cttccgcttc ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg
8641 cgagcggtat cagctcactc aaaggcggta atacggttat ccacagaatc aggggataac
8701 gcaggaaaga acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg
8761 ttgctggcgt ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca
8821 agtcagaggt ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc
8881 tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc
8941 ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta ggtatctcag ttcggtgtag
9001 gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc
9061 ttatccggta actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca
9121 gcagccactg gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg
9181 aagtggtggc ctaactacgg ctacactaga agaacagtat ttggtatctg cgctctgctg
9241 aagccagtta ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct
9301 ggtagcggtg gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa
9361 gaagatcctt tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa
9421 gggattttgg tcatgagatt atcaaaaagg atcttcacct agatcctttt aaattaaaaa
9481 tgaagtttta aatcaatcta aagtatatat gagtaaactt ggtctgacag ttaccaatgc
9541 ttaatcagtg aggcacctat ctcagcgatc tgtctatttc gttcatccat agttgcctga
9601 ctccccgtcg tgtagataac tacgatacgg gagggcttac catctggccc cagtgctgca
9661 atgataccgc gagacccacg ctcaccggct ccagatttat cagcaataaa ccagccagcc
9721 ggaagggccg agcgcagaag tggtcctgca actttatccg cctccatcca gtctattaat
9781 tgttgccggg aagctagagt aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc
9841 attgctacag gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt cagctccggt
9901 tcccaacgat caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc
9961 ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag tgttatcact catggttatg
10021 gcagcactgc ataattctct tactgtcatg ccatccgtaa gatgcttttc tgtgactggt
10081 gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg
10141 gcgtcaatac gggataatac cgcgccacat agcagaactt taaaagtgct catcattgga
10201 aaacgttctt cggggcgaaa actctcaagg atcttaccgc tgttgagatc cagttcgatg
10261 taacccactc gtgcacccaa ctgatcttca gcatctttta ctttcaccag cgtttctggg
10321 tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa taagggcgac acggaaatgt
10381 tgaatactca tactcttcct ttttcaatat tattgaagca tttatcaggg ttattgtctc
10441 atgagcggat acatatttga atgtatttag aaaaataaac aaataggggt tccgcgcaca
10501 tttccccgaa aagtgccacc tgacgtcgac ggatcgggag atctcccgat cccctatggt
10561 gcactctcag tacaatctgc tctgatgccg catagttaag ccagtatctg ctccctgctt
10621 gtgtgttgga ggtcgctgag tagtgcgcga gcaaaattta agctacaaca aggcaaggct
10681 tgaccgacaa ttgcatgaag aatctgctta gggttaggcg ttttgcgctg cttcgcgatg
10741 tacgggccag atatacgcgt tgacattgat tattgactag ttattaatag taatcaatta
10801 cggggtcatt agttcatagc ccatatatgg agttccgcgt tacataactt acggtaaatg
10861 gcccgcctgg ctgaccgccc aacgaccccc gcccattgac gtcaataatg acgtatgttc
10921 ccatagtaac gccaataggg actttccatt gacgtcaatg ggtggagtat ttacggtaaa
10981 ctgcccactt ggcagtacat caagtgtatc atatgccaag tacgccccct attgacgtca
11041 atgacggtaa atggcccgcc tggcattatg cccagtacat gaccttatgg gactttccta
11101 cttggcagta catctacgta ttagtcatcg ctattaccat ggtgatgcgg ttttggcagt
11161 acatcaatgg gcgtggatag cggtttgact cacggggatt tccaagtctc caccccattg
11221 acgtcaatgg gagtttgttt tggcaccaaa atcaacggga ctttccaaaa tgtcgtaaca
11281 actccgcccc attgacgcaa atgggcggta ggcgtgtacg gtgggaggtc tatataagca
11341 gcgcgttttg cctgtactgg gtctctctgg ttagaccaga tctgagcctg ggagctctct
11401 ggctaactag ggaacccact gcttaagcct caataaagct tgccttgagt gcttcaagta
11461 gtgtgtgccc gtctgttgtg tgactctggt aactagagat ccctcagacc cttttagtca
11521 gtgtggaaaa tctctagcag tggcgcccga acagggactt gaaagcgaaa gggaaaccag
11581 aggagctctc tcgacgcagg actcggcttg ctgaagcgcg cacggcaaga ggcgaggggc
11641 ggcgactggt gagtacgcca aaaattttga ctagcggagg ctagaaggag agagatgggt
11701 gcgagagcgt cagtattaag cgggggagaa ttagatcgcg atgggaaaaa attcggttaa
11761 ggccaggggg aaagaaaaaa tataaattaa aacatatagt atgggcaagc agggagctag
11821 aacgattcgc agttaatcct ggcctgttag aaacatcaga aggctgtaga caaatactgg
11881 gacagctaca accatccctt cagacaggat cagaagaact tagatcatta tataatacag
11941 tagcaaccct ctattgtgtg catcaaagga tagagataaa agacaccaag gaagctttag
12001 acaagataga ggaagagcaa aacaaaagta agaccaccgc acagcaagcg gccgctgatc
12061 ttcagacctg gaggaggaga tatgagggac aattggagaa gtgaattata taaatataaa
12121 gtagtaaaaa ttgaaccatt aggagtagca cccaccaagg caaagagaag agtggtgcag
12181 agagaaaaaa gagcagtggg aataggagct ttgttccttg ggttcttggg agcagcagga
12241 agcactatgg gcgcagcgtc aatgacgctg acggtacagg ccagacaatt attgtctggt
12301 atagtgcagc agcagaacaa tttgctgagg gctattgagg cgcaacagca tctgttgcaa
12361 ctcacagtct ggggcatcaa gcagctccag gcaagaatcc tggctgtgga aagataccta
12421 aaggatcaac agctcctggg gatttggggt tgctctggaa aactcatttg caccactgct
12481 gtgccttgga atgctagttg gagtaataaa tctctggaac agatttggaa tcacacgacc
12541 tggatggagt gggacagaga aattaacaat tacacaagct taatacactc cttaattgaa
12601 gaatcgcaaa accagcaaga aaagaatgaa caagaattat tggaattaga taaatgggca
12661 agtttgtgga attggtttaa cataacaaat tggctgtggt atataaaatt attcataatg
12721 atagtaggag gcttggtagg tttaagaata gtttttgctg tactttctat agtgaataga
12781 gttaggcagg gatattcacc attatcgttt cagacccacc tcccaacccc gaggggaccc
12841 gacaggcccg aaggaataga agaagaaggt ggagagagag acagagacag atccattcga
12901 ttagtgaacg gatcggcact gcgtgcgcca attctgcaga caaatggcag tattcatcca
12961 caattttaaa agaaaagggg ggattggggg gtacagtgca ggggaaagaa tagtagacat
13021 aatagcaaca gacatacaaa ctaaagaatt acaaaaacaa attacaaaaa ttcaaaattt
13081 tcgggtttat tacagggaca gcagagatcc agtttggtta attaaggtac c
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