| 质粒名称: | LentiCRISPRv2-Beclin1 |
|---|---|
| 出品公司: | Addgene |
| 目录编号: | 99574 |
| 存储实验室: | Edward Campbell |
|
载体骨架基本信息 |
|
| 载体名称: |
LentiCRISPR V2
|
| 载体类型: | 哺乳动物表达，慢病毒 |
| 载体抗性: | 氨苄青霉素，100μg/mL |
| 生长温度: | 37 ℃ |
| 宿主菌: | NEB Stable |
| 拷贝数: | 未知 |
|
插入基因信息 |
|
| 插入基因名称: | gRNA targeting Beclin1 |
| 物种来源: | H. sapiens (human) |
| 启动子: | U6 |
| 克隆方法: | 限制性内切酶 |
| 5’克隆位点: | 未知（如果被销毁，则未知） |
| 3’克隆位点: | 未知（如果被销毁，则未知） |
| 5’测序引物 | LKO.1 5' |
|
文献引用方法 |
|
| 材料和方法部分: |
LentiCRISPRv2-Beclin1 was a gift from Edward Campbell (Addgene plasmid # 99574 ; http://n2t.net/addgene:99574 ; RRID:Addgene_99574)
|
| 参考文献部分: |
TRIM5alpha degradation via autophagy is not required for retroviral restriction. Imam S, Talley S, Nelson RS, Dharan A, O'Connor C, Hope TJ, Campbell EM. J Virol. 2016 Jan 13. pii: JVI.03033-15. 10.1128/JVI.03033-15 PubMed 26764007
|
买家导航
LentiCRISPRv2-Beclin1载体质粒基本信息
LentiCRISPRv2-Beclin1质粒图谱载体图谱和LentiCRISPRv2-Beclin1载体序列质粒序列多克隆位点信息
LentiCRISPRv2-Beclin1质粒载体简介
LentiCRISPRv2-Beclin1质粒序列载体序列
LOCUS LentiCRISPRv2-Beclin1 13013 bp DNA circular SYN 15-DEC-2025
DEFINITION Expressed Cas9 with sgRNA targeting Beclin1.
ACCESSION .
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 13013)
AUTHORS Imam S, Talley S, Nelson RS, Dharan A, O'Connor C, Hope TJ, Campbell
EM
TITLE TRIM5alpha degradation via autophagy is not required for retroviral
restriction.
JOURNAL J Virol. 2016 Jan 13. pii: JVI.03033-15.
PUBMED 26764007
REFERENCE 2 (bases 1 to 13013)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Dec 15, 2025 from SnapGene Server 8.0.1
https://www.biofeng.com
COMMENT SGRef: number: 1; type: "Journal Article"; journalName: "J Virol.
2016 Jan 13. pii: JVI.03033-15."
COMMENT Sequence Label: LentiCRISPRv2-Beclin1
FEATURES Location/Qualifiers
source 1..13013
/mol_type="other DNA"
/organism="synthetic DNA construct"
primer_bind complement(6..25)
/label=SV40pro-F
/note="SV40 promoter/origin, forward primer"
primer_bind 252..272
/label=pBABE 3'
/note="SV40 enhancer, reverse primer for pBABE vectors"
rep_origin complement(281..709)
/direction=LEFT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
primer_bind complement(391..412)
/label=F1ori-F
/note="F1 origin, forward primer"
primer_bind 603..622
/label=F1ori-R
/note="F1 origin, reverse primer"
polyA_signal 755..979
/label=bGH poly(A) signal
/note="bovine growth hormone polyadenylation signal"
primer_bind 968..985
/label=BGH-rev
/note="Bovine growth hormone terminator, reverse primer.
Also called BGH reverse"
LTR 1011..1244
/label=3' LTR (Delta-U3)
/note="self-inactivating 3' long terminal repeat (LTR) from
HIV-1"
misc_feature 1316..1904
/label=WPRE
/note="woodchuck hepatitis virus posttranscriptional
regulatory element"
CDS 1422..1433
/codon_start=1
/product="Factor Xa recognition and cleavage site"
/label=Factor Xa site
/translation="IEGR"
primer_bind 1831..1851
/label=WPRE-R
/note="WPRE, reverse primer"
CDS complement(1920..2516)
/codon_start=1
/gene="pac from Streptomyces alboniger"
/product="puromycin N-acetyltransferase"
/label=PuroR
/note="confers resistance to puromycin"
/translation="TEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIERV
TELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLAA
QQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETSA
PRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
primer_bind complement(2003..2023)
/label=Puro-F
/note="Puromycin resistance gene, forward primer"
CDS complement(2517..2573)
/codon_start=1
/product="2A peptide from porcine teschovirus-1
polyprotein"
/label=P2A
/note="Eukaryotic ribosomes fail to insert a peptide bond
between the Gly and Pro residues, yielding separate
polypeptides."
/translation="ATNFSLLKQAGDVEENPGP"
CDS complement(2583..2606)
/codon_start=1
/product="FLAG(R) epitope tag, followed by an enterokinase
cleavage site"
/label=FLAG
/translation="DYKDDDDK"
CDS complement(2607..2654)
/codon_start=1
/product="bipartite nuclear localization signal from
nucleoplasmin"
/label=nucleoplasmin NLS
/translation="KRPAATKKAGQAKKKK"
CDS complement(2655..6758)
/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"
CDS complement(2655..6758)
/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"
regulatory 6755..6764
/label=Kozak sequence
/note="vertebrate consensus sequence for strong initiation
of translation (Kozak, 1987)"
/regulatory_class="other"
promoter complement(6783..6994)
/label=EF-1-alpha core promoter
/note="core promoter for human elongation factor
EF-1-alpha"
misc_RNA complement(7056..7131)
/label=gRNA scaffold
/note="guide RNA scaffold for the Streptococcus pyogenes
CRISPR/Cas9 system"
promoter complement(7153..7401)
/label=U6 promoter
/note="RNA polymerase III promoter for human U6 snRNA
(Domitrovich & Kunkel, 2003)"
primer_bind complement(7211..7230)
/label=LKO.1 5'
/note="Human U6 promoter, forward primer"
primer_bind complement(7381..7401)
/label=hU6-F
/note="Human U6 promoter, forward primer"
misc_feature 7452..7569
/label=cPPT/CTS
/note="central polypurine tract and central termination
sequence of HIV-1"
CDS complement(7867..7911)
/codon_start=1
/product="antigenic peptide corresponding to amino acids
655 to 669 of the HIV envelope protein gp41 (Lutje Hulsik
et al., 2013)"
/label=gp41 peptide
/note="recognized by the 2H10 single-chain llama nanobody"
/translation="KNEQELLELDKWASL"
misc_feature 8096..8329
/label=RRE
/note="The Rev response element (RRE) of HIV-1 allows for
Rev-dependent mRNA export from the nucleus to the
cytoplasm."
misc_feature 8822..8947
/label=HIV-1 Psi
/note="packaging signal of human immunodeficiency virus
type 1"
LTR 8994..9174
/label=5' LTR (truncated)
/note="truncated 5' long terminal repeat (LTR) from HIV-1"
promoter complement(9192..9390)
/label=CMV promoter
/note="human cytomegalovirus (CMV) immediate early
promoter"
primer_bind complement(9217..9237)
/label=CMV-F
/note="Human CMV immediate early promoter, forward primer"
enhancer 9392..9771
/label=CMV enhancer
/note="human cytomegalovirus immediate early enhancer"
primer_bind 9943..9962
/label=pRS-marker
/note="pRS vectors, use to sequence yeast selectable
marker"
promoter 10037..10141
/gene="bla"
/label=AmpR promoter
CDS join(10142..10210,10211..11002)
/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(10360..10379)
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
rep_origin 11173..11761
/direction=RIGHT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind 11662..11681
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
primer_bind 11915..11932
/label=L4440
/note="L4440 vector, forward primer"
protein_bind 12049..12070
/label=CAP binding site
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding activates transcription in the presence
of cAMP."
promoter join(12085..12090,12091..12108,12109..12115)
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 12123..12139
/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 12128..12150
/label=M13/pUC Reverse
/note="In lacZ gene"
primer_bind 12147..12163
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
primer_bind 12147..12163
/label=M13 Reverse
/note="In lacZ gene. Also called M13-rev"
polyA_signal 12212..12333
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
primer_bind complement(12223..12242)
/label=EBV-rev
/note="SV40 polyA terminator, reverse primer"
primer_bind 12277..12296
/label=SV40pA-R
/note="SV40 polyA, reverse primer"
CDS complement(12463..12837)
/codon_start=1
/gene="Sh ble from Streptoalloteichus hindustanus"
/product="antibiotic-binding protein"
/label=BleoR
/note="confers resistance to bleomycin, phleomycin, and
Zeocin(TM)"
/translation="MAKLTSAVPVLTARDVAGAVEFWTDRLGFSRDFVEDDFAGVVRDD
VTLFISAVQDQVVPDNTLAWVWVRGLDELYAEWSEVVSTNFRDASGPAMTEIGEQPWGR
EFALRDPAGNCVHFVAEEQD"
promoter complement(12856..12903)
/label=EM7 promoter
/note="synthetic bacterial promoter "
promoter complement(join(12951..13013,1..267))
/label=SV40 promoter
/note="SV40 enhancer and early promoter"
rep_origin join(12965..13013,1..87)
/label=SV40 ori
/note="SV40 origin of replication"
ORIGIN
1 ggcggcctcg gcctctgcat aaataaaaaa aattagtcag ccatggggcg gagaatgggc
61 ggaactgggc ggagttaggg gcgggatggg cggagttagg ggcgggacta tggttgctga
121 ctaattgaga tgcatgcttt gcatacttct gcctgctggg gagcctgggg actttccaca
181 cctggttgct gactaattga gatgcatgct ttgcatactt ctgcctgctg gggagcctgg
241 ggactttcca caccctaact gacacacatt ccacagaatt aattcgcgtt aaatttttgt
301 taaatcagct cattttttaa ccaataggcc gaaatcggca aaatccctta taaatcaaaa
361 gaatagaccg agatagggtt gagtgttgtt ccagtttgga acaagagtcc actattaaag
421 aacgtggact ccaacgtcaa agggcgaaaa accgtctatc agggcgatgg cccactacgt
481 gaaccatcac cctaatcaag ttttttgggg tcgaggtgcc gtaaagcact aaatcggaac
541 cctaaaggga gcccccgatt tagagcttga cggggaaagc cggcgaacgt ggcgagaaag
601 gaagggaaga aagcgaaagg agcgggcgct agggcgctgg caagtgtagc ggtcacgctg
661 cgcgtaacca ccacacccgc cgcgcttaat gcgccgctac agggcgcgtg gggatacccc
721 ctagagcccc agctggttct ttccgcctca gaagccatag agcccaccgc atccccagca
781 tgcctgctat tgtcttccca atcctccccc ttgctgtcct gccccacccc accccccaga
841 atagaatgac acctactcag acaatgcgat gcaatttcct cattttatta ggaaaggaca
901 gtgggagtgg caccttccag ggtcaaggaa ggcacggggg aggggcaaac aacagatggc
961 tggcaactag aaggcacagt cgaggctgat cagcgggttt aaacgggccc tgctagagat
1021 tttccacact gactaaaagg gtctgaggga tctctagtta ccagagtcac acaacagacg
1081 ggcacacact acttgaagca ctcaaggcaa gctttattga ggcttaagca gtgggttccc
1141 tagttagcca gagagctccc aggctcagat ctggtctaac cagagagacc cagtacaagc
1201 aaaaagcaga tcttgtcttc gttgggagtg aattagccct tccagtcccc ccttttcttt
1261 taaaaagtgg ctaagatcta cagctgcctt gtaagtcatt ggtcttaaag tcgacgcggg
1321 gaggcggccc aaagggagat ccgactcgtc tgagggcgaa ggcgaagacg cggaagaggc
1381 cgcagagccg gcagcaggcc gcgggaagga aggtccgctg gattgagggc cgaagggacg
1441 tagcagaagg acgtcccgcg cagaatccag gtggcaacac aggcgagcag ccaaggaaag
1501 gacgatgatt tccccgacaa caccacggaa ttgtcagtgc ccaacagccg agcccctgtc
1561 cagcagcggg caaggcaggc ggcgatgagt tccgccgtgg caatagggag ggggaaagcg
1621 aaagtcccgg aaaggagctg acaggtggtg gcaatgcccc aaccagtggg ggttgcgtca
1681 gcaaacacag tgcacaccac gccacgttgc ctgacaacgg gccacaactc ctcataaaga
1741 gacagcaacc aggatttata caaggaggag aaaatgaaag ccatacggga agcaatagca
1801 tgatacaaag gcattaaagc agcgtatcca catagcgtaa aaggagcaac atagttaaga
1861 ataccagtca atctttcaca aattttgtaa tccagaggtt gattgtcgac ttaacgcgtt
1921 caggcaccgg gcttgcgggt catgcaccag gtgcgcggtc cttcgggcac ctcgacgtcg
1981 gcggtgacgg tgaagccgag ccgctcgtag aaggggaggt tgcggggcgc ggaggtctcc
2041 aggaaggcgg gcaccccggc gcgctcggcc gcctccactc cggggagcac gacggcgctg
2101 cccagaccct tgccctggtg gtcgggcgag actccgacgg tggccaggaa ccacgcgggc
2161 tccttgggcc ggtgcggcgc caggaggcct tccatctgtt gctgcgcggc cagccgggaa
2221 ccgctcaact cggccatgcg cgggccgatc tcggcgaaca ccgcccccgc ttcgacgctc
2281 tccggcgtgg tccagaccgc caccgcggcg ccgtcgtccg cgacccacac cttgccgatg
2341 tcgagcccga cgcgcgtgag gaagagttct tgcagctcgg tgacccgctc gatgtggcgg
2401 tccggatcga cggtgtggcg cgtggcgggg tagtcggcga acgcggcggc gagggtgcgt
2461 acggccctgg ggacgtcgtc gcgggtggcg aggcgcaccg tgggcttgta ctcggtcggt
2521 ccaggattct cttcgacatc tccggcttgt ttcagcagag agaagtttgt tgcgccggat
2581 cccttatcgt catcgtcttt gtaatctttc ttcttcttag cctgtccagc cttctttgtg
2641 gcggcaggtc gcttgtcgcc tcccagctga gacaggtcga tccgtgtctc gtacaggccg
2701 gtgatgctct ggtggatcag ggtggcgtcc agcacctctt tggtgctggt gtacctcttc
2761 cggtcgatgg tggtgtcaaa gtacttgaag gcggcagggg ctcccagatt ggtcagggta
2821 aacaggtgga tgatattctc ggcctgctct ctgatgggct tatcccggtg cttgttgtag
2881 gcggacagca ctttgtccag attagcgtcg gccaggatca ctctcttgga gaactcgctg
2941 atctgctcga tgatctcgtc caggtagtgc ttgtgctgtt ccacaaacag ctgtttctgc
3001 tcattatcct cgggggagcc cttcagcttc tcatagtggc tggccaggta caggaagttc
3061 acatatttgg agggcagggc cagttcgttt cccttctgca gttcgccggc agaggccagc
3121 attctcttcc ggccgttttc cagctcgaac agggagtact taggcagctt gatgatcagg
3181 tcctttttca cttctttgta gcccttggct tccagaaagt cgatgggatt cttctcgaag
3241 ctgcttcttt ccatgatggt gatccccagc agctctttca cactcttcag tttcttggac
3301 ttgccctttt ccactttggc caccaccagc acagaatagg ccacggtggg gctgtcgaag
3361 ccgccgtact tcttagggtc ccagtccttc tttctggcga tcagcttatc gctgttcctc
3421 ttgggcagga tagactcttt gctgaagccg cctgtctgca cctcggtctt tttcacgata
3481 ttcacttggg gcatgctcag cactttccgc acggtggcaa aatcccggcc cttatcccac
3541 acgatctccc cggtttcgcc gtttgtctcg atcagaggcc gcttccggat ctcgccgttg
3601 gccagggtaa tctcggtctt gaaaaagttc atgatgttgc tgtagaagaa gtacttggcg
3661 gtagccttgc cgatttcctg ctcgctcttg gcgatcatct tccgcacgtc gtacaccttg
3721 tagtcgccgt acacgaactc gctttccagc ttagggtact ttttgatcag ggcggttccc
3781 acgacggcgt tcaggtaggc gtcgtgggcg tggtggtagt tgttgatctc gcgcactttg
3841 taaaactgga aatccttccg gaaatcggac accagcttgg acttcagggt gatcactttc
3901 acttcccgga tcagcttgtc attctcgtcg tacttagtgt tcatccggga gtccaggatc
3961 tgtgccacgt gctttgtgat ctgccgggtt tccaccagct gtctcttgat gaagccggcc
4021 ttatccagtt cgctcaggcc gcctctctcg gccttggtca gattgtcgaa ctttctctgg
4081 gtaatcagct tggcgttcag cagctgccgc cagtagttct tcatcttctt cacgacctct
4141 tcggagggca cgttgtcgct cttgccccgg ttcttgtcgc ttctggtcag caccttgttg
4201 tcgatggagt cgtccttcag aaagctctga ggcacgatat ggtccacatc gtagtcggac
4261 agccggttga tgtccagttc ctggtccacg tacatatccc gcccattctg caggtagtac
4321 aggtacagct tctcgttctg cagctgggtg ttttccacgg ggtgttcttt caggatctgg
4381 ctgcccagct ctttgatgcc ctcttcgatc cgcttcattc tctcgcggct gttcttctgt
4441 cccttctggg tggtctggtt ctctctggcc atttcgatca cgatgttctc gggcttgtgc
4501 cggcccatca ctttcacgag ctcgtccacc accttcactg tctgcaggat gcccttctta
4561 atggcggggc tgccggccag attggcaatg tgctcgtgca ggctatcgcc ctggccggac
4621 acctgggctt tctggatgtc ctctttaaag gtcaggctgt cgtcgtggat cagctgcatg
4681 aagtttctgt tggcgaagcc gtcggacttc aggaaatcca ggattgtctt gccggactgc
4741 ttgtcccgga tgccgttgat cagcttccgg ctcagcctgc cccagccggt gtatctccgc
4801 cgcttcagct gcttcatcac tttgtcgtcg aacaggtggg cataggtttt cagccgttcc
4861 tcgatcatct ctctgtcctc aaacagtgtc agggtcagca cgatatcttc cagaatgtcc
4921 tcgttttcct cattgtccag gaagtccttg tccttgataa ttttcagcag atcgtggtat
4981 gtgcccaggg aggcgttgaa ccgatcttcc acgccggaga tttccacgga gtcgaagcac
5041 tcgattttct tgaagtagtc ctctttcagc tgcttcacgg tcactttccg gttggtcttg
5101 aacagcaggt ccacgatggc ctttttctgc tcgccgctca ggaaggcggg ctttctcatt
5161 ccctcggtca cgtatttcac tttggtcagc tcgttataca cggtgaagta ctcgtacagc
5221 aggctgtgct tgggcagcac cttctcgttg ggcaggttct tatcgaagtt ggtcatccgc
5281 tcgatgaagc tctgggcgga agcgcccttg tccaccactt cctcgaagtt ccagggggtg
5341 atggtttcct cgctctttct ggtcatccag gcgaatctgc tgtttcccct ggccagaggg
5401 cccacgtagt aggggatgcg gaaggtcagg atcttctcga tcttttcccg gttgtccttc
5461 aggaatgggt aaaaatcttc ctgccgccgc agaatggcgt gcagctctcc caggtggatc
5521 tggtggggga tgctgccgtt gtcgaaggtc cgctgcttcc gcagcaggtc ctctctgttc
5581 agcttcacga gcagttcctc ggtgccgtcc atcttttcca ggatgggctt gatgaacttg
5641 tagaactctt cctggctggc tccgccgtca atgtagccgg cgtagccgtt cttgctctgg
5701 tcgaagaaaa tctctttgta cttctcaggc agctgctgcc gcacgagagc tttcagcagg
5761 gtcaggtcct ggtggtgctc gtcgtatctc ttgatcatag aggcgctcag gggggccttg
5821 gtgatctcgg tgttcactct caggatgtcg ctcagcagga tggcgtcgga caggttcttg
5881 gcggccagaa acaggtcggc gtactggtcg ccgatctggg ccagcaggtt gtccaggtcg
5941 tcgtcgtagg tgtccttgct cagctgcagt ttggcatcct cggccaggtc gaagttgctc
6001 ttgaagttgg gggtcaggcc caggctcagg gcaatcaggt ttccgaacag gccattcttc
6061 ttctcgccgg gcagctgggc gatcagattt tccagccgtc tgctcttgct cagtctggca
6121 gacaggatgg ccttggcgtc cacgccgctg gcgttgatgg ggttttcctc gaacagctgg
6181 ttgtaggtct gcaccagctg gatgaacagc ttgtccacgt cgctgttgtc ggggttcagg
6241 tcgccctcga tcaggaagtg gccccggaac ttgatcatgt gggccagggc cagatagatc
6301 agccgcaggt cggccttgtc ggtgctgtcc accagtttct ttctcaggtg gtagatggtg
6361 gggtacttct cgtggtaggc cacctcgtcc acgatgttgc cgaagatggg gtgccgctcg
6421 tgcttcttat cctcttccac caggaaggac tcttccagtc tgtggaagaa gctgtcgtcc
6481 accttggcca tctcgttgct gaagatctct tgcagatagc agatccggtt cttccgtctg
6541 gtgtatcttc ttctggcggt tctcttcagc cgggtggcct cggctgtttc gccgctgtcg
6601 aacagcaggg ctccgatcag gttcttcttg atgctgtgcc ggtcggtgtt gcccagcacc
6661 ttgaatttct tgctgggcac cttgtactcg tcggtgatca cggcccagcc cacagagttg
6721 gtgccgatgt ccaggccgat gctgtacttc ttgtccatgg tggcagcgct ctagaaccgg
6781 tcctgtgttc tggcggcaaa cccgttgcga aaaagaacgt tcacggcgac tactgcactt
6841 atatacggtt ctcccccacc ctcgggaaaa aggcggagcc agtacacgac atcactttcc
6901 cagtttaccc cgcgccacct tctctaggca ccggatcaat tgccgacccc tccccccaac
6961 ttctcgggga ctgtgggcga tgtgcgctct gcccactgac gggcaccgga gccaattccc
7021 actcctttca agacctagct agcgaattca aaaaagcacc gactcggtgc cactttttca
7081 agttgataac ggactagcct tattttaact tgctatttct agctctaaaa cggatggtgt
7141 ctctcgcaga tcggtgtttc gtcctttcca caagatatat aaagccaaga aatcgaaata
7201 ctttcaagtt acggtaagca tatgatagtc cattttaaaa cataatttta aaactgcaaa
7261 ctacccaaga aattattact ttctacgtca cgtattttgt actaatatct ttgtgtttac
7321 agtcaaatta attctaatta tctctctaac agccttgtat cgtatatgca aatatgaagg
7381 aatcatggga aataggccct cggtacctta attaaccaaa ctggatctct gctgtccctg
7441 taataaaccc gaaaattttg aatttttgta atttgttttt gtaattcttt agtttgtatg
7501 tctgttgcta ttatgtctac tattctttcc cctgcactgt accccccaat cccccctttt
7561 cttttaaaat tgtggatgaa tactgccatt tgtctgcaga attggcgcac gcagtgccga
7621 tccgttcact aatcgaatgg atctgtctct gtctctctct ccaccttctt cttctattcc
7681 ttcgggcctg tcgggtcccc tcggggttgg gaggtgggtc tgaaacgata atggtgaata
7741 tccctgccta actctattca ctatagaaag tacagcaaaa actattctta aacctaccaa
7801 gcctcctact atcattatga ataattttat ataccacagc caatttgtta tgttaaacca
7861 attccacaaa cttgcccatt tatctaattc caataattct tgttcattct tttcttgctg
7921 gttttgcgat tcttcaatta aggagtgtat taagcttgtg taattgttaa tttctctgtc
7981 ccactccatc caggtcgtgt gattccaaat ctgttccaga gatttattac tccaactagc
8041 attccaaggc acagcagtgg tgcaaatgag ttttccagag caaccccaaa tccccaggag
8101 ctgttgatcc tttaggtatc tttccacagc caggattctt gcctggagct gcttgatgcc
8161 ccagactgtg agttgcaaca gatgctgttg cgcctcaata gccctcagca aattgttctg
8221 ctgctgcact ataccagaca ataattgtct ggcctgtacc gtcagcgtca ttgacgctgc
8281 gcccatagtg cttcctgctg ctcccaagaa cccaaggaac aaagctccta ttcccactgc
8341 tcttttttct ctctgcacca ctcttctctt tgccttggtg ggtgctactc ctaatggttc
8401 aatttttact actttatatt tatataattc acttctccaa ttgtccctca tatctcctcc
8461 tccaggtctg aagatcagcg gccgcttgct gtgcggtggt cttacttttg ttttgctctt
8521 cctctatctt gtctaaagct tccttggtgt cttttatctc tatcctttga tgcacacaat
8581 agagggttgc tactgtatta tataatgatc taagttcttc tgatcctgtc tgaagggatg
8641 gttgtagctg tcccagtatt tgtctacagc cttctgatgt ttctaacagg ccaggattaa
8701 ctgcgaatcg ttctagctcc ctgcttgccc atactatatg ttttaattta tattttttct
8761 ttccccctgg ccttaaccga attttttccc atcgcgatct aattctcccc cgcttaatac
8821 tgacgctctc gcacccatct ctctccttct agcctccgct agtcaaaatt tttggcgtac
8881 tcaccagtcg ccgcccctcg cctcttgccg tgcgcgcttc agcaagccga gtcctgcgtc
8941 gagagagctc ctctggtttc cctttcgctt tcaagtccct gttcgggcgc cactgctaga
9001 gattttccac actgactaaa agggtctgag ggatctctag ttaccagagt cacacaacag
9061 acgggcacac actacttgaa gcactcaagg caagctttat tgaggcttaa gcagtgggtt
9121 ccctagttag ccagagagct cccaggctca gatctggtct aaccagagag acccagtaca
9181 ggcaaaacgc gctgcttata tagacctccc accgtacacg cctaccgccc atttgcgtca
9241 atggggcgga gttgttacga cattttggaa agtcccgttg attttggtgc caaaacaaac
9301 tcccattgac gtcaatgggg tggagacttg gaaatccccg tgagtcaaac cgctatccac
9361 gcccattgat gtactgccaa aaccgcatca ccatggtaat agcgatgact aatacgtaga
9421 tgtactgcca agtaggaaag tcccataagg tcatgtactg ggcataatgc caggcgggcc
9481 atttaccgtc attgacgtca atagggggcg tacttggcat atgatacact tgatgtactg
9541 ccaagtgggc agtttaccgt aaatactcca cccattgacg tcaatggaaa gtccctattg
9601 gcgttactat gggaacatac gtcattattg acgtcaatgg gcgggggtcg ttgggcggtc
9661 agccaggcgg gccatttacc gtaagttatg taacgcggaa ctccatatat gggctatgaa
9721 ctaatgaccc cgtaattgat tactattaat aactagtcaa taatcaatgt caacgcgtat
9781 atctggcccg tacatcgcga agcagcgcaa aacgcctaac cctaagcaga ttcttcatgc
9841 aattgtcggt caagccttgc cttgttgtag cttaaatttt gctcgcgcac tactcagcga
9901 cctccaacac acaagcaggg agcagatact ggcttaacta tgcggcatca gagcagattg
9961 tactgagagt gcaccatagg ggatcgggag atctcccgat ccgtcgacgt caggtggcac
10021 ttttcgggga aatgtgcgcg gaacccctat ttgtttattt ttctaaatac attcaaatat
10081 gtatccgctc atgagacaat aaccctgata aatgcttcaa taatattgaa aaaggaagag
10141 tatgagtatt caacatttcc gtgtcgccct tattcccttt tttgcggcat tttgccttcc
10201 tgtttttgct cacccagaaa cgctggtgaa agtaaaagat gctgaagatc agttgggtgc
10261 acgagtgggt tacatcgaac tggatctcaa cagcggtaag atccttgaga gttttcgccc
10321 cgaagaacgt tttccaatga tgagcacttt taaagttctg ctatgtggcg cggtattatc
10381 ccgtattgac gccgggcaag agcaactcgg tcgccgcata cactattctc agaatgactt
10441 ggttgagtac tcaccagtca cagaaaagca tcttacggat ggcatgacag taagagaatt
10501 atgcagtgct gccataacca tgagtgataa cactgcggcc aacttacttc tgacaacgat
10561 cggaggaccg aaggagctaa ccgctttttt gcacaacatg ggggatcatg taactcgcct
10621 tgatcgttgg gaaccggagc tgaatgaagc cataccaaac gacgagcgtg acaccacgat
10681 gcctgtagca atggcaacaa cgttgcgcaa actattaact ggcgaactac ttactctagc
10741 ttcccggcaa caattaatag actggatgga ggcggataaa gttgcaggac cacttctgcg
10801 ctcggccctt ccggctggct ggtttattgc tgataaatct ggagccggtg agcgtgggtc
10861 tcgcggtatc attgcagcac tggggccaga tggtaagccc tcccgtatcg tagttatcta
10921 cacgacgggg agtcaggcaa ctatggatga acgaaataga cagatcgctg agataggtgc
10981 ctcactgatt aagcattggt aactgtcaga ccaagtttac tcatatatac tttagattga
11041 tttaaaactt catttttaat ttaaaaggat ctaggtgaag atcctttttg ataatctcat
11101 gaccaaaatc ccttaacgtg agttttcgtt ccactgagcg tcagaccccg tagaaaagat
11161 caaaggatct tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa
11221 accaccgcta ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc tttttccgaa
11281 ggtaactggc ttcagcagag cgcagatacc aaatactgtt cttctagtgt agccgtagtt
11341 aggccaccac ttcaagaact ctgtagcacc gcctacatac ctcgctctgc taatcctgtt
11401 accagtggct gctgccagtg gcgataagtc gtgtcttacc gggttggact caagacgata
11461 gttaccggat aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac agcccagctt
11521 ggagcgaacg acctacaccg aactgagata cctacagcgt gagctatgag aaagcgccac
11581 gcttcccgaa gggagaaagg cggacaggta tccggtaagc ggcagggtcg gaacaggaga
11641 gcgcacgagg gagcttccag ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg
11701 ccacctctga cttgagcgtc gatttttgtg atgctcgtca ggggggcgga gcctatggaa
11761 aaacgccagc aacgcggcct ttttacggtt cctggccttt tgctggcctt ttgctcacat
11821 gttctttcct gcgttatccc ctgattctgt ggataaccgt attaccgcct ttgagtgagc
11881 tgataccgct cgccgcagcc gaacgaccga gcgcagcgag tcagtgagcg aggaagcgga
11941 agagcgccca atacgcaaac cgcctctccc cgcgcgttgg ccgattcatt aatgcagctg
12001 gcacgacagg tttcccgact ggaaagcggg cagtgagcgc aacgcaatta atgtgagtta
12061 gctcactcat taggcacccc aggctttaca ctttatgctt ccggctcgta tgttgtgtgg
12121 aattgtgagc ggataacaat ttcacacagg aaacagctat gaccatgatt acgccaagct
12181 ctagctagag gtcgacggta tacagacatg ataagataca ttgatgagtt tggacaaacc
12241 acaactagaa tgcagtgaaa aaaatgcttt atttgtgaaa tttgtgatgc tattgcttta
12301 tttgtaacca ttataagctg caataaacaa gttggggtgg gcgaagaact ccagcatgag
12361 atccccgcgc tggaggatca tccagccggc gtcccggaaa acgattccga agcccaacct
12421 ttcatagaag gcggcggtgg aatcgaaatc tcgtagcacg tgtcagtcct gctcctcggc
12481 cacgaagtgc acgcagttgc cggccgggtc gcgcagggcg aactcccgcc cccacggctg
12541 ctcgccgatc tcggtcatgg ccggcccgga ggcgtcccgg aagttcgtgg acacgacctc
12601 cgaccactcg gcgtacagct cgtccaggcc gcgcacccac acccaggcca gggtgttgtc
12661 cggcaccacc tggtcctgga ccgcgctgat gaacagggtc acgtcgtccc ggaccacacc
12721 ggcgaagtcg tcctccacga agtcccggga gaacccgagc cggtcggtcc agaactcgac
12781 cgctccggcg acgtcgcgcg cggtgagcac cggaacggca ctggtcaact tggccatggt
12841 ttagttcctc accttgtcgt attatactat gccgatatac tatgccgatg attaattgtc
12901 aacacgtgct gatcagatcc gaaaatggat atacaagctc ccgggagctt tttgcaaaag
12961 cctaggcctc caaaaaagcc tcctcactac ttctggaata gctcagaggc aga
//
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