| 质粒名称: | sgCtr- LentiCRISPRv2 |
|---|---|
| 出品公司: | Addgene |
| 目录编号: | 107402 |
| 存储实验室: |
William Kaelin
|
|
载体骨架基本信息 |
|
| 载体名称: | lentiCRISPR v2 |
| 载体出品公司: | Feng Zhang (Addgene plasmid # 52961) |
| 载体类型: | 慢病毒载体 |
| 载体抗性: | 氨苄青霉素，100μg/mL |
| 生长温度: | 30 ℃ |
| 宿主菌: | NEB Stable |
| 拷贝数: | 未知 |
|
插入基因信息 |
|
| 插入基因名称: | control gRNA |
| 物种来源: | H. sapiens (human) |
| 克隆方法: | 限制性内切酶 |
| 5’克隆位点: | BsmBI（销毁后未知） |
| 3’克隆位点: | BsmBI（销毁后未知） |
|
文献引用方法 |
|
| 材料和方法部分: | sgCtr- LentiCRISPRv2 was a gift from William Kaelin (Addgene plasmid # 107402 ; http://n2t.net/addgene:107402 ; RRID:Addgene_107402) |
| 参考文献部分: | Inactivation of the PBRM1 tumor suppressor gene amplifies the HIF-response in VHL-/- clear cell renal carcinoma. Gao W, Li W, Xiao T, Liu XS, Kaelin WG Jr. Proc Natl Acad Sci U S A. 2017 Jan 31;114(5):1027-1032. doi: 10.1073/pnas.1619726114. Epub 2017 Jan 12. 10.1073/pnas.1619726114 PubMed 28082722 |
买家导航
sgCtr- LentiCRISPRv2载体质粒基本信息
sgCtr- LentiCRISPRv2质粒图谱载体图谱和sgCtr- LentiCRISPRv2载体序列质粒序列多克隆位点信息
sgCtr- LentiCRISPRv2质粒载体简介
sgCtr- LentiCRISPRv2质粒序列载体序列
LOCUS sgCtr- LentiCRISPRv2 13011 bp DNA circular SYN 15-DEC-2025
DEFINITION Lentiviral expression of Cas9 and a control gRNA.
ACCESSION .
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 13011)
AUTHORS Gao W, Li W, Xiao T, Liu XS, Kaelin WG Jr
TITLE Inactivation of the PBRM1 tumor suppressor gene amplifies the
HIF-response in VHL-/- clear cell renal carcinoma.
JOURNAL Proc Natl Acad Sci U S A. 2017 Jan 31;114(5):1027-1032. doi:
10.1073/pnas.1619726114. Epub 2017 Jan 12.
PUBMED 28082722
REFERENCE 2 (bases 1 to 13011)
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"; doi:
"10.1073/pnas.1619726114"; journalName: "Proc Natl Acad Sci U S A";
date: "2017-01-31- 31"; volume: "114"; issue: "5"; pages:
"1027-1032"
COMMENT Sequence Label: sgCtr- LentiCRISPRv2
FEATURES Location/Qualifiers
source 1..13011
/mol_type="other DNA"
/organism="synthetic DNA construct"
misc_feature 261..378
/label=cPPT/CTS
/note="central polypurine tract and central termination
sequence of HIV-1"
promoter 429..677
/label=U6 promoter
/note="RNA polymerase III promoter for human U6 snRNA
(Domitrovich & Kunkel, 2003)"
primer_bind 429..449
/label=hU6-F
/note="Human U6 promoter, forward primer"
primer_bind 600..619
/label=LKO.1 5'
/note="Human U6 promoter, forward primer"
misc_RNA 697..772
/label=gRNA scaffold
/note="guide RNA scaffold for the Streptococcus pyogenes
CRISPR/Cas9 system"
promoter 834..1045
/label=EF-1-alpha core promoter
/note="core promoter for human elongation factor
EF-1-alpha"
regulatory 1064..1073
/label=Kozak sequence
/note="vertebrate consensus sequence for strong initiation
of translation (Kozak, 1987)"
/regulatory_class="other"
CDS 1070..5173
/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 1070..5173
/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 5174..5221
/codon_start=1
/product="bipartite nuclear localization signal from
nucleoplasmin"
/label=nucleoplasmin NLS
/translation="KRPAATKKAGQAKKKK"
CDS 5222..5245
/codon_start=1
/product="FLAG(R) epitope tag, followed by an enterokinase
cleavage site"
/label=FLAG
/translation="DYKDDDDK"
CDS 5255..5311
/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 5312..5908
/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 5805..5825
/label=Puro-F
/note="Puromycin resistance gene, forward primer"
misc_feature 5924..6512
/label=WPRE
/note="woodchuck hepatitis virus posttranscriptional
regulatory element"
primer_bind complement(5977..5997)
/label=WPRE-R
/note="WPRE, reverse primer"
CDS complement(6395..6406)
/codon_start=1
/product="Factor Xa recognition and cleavage site"
/label=Factor Xa site
/translation="IEGR"
LTR 6584..6817
/label=3' LTR (Delta-U3)
/note="self-inactivating 3' long terminal repeat (LTR) from
HIV-1"
primer_bind complement(6843..6860)
/label=BGH-rev
/note="Bovine growth hormone terminator, reverse primer.
Also called BGH reverse"
polyA_signal 6849..7073
/label=bGH poly(A) signal
/note="bovine growth hormone polyadenylation signal"
rep_origin 7119..7547
/direction=RIGHT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
primer_bind complement(7206..7225)
/label=F1ori-R
/note="F1 origin, reverse primer"
primer_bind 7416..7437
/label=F1ori-F
/note="F1 origin, forward primer"
primer_bind complement(7556..7576)
/label=pBABE 3'
/note="SV40 enhancer, reverse primer for pBABE vectors"
promoter 7561..7890
/label=SV40 promoter
/note="SV40 enhancer and early promoter"
rep_origin 7741..7876
/label=SV40 ori
/note="SV40 origin of replication"
primer_bind 7803..7822
/label=SV40pro-F
/note="SV40 promoter/origin, forward primer"
promoter 7938..7985
/label=EM7 promoter
/note="synthetic bacterial promoter "
CDS 8004..8378
/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"
polyA_signal 8508..8629
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
primer_bind complement(8545..8564)
/label=SV40pA-R
/note="SV40 polyA, reverse primer"
primer_bind 8599..8618
/label=EBV-rev
/note="SV40 polyA terminator, reverse primer"
primer_bind complement(8678..8694)
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(8678..8694)
/label=M13 Reverse
/note="In lacZ gene. Also called M13-rev"
primer_bind complement(8691..8713)
/label=M13/pUC Reverse
/note="In lacZ gene"
protein_bind 8702..8718
/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)."
promoter complement(join(8726..8732,8733..8750,8751..8756))
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 8771..8792
/label=CAP binding site
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding activates transcription in the presence
of cAMP."
primer_bind complement(8909..8926)
/label=L4440
/note="L4440 vector, forward primer"
rep_origin complement(9080..9668)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind complement(9160..9179)
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
CDS complement(join(9839..10630,10631..10699))
/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 10462..10481
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
promoter complement(10700..10804)
/gene="bla"
/label=AmpR promoter
primer_bind complement(10879..10898)
/label=pRS-marker
/note="pRS vectors, use to sequence yeast selectable
marker"
enhancer 11070..11449
/label=CMV enhancer
/note="human cytomegalovirus immediate early enhancer"
promoter 11451..11649
/label=CMV promoter
/note="human cytomegalovirus (CMV) immediate early
promoter"
primer_bind 11604..11624
/label=CMV-F
/note="Human CMV immediate early promoter, forward primer"
LTR 11667..11847
/label=5' LTR (truncated)
/note="truncated 5' long terminal repeat (LTR) from HIV-1"
misc_feature 11894..12019
/label=HIV-1 Psi
/note="packaging signal of human immunodeficiency virus
type 1"
misc_feature 12512..12745
/label=RRE
/note="The Rev response element (RRE) of HIV-1 allows for
Rev-dependent mRNA export from the nucleus to the
cytoplasm."
CDS 12930..12974
/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"
ORIGIN
1 taaaattatt cataatgata gtaggaggct tggtaggttt aagaatagtt tttgctgtac
61 tttctatagt gaatagagtt aggcagggat attcaccatt atcgtttcag acccacctcc
121 caaccccgag gggacccgac aggcccgaag gaatagaaga agaaggtgga gagagagaca
181 gagacagatc cattcgatta gtgaacggat cggcactgcg tgcgccaatt ctgcagacaa
241 atggcagtat tcatccacaa ttttaaaaga aaagggggga ttggggggta cagtgcaggg
301 gaaagaatag tagacataat agcaacagac atacaaacta aagaattaca aaaacaaatt
361 acaaaaattc aaaattttcg ggtttattac agggacagca gagatccagt ttggttaatt
421 aaggtaccga gggcctattt cccatgattc cttcatattt gcatatacga tacaaggctg
481 ttagagagat aattagaatt aatttgactg taaacacaaa gatattagta caaaatacgt
541 gacgtagaaa gtaataattt cttgggtagt ttgcagtttt aaaattatgt tttaaaatgg
601 actatcatat gcttaccgta acttgaaagt atttcgattt cttggcttta tatatcttgt
661 ggaaaggacg aaacaccgct tccgcggccc gttcaagttt tagagctaga aatagcaagt
721 taaaataagg ctagtccgtt atcaacttga aaaagtggca ccgagtcggt gcttttttga
781 attcgctagc taggtcttga aaggagtggg aattggctcc ggtgcccgtc agtgggcaga
841 gcgcacatcg cccacagtcc ccgagaagtt ggggggaggg gtcggcaatt gatccggtgc
901 ctagagaagg tggcgcgggg taaactggga aagtgatgtc gtgtactggc tccgcctttt
961 tcccgagggt gggggagaac cgtatataag tgcagtagtc gccgtgaacg ttctttttcg
1021 caacgggttt gccgccagaa cacaggaccg gttctagagc gctgccacca tggacaagaa
1081 gtacagcatc ggcctggaca tcggcaccaa ctctgtgggc tgggccgtga tcaccgacga
1141 gtacaaggtg cccagcaaga aattcaaggt gctgggcaac accgaccggc acagcatcaa
1201 gaagaacctg atcggagccc tgctgttcga cagcggcgaa acagccgagg ccacccggct
1261 gaagagaacc gccagaagaa gatacaccag acggaagaac cggatctgct atctgcaaga
1321 gatcttcagc aacgagatgg ccaaggtgga cgacagcttc ttccacagac tggaagagtc
1381 cttcctggtg gaagaggata agaagcacga gcggcacccc atcttcggca acatcgtgga
1441 cgaggtggcc taccacgaga agtaccccac catctaccac ctgagaaaga aactggtgga
1501 cagcaccgac aaggccgacc tgcggctgat ctatctggcc ctggcccaca tgatcaagtt
1561 ccggggccac ttcctgatcg agggcgacct gaaccccgac aacagcgacg tggacaagct
1621 gttcatccag ctggtgcaga cctacaacca gctgttcgag gaaaacccca tcaacgccag
1681 cggcgtggac gccaaggcca tcctgtctgc cagactgagc aagagcagac ggctggaaaa
1741 tctgatcgcc cagctgcccg gcgagaagaa gaatggcctg ttcggaaacc tgattgccct
1801 gagcctgggc ctgaccccca acttcaagag caacttcgac ctggccgagg atgccaaact
1861 gcagctgagc aaggacacct acgacgacga cctggacaac ctgctggccc agatcggcga
1921 ccagtacgcc gacctgtttc tggccgccaa gaacctgtcc gacgccatcc tgctgagcga
1981 catcctgaga gtgaacaccg agatcaccaa ggcccccctg agcgcctcta tgatcaagag
2041 atacgacgag caccaccagg acctgaccct gctgaaagct ctcgtgcggc agcagctgcc
2101 tgagaagtac aaagagattt tcttcgacca gagcaagaac ggctacgccg gctacattga
2161 cggcggagcc agccaggaag agttctacaa gttcatcaag cccatcctgg aaaagatgga
2221 cggcaccgag gaactgctcg tgaagctgaa cagagaggac ctgctgcgga agcagcggac
2281 cttcgacaac ggcagcatcc cccaccagat ccacctggga gagctgcacg ccattctgcg
2341 gcggcaggaa gatttttacc cattcctgaa ggacaaccgg gaaaagatcg agaagatcct
2401 gaccttccgc atcccctact acgtgggccc tctggccagg ggaaacagca gattcgcctg
2461 gatgaccaga aagagcgagg aaaccatcac cccctggaac ttcgaggaag tggtggacaa
2521 gggcgcttcc gcccagagct tcatcgagcg gatgaccaac ttcgataaga acctgcccaa
2581 cgagaaggtg ctgcccaagc acagcctgct gtacgagtac ttcaccgtgt ataacgagct
2641 gaccaaagtg aaatacgtga ccgagggaat gagaaagccc gccttcctga gcggcgagca
2701 gaaaaaggcc atcgtggacc tgctgttcaa gaccaaccgg aaagtgaccg tgaagcagct
2761 gaaagaggac tacttcaaga aaatcgagtg cttcgactcc gtggaaatct ccggcgtgga
2821 agatcggttc aacgcctccc tgggcacata ccacgatctg ctgaaaatta tcaaggacaa
2881 ggacttcctg gacaatgagg aaaacgagga cattctggaa gatatcgtgc tgaccctgac
2941 actgtttgag gacagagaga tgatcgagga acggctgaaa acctatgccc acctgttcga
3001 cgacaaagtg atgaagcagc tgaagcggcg gagatacacc ggctggggca ggctgagccg
3061 gaagctgatc aacggcatcc gggacaagca gtccggcaag acaatcctgg atttcctgaa
3121 gtccgacggc ttcgccaaca gaaacttcat gcagctgatc cacgacgaca gcctgacctt
3181 taaagaggac atccagaaag cccaggtgtc cggccagggc gatagcctgc acgagcacat
3241 tgccaatctg gccggcagcc ccgccattaa gaagggcatc ctgcagacag tgaaggtggt
3301 ggacgagctc gtgaaagtga tgggccggca caagcccgag aacatcgtga tcgaaatggc
3361 cagagagaac cagaccaccc agaagggaca gaagaacagc cgcgagagaa tgaagcggat
3421 cgaagagggc atcaaagagc tgggcagcca gatcctgaaa gaacaccccg tggaaaacac
3481 ccagctgcag aacgagaagc tgtacctgta ctacctgcag aatgggcggg atatgtacgt
3541 ggaccaggaa ctggacatca accggctgtc cgactacgat gtggaccata tcgtgcctca
3601 gagctttctg aaggacgact ccatcgacaa caaggtgctg accagaagcg acaagaaccg
3661 gggcaagagc gacaacgtgc cctccgaaga ggtcgtgaag aagatgaaga actactggcg
3721 gcagctgctg aacgccaagc tgattaccca gagaaagttc gacaatctga ccaaggccga
3781 gagaggcggc ctgagcgaac tggataaggc cggcttcatc aagagacagc tggtggaaac
3841 ccggcagatc acaaagcacg tggcacagat cctggactcc cggatgaaca ctaagtacga
3901 cgagaatgac aagctgatcc gggaagtgaa agtgatcacc ctgaagtcca agctggtgtc
3961 cgatttccgg aaggatttcc agttttacaa agtgcgcgag atcaacaact accaccacgc
4021 ccacgacgcc tacctgaacg ccgtcgtggg aaccgccctg atcaaaaagt accctaagct
4081 ggaaagcgag ttcgtgtacg gcgactacaa ggtgtacgac gtgcggaaga tgatcgccaa
4141 gagcgagcag gaaatcggca aggctaccgc caagtacttc ttctacagca acatcatgaa
4201 ctttttcaag accgagatta ccctggccaa cggcgagatc cggaagcggc ctctgatcga
4261 gacaaacggc gaaaccgggg agatcgtgtg ggataagggc cgggattttg ccaccgtgcg
4321 gaaagtgctg agcatgcccc aagtgaatat cgtgaaaaag accgaggtgc agacaggcgg
4381 cttcagcaaa gagtctatcc tgcccaagag gaacagcgat aagctgatcg ccagaaagaa
4441 ggactgggac cctaagaagt acggcggctt cgacagcccc accgtggcct attctgtgct
4501 ggtggtggcc aaagtggaaa agggcaagtc caagaaactg aagagtgtga aagagctgct
4561 ggggatcacc atcatggaaa gaagcagctt cgagaagaat cccatcgact ttctggaagc
4621 caagggctac aaagaagtga aaaaggacct gatcatcaag ctgcctaagt actccctgtt
4681 cgagctggaa aacggccgga agagaatgct ggcctctgcc ggcgaactgc agaagggaaa
4741 cgaactggcc ctgccctcca aatatgtgaa cttcctgtac ctggccagcc actatgagaa
4801 gctgaagggc tcccccgagg ataatgagca gaaacagctg tttgtggaac agcacaagca
4861 ctacctggac gagatcatcg agcagatcag cgagttctcc aagagagtga tcctggccga
4921 cgctaatctg gacaaagtgc tgtccgccta caacaagcac cgggataagc ccatcagaga
4981 gcaggccgag aatatcatcc acctgtttac cctgaccaat ctgggagccc ctgccgcctt
5041 caagtacttt gacaccacca tcgaccggaa gaggtacacc agcaccaaag aggtgctgga
5101 cgccaccctg atccaccaga gcatcaccgg cctgtacgag acacggatcg acctgtctca
5161 gctgggaggc gacaagcgac ctgccgccac aaagaaggct ggacaggcta agaagaagaa
5221 agattacaaa gacgatgacg ataagggatc cggcgcaaca aacttctctc tgctgaaaca
5281 agccggagat gtcgaagaga atcctggacc gaccgagtac aagcccacgg tgcgcctcgc
5341 cacccgcgac gacgtcccca gggccgtacg caccctcgcc gccgcgttcg ccgactaccc
5401 cgccacgcgc cacaccgtcg atccggaccg ccacatcgag cgggtcaccg agctgcaaga
5461 actcttcctc acgcgcgtcg ggctcgacat cggcaaggtg tgggtcgcgg acgacggcgc
5521 cgcggtggcg gtctggacca cgccggagag cgtcgaagcg ggggcggtgt tcgccgagat
5581 cggcccgcgc atggccgagt tgagcggttc ccggctggcc gcgcagcaac agatggaagg
5641 cctcctggcg ccgcaccggc ccaaggagcc cgcgtggttc ctggccaccg tcggagtctc
5701 gcccgaccac cagggcaagg gtctgggcag cgccgtcgtg ctccccggag tggaggcggc
5761 cgagcgcgcc ggggtgcccg ccttcctgga gacctccgcg ccccgcaacc tccccttcta
5821 cgagcggctc ggcttcaccg tcaccgccga cgtcgaggtg cccgaaggac cgcgcacctg
5881 gtgcatgacc cgcaagcccg gtgcctgaac gcgttaagtc gacaatcaac ctctggatta
5941 caaaatttgt gaaagattga ctggtattct taactatgtt gctcctttta cgctatgtgg
6001 atacgctgct ttaatgcctt tgtatcatgc tattgcttcc cgtatggctt tcattttctc
6061 ctccttgtat aaatcctggt tgctgtctct ttatgaggag ttgtggcccg ttgtcaggca
6121 acgtggcgtg gtgtgcactg tgtttgctga cgcaaccccc actggttggg gcattgccac
6181 cacctgtcag ctcctttccg ggactttcgc tttccccctc cctattgcca cggcggaact
6241 catcgccgcc tgccttgccc gctgctggac aggggctcgg ctgttgggca ctgacaattc
6301 cgtggtgttg tcggggaaat catcgtcctt tccttggctg ctcgcctgtg ttgccacctg
6361 gattctgcgc gggacgtcct tctgctacgt cccttcggcc ctcaatccag cggaccttcc
6421 ttcccgcggc ctgctgccgg ctctgcggcc tcttccgcgt cttcgccttc gccctcagac
6481 gagtcggatc tccctttggg ccgcctcccc gcgtcgactt taagaccaat gacttacaag
6541 gcagctgtag atcttagcca ctttttaaaa gaaaaggggg gactggaagg gctaattcac
6601 tcccaacgaa gacaagatct gctttttgct tgtactgggt ctctctggtt agaccagatc
6661 tgagcctggg agctctctgg ctaactaggg aacccactgc ttaagcctca ataaagcttg
6721 ccttgagtgc ttcaagtagt gtgtgcccgt ctgttgtgtg actctggtaa ctagagatcc
6781 ctcagaccct tttagtcagt gtggaaaatc tctagcaggg cccgtttaaa cccgctgatc
6841 agcctcgact gtgccttcta gttgccagcc atctgttgtt tgcccctccc ccgtgccttc
6901 cttgaccctg gaaggtgcca ctcccactgt cctttcctaa taaaatgagg aaattgcatc
6961 gcattgtctg agtaggtgtc attctattct ggggggtggg gtggggcagg acagcaaggg
7021 ggaggattgg gaagacaata gcaggcatgc tggggatgcg gtgggctcta tggcttctga
7081 ggcggaaaga accagctggg gctctagggg gtatccccac gcgccctgta gcggcgcatt
7141 aagcgcggcg ggtgtggtgg ttacgcgcag cgtgaccgct acacttgcca gcgccctagc
7201 gcccgctcct ttcgctttct tcccttcctt tctcgccacg ttcgccggct ttccccgtca
7261 agctctaaat cgggggctcc ctttagggtt ccgatttagt gctttacggc acctcgaccc
7321 caaaaaactt gattagggtg atggttcacg tagtgggcca tcgccctgat agacggtttt
7381 tcgccctttg acgttggagt ccacgttctt taatagtgga ctcttgttcc aaactggaac
7441 aacactcaac cctatctcgg tctattcttt tgatttataa gggattttgc cgatttcggc
7501 ctattggtta aaaaatgagc tgatttaaca aaaatttaac gcgaattaat tctgtggaat
7561 gtgtgtcagt tagggtgtgg aaagtcccca ggctccccag caggcagaag tatgcaaagc
7621 atgcatctca attagtcagc aaccaggtgt ggaaagtccc caggctcccc agcaggcaga
7681 agtatgcaaa gcatgcatct caattagtca gcaaccatag tcccgcccct aactccgccc
7741 atcccgcccc taactccgcc cagttccgcc cattctccgc cccatggctg actaattttt
7801 tttatttatg cagaggccga ggccgcctct gcctctgagc tattccagaa gtagtgagga
7861 ggcttttttg gaggcctagg cttttgcaaa aagctcccgg gagcttgtat atccattttc
7921 ggatctgatc agcacgtgtt gacaattaat catcggcata gtatatcggc atagtataat
7981 acgacaaggt gaggaactaa accatggcca agttgaccag tgccgttccg gtgctcaccg
8041 cgcgcgacgt cgccggagcg gtcgagttct ggaccgaccg gctcgggttc tcccgggact
8101 tcgtggagga cgacttcgcc ggtgtggtcc gggacgacgt gaccctgttc atcagcgcgg
8161 tccaggacca ggtggtgccg gacaacaccc tggcctgggt gtgggtgcgc ggcctggacg
8221 agctgtacgc cgagtggtcg gaggtcgtgt ccacgaactt ccgggacgcc tccgggccgg
8281 ccatgaccga gatcggcgag cagccgtggg ggcgggagtt cgccctgcgc gacccggccg
8341 gcaactgcgt gcacttcgtg gccgaggagc aggactgaca cgtgctacga gatttcgatt
8401 ccaccgccgc cttctatgaa aggttgggct tcggaatcgt tttccgggac gccggctgga
8461 tgatcctcca gcgcggggat ctcatgctgg agttcttcgc ccaccccaac ttgtttattg
8521 cagcttataa tggttacaaa taaagcaata gcatcacaaa tttcacaaat aaagcatttt
8581 tttcactgca ttctagttgt ggtttgtcca aactcatcaa tgtatcttat catgtctgta
8641 taccgtcgac ctctagctag agcttggcgt aatcatggtc atagctgttt cctgtgtgaa
8701 attgttatcc gctcacaatt ccacacaaca tacgagccgg aagcataaag tgtaaagcct
8761 ggggtgccta atgagtgagc taactcacat taattgcgtt gcgctcactg cccgctttcc
8821 agtcgggaaa cctgtcgtgc cagctgcatt aatgaatcgg ccaacgcgcg gggagaggcg
8881 gtttgcgtat tgggcgctct tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc
8941 ggctgcggcg agcggtatca gctcactcaa aggcggtaat acggttatcc acagaatcag
9001 gggataacgc aggaaagaac atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa
9061 aggccgcgtt gctggcgttt ttccataggc tccgcccccc tgacgagcat cacaaaaatc
9121 gacgctcaag tcagaggtgg cgaaacccga caggactata aagataccag gcgtttcccc
9181 ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga tacctgtccg
9241 cctttctccc ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg tatctcagtt
9301 cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga accccccgtt cagcccgacc
9361 gctgcgcctt atccggtaac tatcgtcttg agtccaaccc ggtaagacac gacttatcgc
9421 cactggcagc agccactggt aacaggatta gcagagcgag gtatgtaggc ggtgctacag
9481 agttcttgaa gtggtggcct aactacggct acactagaag aacagtattt ggtatctgcg
9541 ctctgctgaa gccagttacc ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa
9601 ccaccgctgg tagcggtggt ttttttgttt gcaagcagca gattacgcgc agaaaaaaag
9661 gatctcaaga agatcctttg atcttttcta cggggtctga cgctcagtgg aacgaaaact
9721 cacgttaagg gattttggtc atgagattat caaaaaggat cttcacctag atccttttaa
9781 attaaaaatg aagttttaaa tcaatctaaa gtatatatga gtaaacttgg tctgacagtt
9841 accaatgctt aatcagtgag gcacctatct cagcgatctg tctatttcgt tcatccatag
9901 ttgcctgact ccccgtcgtg tagataacta cgatacggga gggcttacca tctggcccca
9961 gtgctgcaat gataccgcga gacccacgct caccggctcc agatttatca gcaataaacc
10021 agccagccgg aagggccgag cgcagaagtg gtcctgcaac tttatccgcc tccatccagt
10081 ctattaattg ttgccgggaa gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg
10141 ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg gcttcattca
10201 gctccggttc ccaacgatca aggcgagtta catgatcccc catgttgtgc aaaaaagcgg
10261 ttagctcctt cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg ttatcactca
10321 tggttatggc agcactgcat aattctctta ctgtcatgcc atccgtaaga tgcttttctg
10381 tgactggtga gtactcaacc aagtcattct gagaatagtg tatgcggcga ccgagttgct
10441 cttgcccggc gtcaatacgg gataataccg cgccacatag cagaacttta aaagtgctca
10501 tcattggaaa acgttcttcg gggcgaaaac tctcaaggat cttaccgctg ttgagatcca
10561 gttcgatgta acccactcgt gcacccaact gatcttcagc atcttttact ttcaccagcg
10621 tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata agggcgacac
10681 ggaaatgttg aatactcata ctcttccttt ttcaatatta ttgaagcatt tatcagggtt
10741 attgtctcat gagcggatac atatttgaat gtatttagaa aaataaacaa ataggggttc
10801 cgcgcacatt tccccgaaaa gtgccacctg acgtcgacgg atcgggagat ctcccgatcc
10861 cctatggtgc actctcagta caatctgctc tgatgccgca tagttaagcc agtatctgct
10921 ccctgcttgt gtgttggagg tcgctgagta gtgcgcgagc aaaatttaag ctacaacaag
10981 gcaaggcttg accgacaatt gcatgaagaa tctgcttagg gttaggcgtt ttgcgctgct
11041 tcgcgatgta cgggccagat atacgcgttg acattgatta ttgactagtt attaatagta
11101 atcaattacg gggtcattag ttcatagccc atatatggag ttccgcgtta cataacttac
11161 ggtaaatggc ccgcctggct gaccgcccaa cgacccccgc ccattgacgt caataatgac
11221 gtatgttccc atagtaacgc caatagggac tttccattga cgtcaatggg tggagtattt
11281 acggtaaact gcccacttgg cagtacatca agtgtatcat atgccaagta cgccccctat
11341 tgacgtcaat gacggtaaat ggcccgcctg gcattatgcc cagtacatga ccttatggga
11401 ctttcctact tggcagtaca tctacgtatt agtcatcgct attaccatgg tgatgcggtt
11461 ttggcagtac atcaatgggc gtggatagcg gtttgactca cggggatttc caagtctcca
11521 ccccattgac gtcaatggga gtttgttttg gcaccaaaat caacgggact ttccaaaatg
11581 tcgtaacaac tccgccccat tgacgcaaat gggcggtagg cgtgtacggt gggaggtcta
11641 tataagcagc gcgttttgcc tgtactgggt ctctctggtt agaccagatc tgagcctggg
11701 agctctctgg ctaactaggg aacccactgc ttaagcctca ataaagcttg ccttgagtgc
11761 ttcaagtagt gtgtgcccgt ctgttgtgtg actctggtaa ctagagatcc ctcagaccct
11821 tttagtcagt gtggaaaatc tctagcagtg gcgcccgaac agggacttga aagcgaaagg
11881 gaaaccagag gagctctctc gacgcaggac tcggcttgct gaagcgcgca cggcaagagg
11941 cgaggggcgg cgactggtga gtacgccaaa aattttgact agcggaggct agaaggagag
12001 agatgggtgc gagagcgtca gtattaagcg ggggagaatt agatcgcgat gggaaaaaat
12061 tcggttaagg ccagggggaa agaaaaaata taaattaaaa catatagtat gggcaagcag
12121 ggagctagaa cgattcgcag ttaatcctgg cctgttagaa acatcagaag gctgtagaca
12181 aatactggga cagctacaac catcccttca gacaggatca gaagaactta gatcattata
12241 taatacagta gcaaccctct attgtgtgca tcaaaggata gagataaaag acaccaagga
12301 agctttagac aagatagagg aagagcaaaa caaaagtaag accaccgcac agcaagcggc
12361 cgctgatctt cagacctgga ggaggagata tgagggacaa ttggagaagt gaattatata
12421 aatataaagt agtaaaaatt gaaccattag gagtagcacc caccaaggca aagagaagag
12481 tggtgcagag agaaaaaaga gcagtgggaa taggagcttt gttccttggg ttcttgggag
12541 cagcaggaag cactatgggc gcagcgtcaa tgacgctgac ggtacaggcc agacaattat
12601 tgtctggtat agtgcagcag cagaacaatt tgctgagggc tattgaggcg caacagcatc
12661 tgttgcaact cacagtctgg ggcatcaagc agctccaggc aagaatcctg gctgtggaaa
12721 gatacctaaa ggatcaacag ctcctgggga tttggggttg ctctggaaaa ctcatttgca
12781 ccactgctgt gccttggaat gctagttgga gtaataaatc tctggaacag atttggaatc
12841 acacgacctg gatggagtgg gacagagaaa ttaacaatta cacaagctta atacactcct
12901 taattgaaga atcgcaaaac cagcaagaaa agaatgaaca agaattattg gaattagata
12961 aatgggcaag tttgtggaat tggtttaaca taacaaattg gctgtggtat a
//
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sgCtr- LentiCRISPRv2质粒载体应用举例
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