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pCAS9-mCherryempty哺乳CAS9和gRNA表达质粒
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pCAS9-mCherryempty哺乳CAS9和gRNA表达质粒
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基本信息
产品名称:
pCAS9-mCherryempty哺乳CAS9和gRNA表达质粒
产品编号:
P1649
规格:
10μl质粒/300μl菌液
原核抗性:
氨苄青霉素Amp
筛选标记:
红色荧光蛋白mCherry
克隆菌株:
大肠杆菌Stbl3
培养条件:
37℃
质粒简介:
pCAS9-mCherry empty哺乳CAS9和gRNA表达质粒。
质粒图谱:
质粒序列:
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LOCUS Exported 10048 bp ds-DNA circular SYN 30-NOV-2017
DEFINITION synthetic circular DNA
FEATURES Location/Qualifiers
source 1..10048
/organism="synthetic DNA construct"
/mol_type="other DNA"
LTR 8..477
/note="long terminal repeat from Moloney murine leukemia
virus"
CDS 938..1354
/label=gag (truncated)
/note="truncated Moloney murine leukemia virus (MMLV) gag
gene lacking the start codon"
promoter 1365..1605
/label=U6 promoter
/note="RNA polymerase III promoter for human U6 snRNA"
misc_feature 1614..1710
/label=MCS
enhancer 1712..2091
/label=CMV enhancer
/note="human cytomegalovirus immediate early enhancer"
promoter 2092..2295
/label=CMV promoter
/note="human cytomegalovirus (CMV) immediate early
promoter"
CDS 2350..3057
/codon_start=1
/product="monomeric derivative of DsRed fluorescent protein
(Shaner et al., 2004)"
/label=mCherry
/note="mammalian codon-optimized"
/translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEG
TQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNF
EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK
GEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERA
EGRHSTGGMDELYK"
CDS 3082..3135
/codon_start=1
/product="2A peptide from Thosea asigna virus capsid
protein"
/label=T2A
/note="Eukaryotic ribosomes fail to insert a peptide bond
between the Gly and Pro residues, yielding separate
polypeptides."
/translation="EGRGSLLTCGDVEENPGP"
CDS 3136..7239
/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 7252..7272
/codon_start=1
/product="nuclear localization signal of SV40 (simian virus
40) large T antigen"
/label=SV40 NLS
/translation="PKKKRKV"
LTR 7329..7922
/note="long terminal repeat from Moloney murine leukemia
virus"
rep_origin complement(8294..8882)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
CDS complement(9060..9920)
/codon_start=1
/gene="bla"
/product="beta-lactamase"
/label=AmpR
/note="confers resistance to ampicillin, carbenicillin, and
related antibiotics"
/translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
LIKHW"
promoter complement(9921..10025)
/gene="bla"
/label=AmpR promoter
ORIGIN
1 ctgcagcctg aatatgggcc aaacaggata tctgtggtaa gcagttcctg ccccggctca
61 gggccaagaa cagatggaac agctgaatat gggccaaaca ggatatctgt ggtaagcagt
121 tcctgccccg gctcagggcc aagaacagat ggtccccaga tgcggtccag ccctcagcag
181 tttctagaga accatcagat gtttccaggg tgccccaagg acctgaaatg accctgtgcc
241 ttatttgaac taaccaatca gttcgcttct cgcttctgtt cgcgcgcttc tgctccccga
301 gctcaataaa agagcccaca acccctcact cggggcgcca gtcctccgat tgactgagtc
361 gcccgggtac ccgtgtatcc aataaaccct cttgcagttg catccgactt gtggtctcgc
421 tgttccttgg gagggtctcc tctgagtgat tgactacccg tcagcggggg tctttcattt
481 gggggctcgt ccgggatcgg gagacccctg cccagggacc accgacccac caccgggagg
541 taagctggcc agcaacttat ctgtgtctgt ccgattgtct agtgtctatg actgatttta
601 tgcgcctgcg tcggtactag ctagttagct aactagctct gtatctggcg gacccgtggt
661 ggaactgacg agttctgaac acccggccgc aaccctggga gacgtcccag ggactttggg
721 ggccgttttt gtggcccgac ctgaggaagg gagtcgatgt ggaatccgac cccgtcagga
781 tatgtggttc tggtaggaga cgagaaccta aaacagttcc cgcctccgtc tgaatttttg
841 ctttcggttt ggaaccgaag ccgcgcgtct tgtctgctgc agcatcgttc tgtgttgtct
901 ctgtctgact gtgtttctgt atttgtctga aaattagggc cagactgtta ccactccctt
961 aagtttgacc ttagatcact ggaaagatgt cgagcggctc gctcacaacc agtcggtaga
1021 tgtcaagaag agacgttggg ttaccttctg ctctgcagaa tggccaacct ttaacgtcgg
1081 atggccgcga gacggcacct ttaaccgaga cctcatcacc caggttaaga tcaaggtctt
1141 ttcacctggc ccgcatggac acccagacca ggtcccctac atcgtgacct gggaagcctt
1201 ggcttttgac ccccctccct gggtcaagcc ctttgtacac cctaagcctc cgcctcctct
1261 tcttccatcc gcgccgtctc tcccccttga acctcctctt tcgaccccgc ctcaatcctc
1321 cctttatcca gccctcactc cttctctagg cgccggccgg atccgagggc ctatttccca
1381 tgattccttc atatttgcat atacgataca aggctgttag agagataatt ggaattaatt
1441 tgactgtaaa cacaaagata ttagtacaaa atacgtgacg tagaaagtaa taatttcttg
1501 ggtagtttgc agttttaaaa ttatgtttta aaatggacta tcatatgctt accgtaactt
1561 gaaagtattt cgatttcttg gctttatata tcttgtggaa aggacgaaac accgggccct
1621 cgatatcgac tagtccgtta tcaacttgaa aaagtggcac cgagtcggtg ctttttttcg
1681 aattccgatg tacgggccag atatacgcgt tgacattgat tattgtctag ttattaatag
1741 taatcaatta cggggtcatt agttcatagc ccatatatgg agttccgcgt tacataactt
1801 acggtaaatg gcccgcctgg ctgaccgccc aacgaccccc gcccattgac gtcaataatg
1861 acgtatgttc ccatagtaac gccaataggg actttccatt gacgtcaatg ggtggagtat
1921 ttacggtaaa ctgcccactt ggcagtacat caagtgtatc atatgccaag tacgccccct
1981 attgacgtca atgacggtaa atggcccgcc tggcattatg cccagtacat gaccttatgg
2041 gactttccta cttggcagta catctacgta ttagtcatcg ctattaccat ggtgatgcgg
2101 ttttggcagt acatcaatgg gcgtggatag cggtttgact cacggggatt tccaagtctc
2161 caccccattg acgtcaatgg gagtttgttt tggcaccaaa atcaacggga ctttccaaaa
2221 tgtcgtaaca actccgcccc attgacgcaa atgggcggta ggcgtgtacg gtgggaggtc
2281 tatataagca gagctctctg gctaactaga gaacccactg cttactggct tatcgaaatt
2341 aagcttccca tggtgagcaa gggcgaggag gataacatgg ccatcatcaa ggagttcatg
2401 cgcttcaagg tgcacatgga gggctccgtg aacggccacg agttcgagat cgagggcgag
2461 ggcgagggcc gcccctacga gggcacccag accgccaagc tgaaggtgac caagggtggc
2521 cccctgccct tcgcctggga catcctgtcc cctcagttca tgtacggctc caaggcctac
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