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pBK-VRER-BE3哺乳基因编辑质粒
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pBK-VRER-BE3哺乳基因编辑质粒
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基本信息
产品名称:
pBK-VRER-BE3哺乳基因编辑质粒
产品编号:
P1672
规格:
10μl质粒/300μl菌液
原核抗性:
氨苄青霉素Amp
克隆菌株:
大肠杆菌Stbl3
培养条件:
37℃
质粒简介:
pBK-VRER-BE3是一个大鼠基因哺乳过表达质粒。
质粒图谱:
质粒序列:
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LOCUS Exported 8532 bp ds-DNA circular SYN 01-DEC-2017
DEFINITION synthetic circular DNA
FEATURES Location/Qualifiers
source 1..8532
/organism="synthetic DNA construct"
/mol_type="other DNA"
promoter 132..335
/label=CMV promoter
/note="human cytomegalovirus (CMV) immediate early
promoter"
promoter 377..395
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
CDS 409..1095
/codon_start=1
/product="cytidine deaminase (C to U editing enzyme) from
rat"
/label=APOBEC-1
/note="can use ssDNA as a substrate (Komor et al., 2016)"
/translation="MSSETGPVAVDPTLRRRIEPHEFEVFFDPRELRKETCLLYEINWG
GRHSIWRHTSQNTNKHVEVNFIEKFTTERYFCPNTRCSITWFLSWSPCGECSRAITEFL
SRYPHVTLFIYIARLYHHADPRNRQGLRDLISSGVTIQIMTEQESGYCWRNFVNYSPSN
EAHWPRYPHLWVRLYVLELYCIILGLPPCLNILRRKQPQLTFFTIALQSCHYQRLPPHI
LWATGLK"
CDS 1144..5244
/codon_start=1
/product="Streptococcus pyogenes Cas9 endonuclease with the
D1135V/G1218R/R1335E/T1337R mutations"
/label=Cas9 VRER
/note="has an altered recognition specificity for the
protospacer adjacent motif (PAM) (Kleinstiver et al.,
2015)"
/translation="DKKYSIGLAIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKKN
LIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEESF
LVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIKF
RGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRLE
NLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQI
GDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVRQ
QLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLLR
KQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARGN
SRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEYF
TVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFDS
VEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEERL
KTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFMQ
LIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGRH
KPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLYL
YYLQNGRDMYVDQELDINRLSDYDVDHIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVPS
EEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKHV
AQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYLN
AVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKTE
ITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSKE
SILPKRNSDKLIARKKDWDPKKYGGFVSPTVAYSVLVVAKVEKGKSKKLKSVKELLGIT
IMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASARELQKGNEL
ALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILADA
NLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKEYRSTKEVLD
ATLIHQSITGLYETRIDLSQLGGD"
CDS 5257..5505
/codon_start=1
/product="uracil-DNA glycosylase inhibitor from a Bacillus
subtilis bacteriophage (Mol et al., 1995)"
/label=UGI
/translation="TNLSDIIEKETGKQLVIQESILMLPEEVEEVIGNKPESDILVHTA
YDESTDENVMLLTSDAPEYKPWALVIQDSNGENKIKML"
CDS 5518..5538
/codon_start=1
/product="nuclear localization signal of SV40 (simian virus
40) large T antigen"
/label=SV40 NLS
/translation="PKKKRKV"
CDS 5547..5564
/codon_start=1
/product="6xHis affinity tag"
/label=6xHis
/translation="HHHHHH"
polyA_signal 5593..5817
/label=bGH poly(A) signal
/note="bovine growth hormone polyadenylation signal"
rep_origin complement(6290..6878)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
CDS complement(7049..7909)
/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"
promoter complement(7910..8014)
/gene="bla"
/label=AmpR promoter
enhancer 8284..131
/label=CMV enhancer
/note="human cytomegalovirus immediate early enhancer"
ORIGIN
1 atatgccaag tacgccccct attgacgtca atgacggtaa atggcccgcc tggcattatg
61 cccagtacat gaccttatgg gactttccta cttggcagta catctacgta ttagtcatcg
121 ctattaccat ggtgatgcgg ttttggcagt acatcaatgg gcgtggatag cggtttgact
181 cacggggatt tccaagtctc caccccattg acgtcaatgg gagtttgttt tggcaccaaa
241 atcaacggga ctttccaaaa tgtcgtaaca actccgcccc attgacgcaa atgggcggta
301 ggcgtgtacg gtgggaggtc tatataagca gagctggttt agtgaaccgt cagatccgct
361 agagatccgc ggccgctaat acgactcact atagggagag ccgccaccat gagctcagag
421 actggcccag tggctgtgga ccccacattg agacggcgga tcgagcccca tgagtttgag
481 gtattcttcg atccgagaga gctccgcaag gagacctgcc tgctttacga aattaattgg
541 gggggccggc actccatttg gcgacataca tcacagaaca ctaacaagca cgtcgaagtc
601 aacttcatcg agaagttcac gacagaaaga tatttctgtc cgaacacaag gtgcagcatt
661 acctggtttc tcagctggag cccatgcggc gaatgtagta gggccatcac tgaattcctg
721 tcaaggtatc cccacgtcac tctgtttatt tacatcgcaa ggctgtacca ccacgctgac
781 ccccgcaatc gacaaggcct gcgggatttg atctcttcag gtgtgactat ccaaattatg
841 actgagcagg agtcaggata ctgctggaga aactttgtga attatagccc gagtaatgaa
901 gcccactggc ctaggtatcc ccatctgtgg gtacgactgt acgttcttga actgtactgc
961 atcatactgg gcctgcctcc ttgtctcaac attctgagaa ggaagcagcc acagctgaca
1021 ttctttacca tcgctcttca gtcttgtcat taccagcgac tgcccccaca cattctctgg
1081 gccaccgggt tgaaaagcgg cagcgagact cccgggacct cagagtccgc cacacccgaa
1141 agtgataaaa agtattctat tggtttagcc atcggcacta attccgttgg atgggctgtc
1201 ataaccgatg aatacaaagt accttcaaag aaatttaagg tgttggggaa cacagaccgt
1261 cattcgatta aaaagaatct tatcggtgcc ctcctattcg atagtggcga aacggcagag
1321 gcgactcgcc tgaaacgaac cgctcggaga aggtatacac gtcgcaagaa ccgaatatgt
1381 tacttacaag aaatttttag caatgagatg gccaaagttg acgattcttt ctttcaccgt
1441 ttggaagagt ccttccttgt cgaagaggac aagaaacatg aacggcaccc catctttgga
1501 aacatagtag atgaggtggc atatcatgaa aagtacccaa cgatttatca cctcagaaaa
1561 aagctagttg actcaactga taaagcggac ctgaggttaa tctacttggc tcttgcccat
1621 atgataaagt tccgtgggca ctttctcatt gagggtgatc taaatccgga caactcggat
1681 gtcgacaaac tgttcatcca gttagtacaa acctataatc agttgtttga agagaaccct
1741 ataaatgcaa gtggcgtgga tgcgaaggct attcttagcg cccgcctctc taaatcccga
1801 cggctagaaa acctgatcgc acaattaccc ggagagaaga aaaatgggtt gttcggtaac
1861 cttatagcgc tctcactagg cctgacacca aattttaagt cgaacttcga cttagctgaa
1921 gatgccaaat tgcagcttag taaggacacg tacgatgacg atctcgacaa tctactggca
1981 caaattggag atcagtatgc ggacttattt ttggctgcca aaaaccttag cgatgcaatc
2041 ctcctatctg acatactgag agttaatact gagattacca aggcgccgtt atccgcttca
2101 atgatcaaaa ggtacgatga acatcaccaa gacttgacac ttctcaaggc cctagtccgt
2161 cagcaactgc ctgagaaata taaggaaata ttctttgatc agtcgaaaaa cgggtacgca
2221 ggttatattg acggcggagc gagtcaagag gaattctaca agtttatcaa acccatatta
2281 gagaagatgg atgggacgga agagttgctt gtaaaactca atcgcgaaga tctactgcga
2341 aagcagcgga ctttcgacaa cggtagcatt ccacatcaaa tccacttagg cgaattgcat
2401 gctatactta gaaggcagga ggatttttat ccgttcctca aagacaatcg tgaaaagatt
2461 gagaaaatcc taacctttcg cataccttac tatgtgggac ccctggcccg agggaactct
2521 cggttcgcat ggatgacaag aaagtccgaa gaaacgatta ctccatggaa ttttgaggaa
2581 gttgtcgata aaggtgcgtc agctcaatcg ttcatcgaga ggatgaccaa ctttgacaag
2641 aatttaccga acgaaaaagt attgcctaag cacagtttac tttacgagta tttcacagtg
2701 tacaatgaac tcacgaaagt taagtatgtc actgagggca tgcgtaaacc cgcctttcta
2761 agcggagaac agaagaaagc aatagtagat ctgttattca agaccaaccg caaagtgaca
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