质粒名称: | pCAG-Flpe |
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出品公司: | 爱迪基因 |
目录编号: | 13787 |
存储实验室: | Connie Cepko |
载体骨架基本信息 |
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载体名称: | pCAGEN |
载体抗性: | 氨苄霉素 |
空载体大小: | 4779 bp |
完整质粒大小: | 6111 bp |
载体类型: | 哺乳细胞表达 |
高拷贝/低拷贝: | 高拷贝 |
生长条件: | 37 ℃ |
插入基因信息 |
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插入基因名称: | Flpe |
物种来源: | S. cerevisiae (芽殖酵母) |
插入大小: | 1332 bp |
基因又名: | enhanced form of Flp recombinase |
标签/融合蛋白: | Myc (C terminal on insert) |
克隆方法: | 限制性内切酶 |
5' 克隆位点: | EcoRI(未破坏) |
3' 克隆位点: | NotI(未破坏) |
5' 测序引物 | pCAG-F |
3' 测序引物 | Bglob-pA-R |
文献引用方法 |
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材料和方法部分: | pCAG-Flpe was a gift from Connie Cepko (爱迪基因 plasmid # 13787) |
参考文献部分: | Controlled expression of transgenes introduced by in vivo electroporation. Matsuda T, Cepko CL. Proc Natl Acad Sci U S A. 2007 Jan 5. ():. 10.1073/pnas.0610155104 PubMed 17209010 |
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载体基本信息
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载体简介
载体序列
LOCUS pCAG-Flpe 6111 bp ds-DNA circular SYN 10-JAN-2018 DEFINITION Mammalian expression of enhanced form of Flp recombinase. ACCESSION . VERSION . KEYWORDS pCAG-Flpe SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 6111) AUTHORS Matsuda T, Cepko CL TITLE Controlled expression of transgenes introduced by in vivo electroporation. JOURNAL Proc Natl Acad Sci U S A. 2007 Jan 5. ():. PUBMED 17209010 COMMENT This file is created by Vector NTI http://www.biofeng.com/ COMMENT ORIGDB|GenBank COMMENT VNTAUTHORNAME|biofeng.com| COMMENT VNTNAME|pCAG-Flpe| FEATURES Location/Qualifiers source 1..6111 /organism="synthetic DNA construct" /mol_type="other DNA" enhancer 4..383 /label=CMV enhancer /note="human cytomegalovirus immediate early enhancer" promoter 386..661 /label=chicken beta-actin promoter intron 662..1670 /label=chimeric intron /note="chimera between introns from chicken beta-actin and rabbit beta-globin" primer_bind 1594..1616 /label=pCAGGS-5 /note="Chimeric intron in CAG promoter, forward primer" primer_bind 1678..1697 /label=pCAG-F /note="Rabbit beta-globin intron, for pCAG plasmids, forward primer" CDS 1737..3005 /codon_start=1 /product="site-specific recombinase" /label=FLP /note="FLP is a site-specific recombinase from Saccharomyces cerevisiae. Recombination occurs at FRT sequences." /translation="MSQFDILCKTPPKVLVRQFVERFERPSGEKIASCAAELTYLCWMI THNGTAIKRATFMSYNTIISNSLSFDIVNKSLQFKYKTQKATILEASLKKLIPAWEFTI IPYNGQKHQSDITDIVSSLQLQFESSEEADKGNSHSKKMLKALLSEGESIWEITEKILN SFEYTSRFTKTKTLYQFLFLATFINCGRFSDIKNVDPKSFKLVQNKYLGVIIQCLVTET KTSVSRHIYFFSARGRIDPLVYLDEFLRNSEPVLKRVNRTGNSSSNKQEYQLLKDNLVR SYNKALKKNAPYPIFAIKNGPKSHIGRHLMTSFLSMKGLTELTNVVGNWSDKRASAVAR TTYTHQITAIPDHYFALVSRYYAYDPISKEMIALKDETNPIEEWQHIEQLKGSAEGSIR YPAWNGIISQEVLDYLSSYINRRI" CDS 3015..3044 /codon_start=1 /product="Myc (human c-Myc proto-oncogene) epitope tag" /label=Myc /translation="EQKLISEEDL" primer_bind complement(3135..3154) /label=Bglob-pA-R /note="Rabbit beta-globin polyA region, reverse primer" polyA_site 3200..3255 /label=-beta-globin poly(A) signal /note="rabbit beta-globin polyadenylation signal" primer_bind complement(3254..3273) /label=rbglobpA-R /note="Rabbit beta-globin polyA, reverse primer. Also called rb-glob-pA-term-R" primer_bind complement(3614..3630) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind complement(3614..3630) /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" primer_bind complement(3627..3649) /label=M13/pUC Reverse /note="In lacZ gene" protein_bind 3638..3654 /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(3662..3692) /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 3707..3728 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." promoter 3787..3982 /label=SV40 promoter /note="SV40 early promoter" rep_origin 3833..3968 /label=SV40 ori /note="SV40 origin of replication" primer_bind 3895..3914 /label=SV40pro-F /note="SV40 promoter/origin, forward primer" polyA_signal 3988..4122 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(4025..4044) /label=SV40pA-R /note="SV40 polyA, reverse primer" primer_bind 4079..4098 /label=EBV-rev /note="SV40 polyA terminator, reverse primer" primer_bind complement(4190..4207) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(4361..4949) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(4441..4460) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(5120..5980) /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 5743..5762 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(5981..6085) /gene="bla" /label=AmpR promoter ORIGIN 1 gtcgacattg attattgact agttattaat agtaatcaat tacggggtca ttagttcata 61 gcccatatat ggagttccgc gttacataac ttacggtaaa tggcccgcct ggctgaccgc 121 ccaacgaccc ccgcccattg acgtcaataa tgacgtatgt tcccatagta acgccaatag 181 ggactttcca ttgacgtcaa tgggtggact atttacggta aactgcccac ttggcagtac 241 atcaagtgta tcatatgcca agtacgcccc ctattgacgt caatgacggt aaatggcccg 301 cctggcatta tgcccagtac atgaccttat gggactttcc tacttggcag tacatctacg 361 tattagtcat cgctattacc atgggtcgag gtgagcccca cgttctgctt cactctcccc 421 atctcccccc cctccccacc cccaattttg tatttattta ttttttaatt attttgtgca 481 gcgatggggg cggggggggg gggggcgcgc gccaggcggg gcggggcggg gcgaggggcg 541 gggcggggcg aggcggagag gtgcggcggc agccaatcag agcggcgcgc tccgaaagtt 601 tccttttatg gcgaggcggc ggcggcggcg gccctataaa aagcgaagcg cgcggcgggc 661 gggagtcgct gcgttgcctt cgccccgtgc cccgctccgc gccgcctcgc gccgcccgcc 721 ccggctctga ctgaccgcgt tactcccaca ggtgagcggg cgggacggcc cttctcctcc 781 gggctgtaat tagcgcttgg tttaatgacg gctcgtttct tttctgtggc tgcgtgaaag 841 ccttaaaggg ctccgggagg gccctttgtg cgggggggag cggctcgggg ggtgcgtgcg 901 tgtgtgtgtg cgtggggagc gccgcgtgcg gcccgcgctg cccggcggct gtgagcgctg 961 cgggcgcggc gcggggcttt gtgcgctccg cgtgtgcgcg aggggagcgc ggccgggggc 1021 ggtgccccgc ggtgcggggg ggctgcgagg ggaacaaagg ctgcgtgcgg ggtgtgtgcg 1081 tgggggggtg agcagggggt gtgggcgcgg cggtcgggct gtaacccccc cctgcacccc 1141 cctccccgag ttgctgagca cggcccggct tcgggtgcgg ggctccgtgc ggggcgtggc 1201 gcggggctcg ccgtgccggg cggggggtgg cggcaggtgg gggtgccggg cggggcgggg 1261 ccgcctcggg ccggggaggg ctcgggggag gggcgcggcg gccccggagc gccggcggct 1321 gtcgaggcgc ggcgagccgc agccattgcc ttttatggta atcgtgcgag agggcgcagg 1381 gacttccttt gtcccaaatc tggcggagcc gaaatctggg aggcgccgcc gcaccccctc 1441 tagcgggcgc gggcgaagcg gtgcggcgcc ggcaggaagg aaatgggcgg ggagggcctt 1501 cgtgcgtcgc cgcgccgccg tccccttctc catctccagc ctcggggctg ccgcaggggg 1561 acggctgcct tcggggggga cggggcaggg cggggttcgg cttctggcgt gtgaccggcg 1621 gctctagagc ctctgctaac catgttcatg ccttcttctt tttcctacag ctcctgggca 1681 acgtgctggt tattgtgctg tctcatcatt ttggcaaaga attctgagcc gccaccatga 1741 gccaatttga tatattatgt aaaacaccac ctaaggtcct ggttcgtcag tttgtggaaa 1801 ggtttgaaag accttcaggg gaaaaaatag catcatgtgc tgctgaacta acctatttat 1861 gttggatgat tactcataac ggaacagcaa tcaagagagc cacattcatg agctataata 1921 ctatcataag caattcgctg agtttcgata ttgtcaacaa atcactccag tttaaataca 1981 agacgcaaaa agcaacaatt ctggaagcct cattaaagaa attaattcct gcttgggaat 2041 ttacaattat tccttacaat ggacaaaaac atcaatctga tatcactgat attgtaagta 2101 gtttgcaatt acagttcgaa tcatcggaag aagcagataa gggaaatagc cacagtaaaa 2161 aaatgcttaa agcacttcta agtgagggtg aaagcatctg ggagatcact gagaaaatac 2221 taaattcgtt tgagtatacc tcgagattta caaaaacaaa aactttatac caattcctct 2281 tcctagctac tttcatcaat tgtggaagat tcagcgatat taagaacgtt gatccgaaat 2341 catttaaatt agtccaaaat aagtatctgg gagtaataat ccagtgttta gtgacagaga 2401 caaagacaag cgttagtagg cacatatact tctttagcgc aaggggtagg atcgatccac 2461 ttgtatattt ggatgaattt ttgaggaact ctgaaccagt cctaaaacga gtaaatagga 2521 ccggcaattc ttcaagcaac aaacaggaat accaattatt aaaagataac ttagtcagat 2581 cgtacaacaa ggctttgaag aaaaatgcgc cttatccaat ctttgctata aagaatggcc 2641 caaaatctca cattggaaga catttgatga cctcatttct gtcaatgaag ggcctaacgg 2701 agttgactaa tgttgtggga aattggagcg ataagcgtgc ttctgccgtg gccaggacaa 2761 cgtatactca tcagataaca gcaatacctg atcactactt cgcactagtt tctcggtact 2821 atgcatatga tccaatatca aaggaaatga tagcattgaa ggatgagact aatccaattg 2881 aggagtggca gcatatagaa cagctaaagg gtagtgctga aggaagcata cgataccccg 2941 catggaatgg gataatatca caggaggtac tagactacct ttcatcctac ataaatagac 3001 gcataggacc ggtggaacaa aaacttattt ctgaagaaga tctgtgatag cggccgcact 3061 cctcaggtgc aggctgccta tcagaaggtg gtggctggtg tggccaatgc cctggctcac 3121 aaataccact gagatctttt tccctctgcc aaaaattatg gggacatcat gaagcccctt 3181 gagcatctga cttctggcta ataaaggaaa tttattttca ttgcaatagt gtgttggaat 3241 tttttgtgtc tctcactcgg aaggacatat gggagggcaa atcatttaaa acatcagaat 3301 gagtatttgg tttagagttt ggcaacatat gccatatgct ggctgccatg aacaaaggtg 3361 gctataaaga ggtcatcagt atatgaaaca gccccctgct gtccattcct tattccatag 3421 aaaagccttg acttgaggtt agattttttt tatattttgt tttgtgttat ttttttcttt 3481 aacatcccta aaattttcct tacatgtttt actagccaga tttttcctcc tctcctgact 3541 actcccagtc atagctgtcc ctcttctctt atgaagatcc ctcgacctgc agcccaagct 3601 tggcgtaatc atggtcatag ctgtttcctg tgtgaaattg ttatccgctc acaattccac 3661 acaacatacg agccggaagc ataaagtgta aagcctgggg tgcctaatga gtgagctaac 3721 tcacattaat tgcgttgcgc tcactgcccg ctttccagtc gggaaacctg tcgtgccagc 3781 ggatccgcat ctcaattagt cagcaaccat agtcccgccc ctaactccgc ccatcccgcc 3841 cctaactccg cccagttccg cccattctcc gccccatggc tgactaattt tttttattta 3901 tgcagaggcc gaggccgcct cggcctctga gctattccag aagtagtgag gaggcttttt 3961 tggaggccta ggcttttgca aaaagctaac ttgtttattg cagcttataa tggttacaaa 4021 taaagcaata gcatcacaaa tttcacaaat aaagcatttt tttcactgca ttctagttgt 4081 ggtttgtcca aactcatcaa tgtatcttat catgtctgga tccgctgcat taatgaatcg 4141 gccaacgcgc ggggagaggc ggtttgcgta ttgggcgctc ttccgcttcc tcgctcactg 4201 actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc agctcactca aaggcggtaa 4261 tacggttatc cacagaatca ggggataacg caggaaagaa catgtgagca aaaggccagc 4321 aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg ctccgccccc 4381 ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg acaggactat 4441 aaagatacca ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc 4501 cgcttaccgg atacctgtcc gcctttctcc cttcgggaag cgtggcgctt tctcaatgct 4561 cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg 4621 aaccccccgt tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc 4681 cggtaagaca cgacttatcg ccactggcag cagccactgg taacaggatt agcagagcga 4741 ggtatgtagg cggtgctaca gagttcttga agtggtggcc taactacggc tacactagaa 4801 ggacagtatt tggtatctgc gctctgctga agccagttac cttcggaaaa agagttggta 4861 gctcttgatc cggcaaacaa accaccgctg gtagcggtgg tttttttgtt tgcaagcagc 4921 agattacgcg cagaaaaaaa ggatctcaag aagatccttt gatcttttct acggggtctg 4981 acgctcagtg gaacgaaaac tcacgttaag ggattttggt catgagatta tcaaaaagga 5041 tcttcaccta gatcctttta aattaaaaat gaagttttaa atcaatctaa agtatatatg 5101 agtaaacttg gtctgacagt taccaatgct taatcagtga ggcacctatc tcagcgatct 5161 gtctatttcg ttcatccata gttgcctgac tccccgtcgt gtagataact acgatacggg 5221 agggcttacc atctggcccc agtgctgcaa tgataccgcg agacccacgc tcaccggctc 5281 cagatttatc agcaataaac cagccagccg gaagggccga gcgcagaagt ggtcctgcaa 5341 ctttatccgc ctccatccag tctattaatt gttgccggga agctagagta agtagttcgc 5401 cagttaatag tttgcgcaac gttgttgcca ttgctacagg catcgtggtg tcacgctcgt 5461 cgtttggtat ggcttcattc agctccggtt cccaacgatc aaggcgagtt acatgatccc 5521 ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc agaagtaagt 5581 tggccgcagt gttatcactc atggttatgg cagcactgca taattctctt actgtcatgc 5641 catccgtaag atgcttttct gtgactggtg agtactcaac caagtcattc tgagaatagt 5701 gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg ggataatacc gcgccacata 5761 gcagaacttt aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa ctctcaagga 5821 tcttaccgct gttgagatcc agttcgatgt aacccactcg tgcacccaac tgatcttcag 5881 catcttttac tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa 5941 aaaagggaat aagggcgaca cggaaatgtt gaatactcat actcttcctt tttcaatatt 6001 attgaagcat ttatcagggt tattgtctca tgagcggata catatttgaa tgtatttaga 6061 aaaataaaca aataggggtt ccgcgcacat ttccccgaaa agtgccacct g //
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