| 质粒名称: | pCAG-Flpe |
|---|---|
| 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 |
|
| 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 |
|
| 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 |
|
| 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 |
pCAG-Flpe
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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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