| 质粒名称: | pCAG-Flpe:GFP |
|---|---|
| 出品公司: | 爱迪基因 |
| 目录编号: | 13788 |
| 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: | 6803 bp |
| 载体类型: | 哺乳细胞表达，Flp/FRT |
| 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-GFP fusion protein |
| 物种来源: | S. cerevisiae（芽殖酵母） |
| 插入大小: | 2024 bp |
| 标签/融合蛋白: | GFP (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 |
|
文献引用方法 |
|
| 材料和方法部分: | pCAG-Flpe:GFP was a gift from Connie Cepko (爱迪基因 plasmid # 13788) |
| 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:GFP
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LOCUS pCAG-Flpe:GFP 6803 bp ds-DNA circular SYN 10-JAN-2018
DEFINITION Mammalian expression of enhanced form of Flp recombinase fused to
GFP.
ACCESSION .
VERSION .
KEYWORDS pCAG-Flpe:GFP
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 6803)
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:GFP|
FEATURES Location/Qualifiers
source 1..6803
/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"
regulatory 3015..3024
/regulatory_class="other"
/note="vertebrate consensus sequence for strong initiation
of translation (Kozak, 1987)"
CDS 3021..3740
/codon_start=1
/product="enhanced GFP"
/label=EGFP
/note="mammalian codon-optimized"
/translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
EFVTAAGITLGMDELYK"
primer_bind complement(3066..3087)
/label=EGFP-N
/note="EGFP, reverse primer"
primer_bind complement(3327..3346)
/label=EXFP-R
/note="For distinguishing EGFP variants, reverse primer"
primer_bind 3674..3695
/label=EGFP-C
/note="EGFP, forward primer"
primer_bind complement(3827..3846)
/label=Bglob-pA-R
/note="Rabbit beta-globin polyA region, reverse primer"
polyA_site 3892..3947
/label=-beta-globin poly(A) signal
/note="rabbit beta-globin polyadenylation signal"
primer_bind complement(3946..3965)
/label=rbglobpA-R
/note="Rabbit beta-globin polyA, reverse primer. Also
called rb-glob-pA-term-R"
primer_bind complement(4306..4322)
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(4306..4322)
/label=M13 Reverse
/note="In lacZ gene. Also called M13-rev"
primer_bind complement(4319..4341)
/label=M13/pUC Reverse
/note="In lacZ gene"
protein_bind 4330..4346
/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(4354..4384)
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 4399..4420
/label=CAP binding site
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding activates transcription in the presence
of cAMP."
promoter 4479..4674
/label=SV40 promoter
/note="SV40 early promoter"
rep_origin 4525..4660
/label=SV40 ori
/note="SV40 origin of replication"
primer_bind 4587..4606
/label=SV40pro-F
/note="SV40 promoter/origin, forward primer"
polyA_signal 4680..4814
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
primer_bind complement(4717..4736)
/label=SV40pA-R
/note="SV40 polyA, reverse primer"
primer_bind 4771..4790
/label=EBV-rev
/note="SV40 polyA terminator, reverse primer"
primer_bind complement(4882..4899)
/label=L4440
/note="L4440 vector, forward primer"
rep_origin complement(5053..5641)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind complement(5133..5152)
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
CDS complement(5812..6672)
/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 6435..6454
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
promoter complement(6673..6777)
/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 ggtcgccacc atggtgagca agggcgagga gctgttcacc ggggtggtgc
3061 ccatcctggt cgagctggac ggcgacgtaa acggccacaa gttcagcgtg tccggcgagg
3121 gcgagggcga tgccacctac ggcaagctga ccctgaagtt catctgcacc accggcaagc
3181 tgcccgtgcc ctggcccacc ctcgtgacca ccctgaccta cggcgtgcag tgcttcagcc
3241 gctaccccga ccacatgaag cagcacgact tcttcaagtc cgccatgccc gaaggctacg
3301 tccaggagcg caccatcttc ttcaaggacg acggcaacta caagacccgc gccgaggtga
3361 agttcgaggg cgacaccctg gtgaaccgca tcgagctgaa gggcatcgac ttcaaggagg
3421 acggcaacat cctggggcac aagctggagt acaactacaa cagccacaac gtctatatca
3481 tggccgacaa gcagaagaac ggcatcaagg tgaacttcaa gatccgccac aacatcgagg
3541 acggcagcgt gcagctcgcc gaccactacc agcagaacac ccccatcggc gacggccccg
3601 tgctgctgcc cgacaaccac tacctgagca cccagtccgc cctgagcaaa gaccccaacg
3661 agaagcgcga tcacatggtc ctgctggagt tcgtgaccgc cgccgggatc actctcggca
3721 tggacgagct gtacaagtaa agcggccgca ctcctcaggt gcaggctgcc tatcagaagg
3781 tggtggctgg tgtggccaat gccctggctc acaaatacca ctgagatctt tttccctctg
3841 ccaaaaatta tggggacatc atgaagcccc ttgagcatct gacttctggc taataaagga
3901 aatttatttt cattgcaata gtgtgttgga attttttgtg tctctcactc ggaaggacat
3961 atgggagggc aaatcattta aaacatcaga atgagtattt ggtttagagt ttggcaacat
4021 atgccatatg ctggctgcca tgaacaaagg tggctataaa gaggtcatca gtatatgaaa
4081 cagccccctg ctgtccattc cttattccat agaaaagcct tgacttgagg ttagattttt
4141 tttatatttt gttttgtgtt atttttttct ttaacatccc taaaattttc cttacatgtt
4201 ttactagcca gatttttcct cctctcctga ctactcccag tcatagctgt ccctcttctc
4261 ttatgaagat ccctcgacct gcagcccaag cttggcgtaa tcatggtcat agctgtttcc
4321 tgtgtgaaat tgttatccgc tcacaattcc acacaacata cgagccggaa gcataaagtg
4381 taaagcctgg ggtgcctaat gagtgagcta actcacatta attgcgttgc gctcactgcc
4441 cgctttccag tcgggaaacc tgtcgtgcca gcggatccgc atctcaatta gtcagcaacc
4501 atagtcccgc ccctaactcc gcccatcccg cccctaactc cgcccagttc cgcccattct
4561 ccgccccatg gctgactaat tttttttatt tatgcagagg ccgaggccgc ctcggcctct
4621 gagctattcc agaagtagtg aggaggcttt tttggaggcc taggcttttg caaaaagcta
4681 acttgtttat tgcagcttat aatggttaca aataaagcaa tagcatcaca aatttcacaa
4741 ataaagcatt tttttcactg cattctagtt gtggtttgtc caaactcatc aatgtatctt
4801 atcatgtctg gatccgctgc attaatgaat cggccaacgc gcggggagag gcggtttgcg
4861 tattgggcgc tcttccgctt cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg
4921 gcgagcggta tcagctcact caaaggcggt aatacggtta tccacagaat caggggataa
4981 cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc
5041 gttgctggcg tttttccata ggctccgccc ccctgacgag catcacaaaa atcgacgctc
5101 aagtcagagg tggcgaaacc cgacaggact ataaagatac caggcgtttc cccctggaag
5161 ctccctcgtg cgctctcctg ttccgaccct gccgcttacc ggatacctgt ccgcctttct
5221 cccttcggga agcgtggcgc tttctcaatg ctcacgctgt aggtatctca gttcggtgta
5281 ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc gttcagcccg accgctgcgc
5341 cttatccggt aactatcgtc ttgagtccaa cccggtaaga cacgacttat cgccactggc
5401 agcagccact ggtaacagga ttagcagagc gaggtatgta ggcggtgcta cagagttctt
5461 gaagtggtgg cctaactacg gctacactag aaggacagta tttggtatct gcgctctgct
5521 gaagccagtt accttcggaa aaagagttgg tagctcttga tccggcaaac aaaccaccgc
5581 tggtagcggt ggtttttttg tttgcaagca gcagattacg cgcagaaaaa aaggatctca
5641 agaagatcct ttgatctttt ctacggggtc tgacgctcag tggaacgaaa actcacgtta
5701 agggattttg gtcatgagat tatcaaaaag gatcttcacc tagatccttt taaattaaaa
5761 atgaagtttt aaatcaatct aaagtatata tgagtaaact tggtctgaca gttaccaatg
5821 cttaatcagt gaggcaccta tctcagcgat ctgtctattt cgttcatcca tagttgcctg
5881 actccccgtc gtgtagataa ctacgatacg ggagggctta ccatctggcc ccagtgctgc
5941 aatgataccg cgagacccac gctcaccggc tccagattta tcagcaataa accagccagc
6001 cggaagggcc gagcgcagaa gtggtcctgc aactttatcc gcctccatcc agtctattaa
6061 ttgttgccgg gaagctagag taagtagttc gccagttaat agtttgcgca acgttgttgc
6121 cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat tcagctccgg
6181 ttcccaacga tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag cggttagctc
6241 cttcggtcct ccgatcgttg tcagaagtaa gttggccgca gtgttatcac tcatggttat
6301 ggcagcactg cataattctc ttactgtcat gccatccgta agatgctttt ctgtgactgg
6361 tgagtactca accaagtcat tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc
6421 ggcgtcaata cgggataata ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg
6481 aaaacgttct tcggggcgaa aactctcaag gatcttaccg ctgttgagat ccagttcgat
6541 gtaacccact cgtgcaccca actgatcttc agcatctttt actttcacca gcgtttctgg
6601 gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga ataagggcga cacggaaatg
6661 ttgaatactc atactcttcc tttttcaata ttattgaagc atttatcagg gttattgtct
6721 catgagcgga tacatatttg aatgtattta gaaaaataaa caaatagggg ttccgcgcac
6781 atttccccga aaagtgccac ctg
//
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