| 质粒名称: | pCAG-Cre:GFP |
|---|---|
| 出品公司: | 爱迪基因 |
| 目录编号: | 13776 |
| 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: | 6563 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: | Cre-GFP fusion protein |
| 物种来源: | Bacteriophage P1 |
| 插入大小: | 1784 bp |
| 基因又名: | Cre recombinase |
| 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-Cre:GFP was a gift from Connie Cepko (爱迪基因 plasmid # 13776) |
| 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 |
买家导航
载体基本信息
载体图谱质粒图谱和多克隆位点信息
载体简介
载体序列
LOCUS pCAG-Cre:GFP 6569 bp ds-DNA circular SYN 10-JAN-2018
DEFINITION Mammalian expression of Cre recombinase fused to GFP.
ACCESSION .
VERSION .
KEYWORDS pCAG-Cre:GFP
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 6569)
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-Cre:GFP|
FEATURES Location/Qualifiers
source 1..6569
/organism="synthetic DNA construct"
/mol_type="other DNA"
enhancer 5..384
/label=CMV enhancer
/note="human cytomegalovirus immediate early enhancer"
promoter 386..661
/label=chicken beta-actin promoter
intron 662..1674
/label=chimeric intron
/note="chimera between introns from chicken beta-actin and
rabbit beta-globin"
primer_bind 1598..1620
/label=pCAGGS-5
/note="Chimeric intron in CAG promoter, forward primer"
primer_bind 1682..1701
/label=pCAG-F
/note="Rabbit beta-globin intron, for pCAG plasmids,
forward primer"
regulatory 1735..1744
/regulatory_class="other"
/note="vertebrate consensus sequence for strong initiation
of translation (Kozak, 1987)"
CDS 1741..2769
/codon_start=1
/product="site-specific recombinase"
/label=Cre
/note="Cre is a site-specific recombinase from
bacteriophage P1. Recombination occurs at loxP sequences."
/translation="MANLLTVHQNLPALPVDATSDEVRKNLMDMFRDRQAFSEHTWKML
LSVCRSWAAWCKLNNRKWFPAEPEDVRDYLLYLQARGLAVKTIQQHLGQLNMLHRRSGL
PRPSDSNAVSLVMRRIRKENVDAGERAKQALAFERTDFDQVRSLMENSDRCQDIRNLAF
LGIAYNTLLRIAEIARIRVKDISRTDGGRMLIHIGRTKTLVSTAGVEKALSLGVTKLVE
RWISVSGVADDPNNYLFCRVRKNGVAAPSATSQLSTRALEGIFEATHRLIYGAKDDSGQ
RYLAWSGHSARVGAARDMARAGVSIPEIMQAGGWTNVNIVMNYIRNLDSETGAMVRLLE
DGD"
regulatory 2779..2788
/regulatory_class="other"
/note="vertebrate consensus sequence for strong initiation
of translation (Kozak, 1987)"
CDS 2785..3504
/codon_start=1
/product="enhanced GFP"
/label=EGFP
/note="mammalian codon-optimized"
/translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
EFVTAAGITLGMDELYK"
primer_bind complement(2830..2851)
/label=EGFP-N
/note="EGFP, reverse primer"
primer_bind complement(3091..3110)
/label=EXFP-R
/note="For distinguishing EGFP variants, reverse primer"
primer_bind 3438..3459
/label=EGFP-C
/note="EGFP, forward primer"
primer_bind complement(3591..3610)
/label=Bglob-pA-R
/note="Rabbit beta-globin polyA region, reverse primer"
polyA_site 3656..3711
/label=-beta-globin poly(A) signal
/note="rabbit beta-globin polyadenylation signal"
primer_bind complement(3710..3729)
/label=rbglobpA-R
/note="Rabbit beta-globin polyA, reverse primer. Also
called rb-glob-pA-term-R"
primer_bind complement(4072..4088)
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(4072..4088)
/label=M13 Reverse
/note="In lacZ gene. Also called M13-rev"
primer_bind complement(4085..4107)
/label=M13/pUC Reverse
/note="In lacZ gene"
protein_bind 4096..4112
/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(4120..4150)
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 4165..4186
/label=CAP binding site
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding activates transcription in the presence
of cAMP."
promoter 4245..4440
/label=SV40 promoter
/note="SV40 early promoter"
rep_origin 4291..4426
/label=SV40 ori
/note="SV40 origin of replication"
primer_bind 4353..4372
/label=SV40pro-F
/note="SV40 promoter/origin, forward primer"
polyA_signal 4446..4580
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
primer_bind complement(4483..4502)
/label=SV40pA-R
/note="SV40 polyA, reverse primer"
primer_bind 4537..4556
/label=EBV-rev
/note="SV40 polyA terminator, reverse primer"
primer_bind complement(4648..4665)
/label=L4440
/note="L4440 vector, forward primer"
rep_origin complement(4819..5407)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind complement(4899..4918)
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
CDS complement(5578..6438)
/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 6201..6220
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
promoter complement(6439..6543)
/gene="bla"
/label=AmpR promoter
ORIGIN
1 ggtcgacatt gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat
61 agcccatata tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg
121 cccaacgacc cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata
181 gggactttcc attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta
241 catcaagtgt atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc
301 gcctggcatt atgcccagta catgacctta tgggactttc ctacttggca gtacatctac
361 gtattagtca tcgctattac catggtcgag 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 gcccccggag cgccggcggc
1321 tgtcgaggcg cggcgagccg cagccattgc cttttatggt aatcgtgcga gagggcgcag
1381 ggacttcctt tgtcccaaat ctgtgcggag ccgaaatctg ggaggcgccg ccgcaccccc
1441 tctagcgggc gcggggcgaa gcggtgcggc gccggcagga aggaaatggg cggggagggc
1501 cttcgtgcgt cgccgcgccg ccgtcccctt ctccctctcc agcctcgggg ctgtccgcgg
1561 ggggacggct gccttcgggg gggacggggc agggcggggt tcggcttctg gcgtgtgacc
1621 ggcggctcta gagcctctgc taaccatgtt catgccttct tctttttcct acagctcctg
1681 ggcaacgtgc tggttattgt gctgtctcat cattttggca aagaattctg agccgccacc
1741 atggccaatt tactgaccgt acaccaaaat ttgcctgcat taccggtcga tgcaacgagt
1801 gatgaggttc gcaagaacct gatggacatg ttcagggatc gccaggcgtt ttctgagcat
1861 acctggaaaa tgcttctgtc cgtttgccgg tcgtgggcgg catggtgcaa gttgaataac
1921 cggaaatggt ttcccgcaga acctgaagat gttcgcgatt atcttctata tcttcaggcg
1981 cgcggtctgg cagtaaaaac tatccagcaa catttgggcc agctaaacat gcttcatcgt
2041 cggtccgggc tgccacgacc aagtgacagc aatgctgttt cactggttat gcggcggatc
2101 cgaaaagaaa acgttgatgc cggtgaacgt gcaaaacagg ctctagcgtt cgaacgcact
2161 gatttcgacc aggttcgttc actcatggaa aatagcgatc gctgccagga tatacgtaat
2221 ctggcatttc tggggattgc ttataacacc ctgttacgta tagccgaaat tgccaggatc
2281 agggttaaag atatctcacg tactgacggt gggagaatgt taatccatat tggcagaacg
2341 aaaacgctgg ttagcaccgc aggtgtagag aaggcactta gcctgggggt aactaaactg
2401 gtcgagcgat ggatttccgt ctctggtgta gctgatgatc cgaataacta cctgttttgc
2461 cgggtcagaa aaaatggtgt tgccgcgcca tctgccacca gccagctatc aactcgcgcc
2521 ctggaaggga tttttgaagc aactcatcga ttgatttacg gcgctaagga tgactctggt
2581 cagagatacc tggcctggtc tggacacagt gcccgtgtcg gagccgcgcg agatatggcc
2641 cgcgctggag tttcaatacc ggagatcatg caagctggtg gctggaccaa tgtaaatatt
2701 gtcatgaact atatccgtaa cctggatagt gaaacagggg caatggtgcg cctgctggaa
2761 gatggcgatg gaccggtcgc caccatggtg agcaagggcg aggagctgtt caccggggtg
2821 gtgcccatcc tggtcgagct ggacggcgac gtaaacggcc acaagttcag cgtgtccggc
2881 gagggcgagg gcgatgccac ctacggcaag ctgaccctga agttcatctg caccaccggc
2941 aagctgcccg tgccctggcc caccctcgtg accaccctga cctacggcgt gcagtgcttc
3001 agccgctacc ccgaccacat gaagcagcac gacttcttca agtccgccat gcccgaaggc
3061 tacgtccagg agcgcaccat cttcttcaag gacgacggca actacaagac ccgcgccgag
3121 gtgaagttcg agggcgacac cctggtgaac cgcatcgagc tgaagggcat cgacttcaag
3181 gaggacggca acatcctggg gcacaagctg gagtacaact acaacagcca caacgtctat
3241 atcatggccg acaagcagaa gaacggcatc aaggtgaact tcaagatccg ccacaacatc
3301 gaggacggca gcgtgcagct cgccgaccac taccagcaga acacccccat cggcgacggc
3361 cccgtgctgc tgcccgacaa ccactacctg agcacccagt ccgccctgag caaagacccc
3421 aacgagaagc gcgatcacat ggtcctgctg gagttcgtga ccgccgccgg gatcactctc
3481 ggcatggacg agctgtacaa gtaaagcggc cgcactcctc aggtgcaggc tgcctatcag
3541 aaggtggtgg ctggtgtggc caatgccctg gctcacaaat accactgaga tctttttccc
3601 tctgccaaaa attatgggga catcatgaag ccccttgagc atctgacttc tggctaataa
3661 aggaaattta ttttcattgc aatagtgtgt tggaattttt tgtgtctctc actcggaagg
3721 acatatggga gggcaaatca tttaaaacat cagaatgagt atttggttta gagtttggca
3781 acatatgccc atatgctggc tgccatgaac aaaggttggc tataaagagg tcatcagtat
3841 atgaaacagc cccctgctgt ccattcctta ttccatagaa aagccttgac ttgaggttag
3901 atttttttta tattttgttt tgtgttattt ttttctttaa catccctaaa attttcctta
3961 catgttttac tagccagatt tttcctcctc tcctgactac tcccagtcat agctgtccct
4021 cttctcttat ggagatccct cgacctgcag cccaagcttg gcgtaatcat ggtcatagct
4081 gtttcctgtg tgaaattgtt atccgctcac aattccacac aacatacgag ccggaagcat
4141 aaagtgtaaa gcctggggtg cctaatgagt gagctaactc acattaattg cgttgcgctc
4201 actgcccgct ttccagtcgg gaaacctgtc gtgccagcgg atccgcatct caattagtca
4261 gcaaccatag tcccgcccct aactccgccc atcccgcccc taactccgcc cagttccgcc
4321 cattctccgc cccatggctg actaattttt tttatttatg cagaggccga ggccgcctcg
4381 gcctctgagc tattccagaa gtagtgagga ggcttttttg gaggcctagg cttttgcaaa
4441 aagctaactt gtttattgca gcttataatg gttacaaata aagcaatagc atcacaaatt
4501 tcacaaataa agcatttttt tcactgcatt ctagttgtgg tttgtccaaa ctcatcaatg
4561 tatcttatca tgtctggatc cgctgcatta atgaatcggc caacgcgcgg ggagaggcgg
4621 tttgcgtatt gggcgctctt ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg
4681 gctgcggcga gcggtatcag ctcactcaaa ggcggtaata cggttatcca cagaatcagg
4741 ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa
4801 ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc acaaaaatcg
4861 acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg cgtttccccc
4921 tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat acctgtccgc
4981 ctttctccct tcgggaagcg tggcgctttc tcatagctca cgctgtaggt atctcagttc
5041 ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc agcccgaccg
5101 ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg acttatcgcc
5161 actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg gtgctacaga
5221 gttcttgaag tggtggccta actacggcta cactagaaga acagtatttg gtatctgcgc
5281 tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg gcaaacaaac
5341 caccgctggt agcggtggtt tttttgtttg caagcagcag attacgcgca gaaaaaaagg
5401 atctcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga acgaaaactc
5461 acgttaaggg attttggtca tgagattatc aaaaaggatc ttcacctaga tccttttaaa
5521 ttaaaaatga agttttaaat caatctaaag tatatatgag taaacttggt ctgacagtta
5581 ccaatgctta atcagtgagg cacctatctc agcgatctgt ctatttcgtt catccatagt
5641 tgcctgactc cccgtcgtgt agataactac gatacgggag ggcttaccat ctggccccag
5701 tgctgcaatg ataccgcgag acccacgctc accggctcca gatttatcag caataaacca
5761 gccagccgga agggccgagc gcagaagtgg tcctgcaact ttatccgcct ccatccagtc
5821 tattaattgt tgccgggaag ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt
5881 tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag
5941 ctccggttcc caacgatcaa ggcgagttac atgatccccc atgttgtgca aaaaagcggt
6001 tagctccttc ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt tatcactcat
6061 ggttatggca gcactgcata attctcttac tgtcatgcca tccgtaagat gcttttctgt
6121 gactggtgag tactcaacca agtcattctg agaatagtgt atgcggcgac cgagttgctc
6181 ttgcccggcg tcaatacggg ataataccgc gccacatagc agaactttaa aagtgctcat
6241 cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag
6301 ttcgatgtaa cccactcgtg cacccaactg atcttcagca tcttttactt tcaccagcgt
6361 ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg
6421 gaaatgttga atactcatac tcttcctttt tcaatattat tgaagcattt atcagggtta
6481 ttgtctcatg agcggataca tatttgaatg tatttagaaa aataaacaaa taggggttcc
6541 gcgcacattt ccccgaaaag tgccacctg
//
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