| 质粒名称: | pCAG-GFP |
|---|---|
| 出品公司: | Addgene |
| 目录编号: | 11150 |
| 存储实验室: | Connie Cepko |
|
载体骨架基本信息 |
|
| 载体名称: | pCAGEN |
| 载体出品公司: | Available at Addgene (#11160) |
| 载体抗性: | 氨苄青霉素 |
| 空载体大小: | 4779 bp |
| 完整质粒大小: | -- |
| 载体修饰: | -- |
| 载体类型: | -- |
| 筛选标记: | -- |
| 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: | EGFP from Aequorea victoria,GFP，green fluorescent protein |
| 物种来源: | -- |
| 插入大小: | 772 bp |
| 基因突变: | -- |
| GenBank编号 | -- |
| 启动子 | -- |
| 标签/融合蛋白: | -- |
| 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 | EGFP-N |
|
文献引用方法 |
|
| 材料和方法部分: | pCAG-GFP was a gift from Connie Cepko (Addgene plasmid # 11150 ; http://n2t.net/addgene:11150 ; RRID:Addgene_11150) |
| 参考文献部分: | Electroporation and RNA interference in the rodent retina in vivo and in vitro. Matsuda T, Cepko CL. Proc Natl Acad Sci U S A. 2004 Jan 6. 101(1):16-22. 10.1073/pnas.2235688100 PubMed 14603031 |
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pCAG-GFP载体 pCAGGFP质粒基本信息
pCAG-GFP质粒图谱 pCAGGFP载体图谱和pCAG-GFP载体序列 pCAGGFP质粒序列多克隆位点信息
pCAG-GFP质粒 pCAGGFP载体简介
pCAG-GFP质粒序列 pCAGGFP载体序列
LOCUS pCAG-GFP 5561 bp ds-DNA circular SYN 25-NOV-2019
DEFINITION Mammalian expression vector for expression of GFP (CAG promoter).
ACCESSION .
VERSION .
KEYWORDS pCAG-GFP
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 5561)
AUTHORS Matsuda T, Cepko CL
TITLE Electroporation and RNA interference in the rodent retina in vivo
and in vitro.
JOURNAL Proc Natl Acad Sci U S A. 2004 Jan 6. 101(1):16-22.
PUBMED 14603031
REFERENCE 2 (bases 1 to 5561)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Nov 25, 2019 from SnapGene Server 1.1.58
http://www.biofeng.com/
FEATURES Location/Qualifiers
source 1..5561
/organism="synthetic DNA construct"
/mol_type="other DNA"
enhancer 5..384
/label=CMV enhancer
/note="human cytomegalovirus immediate early enhancer"
intron 662..1678
/label=chimeric intron
/note="chimera between introns from chicken beta-actin and
rabbit beta-globin"
primer_bind 1602..1624
/label=pCAGGS-5
/note="Chimeric intron in CAG promoter, forward primer"
primer_bind 1686..1705
/label=pCAG-F
/note="Rabbit beta-globin intron, for pCAG plasmids,
forward primer"
regulatory 1771..1780
/regulatory_class="other"
/note="vertebrate consensus sequence for strong initiation
of translation (Kozak, 1987)"
CDS 1777..2496
/codon_start=1
/product="enhanced GFP"
/label=EGFP
/note="mammalian codon-optimized"
/translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
EFVTAAGITLGMDELYK"
primer_bind complement(1822..1843)
/label=EGFP-N
/note="EGFP, reverse primer"
primer_bind complement(2083..2102)
/label=EXFP-R
/note="For distinguishing EGFP variants, reverse primer"
primer_bind 2430..2451
/label=EGFP-C
/note="EGFP, forward primer"
primer_bind complement(2583..2602)
/label=Bglob-pA-R
/note="Rabbit beta-globin polyA region, reverse primer"
polyA_site 2648..2703
/label=-beta-globin poly(A) signal
/note="rabbit beta-globin polyadenylation signal"
primer_bind complement(2702..2721)
/label=rbglobpA-R
/note="Rabbit beta-globin polyA, reverse primer. Also
called rb-glob-pA-term-R"
primer_bind complement(3064..3080)
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(3064..3080)
/label=M13 Reverse
/note="In lacZ gene. Also called M13-rev"
primer_bind complement(3077..3099)
/label=M13/pUC Reverse
/note="In lacZ gene"
protein_bind 3088..3104
/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(3112..3142)
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 3157..3178
/label=CAP binding site
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding activates transcription in the presence
of cAMP."
promoter 3237..3432
/label=SV40 promoter
/note="SV40 early promoter"
rep_origin 3283..3418
/label=SV40 ori
/note="SV40 origin of replication"
primer_bind 3345..3364
/label=SV40pro-F
/note="SV40 promoter/origin, forward primer"
polyA_signal 3438..3572
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
primer_bind complement(3475..3494)
/label=SV40pA-R
/note="SV40 polyA, reverse primer"
primer_bind 3529..3548
/label=EBV-rev
/note="SV40 polyA terminator, reverse primer"
primer_bind complement(3640..3657)
/label=L4440
/note="L4440 vector, forward primer"
rep_origin complement(3811..4399)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind complement(3891..3910)
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
CDS complement(4570..5430)
/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 5193..5212
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
promoter complement(5431..5535)
/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 gggggsgcgc gccrggsggg gsggggsggg gsgrggggsg
541 gggsggggsg rggcggagag gtgcggcggc agccaatcag agcggcgcgc tccgaaagtt
601 tccttttatg gcgaggcggc ggcggcggcg gccctataaa aagcgaagcg cgcggcgggc
661 gggagtcgct gcgcgctgcc ttcgccccgt gccccgctcc gccgccgcct cgcgccgccc
721 gccccggctc tgactgaccg cgttactccc acaggtgagc gggcgggacg gcccttctcc
781 tccgggctgt aattagcgct tggtttaatg acggcttgtt tcttttctgt ggctgcgtga
841 aagccttgag gggctccggg agggcccttt gtgcgggggg agcggctcgg ggggtgcgtg
901 cgtgtgtgtg tgcgtgggga gcgccgcgtg cggctccgcg ctgcccggcg gctgtgagcg
961 ctgcgggcgc ggcgcggggc tttgtgcgct ccgcagtgtg cgcgagggga gcgcggccgg
1021 gggcggtgcc ccgcggtgcg gggggggctg cgaggggaac aaaggctgcg tgcggggtgt
1081 gtgcgtgggg gggtgagcag ggggtgtggg cgcgtcggtc gggctgcaac cccccctgca
1141 cccccctccc cgagttgctg agcacggccc ggcttcgggt gcggggctcc gtacggggcg
1201 tggcgcgggg ctcgccgtgc cgggcggggg gtggcggcag gtgggggtgc cgggcggggc
1261 ggggccgcct cgggccgggg rgggctcggg ggaggggcgc ggcggccccc ggagcgccgg
1321 cggctgtcga ggcgcggcga gccgcagcca ttgcctttta tggtaatcgt gcgagagggc
1381 gcagggactt cctttgtccc aaatctgtgc ggagccgaaa tctgggaggc gccgccgcac
1441 cccctctagc gggcgcgggg cgaagcggtg cggcgccggc aggaaggaaa tgggcgggga
1501 gggccttcgt gcgtcgccgc gccgccgtcc ccttctccct ctccagcctc ggggctgtcc
1561 gcggggggac ggctgccttc gggggggacg gggcagggcg gggttcggct tctggcgtgt
1621 gaccggcggc tctagagcct ctgctaacca tgttcatgcc ttcttctttt tcctacagct
1681 cctgggcaac gtgctggtta ttgtgctgtc tcatcatttt ggcaaagaat tctgcagtcg
1741 acggtaccgc gggcccggga tccaccggtc gccaccatgg tgagcaaggg cgaggagctg
1801 ttcaccgggg tggtgcccat cctggtcgag ctggacggcg acgtaaacgg ccacaagttc
1861 agcgtgtccg gcgagggcga gggcgatgcc acctacggca agctgaccct gaagttcatc
1921 tgcaccaccg gcaagctgcc cgtgccctgg cccaccctcg tgaccaccct gacctacggc
1981 gtgcagtgct tcagccgcta ccccgaccac atgaagcagc acgacttctt caagtccgcc
2041 atgcccgaag gctacgtcca ggagcgcacc atcttcttca aggacgacgg caactacaag
2101 acccgcgccg aggtgaagtt cgagggcgac accctggtga accgcatcga gctgaagggc
2161 atcgacttca aggaggacgg caacatcctg gggcacaagc tggagtacaa ctacaacagc
2221 cacaacgtct atatcatggc cgacaagcag aagaacggca tcaaggtgaa cttcaagatc
2281 cgccacaaca tcgaggacgg cagcgtgcag ctcgccgacc actaccagca gaacaccccc
2341 atcggcgacg gccccgtgct gctgcccgac aaccactacc tgagcaccca gtccgccctg
2401 agcaaagacc ccaacgagaa gcgcgatcac atggtcctgc tggagttcgt gaccgccgcc
2461 gggatcactc tcggcatgga cgagctgtac aagtaaagcg gccgcactcc tcaggtgcag
2521 gctgcctatc agaaggtggt ggctggtgtg gccaatgccc tggctcacaa ataccactga
2581 gatctttttc cctctgccaa aaattatggg gacatcatga agccccttga gcatctgact
2641 tctggctaat aaaggaaatt tattttcatt gcaatagtgt gttggaattt tttgtgtctc
2701 tcactcggaa ggacatatgg gagggcaaat catttaaaac atcagaatga gtatttggtt
2761 tagagtttgg caacatatgc ccatatgctg gctgccatga acaaaggttg gctataaaga
2821 ggtcatcagt atatgaaaca gccccctgct gtccattcct tattccatag aaaagccttg
2881 acttgaggtt agattttttt tatattttgt tttgtgttat ttttttcttt aacatcccta
2941 aaattttcct tacatgtttt actagccaga tttttcctcc tctcctgact actcccagtc
3001 atagctgtcc ctcttctctt atggagatcc ctcgacctgc agcccaagct tggcgtaatc
3061 atggtcatag ctgtttcctg tgtgaaattg ttatccgctc acaattccac acaacatacg
3121 agccggaagc ataaagtgta aagcctgggg tgcctaatga gtgagctaac tcacattaat
3181 tgcgttgcgc tcactgcccg ctttccagtc gggaaacctg tcgtgccagc ggatccgcat
3241 ctcaattagt cagcaaccat agtcccgccc ctaactccgc ccatcccgcc cctaactccg
3301 cccagttccg cccattctcc gccccatggc tgactaattt tttttattta tgcagaggcc
3361 gaggccgcct cggcctctga gctattccag aagtagtgag gaggcttttt tggaggccta
3421 ggcttttgca aaaagctaac ttgtttattg cagcttataa tggttacaaa taaagcaata
3481 gcatcacaaa tttcacaaat aaagcatttt tttcactgca ttctagttgt ggtttgtcca
3541 aactcatcaa tgtatcttat catgtctgga tccgctgcat taatgaatcg gccaacgcgc
3601 ggggagaggc ggtttgcgta ttgggcgctc ttccgcttcc tcgctcactg actcgctgcg
3661 ctcggtcgtt cggctgcggc gagcggtatc agctcactca aaggcggtaa tacggttatc
3721 cacagaatca ggggataacg caggaaagaa catgtgagca aaaggccagc aaaaggccag
3781 gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg ctccgccccc ctgacgagca
3841 tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg acaggactat aaagatacca
3901 ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg
3961 atacctgtcc gcctttctcc cttcgggaag cgtggcgctt tctcatagct cacgctgtag
4021 gtatctcagt tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt
4081 tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggtaagaca
4141 cgacttatcg ccactggcag cagccactgg taacaggatt agcagagcga ggtatgtagg
4201 cggtgctaca gagttcttga agtggtggcc taactacggc tacactagaa gaacagtatt
4261 tggtatctgc gctctgctga agccagttac cttcggaaaa agagttggta gctcttgatc
4321 cggcaaacaa accaccgctg gtagcggtgg tttttttgtt tgcaagcagc agattacgcg
4381 cagaaaaaaa ggatctcaag aagatccttt gatcttttct acggggtctg acgctcagtg
4441 gaacgaaaac tcacgttaag ggattttggt catgagatta tcaaaaagga tcttcaccta
4501 gatcctttta aattaaaaat gaagttttaa atcaatctaa agtatatatg agtaaacttg
4561 gtctgacagt taccaatgct taatcagtga ggcacctatc tcagcgatct gtctatttcg
4621 ttcatccata gttgcctgac tccccgtcgt gtagataact acgatacggg agggcttacc
4681 atctggcccc agtgctgcaa tgataccgcg agacccacgc tcaccggctc cagatttatc
4741 agcaataaac cagccagccg gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc
4801 ctccatccag tctattaatt gttgccggga agctagagta agtagttcgc cagttaatag
4861 tttgcgcaac gttgttgcca ttgctacagg catcgtggtg tcacgctcgt cgtttggtat
4921 ggcttcattc agctccggtt cccaacgatc aaggcgagtt acatgatccc ccatgttgtg
4981 caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt
5041 gttatcactc atggttatgg cagcactgca taattctctt actgtcatgc catccgtaag
5101 atgcttttct gtgactggtg agtactcaac caagtcattc tgagaatagt gtatgcggcg
5161 accgagttgc tcttgcccgg cgtcaatacg ggataatacc gcgccacata gcagaacttt
5221 aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct
5281 gttgagatcc agttcgatgt aacccactcg tgcacccaac tgatcttcag catcttttac
5341 tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat
5401 aagggcgaca cggaaatgtt gaatactcat actcttcctt tttcaatatt attgaagcat
5461 ttatcagggt tattgtctca tgagcggata catatttgaa tgtatttaga aaaataaaca
5521 aataggggtt ccgcgcacat ttccccgaaa agtgccacct g
//
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