| 质粒名称: | pEF-GFP |
|---|---|
| 出品公司: | Addgene |
| 目录编号: | 11154 |
| 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: | -- |
| 载体抗性: | 氨苄青霉素 |
| 空载体大小: | 3086 bp |
| 完整质粒大小: | -- |
| 载体修饰: | -- |
| 载体类型: | 哺乳细胞表达 |
| 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: | EF1 alpha promoter,EF1A1 promoter,eukaryotic translation elongation factor 1 alpha 1，EF1A1 |
| 物种来源: | H. sapiens (human) |
| 插入大小: | 1188 bp |
| 基因突变: | -- |
| GenBank编号 | -- |
| 启动子 | -- |
| 标签/融合蛋白: | EGFP（主干上的C端） |
| 克隆方法: | 限制性内切酶 |
| 5' 克隆位点: | SpeI（在克隆过程中销毁） |
| 3' 克隆位点: | NotI(未破坏) |
| 5' 测序引物 | -- |
| 3' 测序引物 | EGFP-N |
|
文献引用方法 |
|
| 材料和方法部分: | pEF-GFP was a gift from Connie Cepko (Addgene plasmid # 11154 ; http://n2t.net/addgene:11154 ; RRID:Addgene_11154) |
| 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 |
买家导航
pEFGFP载体基本信息
pEFGFP载体图谱质粒图谱和多克隆位点信息
pEFGFP载体简介
pEFGFP载体序列
LOCUS pEF-GFP 5054 bp ds-DNA circular SYN 12-SEP-2018
DEFINITION Mammalian expression vector for expression of GFP (EF1a promoter).
ACCESSION .
VERSION .
KEYWORDS pEF-GFP
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 5054)
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 5054)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Sep 12, 2018 from SnapGene Server 1.1.58
http://www.biofeng.com/
FEATURES Location/Qualifiers
source 1..5054
/organism="synthetic DNA construct"
/mol_type="other DNA"
promoter 36..1217
/label=EF-1-alpha promoter
/note="strong constitutive promoter for human elongation
factor EF-1-alpha"
intron 266..1208
/label=EF-1-alpha intron A
/note="intron upstream of the start codon of human
EF-1-alpha"
primer_bind 1165..1185
/label=EF1a-F
/note="Human elongation factor-1a promoter, forward primer"
regulatory 1263..1272
/regulatory_class="other"
/note="vertebrate consensus sequence for strong initiation
of translation (Kozak, 1987)"
CDS 1269..1988
/codon_start=1
/product="enhanced GFP"
/label=EGFP
/note="mammalian codon-optimized"
/translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
EFVTAAGITLGMDELYK"
primer_bind complement(1314..1335)
/label=EGFP-N
/note="EGFP, reverse primer"
primer_bind complement(1575..1594)
/label=EXFP-R
/note="For distinguishing EGFP variants, reverse primer"
primer_bind 1922..1943
/label=EGFP-C
/note="EGFP, forward primer"
primer_bind complement(2075..2094)
/label=Bglob-pA-R
/note="Rabbit beta-globin polyA region, reverse primer"
polyA_site 2140..2195
/label=-beta-globin poly(A) signal
/note="rabbit beta-globin polyadenylation signal"
primer_bind complement(2194..2213)
/label=rbglobpA-R
/note="Rabbit beta-globin polyA, reverse primer. Also
called rb-glob-pA-term-R"
primer_bind complement(2556..2572)
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(2556..2572)
/label=M13 Reverse
/note="In lacZ gene. Also called M13-rev"
primer_bind complement(2569..2591)
/label=M13/pUC Reverse
/note="In lacZ gene"
protein_bind 2580..2596
/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(2604..2634)
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 2649..2670
/label=CAP binding site
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding activates transcription in the presence
of cAMP."
promoter 2729..2924
/label=SV40 promoter
/note="SV40 early promoter"
rep_origin 2775..2910
/label=SV40 ori
/note="SV40 origin of replication"
primer_bind 2837..2856
/label=SV40pro-F
/note="SV40 promoter/origin, forward primer"
polyA_signal 2930..3064
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
primer_bind complement(2967..2986)
/label=SV40pA-R
/note="SV40 polyA, reverse primer"
primer_bind 3021..3040
/label=EBV-rev
/note="SV40 polyA terminator, reverse primer"
primer_bind complement(3132..3149)
/label=L4440
/note="L4440 vector, forward primer"
rep_origin complement(3303..3891)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind complement(3383..3402)
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
CDS complement(4062..4922)
/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 4685..4704
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
promoter complement(4923..5027)
/gene="bla"
/label=AmpR promoter
ORIGIN
1 gtcgacattg attattgact agatcatcgc gtgaggctcc ggtgcccgtc agtgggcaga
61 gcgcacatcg cccacagtcc ccgagaagtt ggggggaggg gtcggcaatt gaaccggtgc
121 ctagagaagg tggcgcgggg taaactggga aagtgatgtc gtgtactggc tccgcctttt
181 tcccgagggt gggggagaac cgtatataag tgcagtagtc gccgtgaacg ttctttttcg
241 caacgggttt gccgccagaa cacaggtaag tgccgtgtgt ggttcccgcg ggcctggcct
301 ctttacgggt tatggccctt gcgtgccttg aattacttcc acgcccctgg ctgcagtacg
361 tgattcttga tcccgagctt cgggttggaa gtgggtggga gagttcgagg ccttgcgctt
421 aaggagcccc ttcgcctcgt gcttgagttg aggcctggcc tgggcgctgg ggccgccgcg
481 tgcgaatctg gtggcacctt cgcgcctgtc tcgctgcttt cgataagtct ctagccattt
541 aaaatttttg atgacctgct gcgacgcttt ttttctggca agatagtctt gtaaatgcgg
601 gccaagatct gcacactggt atttcggttt ttggggccgc gggcggcgac ggggcccgtg
661 cgtcccagcg cacatgttcg gcgaggcggg gcctgcgagc gcggccaccg agaatcggac
721 gggggtagtc tcaagctggc cggcctgctc tggtgcctgg cctcgcgccg ccgtgtatcg
781 ccccgccctg ggcggcaagg ctggcccggt cggcaccagt tgcgtgagcg gaaagatggc
841 cgcttcccgg ccctgctgca gggagctcaa aatggaggac gcggcgctcg ggagagcggg
901 cgggtgagtc acccacacaa aggaaaaggg cctttccgtc ctcagccgtc gcttcatgtg
961 actccacgga gtaccgggcg ccgtccaggc acctcgatta gttctcgagc ttttggagta
1021 cgtcgtcttt aggttggggg gaggggtttt atgcgatgga gtttccccac actgagtggg
1081 tggagactga agttaggcca gcttggcact tgatgtaatt ctccttggaa tttgcccttt
1141 ttgagtttgg atcttggttc attctcaagc ctcagacagt ggttcaaagt ttttttcttc
1201 catttcaggt gtcgtgagga attctgcagt cgacggtacc gcgggcccgg gatccaccgg
1261 tcgccaccat ggtgagcaag ggcgaggagc tgttcaccgg ggtggtgccc atcctggtcg
1321 agctggacgg cgacgtaaac ggccacaagt tcagcgtgtc cggcgagggc gagggcgatg
1381 ccacctacgg caagctgacc ctgaagttca tctgcaccac cggcaagctg cccgtgccct
1441 ggcccaccct cgtgaccacc ctgacctacg gcgtgcagtg cttcagccgc taccccgacc
1501 acatgaagca gcacgacttc ttcaagtccg ccatgcccga aggctacgtc caggagcgca
1561 ccatcttctt caaggacgac ggcaactaca agacccgcgc cgaggtgaag ttcgagggcg
1621 acaccctggt gaaccgcatc gagctgaagg gcatcgactt caaggaggac ggcaacatcc
1681 tggggcacaa gctggagtac aactacaaca gccacaacgt ctatatcatg gccgacaagc
1741 agaagaacgg catcaaggtg aacttcaaga tccgccacaa catcgaggac ggcagcgtgc
1801 agctcgccga ccactaccag cagaacaccc ccatcggcga cggccccgtg ctgctgcccg
1861 acaaccacta cctgagcacc cagtccgccc tgagcaaaga ccccaacgag aagcgcgatc
1921 acatggtcct gctggagttc gtgaccgccg ccgggatcac tctcggcatg gacgagctgt
1981 acaagtaaag cggccgcact cctcaggtgc aggctgccta tcagaaggtg gtggctggtg
2041 tggccaatgc cctggctcac aaataccact gagatctttt tccctctgcc aaaaattatg
2101 gggacatcat gaagcccctt gagcatctga cttctggcta ataaaggaaa tttattttca
2161 ttgcaatagt gtgttggaat tttttgtgtc tctcactcgg aaggacatat gggagggcaa
2221 atcatttaaa acatcagaat gagtatttgg tttagagttt ggcaacatat gcccatatgc
2281 tggctgccat gaacaaaggt tggctataaa gaggtcatca gtatatgaaa cagccccctg
2341 ctgtccattc cttattccat agaaaagcct tgacttgagg ttagattttt tttatatttt
2401 gttttgtgtt atttttttct ttaacatccc taaaattttc cttacatgtt ttactagcca
2461 gatttttcct cctctcctga ctactcccag tcatagctgt ccctcttctc ttatggagat
2521 ccctcgacct gcagcccaag cttggcgtaa tcatggtcat agctgtttcc tgtgtgaaat
2581 tgttatccgc tcacaattcc acacaacata cgagccggaa gcataaagtg taaagcctgg
2641 ggtgcctaat gagtgagcta actcacatta attgcgttgc gctcactgcc cgctttccag
2701 tcgggaaacc tgtcgtgcca gcggatccgc atctcaatta gtcagcaacc atagtcccgc
2761 ccctaactcc gcccatcccg cccctaactc cgcccagttc cgcccattct ccgccccatg
2821 gctgactaat tttttttatt tatgcagagg ccgaggccgc ctcggcctct gagctattcc
2881 agaagtagtg aggaggcttt tttggaggcc taggcttttg caaaaagcta acttgtttat
2941 tgcagcttat aatggttaca aataaagcaa tagcatcaca aatttcacaa ataaagcatt
3001 tttttcactg cattctagtt gtggtttgtc caaactcatc aatgtatctt atcatgtctg
3061 gatccgctgc attaatgaat cggccaacgc gcggggagag gcggtttgcg tattgggcgc
3121 tcttccgctt cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta
3181 tcagctcact caaaggcggt aatacggtta tccacagaat caggggataa cgcaggaaag
3241 aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg
3301 tttttccata ggctccgccc ccctgacgag catcacaaaa atcgacgctc aagtcagagg
3361 tggcgaaacc cgacaggact ataaagatac caggcgtttc cccctggaag ctccctcgtg
3421 cgctctcctg ttccgaccct gccgcttacc ggatacctgt ccgcctttct cccttcggga
3481 agcgtggcgc tttctcatag ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc
3541 tccaagctgg gctgtgtgca cgaacccccc gttcagcccg accgctgcgc cttatccggt
3601 aactatcgtc ttgagtccaa cccggtaaga cacgacttat cgccactggc agcagccact
3661 ggtaacagga ttagcagagc gaggtatgta ggcggtgcta cagagttctt gaagtggtgg
3721 cctaactacg gctacactag aagaacagta tttggtatct gcgctctgct gaagccagtt
3781 accttcggaa aaagagttgg tagctcttga tccggcaaac aaaccaccgc tggtagcggt
3841 ggtttttttg tttgcaagca gcagattacg cgcagaaaaa aaggatctca agaagatcct
3901 ttgatctttt ctacggggtc tgacgctcag tggaacgaaa actcacgtta agggattttg
3961 gtcatgagat tatcaaaaag gatcttcacc tagatccttt taaattaaaa atgaagtttt
4021 aaatcaatct aaagtatata tgagtaaact tggtctgaca gttaccaatg cttaatcagt
4081 gaggcaccta tctcagcgat ctgtctattt cgttcatcca tagttgcctg actccccgtc
4141 gtgtagataa ctacgatacg ggagggctta ccatctggcc ccagtgctgc aatgataccg
4201 cgagacccac gctcaccggc tccagattta tcagcaataa accagccagc cggaagggcc
4261 gagcgcagaa gtggtcctgc aactttatcc gcctccatcc agtctattaa ttgttgccgg
4321 gaagctagag taagtagttc gccagttaat agtttgcgca acgttgttgc cattgctaca
4381 ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat tcagctccgg ttcccaacga
4441 tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag cggttagctc cttcggtcct
4501 ccgatcgttg tcagaagtaa gttggccgca gtgttatcac tcatggttat ggcagcactg
4561 cataattctc ttactgtcat gccatccgta agatgctttt ctgtgactgg tgagtactca
4621 accaagtcat tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata
4681 cgggataata ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg aaaacgttct
4741 tcggggcgaa aactctcaag gatcttaccg ctgttgagat ccagttcgat gtaacccact
4801 cgtgcaccca actgatcttc agcatctttt actttcacca gcgtttctgg gtgagcaaaa
4861 acaggaaggc aaaatgccgc aaaaaaggga ataagggcga cacggaaatg ttgaatactc
4921 atactcttcc tttttcaata ttattgaagc atttatcagg gttattgtct catgagcgga
4981 tacatatttg aatgtattta gaaaaataaa caaatagggg ttccgcgcac atttccccga
5041 aaagtgccac ctgg
//
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