| 质粒名称: | pcDNA3-EGFP |
|---|---|
| 出品公司: | 爱迪基因 |
| 目录编号: | 13031 |
| 存储实验室: | Doug Golenbock |
|
载体骨架基本信息 |
|
| 载体名称: | pcDNA3 |
| 载体出品公司: | -- |
| 载体抗性: | 氨苄霉素 |
| 空载体大小: | 5446 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: | 新霉素（G418） |
| 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: | Enhanced Green Fluorescent Protein |
| 物种来源: | -- |
| 插入大小: | 700 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: | XhoI(未破坏) |
| 3' 克隆位点: | XbaI (未破坏) |
| 5' 测序引物 | T7 |
| 3' 测序引物 | GTCTTGTAGTTGCCGTCGTC |
|
文献引用方法 |
|
| 材料和方法部分: | pcDNA3-EGFP was a gift from Doug Golenbock (Addgene plasmid # 13031 ; http://n2t.net/addgene:13031 ; RRID:Addgene_13031) |
买家导航
pcDNA3EGFP载体基本信息
pcDNA3EGFP载体图谱质粒图谱和多克隆位点信息
pcDNA3EGFP载体简介
pcDNA3EGFP载体序列
LOCUS pcDNA3-EGFP 6159 bp ds-DNA circular SYN 19-MAR-2019
DEFINITION Mammalian expression vector for cloning your gene fused to EGFP.
ACCESSION .
VERSION .
KEYWORDS pcDNA3-EGFP
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 6159)
TITLE Fluorescent Protein Plasmids
REFERENCE 2 (bases 1 to 6159)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Mar 19, 2019 from SnapGene Server 1.1.58
http://www.snapgene.com
FEATURES Location/Qualifiers
source 1..6159
/organism="synthetic DNA construct"
/mol_type="other DNA"
primer_bind complement(42..61)
/label=pRS-marker
/note="pRS vectors, use to sequence yeast selectable
marker"
enhancer 233..612
/label=CMV enhancer
/note="human cytomegalovirus immediate early enhancer"
promoter 613..816
/label=CMV promoter
/note="human cytomegalovirus (CMV) immediate early
promoter"
primer_bind 767..787
/label=CMV-F
/note="Human CMV immediate early promoter, forward primer"
primer_bind 861..880
/label=T7
/note="T7 promoter, forward primer"
promoter 861..879
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
CDS 975..1694
/codon_start=1
/product="enhanced GFP"
/label=EGFP
/note="mammalian codon-optimized"
/translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
EFVTAAGITLGMDELYK"
primer_bind complement(1020..1041)
/label=EGFP-N
/note="EGFP, reverse primer"
primer_bind complement(1281..1300)
/label=EXFP-R
/note="For distinguishing EGFP variants, reverse primer"
primer_bind 1628..1649
/label=EGFP-C
/note="EGFP, forward primer"
promoter complement(1710..1728)
/label=SP6 promoter
/note="promoter for bacteriophage SP6 RNA polymerase"
primer_bind complement(1711..1728)
/label=SP6
/note="SP6 promoter, forward primer"
primer_bind complement(1748..1765)
/label=BGH-rev
/note="Bovine growth hormone terminator, reverse primer.
Also called BGH reverse"
polyA_signal 1754..1978
/label=bGH poly(A) signal
/note="bovine growth hormone polyadenylation signal"
rep_origin 2024..2452
/direction=RIGHT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
primer_bind complement(2111..2130)
/label=F1ori-R
/note="F1 origin, reverse primer"
primer_bind 2321..2342
/label=F1ori-F
/note="F1 origin, forward primer"
primer_bind complement(2461..2481)
/label=pBABE 3'
/note="SV40 enhancer, reverse primer for pBABE vectors"
promoter 2466..2795
/label=SV40 promoter
/note="SV40 enhancer and early promoter"
rep_origin 2646..2781
/label=SV40 ori
/note="SV40 origin of replication"
primer_bind 2708..2727
/label=SV40pro-F
/note="SV40 promoter/origin, forward primer"
CDS 2862..3656
/codon_start=1
/gene="aph(3')-II (or nptII)"
/product="aminoglycoside phosphotransferase from Tn5"
/label=NeoR/KanR
/note="confers resistance to neomycin, kanamycin, and G418
(Geneticin(R))"
/translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
primer_bind complement(2916..2935)
/label=Neo-R
/note="Neomycin resistance gene, reverse primer"
primer_bind 3526..3545
/label=Neo-F
/note="Neomycin resistance gene, forward primer"
polyA_signal 3830..3951
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
primer_bind complement(3867..3886)
/label=SV40pA-R
/note="SV40 polyA, reverse primer"
primer_bind 3921..3940
/label=EBV-rev
/note="SV40 polyA terminator, reverse primer"
primer_bind complement(4000..4016)
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(4000..4016)
/label=M13 Reverse
/note="In lacZ gene. Also called M13-rev"
primer_bind complement(4013..4035)
/label=M13/pUC Reverse
/note="In lacZ gene"
protein_bind 4024..4040
/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(4048..4078)
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 4093..4114
/label=CAP binding site
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding activates transcription in the presence
of cAMP."
primer_bind complement(4231..4248)
/label=L4440
/note="L4440 vector, forward primer"
rep_origin complement(4402..4990)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind complement(4482..4501)
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
CDS complement(5161..6021)
/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 5784..5803
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
promoter complement(6022..6126)
/gene="bla"
/label=AmpR promoter
ORIGIN
1 cggatcggga gatctcccga tcccctatgg tgcactctca gtacaatctg ctctgatgcc
61 gcatagttaa gccagtatct gctccctgct tgtgtgttgg aggtcgctga gtagtgcgcg
121 agcaaaattt aagctacaac aaggcaaggc ttgaccgaca attgcatgaa gaatctgctt
181 agggttaggc gttttgcgct gcttcgcgat gtacgggcca gatatacgcg ttgacattga
241 ttattgacta gttattaata gtaatcaatt acggggtcat tagttcatag cccatatatg
301 gagttccgcg ttacataact tacggtaaat ggcccgcctg gctgaccgcc caacgacccc
361 cgcccattga cgtcaataat gacgtatgtt cccatagtaa cgccaatagg gactttccat
421 tgacgtcaat gggtggagta tttacggtaa actgcccact tggcagtaca tcaagtgtat
481 catatgccaa gtacgccccc tattgacgtc aatgacggta aatggcccgc ctggcattat
541 gcccagtaca tgaccttatg ggactttcct acttggcagt acatctacgt attagtcatc
601 gctattacca tggtgatgcg gttttggcag tacatcaatg ggcgtggata gcggtttgac
661 tcacggggat ttccaagtct ccaccccatt gacgtcaatg ggagtttgtt ttggcaccaa
721 aatcaacggg actttccaaa atgtcgtaac aactccgccc cattgacgca aatgggcggt
781 aggcgtgtac ggtgggaggt ctatataagc agagctctct ggctaactag agaacccact
841 gcttactggc ttatcgaaat taatacgact cactataggg agacccaagc ttggtaccga
901 gctcggatcc actagtaacg gccgccagtg tgctggaatt ctgcagatat ccatcacact
961 ggcggccgct cgagatggtg agcaagggcg aggagctgtt caccggggtg gtgcccatcc
1021 tggtcgagct ggacggcgac gtaaacggcc acaagttcag cgtgtccggc gagggcgagg
1081 gcgatgccac ctacggcaag ctgaccctga agttcatctg caccaccggc aagctgcccg
1141 tgccctggcc caccctcgtg accaccctga cctacggcgt gcagtgcttc agccgctacc
1201 ccgaccacat gaagcagcac gacttcttca agtccgccat gcccgaaggc tacgtccagg
1261 agcgcaccat cttcttcaag gacgacggca actacaagac ccgcgccgag gtgaagttcg
1321 agggcgacac cctggtgaac cgcatcgagc tgaagggcat cgacttcaag gaggacggca
1381 acatcctggg gcacaagctg gagtacaact acaacagcca caacgtctat atcatggccg
1441 acaagcagaa gaacggcatc aaggtgaact tcaagatccg ccacaacatc gaggacggca
1501 gcgtgcagct cgccgaccac taccagcaga acacccccat cggcgacggc cccgtgctgc
1561 tgcccgacaa ccactacctg agcacccagt ccgccctgag caaagacccc aacgagaagc
1621 gcgatcacat ggtcctgctg gagttcgtga ccgccgccgg gatcactctc ggcatggacg
1681 agctgtacaa gtaatctaga gggccctatt ctatagtgtc acctaaatgc tagagctcgc
1741 tgatcagcct cgactgtgcc ttctagttgc cagccatctg ttgtttgccc ctcccccgtg
1801 ccttccttga ccctggaagg tgccactccc actgtccttt cctaataaaa tgaggaaatt
1861 gcatcgcatt gtctgagtag gtgtcattct attctggggg gtggggtggg gcaggacagc
1921 aagggggagg attgggaaga caatagcagg catgctgggg atgcggtggg ctctatggct
1981 tctgaggcgg aaagaaccag ctggggctct agggggtatc cccacgcgcc ctgtagcggc
2041 gcattaagcg cggcgggtgt ggtggttacg cgcagcgtga ccgctacact tgccagcgcc
2101 ctagcgcccg ctcctttcgc tttcttccct tcctttctcg ccacgttcgc cggctttccc
2161 cgtcaagctc taaatcgggg gctcccttta gggttccgat ttagtgcttt acggcacctc
2221 gaccccaaaa aacttgatta gggtgatggt tcacgtagtg ggccatcgcc ctgatagacg
2281 gtttttcgcc ctttgacgtt ggagtccacg ttctttaata gtggactctt gttccaaact
2341 ggaacaacac tcaaccctat ctcggtctat tcttttgatt tataagggat tttgccgatt
2401 tcggcctatt ggttaaaaaa tgagctgatt taacaaaaat ttaacgcgaa ttaattctgt
2461 ggaatgtgtg tcagttaggg tgtggaaagt ccccaggctc cccagcaggc agaagtatgc
2521 aaagcatgca tctcaattag tcagcaacca ggtgtggaaa gtccccaggc tccccagcag
2581 gcagaagtat gcaaagcatg catctcaatt agtcagcaac catagtcccg cccctaactc
2641 cgcccatccc gcccctaact ccgcccagtt ccgcccattc tccgccccat ggctgactaa
2701 ttttttttat ttatgcagag gccgaggccg cctctgcctc tgagctattc cagaagtagt
2761 gaggaggctt ttttggaggc ctaggctttt gcaaaaagct cccgggagct tgtatatcca
2821 ttttcggatc tgatcaagag acaggatgag gatcgtttcg catgattgaa caagatggat
2881 tgcacgcagg ttctccggcc gcttgggtgg agaggctatt cggctatgac tgggcacaac
2941 agacaatcgg ctgctctgat gccgccgtgt tccggctgtc agcgcagggg cgcccggttc
3001 tttttgtcaa gaccgacctg tccggtgccc tgaatgaact gcaggacgag gcagcgcggc
3061 tatcgtggct ggccacgacg ggcgttcctt gcgcagctgt gctcgacgtt gtcactgaag
3121 cgggaaggga ctggctgcta ttgggcgaag tgccggggca ggatctcctg tcatctcacc
3181 ttgctcctgc cgagaaagta tccatcatgg ctgatgcaat gcggcggctg catacgcttg
3241 atccggctac ctgcccattc gaccaccaag cgaaacatcg catcgagcga gcacgtactc
3301 ggatggaagc cggtcttgtc gatcaggatg atctggacga agagcatcag gggctcgcgc
3361 cagccgaact gttcgccagg ctcaaggcgc gcatgcccga cggcgaggat ctcgtcgtga
3421 cccatggcga tgcctgcttg ccgaatatca tggtggaaaa tggccgcttt tctggattca
3481 tcgactgtgg ccggctgggt gtggcggacc gctatcagga catagcgttg gctacccgtg
3541 atattgctga agagcttggc ggcgaatggg ctgaccgctt cctcgtgctt tacggtatcg
3601 ccgctcccga ttcgcagcgc atcgccttct atcgccttct tgacgagttc ttctgagcgg
3661 gactctgggg ttcgaaatga ccgaccaagc gacgcccaac ctgccatcac gagatttcga
3721 ttccaccgcc gccttctatg aaaggttggg cttcggaatc gttttccggg acgccggctg
3781 gatgatcctc cagcgcgggg atctcatgct ggagttcttc gcccacccca acttgtttat
3841 tgcagcttat aatggttaca aataaagcaa tagcatcaca aatttcacaa ataaagcatt
3901 tttttcactg cattctagtt gtggtttgtc caaactcatc aatgtatctt atcatgtctg
3961 tataccgtcg acctctagct agagcttggc gtaatcatgg tcatagctgt ttcctgtgtg
4021 aaattgttat ccgctcacaa ttccacacaa catacgagcc ggaagcataa agtgtaaagc
4081 ctggggtgcc taatgagtga gctaactcac attaattgcg ttgcgctcac tgcccgcttt
4141 ccagtcggga aacctgtcgt gccagctgca ttaatgaatc ggccaacgcg cggggagagg
4201 cggtttgcgt attgggcgct cttccgcttc ctcgctcact gactcgctgc gctcggtcgt
4261 tcggctgcgg cgagcggtat cagctcactc aaaggcggta atacggttat ccacagaatc
4321 aggggataac gcaggaaaga acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa
4381 aaaggccgcg ttgctggcgt ttttccatag gctccgcccc cctgacgagc atcacaaaaa
4441 tcgacgctca agtcagaggt ggcgaaaccc gacaggacta taaagatacc aggcgtttcc
4501 ccctggaagc tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg gatacctgtc
4561 cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta ggtatctcag
4621 ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga
4681 ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggtaagac acgacttatc
4741 gccactggca gcagccactg gtaacaggat tagcagagcg aggtatgtag gcggtgctac
4801 agagttcttg aagtggtggc ctaactacgg ctacactaga agaacagtat ttggtatctg
4861 cgctctgctg aagccagtta ccttcggaaa aagagttggt agctcttgat ccggcaaaca
4921 aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa
4981 aggatctcaa gaagatcctt tgatcttttc tacggggtct gacgctcagt ggaacgaaaa
5041 ctcacgttaa gggattttgg tcatgagatt atcaaaaagg atcttcacct agatcctttt
5101 aaattaaaaa tgaagtttta aatcaatcta aagtatatat gagtaaactt ggtctgacag
5161 ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc tgtctatttc gttcatccat
5221 agttgcctga ctccccgtcg tgtagataac tacgatacgg gagggcttac catctggccc
5281 cagtgctgca atgataccgc gagacccacg ctcaccggct ccagatttat cagcaataaa
5341 ccagccagcc ggaagggccg agcgcagaag tggtcctgca actttatccg cctccatcca
5401 gtctattaat tgttgccggg aagctagagt aagtagttcg ccagttaata gtttgcgcaa
5461 cgttgttgcc attgctacag gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt
5521 cagctccggt tcccaacgat caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc
5581 ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag tgttatcact
5641 catggttatg gcagcactgc ataattctct tactgtcatg ccatccgtaa gatgcttttc
5701 tgtgactggt gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg
5761 ctcttgcccg gcgtcaatac gggataatac cgcgccacat agcagaactt taaaagtgct
5821 catcattgga aaacgttctt cggggcgaaa actctcaagg atcttaccgc tgttgagatc
5881 cagttcgatg taacccactc gtgcacccaa ctgatcttca gcatctttta ctttcaccag
5941 cgtttctggg tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa taagggcgac
6001 acggaaatgt tgaatactca tactcttcct ttttcaatat tattgaagca tttatcaggg
6061 ttattgtctc atgagcggat acatatttga atgtatttag aaaaataaac aaataggggt
6121 tccgcgcaca tttccccgaa aagtgccacc tgacgtcga
//
