| 质粒名称: | pcDNA-GFP |
|---|---|
| 出品公司: | Addgene |
| 目录编号: | 160697 |
| 存储实验室: | Christine Mayr |
|
载体骨架基本信息 |
|
| 载体名称: | pcDNA3.1 |
| 载体出品公司: | Clontech |
| 完整质粒大小: | 6188bp |
| 载体类型: | 哺乳细胞表达 |
| 载体抗性: | 氨苄霉素 |
| 生长温度: | 37 ℃ |
| 宿主菌: | DH5a |
| 拷贝数: | 未知 |
|
插入基因信息 |
|
| 插入基因名称: | GFP |
|
文献引用方法 |
|
| 材料和方法部分: |
pcDNA-GFP was a gift from Christine Mayr (Addgene plasmid # 160697 ; http://n2t.net/addgene:160697 ; RRID:Addgene_160697)
|
| 参考文献部分: | Gain of Additional BIRC3 Protein Functions through 3'-UTR-Mediated Protein Complex Formation. Lee SH, Mayr C. Mol Cell. 2019 May 16;74(4):701-712.e9. doi: 10.1016/j.molcel.2019.03.006. Epub 2019 Apr 1. 10.1016/j.molcel.2019.03.006 PubMed 30948266 |
买家导航
pcDNA-GFP载体质粒基本信息
pcDNA-GFP质粒图谱载体图谱和pcDNA-GFP载体序列质粒序列多克隆位点信息
pcDNA-GFP质粒载体简介
pcDNA-GFP质粒序列载体序列
LOCUS pcDNA-GFP 6108 bp ds-DNA circular SYN 12-MAY-2021
DEFINITION Expresses GFP in mammalian cells.
ACCESSION .
VERSION .
KEYWORDS pcDNA-GFP
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 6108)
AUTHORS Lee SH, Mayr C
TITLE Gain of Additional BIRC3 Protein Functions through 3'-UTR-Mediated
Protein Complex Formation.
JOURNAL Mol Cell. 2019 May 16;74(4):701-712.e9. doi:
10.1016/j.molcel.2019.03.006. Epub 2019 Apr 1.
PUBMED 30948266
REFERENCE 2 (bases 1 to 6108)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58
http://www.biofeng.com
FEATURES Location/Qualifiers
source 1..6108
/organism="synthetic DNA construct"
/mol_type="other DNA"
enhancer 1..380
/label=CMV enhancer
/note="human cytomegalovirus immediate early enhancer"
promoter 381..584
/label=CMV promoter
/note="human cytomegalovirus (CMV) immediate early
promoter"
primer_bind 535..555
/label=CMV-F
/note="Human CMV immediate early promoter, forward primer"
primer_bind 629..648
/label=T7
/note="T7 promoter, forward primer"
promoter 629..647
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
CDS 701..1420
/codon_start=1
/product="the original enhanced GFP (Yang et al., 1996)"
/label=EGFP
/note="mammalian codon-optimized"
/translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
ANFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
EFVTAAGITLGMDELYK"
primer_bind complement(746..767)
/label=EGFP-N
/note="EGFP, reverse primer"
primer_bind complement(1007..1026)
/label=EXFP-R
/note="For distinguishing EGFP variants, reverse primer"
primer_bind 1354..1375
/label=EGFP-C
/note="EGFP, forward primer"
primer_bind complement(1465..1482)
/label=BGH-rev
/note="Bovine growth hormone terminator, reverse primer.
Also called BGH reverse"
polyA_signal 1471..1695
/label=bGH poly(A) signal
/note="bovine growth hormone polyadenylation signal"
rep_origin 1741..2169
/direction=RIGHT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
primer_bind complement(1828..1847)
/label=F1ori-R
/note="F1 origin, reverse primer"
primer_bind 2038..2059
/label=F1ori-F
/note="F1 origin, forward primer"
primer_bind complement(2178..2198)
/label=pBABE 3'
/note="SV40 enhancer, reverse primer for pBABE vectors"
promoter 2183..2512
/label=SV40 promoter
/note="SV40 enhancer and early promoter"
rep_origin 2363..2498
/label=SV40 ori
/note="SV40 origin of replication"
primer_bind 2425..2444
/label=SV40pro-F
/note="SV40 promoter/origin, forward primer"
CDS 2579..3373
/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(2633..2652)
/label=Neo-R
/note="Neomycin resistance gene, reverse primer"
primer_bind 3243..3262
/label=Neo-F
/note="Neomycin resistance gene, forward primer"
polyA_signal 3547..3668
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
primer_bind complement(3584..3603)
/label=SV40pA-R
/note="SV40 polyA, reverse primer"
primer_bind 3638..3657
/label=EBV-rev
/note="SV40 polyA terminator, reverse primer"
primer_bind complement(3717..3733)
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(3717..3733)
/label=M13 Reverse
/note="In lacZ gene. Also called M13-rev"
primer_bind complement(3730..3752)
/label=M13/pUC Reverse
/note="In lacZ gene"
protein_bind 3741..3757
/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(3765..3795)
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 3810..3831
/label=CAP binding site
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding activates transcription in the presence
of cAMP."
primer_bind complement(3948..3965)
/label=L4440
/note="L4440 vector, forward primer"
rep_origin complement(4119..4707)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind complement(4199..4218)
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
CDS complement(4878..5738)
/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 5501..5520
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
promoter complement(5739..5843)
/gene="bla"
/label=AmpR promoter
primer_bind complement(5918..5937)
/label=pRS-marker
/note="pRS vectors, use to sequence yeast selectable
marker"
ORIGIN
1 gacattgatt attgactagt tattaatagt aatcaattac ggggtcatta gttcatagcc
61 catatatgga gttccgcgtt acataactta cggtaaatgg cccgcctggc tgaccgccca
121 acgacccccg cccattgacg tcaataatga cgtatgttcc catagtaacg ccaataggga
181 ctttccattg acgtcaatgg gtggagtatt tacggtaaac tgcccacttg gcagtacatc
241 aagtgtatca tatgccaagt acgcccccta ttgacgtcaa tgacggtaaa tggcccgcct
301 ggcattatgc ccagtacatg accttatggg actttcctac ttggcagtac atctacgtat
361 tagtcatcgc tattaccatg gtgatgcggt tttggcagta catcaatggg cgtggatagc
421 ggtttgactc acggggattt ccaagtctcc accccattga cgtcaatggg agtttgtttt
481 ggcaccaaaa tcaacgggac tttccaaaat gtcgtaacaa ctccgcccca ttgacgcaaa
541 tgggcggtag gcgtgtacgg tgggaggtct atataagcag agctctctgg ctaactagag
601 aacccactgc ttactggctt atcgaaatta atacgactca ctatagggag acccaagctg
661 gctagcgttt aaacttaagc ttggtaccga gctcggatcc atggtgagca agggcgagga
721 gctgttcacc ggggtggtgc ccatcctggt cgagctggac ggcgacgtaa acggccacaa
781 gttcagcgtg tctggcgagg gcgagggcga tgccacctac ggcaagctga ccctgaagtt
841 catctgcacc accggcaagc tgcccgtgcc ctggcccacc ctcgtgacca ccctgaccta
901 cggcgtgcag tgcttcagcc gctaccccga ccacatgaag cagcacgact tcttcaagtc
961 cgccatgccc gaaggctacg tccaggagcg caccatcttc ttcaaggacg acggcaacta
1021 caagacccgc gccgaggtga agttcgaggg cgacaccctg gtgaaccgca tcgagctgaa
1081 gggcatcgac ttcaaggagg acggcaacat cctggggcac aagctggagt acaactacaa
1141 cagccacaac gtctatatca tggccgacaa gcagaagaac ggcatcaagg cgaacttcaa
1201 gatccgccac aacatcgagg acggcagcgt gcagctcgcc gaccactacc agcagaacac
1261 ccccatcggc gacggccccg tgctgctgcc cgacaaccac tacctgagca cccagtccgc
1321 cctgagcaaa gaccccaacg agaagcgcga tcacatggtc ctgctggagt tcgtgaccgc
1381 cgccgggatc actctcggca tggacgagct gtacaagtaa gcggccgctc gagtctagag
1441 ggcccgttta aacccgctga tcagcctcga ctgtgccttc tagttgccag ccatctgttg
1501 tttgcccctc ccccgtgcct tccttgaccc tggaaggtgc cactcccact gtcctttcct
1561 aataaaatga ggaaattgca tcgcattgtc tgagtaggtg tcattctatt ctggggggtg
1621 gggtggggca ggacagcaag ggggaggatt gggaagacaa tagcaggcat gctggggatg
1681 cggtgggctc tatggcttct gaggcggaaa gaaccagctg gggctctagg gggtatcccc
1741 acgcgccctg tagcggcgca ttaagcgcgg cgggtgtggt ggttacgcgc agcgtgaccg
1801 ctacacttgc cagcgcccta gcgcccgctc ctttcgcttt cttcccttcc tttctcgcca
1861 cgttcgccgg ctttccccgt caagctctaa atcgggggct ccctttaggg ttccgattta
1921 gtgctttacg gcacctcgac cccaaaaaac ttgattaggg tgatggttca cgtagtgggc
1981 catcgccctg atagacggtt tttcgccctt tgacgttgga gtccacgttc tttaatagtg
2041 gactcttgtt ccaaactgga acaacactca accctatctc ggtctattct tttgatttat
2101 aagggatttt gccgatttcg gcctattggt taaaaaatga gctgatttaa caaaaattta
2161 acgcgaatta attctgtgga atgtgtgtca gttagggtgt ggaaagtccc caggctcccc
2221 agcaggcaga agtatgcaaa gcatgcatct caattagtca gcaaccaggt gtggaaagtc
2281 cccaggctcc ccagcaggca gaagtatgca aagcatgcat ctcaattagt cagcaaccat
2341 agtcccgccc ctaactccgc ccatcccgcc cctaactccg cccagttccg cccattctcc
2401 gccccatggc tgactaattt tttttattta tgcagaggcc gaggccgcct ctgcctctga
2461 gctattccag aagtagtgag gaggcttttt tggaggccta ggcttttgca aaaagctccc
2521 gggagcttgt atatccattt tcggatctga tcaagagaca ggatgaggat cgtttcgcat
2581 gattgaacaa gatggattgc acgcaggttc tccggccgct tgggtggaga ggctattcgg
2641 ctatgactgg gcacaacaga caatcggctg ctctgatgcc gccgtgttcc ggctgtcagc
2701 gcaggggcgc ccggttcttt ttgtcaagac cgacctgtcc ggtgccctga atgaactgca
2761 ggacgaggca gcgcggctat cgtggctggc cacgacgggc gttccttgcg cagctgtgct
2821 cgacgttgtc actgaagcgg gaagggactg gctgctattg ggcgaagtgc cggggcagga
2881 tctcctgtca tctcaccttg ctcctgccga gaaagtatcc atcatggctg atgcaatgcg
2941 gcggctgcat acgcttgatc cggctacctg cccattcgac caccaagcga aacatcgcat
3001 cgagcgagca cgtactcgga tggaagccgg tcttgtcgat caggatgatc tggacgaaga
3061 gcatcagggg ctcgcgccag ccgaactgtt cgccaggctc aaggcgcgca tgcccgacgg
3121 cgaggatctc gtcgtgaccc atggcgatgc ctgcttgccg aatatcatgg tggaaaatgg
3181 ccgcttttct ggattcatcg actgtggccg gctgggtgtg gcggaccgct atcaggacat
3241 agcgttggct acccgtgata ttgctgaaga gcttggcggc gaatgggctg accgcttcct
3301 cgtgctttac ggtatcgccg ctcccgattc gcagcgcatc gccttctatc gccttcttga
3361 cgagttcttc tgagcgggac tctggggttc gaaatgaccg accaagcgac gcccaacctg
3421 ccatcacgag atttcgattc caccgccgcc ttctatgaaa ggttgggctt cggaatcgtt
3481 ttccgggacg ccggctggat gatcctccag cgcggggatc tcatgctgga gttcttcgcc
3541 caccccaact tgtttattgc agcttataat ggttacaaat aaagcaatag catcacaaat
3601 ttcacaaata aagcattttt ttcactgcat tctagttgtg gtttgtccaa actcatcaat
3661 gtatcttatc atgtctgtat accgtcgacc tctagctaga gcttggcgta atcatggtca
3721 tagctgtttc ctgtgtgaaa ttgttatccg ctcacaattc cacacaacat acgagccgga
3781 agcataaagt gtaaagcctg gggtgcctaa tgagtgagct aactcacatt aattgcgttg
3841 cgctcactgc ccgctttcca gtcgggaaac ctgtcgtgcc agctgcatta atgaatcggc
3901 caacgcgcgg ggagaggcgg tttgcgtatt gggcgctctt ccgcttcctc gctcactgac
3961 tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa ggcggtaata
4021 cggttatcca cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa
4081 aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt tccataggct ccgcccccct
4141 gacgagcatc acaaaaatcg acgctcaagt cagaggtggc gaaacccgac aggactataa
4201 agataccagg cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg
4261 cttaccggat acctgtccgc ctttctccct tcgggaagcg tggcgctttc tcatagctca
4321 cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa
4381 ccccccgttc agcccgaccg ctgcgcctta tccggtaact atcgtcttga gtccaacccg
4441 gtaagacacg acttatcgcc actggcagca gccactggta acaggattag cagagcgagg
4501 tatgtaggcg gtgctacaga gttcttgaag tggtggccta actacggcta cactagaaga
4561 acagtatttg gtatctgcgc tctgctgaag ccagttacct tcggaaaaag agttggtagc
4621 tcttgatccg gcaaacaaac caccgctggt agcggtggtt tttttgtttg caagcagcag
4681 attacgcgca gaaaaaaagg atctcaagaa gatcctttga tcttttctac ggggtctgac
4741 gctcagtgga acgaaaactc acgttaaggg attttggtca tgagattatc aaaaaggatc
4801 ttcacctaga tccttttaaa ttaaaaatga agttttaaat caatctaaag tatatatgag
4861 taaacttggt ctgacagtta ccaatgctta atcagtgagg cacctatctc agcgatctgt
4921 ctatttcgtt catccatagt tgcctgactc cccgtcgtgt agataactac gatacgggag
4981 ggcttaccat ctggccccag tgctgcaatg ataccgcgag acccacgctc accggctcca
5041 gatttatcag caataaacca gccagccgga agggccgagc gcagaagtgg tcctgcaact
5101 ttatccgcct ccatccagtc tattaattgt tgccgggaag ctagagtaag tagttcgcca
5161 gttaatagtt tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg
5221 tttggtatgg cttcattcag ctccggttcc caacgatcaa ggcgagttac atgatccccc
5281 atgttgtgca aaaaagcggt tagctccttc ggtcctccga tcgttgtcag aagtaagttg
5341 gccgcagtgt tatcactcat ggttatggca gcactgcata attctcttac tgtcatgcca
5401 tccgtaagat gcttttctgt gactggtgag tactcaacca agtcattctg agaatagtgt
5461 atgcggcgac cgagttgctc ttgcccggcg tcaatacggg ataataccgc gccacatagc
5521 agaactttaa aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc
5581 ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg cacccaactg atcttcagca
5641 tcttttactt tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa
5701 aagggaataa gggcgacacg gaaatgttga atactcatac tcttcctttt tcaatattat
5761 tgaagcattt atcagggtta ttgtctcatg agcggataca tatttgaatg tatttagaaa
5821 aataaacaaa taggggttcc gcgcacattt ccccgaaaag tgccacctga cgtcgacgga
5881 tcgggagatc tcccgatccc ctatggtgca ctctcagtac aatctgctct gatgccgcat
5941 agttaagcca gtatctgctc cctgcttgtg tgttggaggt cgctgagtag tgcgcgagca
6001 aaatttaagc tacaacaagg caaggcttga ccgacaattg catgaagaat ctgcttaggg
6061 ttaggcgttt tgcgctgctt cgcgatgtac gggccagata tacgcgtt
//
