| 质粒名称: | pcDNA3-CFP |
|---|---|
| 出品公司: | Addgene |
| 目录编号: | 13030 |
| 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: | Invitrogen |
| 载体抗性: | 氨苄青霉素 |
| 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: | cyan fluorescent protein,CFP,ECFP |
| 物种来源: | -- |
| 插入大小: | 720 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-CFP was a gift from Doug Golenbock (Addgene plasmid # 13030 ; http://n2t.net/addgene:13030 ; RRID:Addgene_13030) |
买家导航
pcDNA3CFP载体基本信息
pcDNA3CFP载体图谱质粒图谱和多克隆位点信息
pcDNA3CFP载体简介
pcDNA3CFP载体序列
LOCUS pcDNA3-CFP 6159 bp ds-DNA circular SYN 06-AUG-2018
DEFINITION Mammalian expression vector for cloning your gene fused to CFP.
ACCESSION .
VERSION .
KEYWORDS pcDNA3-CFP
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 Aug 6, 2018 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(32..51)
/label=pRS-marker
/note="pRS vectors, use to sequence yeast selectable
marker"
enhancer 223..602
/label=CMV enhancer
/note="human cytomegalovirus immediate early enhancer"
promoter 603..806
/label=CMV promoter
/note="human cytomegalovirus (CMV) immediate early
promoter"
primer_bind 757..777
/label=CMV-F
/note="Human CMV immediate early promoter, forward primer"
primer_bind 851..870
/label=T7
/note="T7 promoter, forward primer"
promoter 851..869
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
CDS 965..1684
/codon_start=1
/product="cyan variant of GFP"
/label=CFP
/note="mammalian codon-optimized"
/translation="MVSKGEELFTGVVPILVELDGDVNGHRFSVSGEGEGDATYGKLTL
KFICTTGKLPVPWPTLVTTLTWGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYISHNVYITADKQKNGIK
AHFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
EFVTAAGITLGMDELYK"
primer_bind complement(1010..1031)
/label=EGFP-N
/note="EGFP, reverse primer"
primer_bind complement(1271..1290)
/label=EXFP-R
/note="For distinguishing EGFP variants, reverse primer"
primer_bind 1618..1639
/label=EGFP-C
/note="EGFP, forward primer"
promoter complement(1700..1718)
/label=SP6 promoter
/note="promoter for bacteriophage SP6 RNA polymerase"
primer_bind complement(1701..1718)
/label=SP6
/note="SP6 promoter, forward primer"
primer_bind complement(1738..1755)
/label=BGH-rev
/note="Bovine growth hormone terminator, reverse primer.
Also called BGH reverse"
polyA_signal 1744..1968
/label=bGH poly(A) signal
/note="bovine growth hormone polyadenylation signal"
rep_origin 2014..2442
/direction=RIGHT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
primer_bind complement(2101..2120)
/label=F1ori-R
/note="F1 origin, reverse primer"
primer_bind 2311..2332
/label=F1ori-F
/note="F1 origin, forward primer"
primer_bind complement(2451..2471)
/label=pBABE 3'
/note="SV40 enhancer, reverse primer for pBABE vectors"
promoter 2456..2785
/label=SV40 promoter
/note="SV40 enhancer and early promoter"
rep_origin 2636..2771
/label=SV40 ori
/note="SV40 origin of replication"
primer_bind 2698..2717
/label=SV40pro-F
/note="SV40 promoter/origin, forward primer"
CDS 2852..3646
/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(2906..2925)
/label=Neo-R
/note="Neomycin resistance gene, reverse primer"
primer_bind 3516..3535
/label=Neo-F
/note="Neomycin resistance gene, forward primer"
polyA_signal 3820..3941
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
primer_bind complement(3857..3876)
/label=SV40pA-R
/note="SV40 polyA, reverse primer"
primer_bind 3911..3930
/label=EBV-rev
/note="SV40 polyA terminator, reverse primer"
primer_bind complement(3990..4006)
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(3990..4006)
/label=M13 Reverse
/note="In lacZ gene. Also called M13-rev"
primer_bind complement(4003..4025)
/label=M13/pUC Reverse
/note="In lacZ gene"
protein_bind 4014..4030
/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(4038..4068)
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 4083..4104
/label=CAP binding site
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding activates transcription in the presence
of cAMP."
primer_bind complement(4221..4238)
/label=L4440
/note="L4440 vector, forward primer"
rep_origin complement(4392..4980)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind complement(4472..4491)
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
CDS complement(5151..6011)
/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 5774..5793
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
promoter complement(6012..6116)
/gene="bla"
/label=AmpR promoter
ORIGIN
1 gatctcccga tcccctatgg tgcactctca gtacaatctg ctctgatgcc gcatagttaa
61 gccagtatct gctccctgct tgtgtgttgg aggtcgctga gtagtgcgcg agcaaaattt
121 aagctacaac aaggcaaggc ttgaccgaca attgcatgaa gaatctgctt agggttaggc
181 gttttgcgct gcttcgcgat gtacgggcca gatatacgcg ttgacattga ttattgacta
241 gttattaata gtaatcaatt acggggtcat tagttcatag cccatatatg gagttccgcg
301 ttacataact tacggtaaat ggcccgcctg gctgaccgcc caacgacccc cgcccattga
361 cgtcaataat gacgtatgtt cccatagtaa cgccaatagg gactttccat tgacgtcaat
421 gggtggagta tttacggtaa actgcccact tggcagtaca tcaagtgtat catatgccaa
481 gtacgccccc tattgacgtc aatgacggta aatggcccgc ctggcattat gcccagtaca
541 tgaccttatg ggactttcct acttggcagt acatctacgt attagtcatc gctattacca
601 tggtgatgcg gttttggcag tacatcaatg ggcgtggata gcggtttgac tcacggggat
661 ttccaagtct ccaccccatt gacgtcaatg ggagtttgtt ttggcaccaa aatcaacggg
721 actttccaaa atgtcgtaac aactccgccc cattgacgca aatgggcggt aggcgtgtac
781 ggtgggaggt ctatataagc agagctctct ggctaactag agaacccact gcttactggc
841 ttatcgaaat taatacgact cactataggg agacccaagc ttggtaccga gctcggatcc
901 actagtaacg gccgccagtg tgctggaatt ctgcagatat ccatcacact ggcggccgct
961 cgagatggtg agcaagggcg aggagctgtt caccggggtg gtgcccatcc tggtcgagct
1021 ggacggcgac gtaaacggcc acaggttcag cgtgtccggc gagggcgagg gcgatgccac
1081 ctacggcaag ctgaccctga agttcatctg caccaccggc aagctgcccg tgccctggcc
1141 caccctcgtg accaccctga cctggggcgt gcagtgcttc agccgctacc ccgaccacat
1201 gaagcagcac gacttcttca agtccgccat gcccgaaggc tacgtccagg agcgtaccat
1261 cttcttcaag gacgacggca actacaagac ccgcgccgag gtgaagttcg agggcgacac
1321 cctggtgaac cgcatcgagc tgaagggcat cgacttcaag gaggacggca acatcctggg
1381 gcacaagctg gagtacaact acatcagcca caacgtctat atcaccgccg acaagcagaa
1441 gaacggcatc aaggcccact tcaagatccg ccacaacatc gaggacggca gcgtgcagct
1501 cgccgaccac taccagcaga acacccccat cggcgacggc cccgtgctgc tgcccgacaa
1561 ccactacctg agcacccagt ccgccctgag caaagacccc aacgagaagc gcgatcacat
1621 ggtcctgctg gagttcgtga ccgccgccgg gatcactctc ggcatggacg agctgtacaa
1681 gtaatctaga gggccctatt ctatagtgtc acctaaatgc tagagctcgc tgatcagcct
1741 cgactgtgcc ttctagttgc cagccatctg ttgtttgccc ctcccccgtg ccttccttga
1801 ccctggaagg tgccactccc actgtccttt cctaataaaa tgaggaaatt gcatcgcatt
1861 gtctgagtag gtgtcattct attctggggg gtggggtggg gcaggacagc aagggggagg
1921 attgggaaga caatagcagg catgctgggg atgcggtggg ctctatggct tctgaggcgg
1981 aaagaaccag ctggggctct agggggtatc cccacgcgcc ctgtagcggc gcattaagcg
2041 cggcgggtgt ggtggttacg cgcagcgtga ccgctacact tgccagcgcc ctagcgcccg
2101 ctcctttcgc tttcttccct tcctttctcg ccacgttcgc cggctttccc cgtcaagctc
2161 taaatcgggg gctcccttta gggttccgat ttagtgcttt acggcacctc gaccccaaaa
2221 aacttgatta gggtgatggt tcacgtagtg ggccatcgcc ctgatagacg gtttttcgcc
2281 ctttgacgtt ggagtccacg ttctttaata gtggactctt gttccaaact ggaacaacac
2341 tcaaccctat ctcggtctat tcttttgatt tataagggat tttgccgatt tcggcctatt
2401 ggttaaaaaa tgagctgatt taacaaaaat ttaacgcgaa ttaattctgt ggaatgtgtg
2461 tcagttaggg tgtggaaagt ccccaggctc cccagcaggc agaagtatgc aaagcatgca
2521 tctcaattag tcagcaacca ggtgtggaaa gtccccaggc tccccagcag gcagaagtat
2581 gcaaagcatg catctcaatt agtcagcaac catagtcccg cccctaactc cgcccatccc
2641 gcccctaact ccgcccagtt ccgcccattc tccgccccat ggctgactaa ttttttttat
2701 ttatgcagag gccgaggccg cctctgcctc tgagctattc cagaagtagt gaggaggctt
2761 ttttggaggc ctaggctttt gcaaaaagct cccgggagct tgtatatcca ttttcggatc
2821 tgatcaagag acaggatgag gatcgtttcg catgattgaa caagatggat tgcacgcagg
2881 ttctccggcc gcttgggtgg agaggctatt cggctatgac tgggcacaac agacaatcgg
2941 ctgctctgat gccgccgtgt tccggctgtc agcgcagggg cgcccggttc tttttgtcaa
3001 gaccgacctg tccggtgccc tgaatgaact gcaggacgag gcagcgcggc tatcgtggct
3061 ggccacgacg ggcgttcctt gcgcagctgt gctcgacgtt gtcactgaag cgggaaggga
3121 ctggctgcta ttgggcgaag tgccggggca ggatctcctg tcatctcacc ttgctcctgc
3181 cgagaaagta tccatcatgg ctgatgcaat gcggcggctg catacgcttg atccggctac
3241 ctgcccattc gaccaccaag cgaaacatcg catcgagcga gcacgtactc ggatggaagc
3301 cggtcttgtc gatcaggatg atctggacga agagcatcag gggctcgcgc cagccgaact
3361 gttcgccagg ctcaaggcgc gcatgcccga cggcgaggat ctcgtcgtga cccatggcga
3421 tgcctgcttg ccgaatatca tggtggaaaa tggccgcttt tctggattca tcgactgtgg
3481 ccggctgggt gtggcggacc gctatcagga catagcgttg gctacccgtg atattgctga
3541 agagcttggc ggcgaatggg ctgaccgctt cctcgtgctt tacggtatcg ccgctcccga
3601 ttcgcagcgc atcgccttct atcgccttct tgacgagttc ttctgagcgg gactctgggg
3661 ttcgaaatga ccgaccaagc gacgcccaac ctgccatcac gagatttcga ttccaccgcc
3721 gccttctatg aaaggttggg cttcggaatc gttttccggg acgccggctg gatgatcctc
3781 cagcgcgggg atctcatgct ggagttcttc gcccacccca acttgtttat tgcagcttat
3841 aatggttaca aataaagcaa tagcatcaca aatttcacaa ataaagcatt tttttcactg
3901 cattctagtt gtggtttgtc caaactcatc aatgtatctt atcatgtctg tataccgtcg
3961 acctctagct agagcttggc gtaatcatgg tcatagctgt ttcctgtgtg aaattgttat
4021 ccgctcacaa ttccacacaa catacgagcc ggaagcataa agtgtaaagc ctggggtgcc
4081 taatgagtga gctaactcac attaattgcg ttgcgctcac tgcccgcttt ccagtcggga
4141 aacctgtcgt gccagctgca ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt
4201 attgggcgct cttccgcttc ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg
4261 cgagcggtat cagctcactc aaaggcggta atacggttat ccacagaatc aggggataac
4321 gcaggaaaga acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg
4381 ttgctggcgt ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca
4441 agtcagaggt ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc
4501 tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc
4561 ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta ggtatctcag ttcggtgtag
4621 gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc
4681 ttatccggta actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca
4741 gcagccactg gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg
4801 aagtggtggc ctaactacgg ctacactaga agaacagtat ttggtatctg cgctctgctg
4861 aagccagtta ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct
4921 ggtagcggtg gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa
4981 gaagatcctt tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa
5041 gggattttgg tcatgagatt atcaaaaagg atcttcacct agatcctttt aaattaaaaa
5101 tgaagtttta aatcaatcta aagtatatat gagtaaactt ggtctgacag ttaccaatgc
5161 ttaatcagtg aggcacctat ctcagcgatc tgtctatttc gttcatccat agttgcctga
5221 ctccccgtcg tgtagataac tacgatacgg gagggcttac catctggccc cagtgctgca
5281 atgataccgc gagacccacg ctcaccggct ccagatttat cagcaataaa ccagccagcc
5341 ggaagggccg agcgcagaag tggtcctgca actttatccg cctccatcca gtctattaat
5401 tgttgccggg aagctagagt aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc
5461 attgctacag gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt cagctccggt
5521 tcccaacgat caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc
5581 ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag tgttatcact catggttatg
5641 gcagcactgc ataattctct tactgtcatg ccatccgtaa gatgcttttc tgtgactggt
5701 gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg
5761 gcgtcaatac gggataatac cgcgccacat agcagaactt taaaagtgct catcattgga
5821 aaacgttctt cggggcgaaa actctcaagg atcttaccgc tgttgagatc cagttcgatg
5881 taacccactc gtgcacccaa ctgatcttca gcatctttta ctttcaccag cgtttctggg
5941 tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa taagggcgac acggaaatgt
6001 tgaatactca tactcttcct ttttcaatat tattgaagca tttatcaggg ttattgtctc
6061 atgagcggat acatatttga atgtatttag aaaaataaac aaataggggt tccgcgcaca
6121 tttccccgaa aagtgccacc tgacgtcgac ggatcggga
//
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