| 质粒名称: | pcDNA3-YFP |
|---|---|
| 出品公司: | Addgene |
| 目录编号: | 13033 |
| 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: | Yellow fluorescent protein,YFP,EYFP |
| 物种来源: | -- |
| 插入大小: | 720bp |
| 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-YFP was a gift from Doug Golenbock (Addgene plasmid # 13033 ; http://n2t.net/addgene:13033 ; RRID:Addgene_13033) |
买家导航
pcDNA3YFP载体基本信息
pcDNA3YFP载体图谱质粒图谱和多克隆位点信息
pcDNA3YFP载体简介
pcDNA3YFP载体序列
LOCUS pcDNA3-YFP 6159 bp ds-DNA circular SYN 06-AUG-2018
DEFINITION Mammalian expression vector for cloning your gene fused to YFP.
ACCESSION .
VERSION .
KEYWORDS pcDNA3-YFP
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(46..65)
/label=pRS-marker
/note="pRS vectors, use to sequence yeast selectable
marker"
enhancer 237..616
/label=CMV enhancer
/note="human cytomegalovirus immediate early enhancer"
promoter 617..820
/label=CMV promoter
/note="human cytomegalovirus (CMV) immediate early
promoter"
primer_bind 771..791
/label=CMV-F
/note="Human CMV immediate early promoter, forward primer"
primer_bind 865..884
/label=T7
/note="T7 promoter, forward primer"
promoter 865..883
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
CDS 979..1698
/codon_start=1
/product="enhanced YFP"
/label=EYFP
/note="mammalian codon-optimized"
/translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
KFICTTGKLPVPWPTLVTTFGYGLKCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALSKDPNEKRDHMVLL
EFVTAAGITLGMDELYK"
primer_bind complement(1024..1045)
/label=EGFP-N
/note="EGFP, reverse primer"
primer_bind complement(1285..1304)
/label=EXFP-R
/note="For distinguishing EGFP variants, reverse primer"
primer_bind 1632..1653
/label=EGFP-C
/note="EGFP, forward primer"
promoter complement(1714..1732)
/label=SP6 promoter
/note="promoter for bacteriophage SP6 RNA polymerase"
primer_bind complement(1715..1732)
/label=SP6
/note="SP6 promoter, forward primer"
primer_bind complement(1752..1769)
/label=BGH-rev
/note="Bovine growth hormone terminator, reverse primer.
Also called BGH reverse"
polyA_signal 1758..1982
/label=bGH poly(A) signal
/note="bovine growth hormone polyadenylation signal"
rep_origin 2028..2456
/direction=RIGHT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
primer_bind complement(2115..2134)
/label=F1ori-R
/note="F1 origin, reverse primer"
primer_bind 2325..2346
/label=F1ori-F
/note="F1 origin, forward primer"
primer_bind complement(2465..2485)
/label=pBABE 3'
/note="SV40 enhancer, reverse primer for pBABE vectors"
promoter 2470..2799
/label=SV40 promoter
/note="SV40 enhancer and early promoter"
rep_origin 2650..2785
/label=SV40 ori
/note="SV40 origin of replication"
primer_bind 2712..2731
/label=SV40pro-F
/note="SV40 promoter/origin, forward primer"
CDS 2866..3660
/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(2920..2939)
/label=Neo-R
/note="Neomycin resistance gene, reverse primer"
primer_bind 3530..3549
/label=Neo-F
/note="Neomycin resistance gene, forward primer"
polyA_signal 3834..3955
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
primer_bind complement(3871..3890)
/label=SV40pA-R
/note="SV40 polyA, reverse primer"
primer_bind 3925..3944
/label=EBV-rev
/note="SV40 polyA terminator, reverse primer"
primer_bind complement(4004..4020)
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(4004..4020)
/label=M13 Reverse
/note="In lacZ gene. Also called M13-rev"
primer_bind complement(4017..4039)
/label=M13/pUC Reverse
/note="In lacZ gene"
protein_bind 4028..4044
/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(4052..4082)
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 4097..4118
/label=CAP binding site
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding activates transcription in the presence
of cAMP."
primer_bind complement(4235..4252)
/label=L4440
/note="L4440 vector, forward primer"
rep_origin complement(4406..4994)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind complement(4486..4505)
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
CDS complement(5165..6025)
/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 5788..5807
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
promoter complement(6026..6130)
/gene="bla"
/label=AmpR promoter
ORIGIN
1 tcgacggatc gggagatctc ccgatcccct atggtgcact ctcagtacaa tctgctctga
61 tgccgcatag ttaagccagt atctgctccc tgcttgtgtg ttggaggtcg ctgagtagtg
121 cgcgagcaaa atttaagcta caacaaggca aggcttgacc gacaattgca tgaagaatct
181 gcttagggtt aggcgttttg cgctgcttcg cgatgtacgg gccagatata cgcgttgaca
241 ttgattattg actagttatt aatagtaatc aattacgggg tcattagttc atagcccata
301 tatggagttc cgcgttacat aacttacggt aaatggcccg cctggctgac cgcccaacga
361 cccccgccca ttgacgtcaa taatgacgta tgttcccata gtaacgccaa tagggacttt
421 ccattgacgt caatgggtgg agtatttacg gtaaactgcc cacttggcag tacatcaagt
481 gtatcatatg ccaagtacgc cccctattga cgtcaatgac ggtaaatggc ccgcctggca
541 ttatgcccag tacatgacct tatgggactt tcctacttgg cagtacatct acgtattagt
601 catcgctatt accatggtga tgcggttttg gcagtacatc aatgggcgtg gatagcggtt
661 tgactcacgg ggatttccaa gtctccaccc cattgacgtc aatgggagtt tgttttggca
721 ccaaaatcaa cgggactttc caaaatgtcg taacaactcc gccccattga cgcaaatggg
781 cggtaggcgt gtacggtggg aggtctatat aagcagagct ctctggctaa ctagagaacc
841 cactgcttac tggcttatcg aaattaatac gactcactat agggagaccc aagcttggta
901 ccgagctcgg atccactagt aacggccgcc agtgtgctgg aattctgcag atatccatca
961 cactggcggc cgctcgagat ggtgagcaag ggcgaggagc tgttcaccgg ggtggtgccc
1021 atcctggtcg agctggacgg cgacgtaaac ggccacaagt tcagcgtgtc cggcgagggc
1081 gagggcgatg ccacctacgg caagctgacc ctgaagttca tctgcaccac cggcaagctg
1141 cccgtgccct ggcccaccct cgtgaccacc ttcggctacg gcctgaagtg cttcgcccgc
1201 taccccgacc acatgaagca gcacgacttc ttcaagtccg ccatgcccga aggctacgtc
1261 caggagcgca ccatcttctt caaggacgac ggcaactaca agacccgcgc cgaggtgaag
1321 ttcgagggcg acaccctggt gaaccgcatc gagctgaagg gcatcgactt caaggaggac
1381 ggcaacatcc tggggcacaa gctggagtac aactacaaca gccacaacgt ctatatcatg
1441 gccgacaagc agaagaacgg catcaaggtg aacttcaaga tccgccacaa catcgaggac
1501 ggcagcgtgc agctcgccga ccactaccag cagaacaccc ccatcggcga cggccccgtg
1561 ctgctgcccg acaaccacta cctgagctac cagtccgccc tgagcaaaga ccccaacgag
1621 aagcgcgatc acatggtcct gctggagttc gtgaccgccg ccgggatcac tctcggcatg
1681 gacgagctgt acaagtaatc tagagggccc tattctatag tgtcacctaa atgctagagc
1741 tcgctgatca gcctcgactg tgccttctag ttgccagcca tctgttgttt gcccctcccc
1801 cgtgccttcc ttgaccctgg aaggtgccac tcccactgtc ctttcctaat aaaatgagga
1861 aattgcatcg cattgtctga gtaggtgtca ttctattctg gggggtgggg tggggcagga
1921 cagcaagggg gaggattggg aagacaatag caggcatgct ggggatgcgg tgggctctat
1981 ggcttctgag gcggaaagaa ccagctgggg ctctaggggg tatccccacg cgccctgtag
2041 cggcgcatta agcgcggcgg gtgtggtggt tacgcgcagc gtgaccgcta cacttgccag
2101 cgccctagcg cccgctcctt tcgctttctt cccttccttt ctcgccacgt tcgccggctt
2161 tccccgtcaa gctctaaatc gggggctccc tttagggttc cgatttagtg ctttacggca
2221 cctcgacccc aaaaaacttg attagggtga tggttcacgt agtgggccat cgccctgata
2281 gacggttttt cgccctttga cgttggagtc cacgttcttt aatagtggac tcttgttcca
2341 aactggaaca acactcaacc ctatctcggt ctattctttt gatttataag ggattttgcc
2401 gatttcggcc tattggttaa aaaatgagct gatttaacaa aaatttaacg cgaattaatt
2461 ctgtggaatg tgtgtcagtt agggtgtgga aagtccccag gctccccagc aggcagaagt
2521 atgcaaagca tgcatctcaa ttagtcagca accaggtgtg gaaagtcccc aggctcccca
2581 gcaggcagaa gtatgcaaag catgcatctc aattagtcag caaccatagt cccgccccta
2641 actccgccca tcccgcccct aactccgccc agttccgccc attctccgcc ccatggctga
2701 ctaatttttt ttatttatgc agaggccgag gccgcctctg cctctgagct attccagaag
2761 tagtgaggag gcttttttgg aggcctaggc ttttgcaaaa agctcccggg agcttgtata
2821 tccattttcg gatctgatca agagacagga tgaggatcgt ttcgcatgat tgaacaagat
2881 ggattgcacg caggttctcc ggccgcttgg gtggagaggc tattcggcta tgactgggca
2941 caacagacaa tcggctgctc tgatgccgcc gtgttccggc tgtcagcgca ggggcgcccg
3001 gttctttttg tcaagaccga cctgtccggt gccctgaatg aactgcagga cgaggcagcg
3061 cggctatcgt ggctggccac gacgggcgtt ccttgcgcag ctgtgctcga cgttgtcact
3121 gaagcgggaa gggactggct gctattgggc gaagtgccgg ggcaggatct cctgtcatct
3181 caccttgctc ctgccgagaa agtatccatc atggctgatg caatgcggcg gctgcatacg
3241 cttgatccgg ctacctgccc attcgaccac caagcgaaac atcgcatcga gcgagcacgt
3301 actcggatgg aagccggtct tgtcgatcag gatgatctgg acgaagagca tcaggggctc
3361 gcgccagccg aactgttcgc caggctcaag gcgcgcatgc ccgacggcga ggatctcgtc
3421 gtgacccatg gcgatgcctg cttgccgaat atcatggtgg aaaatggccg cttttctgga
3481 ttcatcgact gtggccggct gggtgtggcg gaccgctatc aggacatagc gttggctacc
3541 cgtgatattg ctgaagagct tggcggcgaa tgggctgacc gcttcctcgt gctttacggt
3601 atcgccgctc ccgattcgca gcgcatcgcc ttctatcgcc ttcttgacga gttcttctga
3661 gcgggactct ggggttcgaa atgaccgacc aagcgacgcc caacctgcca tcacgagatt
3721 tcgattccac cgccgccttc tatgaaaggt tgggcttcgg aatcgttttc cgggacgccg
3781 gctggatgat cctccagcgc ggggatctca tgctggagtt cttcgcccac cccaacttgt
3841 ttattgcagc ttataatggt tacaaataaa gcaatagcat cacaaatttc acaaataaag
3901 catttttttc actgcattct agttgtggtt tgtccaaact catcaatgta tcttatcatg
3961 tctgtatacc gtcgacctct agctagagct tggcgtaatc atggtcatag ctgtttcctg
4021 tgtgaaattg ttatccgctc acaattccac acaacatacg agccggaagc ataaagtgta
4081 aagcctgggg tgcctaatga gtgagctaac tcacattaat tgcgttgcgc tcactgcccg
4141 ctttccagtc gggaaacctg tcgtgccagc tgcattaatg aatcggccaa cgcgcgggga
4201 gaggcggttt gcgtattggg cgctcttccg cttcctcgct cactgactcg ctgcgctcgg
4261 tcgttcggct gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg ttatccacag
4321 aatcagggga taacgcagga aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc
4381 gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg cccccctgac gagcatcaca
4441 aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg actataaaga taccaggcgt
4501 ttccccctgg aagctccctc gtgcgctctc ctgttccgac cctgccgctt accggatacc
4561 tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc
4621 tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc
4681 ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc caacccggta agacacgact
4741 tatcgccact ggcagcagcc actggtaaca ggattagcag agcgaggtat gtaggcggtg
4801 ctacagagtt cttgaagtgg tggcctaact acggctacac tagaagaaca gtatttggta
4861 tctgcgctct gctgaagcca gttaccttcg gaaaaagagt tggtagctct tgatccggca
4921 aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa
4981 aaaaaggatc tcaagaagat cctttgatct tttctacggg gtctgacgct cagtggaacg
5041 aaaactcacg ttaagggatt ttggtcatga gattatcaaa aaggatcttc acctagatcc
5101 ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa acttggtctg
5161 acagttacca atgcttaatc agtgaggcac ctatctcagc gatctgtcta tttcgttcat
5221 ccatagttgc ctgactcccc gtcgtgtaga taactacgat acgggagggc ttaccatctg
5281 gccccagtgc tgcaatgata ccgcgagacc cacgctcacc ggctccagat ttatcagcaa
5341 taaaccagcc agccggaagg gccgagcgca gaagtggtcc tgcaacttta tccgcctcca
5401 tccagtctat taattgttgc cgggaagcta gagtaagtag ttcgccagtt aatagtttgc
5461 gcaacgttgt tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt
5521 cattcagctc cggttcccaa cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa
5581 aagcggttag ctccttcggt cctccgatcg ttgtcagaag taagttggcc gcagtgttat
5641 cactcatggt tatggcagca ctgcataatt ctcttactgt catgccatcc gtaagatgct
5701 tttctgtgac tggtgagtac tcaaccaagt cattctgaga atagtgtatg cggcgaccga
5761 gttgctcttg cccggcgtca atacgggata ataccgcgcc acatagcaga actttaaaag
5821 tgctcatcat tggaaaacgt tcttcggggc gaaaactctc aaggatctta ccgctgttga
5881 gatccagttc gatgtaaccc actcgtgcac ccaactgatc ttcagcatct tttactttca
5941 ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg
6001 cgacacggaa atgttgaata ctcatactct tcctttttca atattattga agcatttatc
6061 agggttattg tctcatgagc ggatacatat ttgaatgtat ttagaaaaat aaacaaatag
6121 gggttccgcg cacatttccc cgaaaagtgc cacctgacg
//
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