| 质粒名称: | pcDNA3 HA Arf6 |
|---|---|
| 出品公司: | 爱迪基因 |
| 目录编号: | 10834 |
| 存储实验室: | Thomas Roberts |
|
载体骨架基本信息 |
|
| 载体名称: | pcDNA3.1 |
| 载体出品公司: | Invitrogen |
| 载体抗性: | 氨苄青霉素 |
| 空载体大小: | 5428 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: | Arf6 |
| 物种来源: | H. sapiens (human) |
| 插入大小: | 525 bp |
| 基因突变: | -- |
| GenBank编号 | -- |
| 启动子 | -- |
| 标签/融合蛋白: | HA（插入件上的C端子） |
| 克隆方法: | 限制性内切酶 |
| 5' 克隆位点: | EcoRI（未销毁） |
| 3' 克隆位点: | XhoI（未销毁） |
| 5' 测序引物 | T7 |
| 3' 测序引物 | bGH PA terminator |
|
文献引用方法 |
|
| 材料和方法部分: | pcDNA3 HA Arf6 was a gift from Thomas Roberts (Addgene plasmid # 10834 ; http://n2t.net/addgene:10834 ; RRID:Addgene_10834) |
| 参考文献部分: | DEF-1/ASAP1 is a GTPase-activating protein (GAP) for ARF1 that enhances cell motility through a GAP-dependent mechanism. Furman C, Short SM, Subramanian RR, Zetter BR, Roberts TM. J Biol Chem. 2002 Mar 8. 277(10):7962-9. 10.1074/jbc.M109149200 PubMed 11773070 |
买家导航
载体基本信息
载体图谱质粒图谱和多克隆位点信息
载体简介
载体序列
LOCUS pcDNA3 HA Arf6 6017 bp ds-DNA circular SYN 06-AUG-2018
DEFINITION synthetic circular DNA
ACCESSION .
VERSION .
KEYWORDS pcDNA3 HA Arf6
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 6017)
AUTHORS Furman C, Short SM, Subramanian RR, Zetter BR, Roberts TM
TITLE DEF-1/ASAP1 is a GTPase-activating protein (GAP) for ARF1 that
enhances cell motility through a GAP-dependent mechanism.
JOURNAL J Biol Chem. 2002 Mar 8. 277(10):7962-9.
PUBMED 11773070
REFERENCE 2 (bases 1 to 6017)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Aug 6, 2018 from SnapGene Server 1.1.58
http://www.snapgene.com
FEATURES Location/Qualifiers
source 1..6017
/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 1471..1497
/codon_start=1
/product="HA (human influenza hemagglutinin) epitope tag"
/label=HA
/translation="YPYDVPDYA"
promoter complement(1572..1590)
/label=SP6 promoter
/note="promoter for bacteriophage SP6 RNA polymerase"
primer_bind complement(1573..1590)
/label=SP6
/note="SP6 promoter, forward primer"
primer_bind complement(1610..1627)
/label=BGH-rev
/note="Bovine growth hormone terminator, reverse primer.
Also called BGH reverse"
polyA_signal 1616..1840
/label=bGH poly(A) signal
/note="bovine growth hormone polyadenylation signal"
rep_origin 1886..2314
/direction=RIGHT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
primer_bind complement(1973..1992)
/label=F1ori-R
/note="F1 origin, reverse primer"
primer_bind 2183..2204
/label=F1ori-F
/note="F1 origin, forward primer"
primer_bind complement(2323..2343)
/label=pBABE 3'
/note="SV40 enhancer, reverse primer for pBABE vectors"
promoter 2328..2657
/label=SV40 promoter
/note="SV40 enhancer and early promoter"
rep_origin 2508..2643
/label=SV40 ori
/note="SV40 origin of replication"
primer_bind 2570..2589
/label=SV40pro-F
/note="SV40 promoter/origin, forward primer"
CDS 2724..3518
/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(2778..2797)
/label=Neo-R
/note="Neomycin resistance gene, reverse primer"
primer_bind 3388..3407
/label=Neo-F
/note="Neomycin resistance gene, forward primer"
polyA_signal 3692..3813
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
primer_bind complement(3729..3748)
/label=SV40pA-R
/note="SV40 polyA, reverse primer"
primer_bind 3783..3802
/label=EBV-rev
/note="SV40 polyA terminator, reverse primer"
primer_bind complement(3862..3878)
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(3862..3878)
/label=M13 Reverse
/note="In lacZ gene. Also called M13-rev"
primer_bind complement(3875..3897)
/label=M13/pUC Reverse
/note="In lacZ gene"
protein_bind 3886..3902
/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(3910..3940)
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 3955..3976
/label=CAP binding site
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding activates transcription in the presence
of cAMP."
primer_bind complement(4093..4110)
/label=L4440
/note="L4440 vector, forward primer"
rep_origin complement(4264..4852)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind complement(4344..4363)
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
CDS complement(5023..5883)
/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 5646..5665
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
promoter complement(5884..5988)
/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 aattcatggg gaaggtgcta
961 tccaaaatct tcgggaacaa ggaaatgcgg atcctcatgt tgggcctgga cgcggccggc
1021 aagacaacaa tcctgtacaa gttgaagctg ggccagtcgg tgaccaccat tcccactgtg
1081 ggtttcaacg tggagacggt gacttacaaa aatgtcaagt tcaacgtatg ggatgtgggc
1141 ggccaggaca agatccggcc gctctggcgg cattactaca ctgggaccca aggtctcatc
1201 ttcgtagtgg actgcgccga ccgcgaccgc atcgatgagg ctcgccagga gctgcaccgc
1261 attatcaatg accgggagat gagggacgcc ataatcctca tcttcgccaa caagcaggac
1321 ctgcccgatg ccatgaaacc ccacgagatc caggagaaac tgggcctgac ccggattcgg
1381 gacaggaact ggtatgtgca gccctcctgt gccacctcag gggacggact ctatgagggg
1441 ctcacatggt taacctctaa ctacaaatct tacccatacg atgttccaga ttacgcttaa
1501 tagtctagac agctgagatc tgatatccat cacactggcg gccgctcgag catgcatcta
1561 gagggcccta ttctatagtg tcacctaaat gctagagctc gctgatcagc ctcgactgtg
1621 ccttctagtt gccagccatc tgttgtttgc ccctcccccg tgccttcctt gaccctggaa
1681 ggtgccactc ccactgtcct ttcctaataa aatgaggaaa ttgcatcgca ttgtctgagt
1741 aggtgtcatt ctattctggg gggtggggtg gggcaggaca gcaaggggga ggattgggaa
1801 gacaatagca ggcatgctgg ggatgcggtg ggctctatgg cttctgaggc ggaaagaacc
1861 agctggggct ctagggggta tccccacgcg ccctgtagcg gcgcattaag cgcggcgggt
1921 gtggtggtta cgcgcagcgt gaccgctaca cttgccagcg ccctagcgcc cgctcctttc
1981 gctttcttcc cttcctttct cgccacgttc gccggctttc cccgtcaagc tctaaatcgg
2041 gggctccctt tagggttccg atttagtgct ttacggcacc tcgaccccaa aaaacttgat
2101 tagggtgatg gttcacgtag tgggccatcg ccctgataga cggtttttcg ccctttgacg
2161 ttggagtcca cgttctttaa tagtggactc ttgttccaaa ctggaacaac actcaaccct
2221 atctcggtct attcttttga tttataaggg attttgccga tttcggccta ttggttaaaa
2281 aatgagctga tttaacaaaa atttaacgcg aattaattct gtggaatgtg tgtcagttag
2341 ggtgtggaaa gtccccaggc tccccagcag gcagaagtat gcaaagcatg catctcaatt
2401 agtcagcaac caggtgtgga aagtccccag gctccccagc aggcagaagt atgcaaagca
2461 tgcatctcaa ttagtcagca accatagtcc cgcccctaac tccgcccatc ccgcccctaa
2521 ctccgcccag ttccgcccat tctccgcccc atggctgact aatttttttt atttatgcag
2581 aggccgaggc cgcctctgcc tctgagctat tccagaagta gtgaggaggc ttttttggag
2641 gcctaggctt ttgcaaaaag ctcccgggag cttgtatatc cattttcgga tctgatcaag
2701 agacaggatg aggatcgttt cgcatgattg aacaagatgg attgcacgca ggttctccgg
2761 ccgcttgggt ggagaggcta ttcggctatg actgggcaca acagacaatc ggctgctctg
2821 atgccgccgt gttccggctg tcagcgcagg ggcgcccggt tctttttgtc aagaccgacc
2881 tgtccggtgc cctgaatgaa ctgcaggacg aggcagcgcg gctatcgtgg ctggccacga
2941 cgggcgttcc ttgcgcagct gtgctcgacg ttgtcactga agcgggaagg gactggctgc
3001 tattgggcga agtgccgggg caggatctcc tgtcatctca ccttgctcct gccgagaaag
3061 tatccatcat ggctgatgca atgcggcggc tgcatacgct tgatccggct acctgcccat
3121 tcgaccacca agcgaaacat cgcatcgagc gagcacgtac tcggatggaa gccggtcttg
3181 tcgatcagga tgatctggac gaagagcatc aggggctcgc gccagccgaa ctgttcgcca
3241 ggctcaaggc gcgcatgccc gacggcgagg atctcgtcgt gacccatggc gatgcctgct
3301 tgccgaatat catggtggaa aatggccgct tttctggatt catcgactgt ggccggctgg
3361 gtgtggcgga ccgctatcag gacatagcgt tggctacccg tgatattgct gaagagcttg
3421 gcggcgaatg ggctgaccgc ttcctcgtgc tttacggtat cgccgctccc gattcgcagc
3481 gcatcgcctt ctatcgcctt cttgacgagt tcttctgagc gggactctgg ggttcgaaat
3541 gaccgaccaa gcgacgccca acctgccatc acgagatttc gattccaccg ccgccttcta
3601 tgaaaggttg ggcttcggaa tcgttttccg ggacgccggc tggatgatcc tccagcgcgg
3661 ggatctcatg ctggagttct tcgcccaccc caacttgttt attgcagctt ataatggtta
3721 caaataaagc aatagcatca caaatttcac aaataaagca tttttttcac tgcattctag
3781 ttgtggtttg tccaaactca tcaatgtatc ttatcatgtc tgtataccgt cgacctctag
3841 ctagagcttg gcgtaatcat ggtcatagct gtttcctgtg tgaaattgtt atccgctcac
3901 aattccacac aacatacgag ccggaagcat aaagtgtaaa gcctggggtg cctaatgagt
3961 gagctaactc acattaattg cgttgcgctc actgcccgct ttccagtcgg gaaacctgtc
4021 gtgccagctg cattaatgaa tcggccaacg cgcggggaga ggcggtttgc gtattgggcg
4081 ctcttccgct tcctcgctca ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt
4141 atcagctcac tcaaaggcgg taatacggtt atccacagaa tcaggggata acgcaggaaa
4201 gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc
4261 gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag
4321 gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt
4381 gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg
4441 aagcgtggcg ctttctcata gctcacgctg taggtatctc agttcggtgt aggtcgttcg
4501 ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg
4561 taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac
4621 tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg
4681 gcctaactac ggctacacta gaagaacagt atttggtatc tgcgctctgc tgaagccagt
4741 taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg
4801 tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc
4861 tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt
4921 ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa aatgaagttt
4981 taaatcaatc taaagtatat atgagtaaac ttggtctgac agttaccaat gcttaatcag
5041 tgaggcacct atctcagcga tctgtctatt tcgttcatcc atagttgcct gactccccgt
5101 cgtgtagata actacgatac gggagggctt accatctggc cccagtgctg caatgatacc
5161 gcgagaccca cgctcaccgg ctccagattt atcagcaata aaccagccag ccggaagggc
5221 cgagcgcaga agtggtcctg caactttatc cgcctccatc cagtctatta attgttgccg
5281 ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg ccattgctac
5341 aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg gttcccaacg
5401 atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc
5461 tccgatcgtt gtcagaagta agttggccgc agtgttatca ctcatggtta tggcagcact
5521 gcataattct cttactgtca tgccatccgt aagatgcttt tctgtgactg gtgagtactc
5581 aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat
5641 acgggataat accgcgccac atagcagaac tttaaaagtg ctcatcattg gaaaacgttc
5701 ttcggggcga aaactctcaa ggatcttacc gctgttgaga tccagttcga tgtaacccac
5761 tcgtgcaccc aactgatctt cagcatcttt tactttcacc agcgtttctg ggtgagcaaa
5821 aacaggaagg caaaatgccg caaaaaaggg aataagggcg acacggaaat gttgaatact
5881 catactcttc ctttttcaat attattgaag catttatcag ggttattgtc tcatgagcgg
5941 atacatattt gaatgtattt agaaaaataa acaaataggg gttccgcgca catttccccg
6001 aaaagtgcca cctgacg
//
