| 质粒名称: | pcDNA3 HA Arf6 ActQ67L |
|---|---|
| 出品公司: | 爱迪基因 |
| 目录编号: | 10835 |
| 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 Q67L,Arf6 |
| 物种来源: | H. sapiens (human) |
| 插入大小: | 525 bp |
| 基因突变: | Q67L. Defective in GTP hydrolysis. Constitutively active. |
| 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(未破坏) |
| 3' 克隆位点: | XhoI (未破坏) |
| 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 ActQ67L was a gift from Thomas Roberts (Addgene plasmid # 10835 ; http://n2t.net/addgene:10835 ; RRID:Addgene_10835) |
| 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 ActQ67L 6017 bp ds-DNA circular SYN 06-AUG-2018
DEFINITION synthetic circular DNA
ACCESSION .
VERSION .
KEYWORDS pcDNA3 HA Arf6 ActQ67L
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"
enhancer 76..455
/label=CMV enhancer
/note="human cytomegalovirus immediate early enhancer"
promoter 456..659
/label=CMV promoter
/note="human cytomegalovirus (CMV) immediate early
promoter"
primer_bind 610..630
/label=CMV-F
/note="Human CMV immediate early promoter, forward primer"
primer_bind 704..723
/label=T7
/note="T7 promoter, forward primer"
promoter 704..722
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
CDS 1310..1336
/codon_start=1
/product="HA (human influenza hemagglutinin) epitope tag"
/label=HA
/translation="YPYDVPDYA"
promoter complement(1411..1429)
/label=SP6 promoter
/note="promoter for bacteriophage SP6 RNA polymerase"
primer_bind complement(1412..1429)
/label=SP6
/note="SP6 promoter, forward primer"
primer_bind complement(1449..1466)
/label=BGH-rev
/note="Bovine growth hormone terminator, reverse primer.
Also called BGH reverse"
polyA_signal 1455..1679
/label=bGH poly(A) signal
/note="bovine growth hormone polyadenylation signal"
rep_origin 1725..2153
/direction=RIGHT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
primer_bind complement(1812..1831)
/label=F1ori-R
/note="F1 origin, reverse primer"
primer_bind 2022..2043
/label=F1ori-F
/note="F1 origin, forward primer"
primer_bind complement(2162..2182)
/label=pBABE 3'
/note="SV40 enhancer, reverse primer for pBABE vectors"
promoter 2167..2496
/label=SV40 promoter
/note="SV40 enhancer and early promoter"
rep_origin 2347..2482
/label=SV40 ori
/note="SV40 origin of replication"
primer_bind 2409..2428
/label=SV40pro-F
/note="SV40 promoter/origin, forward primer"
CDS 2563..3357
/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(2617..2636)
/label=Neo-R
/note="Neomycin resistance gene, reverse primer"
primer_bind 3227..3246
/label=Neo-F
/note="Neomycin resistance gene, forward primer"
polyA_signal 3531..3652
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
primer_bind complement(3568..3587)
/label=SV40pA-R
/note="SV40 polyA, reverse primer"
primer_bind 3622..3641
/label=EBV-rev
/note="SV40 polyA terminator, reverse primer"
primer_bind complement(3701..3717)
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(3701..3717)
/label=M13 Reverse
/note="In lacZ gene. Also called M13-rev"
primer_bind complement(3714..3736)
/label=M13/pUC Reverse
/note="In lacZ gene"
protein_bind 3725..3741
/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(3749..3779)
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 3794..3815
/label=CAP binding site
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding activates transcription in the presence
of cAMP."
primer_bind complement(3932..3949)
/label=L4440
/note="L4440 vector, forward primer"
rep_origin complement(4103..4691)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind complement(4183..4202)
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
CDS complement(4862..5722)
/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 5485..5504
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
promoter complement(5723..5827)
/gene="bla"
/label=AmpR promoter
primer_bind complement(5902..5921)
/label=pRS-marker
/note="pRS vectors, use to sequence yeast selectable
marker"
ORIGIN
1 acaattgcat gaagaatctg cttagggtta ggcgttttgc gctgcttcgc gatgtacggg
61 ccagatatac gcgttgacat tgattattga ctagttatta atagtaatca attacggggt
121 cattagttca tagcccatat atggagttcc gcgttacata acttacggta aatggcccgc
181 ctggctgacc gcccaacgac ccccgcccat tgacgtcaat aatgacgtat gttcccatag
241 taacgccaat agggactttc cattgacgtc aatgggtgga gtatttacgg taaactgccc
301 acttggcagt acatcaagtg tatcatatgc caagtacgcc ccctattgac gtcaatgacg
361 gtaaatggcc cgcctggcat tatgcccagt acatgacctt atgggacttt cctacttggc
421 agtacatcta cgtattagtc atcgctatta ccatggtgat gcggttttgg cagtacatca
481 atgggcgtgg atagcggttt gactcacggg gatttccaag tctccacccc attgacgtca
541 atgggagttt gttttggcac caaaatcaac gggactttcc aaaatgtcgt aacaactccg
601 ccccattgac gcaaatgggc ggtaggcgtg tacggtggga ggtctatata agcagagctc
661 tctggctaac tagagaaccc actgcttact ggcttatcga aattaatacg actcactata
721 gggagaccca agcttggtac cgagctcgga tccactagta acggccgcca gtgtgctgga
781 attcatgggg aaggtgctat ccaaaatctt cgggaacaag gaaatgcgga tcctcatgtt
841 gggcctggac gcggccggca agacaacaat cctgtacaag ttgaagctgg gccagtcggt
901 gaccaccatt cccactgtgg gtttcaacgt ggagacggtg acttacaaaa atgtcaagtt
961 caacgtatgg gatgtgggcg gtctagacaa gatccggccg ctctggcggc attactacac
1021 tgggacccaa ggtctcatct tcgtagtgga ctgcgccgac cgcgaccgca tcgatgaggc
1081 tcgccaggag ctgcaccgca ttatcaatga ccgggagatg agggacgcca taatcctcat
1141 cttcgccaac aagcaggacc tgcccgatgc catgaaaccc cacgagatcc aggagaaact
1201 gggcctgacc cggattcggg acaggaactg gtatgtgcag ccctcctgtg ccacctcagg
1261 ggacggactc tatgaggggc tcacatggtt aacctctaac tacaaatctt acccatacga
1321 tgttccagat tacgcttaat agtctagaca gctgagatct gatatccatc acactggcgg
1381 ccgctcgagc atgcatctag agggccctat tctatagtgt cacctaaatg ctagagctcg
1441 ctgatcagcc tcgactgtgc cttctagttg ccagccatct gttgtttgcc cctcccccgt
1501 gccttccttg accctggaag gtgccactcc cactgtcctt tcctaataaa atgaggaaat
1561 tgcatcgcat tgtctgagta ggtgtcattc tattctgggg ggtggggtgg ggcaggacag
1621 caagggggag gattgggaag acaatagcag gcatgctggg gatgcggtgg gctctatggc
1681 ttctgaggcg gaaagaacca gctggggctc tagggggtat ccccacgcgc cctgtagcgg
1741 cgcattaagc gcggcgggtg tggtggttac gcgcagcgtg accgctacac ttgccagcgc
1801 cctagcgccc gctcctttcg ctttcttccc ttcctttctc gccacgttcg ccggctttcc
1861 ccgtcaagct ctaaatcggg ggctcccttt agggttccga tttagtgctt tacggcacct
1921 cgaccccaaa aaacttgatt agggtgatgg ttcacgtagt gggccatcgc cctgatagac
1981 ggtttttcgc cctttgacgt tggagtccac gttctttaat agtggactct tgttccaaac
2041 tggaacaaca ctcaacccta tctcggtcta ttcttttgat ttataaggga ttttgccgat
2101 ttcggcctat tggttaaaaa atgagctgat ttaacaaaaa tttaacgcga attaattctg
2161 tggaatgtgt gtcagttagg gtgtggaaag tccccaggct ccccagcagg cagaagtatg
2221 caaagcatgc atctcaatta gtcagcaacc aggtgtggaa agtccccagg ctccccagca
2281 ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa ccatagtccc gcccctaact
2341 ccgcccatcc cgcccctaac tccgcccagt tccgcccatt ctccgcccca tggctgacta
2401 atttttttta tttatgcaga ggccgaggcc gcctctgcct ctgagctatt ccagaagtag
2461 tgaggaggct tttttggagg cctaggcttt tgcaaaaagc tcccgggagc ttgtatatcc
2521 attttcggat ctgatcaaga gacaggatga ggatcgtttc gcatgattga acaagatgga
2581 ttgcacgcag gttctccggc cgcttgggtg gagaggctat tcggctatga ctgggcacaa
2641 cagacaatcg gctgctctga tgccgccgtg ttccggctgt cagcgcaggg gcgcccggtt
2701 ctttttgtca agaccgacct gtccggtgcc ctgaatgaac tgcaggacga ggcagcgcgg
2761 ctatcgtggc tggccacgac gggcgttcct tgcgcagctg tgctcgacgt tgtcactgaa
2821 gcgggaaggg actggctgct attgggcgaa gtgccggggc aggatctcct gtcatctcac
2881 cttgctcctg ccgagaaagt atccatcatg gctgatgcaa tgcggcggct gcatacgctt
2941 gatccggcta cctgcccatt cgaccaccaa gcgaaacatc gcatcgagcg agcacgtact
3001 cggatggaag ccggtcttgt cgatcaggat gatctggacg aagagcatca ggggctcgcg
3061 ccagccgaac tgttcgccag gctcaaggcg cgcatgcccg acggcgagga tctcgtcgtg
3121 acccatggcg atgcctgctt gccgaatatc atggtggaaa atggccgctt ttctggattc
3181 atcgactgtg gccggctggg tgtggcggac cgctatcagg acatagcgtt ggctacccgt
3241 gatattgctg aagagcttgg cggcgaatgg gctgaccgct tcctcgtgct ttacggtatc
3301 gccgctcccg attcgcagcg catcgccttc tatcgccttc ttgacgagtt cttctgagcg
3361 ggactctggg gttcgaaatg accgaccaag cgacgcccaa cctgccatca cgagatttcg
3421 attccaccgc cgccttctat gaaaggttgg gcttcggaat cgttttccgg gacgccggct
3481 ggatgatcct ccagcgcggg gatctcatgc tggagttctt cgcccacccc aacttgttta
3541 ttgcagctta taatggttac aaataaagca atagcatcac aaatttcaca aataaagcat
3601 ttttttcact gcattctagt tgtggtttgt ccaaactcat caatgtatct tatcatgtct
3661 gtataccgtc gacctctagc tagagcttgg cgtaatcatg gtcatagctg tttcctgtgt
3721 gaaattgtta tccgctcaca attccacaca acatacgagc cggaagcata aagtgtaaag
3781 cctggggtgc ctaatgagtg agctaactca cattaattgc gttgcgctca ctgcccgctt
3841 tccagtcggg aaacctgtcg tgccagctgc attaatgaat cggccaacgc gcggggagag
3901 gcggtttgcg tattgggcgc tcttccgctt cctcgctcac tgactcgctg cgctcggtcg
3961 ttcggctgcg gcgagcggta tcagctcact caaaggcggt aatacggtta tccacagaat
4021 caggggataa cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta
4081 aaaaggccgc gttgctggcg tttttccata ggctccgccc ccctgacgag catcacaaaa
4141 atcgacgctc aagtcagagg tggcgaaacc cgacaggact ataaagatac caggcgtttc
4201 cccctggaag ctccctcgtg cgctctcctg ttccgaccct gccgcttacc ggatacctgt
4261 ccgcctttct cccttcggga agcgtggcgc tttctcatag ctcacgctgt aggtatctca
4321 gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc gttcagcccg
4381 accgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggtaaga cacgacttat
4441 cgccactggc agcagccact ggtaacagga ttagcagagc gaggtatgta ggcggtgcta
4501 cagagttctt gaagtggtgg cctaactacg gctacactag aagaacagta tttggtatct
4561 gcgctctgct gaagccagtt accttcggaa aaagagttgg tagctcttga tccggcaaac
4621 aaaccaccgc tggtagcggt ggtttttttg tttgcaagca gcagattacg cgcagaaaaa
4681 aaggatctca agaagatcct ttgatctttt ctacggggtc tgacgctcag tggaacgaaa
4741 actcacgtta agggattttg gtcatgagat tatcaaaaag gatcttcacc tagatccttt
4801 taaattaaaa atgaagtttt aaatcaatct aaagtatata tgagtaaact tggtctgaca
4861 gttaccaatg cttaatcagt gaggcaccta tctcagcgat ctgtctattt cgttcatcca
4921 tagttgcctg actccccgtc gtgtagataa ctacgatacg ggagggctta ccatctggcc
4981 ccagtgctgc aatgataccg cgagacccac gctcaccggc tccagattta tcagcaataa
5041 accagccagc cggaagggcc gagcgcagaa gtggtcctgc aactttatcc gcctccatcc
5101 agtctattaa ttgttgccgg gaagctagag taagtagttc gccagttaat agtttgcgca
5161 acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat
5221 tcagctccgg ttcccaacga tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag
5281 cggttagctc cttcggtcct ccgatcgttg tcagaagtaa gttggccgca gtgttatcac
5341 tcatggttat ggcagcactg cataattctc ttactgtcat gccatccgta agatgctttt
5401 ctgtgactgg tgagtactca accaagtcat tctgagaata gtgtatgcgg cgaccgagtt
5461 gctcttgccc ggcgtcaata cgggataata ccgcgccaca tagcagaact ttaaaagtgc
5521 tcatcattgg aaaacgttct tcggggcgaa aactctcaag gatcttaccg ctgttgagat
5581 ccagttcgat gtaacccact cgtgcaccca actgatcttc agcatctttt actttcacca
5641 gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga ataagggcga
5701 cacggaaatg ttgaatactc atactcttcc tttttcaata ttattgaagc atttatcagg
5761 gttattgtct catgagcgga tacatatttg aatgtattta gaaaaataaa caaatagggg
5821 ttccgcgcac atttccccga aaagtgccac ctgacgtcga cggatcggga gatctcccga
5881 tcccctatgg tgcactctca gtacaatctg ctctgatgcc gcatagttaa gccagtatct
5941 gctccctgct tgtgtgttgg aggtcgctga gtagtgcgcg agcaaaattt aagctacaac
6001 aaggcaaggc ttgaccg
//
