| 质粒名称: | pcDNA3 HA Arf6 DN T27N |
|---|---|
| 出品公司: | 爱迪基因 |
| 目录编号: | 10831 |
| 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 T27N,Arf6 |
| 物种来源: | H. sapiens (human) |
| 插入大小: | 525 bp |
| 基因突变: | T27N. Defective in GTP binding. Dominant negative. |
| 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 DN T27N was a gift from Thomas Roberts (Addgene plasmid # 10831 ; http://n2t.net/addgene:10831 ; RRID:Addgene_10831) |
| 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 DN T27N 6017 bp ds-DNA circular SYN 06-AUG-2018
DEFINITION synthetic circular DNA
ACCESSION .
VERSION .
KEYWORDS pcDNA3 HA Arf6 DN T27N
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 90..469
/label=CMV enhancer
/note="human cytomegalovirus immediate early enhancer"
promoter 470..673
/label=CMV promoter
/note="human cytomegalovirus (CMV) immediate early
promoter"
primer_bind 624..644
/label=CMV-F
/note="Human CMV immediate early promoter, forward primer"
primer_bind 718..737
/label=T7
/note="T7 promoter, forward primer"
promoter 718..736
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
CDS 1324..1350
/codon_start=1
/product="HA (human influenza hemagglutinin) epitope tag"
/label=HA
/translation="YPYDVPDYA"
promoter complement(1425..1443)
/label=SP6 promoter
/note="promoter for bacteriophage SP6 RNA polymerase"
primer_bind complement(1426..1443)
/label=SP6
/note="SP6 promoter, forward primer"
primer_bind complement(1463..1480)
/label=BGH-rev
/note="Bovine growth hormone terminator, reverse primer.
Also called BGH reverse"
polyA_signal 1469..1693
/label=bGH poly(A) signal
/note="bovine growth hormone polyadenylation signal"
rep_origin 1739..2167
/direction=RIGHT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
primer_bind complement(1826..1845)
/label=F1ori-R
/note="F1 origin, reverse primer"
primer_bind 2036..2057
/label=F1ori-F
/note="F1 origin, forward primer"
primer_bind complement(2176..2196)
/label=pBABE 3'
/note="SV40 enhancer, reverse primer for pBABE vectors"
promoter 2181..2510
/label=SV40 promoter
/note="SV40 enhancer and early promoter"
rep_origin 2361..2496
/label=SV40 ori
/note="SV40 origin of replication"
primer_bind 2423..2442
/label=SV40pro-F
/note="SV40 promoter/origin, forward primer"
CDS 2577..3371
/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(2631..2650)
/label=Neo-R
/note="Neomycin resistance gene, reverse primer"
primer_bind 3241..3260
/label=Neo-F
/note="Neomycin resistance gene, forward primer"
polyA_signal 3545..3666
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
primer_bind complement(3582..3601)
/label=SV40pA-R
/note="SV40 polyA, reverse primer"
primer_bind 3636..3655
/label=EBV-rev
/note="SV40 polyA terminator, reverse primer"
primer_bind complement(3715..3731)
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(3715..3731)
/label=M13 Reverse
/note="In lacZ gene. Also called M13-rev"
primer_bind complement(3728..3750)
/label=M13/pUC Reverse
/note="In lacZ gene"
protein_bind 3739..3755
/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(3763..3793)
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 3808..3829
/label=CAP binding site
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding activates transcription in the presence
of cAMP."
primer_bind complement(3946..3963)
/label=L4440
/note="L4440 vector, forward primer"
rep_origin complement(4117..4705)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind complement(4197..4216)
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
CDS complement(4876..5736)
/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 5499..5518
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
promoter complement(5737..5841)
/gene="bla"
/label=AmpR promoter
primer_bind complement(5916..5935)
/label=pRS-marker
/note="pRS vectors, use to sequence yeast selectable
marker"
ORIGIN
1 gcaaggcttg accgacaatt gcatgaagaa tctgcttagg gttaggcgtt ttgcgctgct
61 tcgcgatgta cgggccagat atacgcgttg acattgatta ttgactagtt attaatagta
121 atcaattacg gggtcattag ttcatagccc atatatggag ttccgcgtta cataacttac
181 ggtaaatggc ccgcctggct gaccgcccaa cgacccccgc ccattgacgt caataatgac
241 gtatgttccc atagtaacgc caatagggac tttccattga cgtcaatggg tggagtattt
301 acggtaaact gcccacttgg cagtacatca agtgtatcat atgccaagta cgccccctat
361 tgacgtcaat gacggtaaat ggcccgcctg gcattatgcc cagtacatga ccttatggga
421 ctttcctact tggcagtaca tctacgtatt agtcatcgct attaccatgg tgatgcggtt
481 ttggcagtac atcaatgggc gtggatagcg gtttgactca cggggatttc caagtctcca
541 ccccattgac gtcaatggga gtttgttttg gcaccaaaat caacgggact ttccaaaatg
601 tcgtaacaac tccgccccat tgacgcaaat gggcggtagg cgtgtacggt gggaggtcta
661 tataagcaga gctctctggc taactagaga acccactgct tactggctta tcgaaattaa
721 tacgactcac tatagggaga cccaagcttg gtaccgagct cggatccact agtaacggcc
781 gccagtgtgc tggaattcat ggggaaggtg ctatccaaaa tcttcgggaa caaggaaatg
841 cggatcctca tgttgggcct ggacgcggcc ggcaagaaca caatcctgta caaattgaag
901 cttggccagt cggtgaccac cattcccact gtgggtttca acgtggagac ggtgacttac
961 aaaaatgtca agttcaacgt atgggatgtg ggcggccagg acaagatccg gccgctctgg
1021 cggcattact acactgggac ccaaggtctc atcttcgtag tggactgcgc cgaccgcgac
1081 cgcatcgatg aggctcgcca ggagctgcac cgcattatca atgaccggga gatgagggac
1141 gccataatcc tcatcttcgc caacaagcag gacctgcccg atgccatgaa accccacgag
1201 atccaggaga aactgggcct gacccggatt cgggacagga actggtatgt gcagccctcc
1261 tgtgccacct caggggacgg actctatgag gggctcacat ggttaacctc taactacaaa
1321 tcttacccat acgatgttcc agattacgct taatagtcta gacagctgag atctgatatc
1381 catcacactg gcggccgctc gagcatgcat ctagagggcc ctattctata gtgtcaccta
1441 aatgctagag ctcgctgatc agcctcgact gtgccttcta gttgccagcc atctgttgtt
1501 tgcccctccc ccgtgccttc cttgaccctg gaaggtgcca ctcccactgt cctttcctaa
1561 taaaatgagg aaattgcatc gcattgtctg agtaggtgtc attctattct ggggggtggg
1621 gtggggcagg acagcaaggg ggaggattgg gaagacaata gcaggcatgc tggggatgcg
1681 gtgggctcta tggcttctga ggcggaaaga accagctggg gctctagggg gtatccccac
1741 gcgccctgta gcggcgcatt aagcgcggcg ggtgtggtgg ttacgcgcag cgtgaccgct
1801 acacttgcca gcgccctagc gcccgctcct ttcgctttct tcccttcctt tctcgccacg
1861 ttcgccggct ttccccgtca agctctaaat cgggggctcc ctttagggtt ccgatttagt
1921 gctttacggc acctcgaccc caaaaaactt gattagggtg atggttcacg tagtgggcca
1981 tcgccctgat agacggtttt tcgccctttg acgttggagt ccacgttctt taatagtgga
2041 ctcttgttcc aaactggaac aacactcaac cctatctcgg tctattcttt tgatttataa
2101 gggattttgc cgatttcggc ctattggtta aaaaatgagc tgatttaaca aaaatttaac
2161 gcgaattaat tctgtggaat gtgtgtcagt tagggtgtgg aaagtcccca ggctccccag
2221 caggcagaag tatgcaaagc atgcatctca attagtcagc aaccaggtgt ggaaagtccc
2281 caggctcccc agcaggcaga agtatgcaaa gcatgcatct caattagtca gcaaccatag
2341 tcccgcccct aactccgccc atcccgcccc taactccgcc cagttccgcc cattctccgc
2401 cccatggctg actaattttt tttatttatg cagaggccga ggccgcctct gcctctgagc
2461 tattccagaa gtagtgagga ggcttttttg gaggcctagg cttttgcaaa aagctcccgg
2521 gagcttgtat atccattttc ggatctgatc aagagacagg atgaggatcg tttcgcatga
2581 ttgaacaaga tggattgcac gcaggttctc cggccgcttg ggtggagagg ctattcggct
2641 atgactgggc acaacagaca atcggctgct ctgatgccgc cgtgttccgg ctgtcagcgc
2701 aggggcgccc ggttcttttt gtcaagaccg acctgtccgg tgccctgaat gaactgcagg
2761 acgaggcagc gcggctatcg tggctggcca cgacgggcgt tccttgcgca gctgtgctcg
2821 acgttgtcac tgaagcggga agggactggc tgctattggg cgaagtgccg gggcaggatc
2881 tcctgtcatc tcaccttgct cctgccgaga aagtatccat catggctgat gcaatgcggc
2941 ggctgcatac gcttgatccg gctacctgcc cattcgacca ccaagcgaaa catcgcatcg
3001 agcgagcacg tactcggatg gaagccggtc ttgtcgatca ggatgatctg gacgaagagc
3061 atcaggggct cgcgccagcc gaactgttcg ccaggctcaa ggcgcgcatg cccgacggcg
3121 aggatctcgt cgtgacccat ggcgatgcct gcttgccgaa tatcatggtg gaaaatggcc
3181 gcttttctgg attcatcgac tgtggccggc tgggtgtggc ggaccgctat caggacatag
3241 cgttggctac ccgtgatatt gctgaagagc ttggcggcga atgggctgac cgcttcctcg
3301 tgctttacgg tatcgccgct cccgattcgc agcgcatcgc cttctatcgc cttcttgacg
3361 agttcttctg agcgggactc tggggttcga aatgaccgac caagcgacgc ccaacctgcc
3421 atcacgagat ttcgattcca ccgccgcctt ctatgaaagg ttgggcttcg gaatcgtttt
3481 ccgggacgcc ggctggatga tcctccagcg cggggatctc atgctggagt tcttcgccca
3541 ccccaacttg tttattgcag cttataatgg ttacaaataa agcaatagca tcacaaattt
3601 cacaaataaa gcattttttt cactgcattc tagttgtggt ttgtccaaac tcatcaatgt
3661 atcttatcat gtctgtatac cgtcgacctc tagctagagc ttggcgtaat catggtcata
3721 gctgtttcct gtgtgaaatt gttatccgct cacaattcca cacaacatac gagccggaag
3781 cataaagtgt aaagcctggg gtgcctaatg agtgagctaa ctcacattaa ttgcgttgcg
3841 ctcactgccc gctttccagt cgggaaacct gtcgtgccag ctgcattaat gaatcggcca
3901 acgcgcgggg agaggcggtt tgcgtattgg gcgctcttcc gcttcctcgc tcactgactc
3961 gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg cggtaatacg
4021 gttatccaca gaatcagggg ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa
4081 ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc gcccccctga
4141 cgagcatcac aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag
4201 ataccaggcg tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct
4261 taccggatac ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc atagctcacg
4321 ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc
4381 ccccgttcag cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt
4441 aagacacgac ttatcgccac tggcagcagc cactggtaac aggattagca gagcgaggta
4501 tgtaggcggt gctacagagt tcttgaagtg gtggcctaac tacggctaca ctagaagaac
4561 agtatttggt atctgcgctc tgctgaagcc agttaccttc ggaaaaagag ttggtagctc
4621 ttgatccggc aaacaaacca ccgctggtag cggtggtttt tttgtttgca agcagcagat
4681 tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc
4741 tcagtggaac gaaaactcac gttaagggat tttggtcatg agattatcaa aaaggatctt
4801 cacctagatc cttttaaatt aaaaatgaag ttttaaatca atctaaagta tatatgagta
4861 aacttggtct gacagttacc aatgcttaat cagtgaggca cctatctcag cgatctgtct
4921 atttcgttca tccatagttg cctgactccc cgtcgtgtag ataactacga tacgggaggg
4981 cttaccatct ggccccagtg ctgcaatgat accgcgagac ccacgctcac cggctccaga
5041 tttatcagca ataaaccagc cagccggaag ggccgagcgc agaagtggtc ctgcaacttt
5101 atccgcctcc atccagtcta ttaattgttg ccgggaagct agagtaagta gttcgccagt
5161 taatagtttg cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt
5221 tggtatggct tcattcagct ccggttccca acgatcaagg cgagttacat gatcccccat
5281 gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc gttgtcagaa gtaagttggc
5341 cgcagtgtta tcactcatgg ttatggcagc actgcataat tctcttactg tcatgccatc
5401 cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag tcattctgag aatagtgtat
5461 gcggcgaccg agttgctctt gcccggcgtc aatacgggat aataccgcgc cacatagcag
5521 aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct caaggatctt
5581 accgctgttg agatccagtt cgatgtaacc cactcgtgca cccaactgat cttcagcatc
5641 ttttactttc accagcgttt ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa
5701 gggaataagg gcgacacgga aatgttgaat actcatactc ttcctttttc aatattattg
5761 aagcatttat cagggttatt gtctcatgag cggatacata tttgaatgta tttagaaaaa
5821 taaacaaata ggggttccgc gcacatttcc ccgaaaagtg ccacctgacg tcgacggatc
5881 gggagatctc ccgatcccct atggtgcact ctcagtacaa tctgctctga tgccgcatag
5941 ttaagccagt atctgctccc tgcttgtgtg ttggaggtcg ctgagtagtg cgcgagcaaa
6001 atttaagcta caacaag
//
