| 质粒名称: | EGFP-Rab7A T22N |
|---|---|
| 出品公司: | Addgene |
| 目录编号: | 28048 |
| 存储实验室: | Qing Zhong |
|
载体骨架基本信息 |
|
| 载体名称: | pcDNA3 |
| 载体出品公司: | Invitrogen |
| 空载体大小: | 5446bp |
| 完整质粒大小: | 6780 bp |
| 载体类型: | 哺乳细胞表达 |
| 筛选标记: | Gentamicin |
| 载体抗性: | 氨苄霉素 |
| 生长温度: | 37 ℃ |
| 宿主菌: | DH5a |
| 拷贝数: | 高拷贝 |
|
插入基因信息 |
|
| 插入基因名称: | Rab7A |
| 物种来源: | H. sapiens (human) |
| 插入大小: | 623bp |
| 基因突变: | T22N |
| 标签/融合蛋白: |
EGFP
2xHA (N terminal on insert)
|
| 克隆方法: | 限制性内切酶 |
| 5’测序引物 | CMV-F |
| 3’测序引物 | BGH-rev |
|
文献引用方法 |
|
| 材料和方法部分: |
EGFP-Rab7A T22N was a gift from Qing Zhong (Addgene plasmid # 28048 ; http://n2t.net/addgene:28048 ; RRID:Addgene_28048)
|
| 参考文献部分: | Rubicon controls endosome maturation as a Rab7 effector. Sun Q, Westphal W, Wong KN, Tan I, Zhong Q. Proc Natl Acad Sci U S A. 2010 Nov 9. 107(45):19338-43. 10.1073/pnas.1010554107 PubMed 20974968 |
买家导航
EGFP-Rab7A T22N载体质粒基本信息
EGFP-Rab7A T22N质粒图谱载体图谱和EGFP-Rab7A T22N载体序列质粒序列多克隆位点信息
EGFP-Rab7A T22N质粒载体简介
EGFP-Rab7A T22N质粒序列载体序列
LOCUS EGFP-Rab7A T22N 6780 bp ds-DNA circular SYN 12-MAY-2021
DEFINITION synthetic circular DNA
ACCESSION .
VERSION .
KEYWORDS EGFP-Rab7A T22N
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 6780)
AUTHORS Sun Q, Westphal W, Wong KN, Tan I, Zhong Q
TITLE Rubicon controls endosome maturation as a Rab7 effector.
JOURNAL Proc Natl Acad Sci U S A. 2010 Nov 9. 107(45):19338-43.
PUBMED 20974968
REFERENCE 2 (bases 1 to 6780)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58
http://www.biofeng.com
FEATURES Location/Qualifiers
source 1..6780
/organism="synthetic DNA construct"
/mol_type="other DNA"
primer_bind complement(42..61)
/label=pRS-marker
/note="pRS vectors, use to sequence yeast selectable
marker"
enhancer 233..612
/label=CMV enhancer
/note="human cytomegalovirus immediate early enhancer"
promoter 613..816
/label=CMV promoter
/note="human cytomegalovirus (CMV) immediate early
promoter"
primer_bind 767..787
/label=CMV-F
/note="Human CMV immediate early promoter, forward primer"
primer_bind 861..880
/label=T7
/note="T7 promoter, forward primer"
promoter 861..879
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
CDS 893..1609
/codon_start=1
/product="enhanced GFP"
/label=EGFP
/translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
EFVTAAGITLGMDELYK"
CDS 893..1609
/codon_start=1
/product="the original enhanced GFP (Yang et al., 1996)"
/label=EGFP
/note="mammalian codon-optimized"
/translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
EFVTAAGITLGMDELYK"
primer_bind complement(938..959)
/label=EGFP-N
/note="EGFP, reverse primer"
primer_bind complement(1199..1218)
/label=EXFP-R
/note="For distinguishing EGFP variants, reverse primer"
primer_bind 1546..1567
/label=EGFP-C
/note="EGFP, forward primer"
CDS 1619..1645
/codon_start=1
/product="HA (human influenza hemagglutinin) epitope tag"
/label=HA
/translation="YPYDVPDYA"
CDS 1646..1672
/codon_start=1
/product="HA (human influenza hemagglutinin) epitope tag"
/label=HA
/translation="YPYDVPDYA"
promoter complement(2331..2349)
/label=SP6 promoter
/note="promoter for bacteriophage SP6 RNA polymerase"
primer_bind complement(2332..2349)
/label=SP6
/note="SP6 promoter, forward primer"
primer_bind complement(2369..2386)
/label=BGH-rev
/note="Bovine growth hormone terminator, reverse primer.
Also called BGH reverse"
polyA_signal 2375..2599
/label=bGH poly(A) signal
/note="bovine growth hormone polyadenylation signal"
rep_origin 2645..3073
/direction=RIGHT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
primer_bind complement(2732..2751)
/label=F1ori-R
/note="F1 origin, reverse primer"
primer_bind 2942..2963
/label=F1ori-F
/note="F1 origin, forward primer"
primer_bind complement(3082..3102)
/label=pBABE 3'
/note="SV40 enhancer, reverse primer for pBABE vectors"
promoter 3087..3416
/label=SV40 promoter
/note="SV40 enhancer and early promoter"
rep_origin 3267..3402
/label=SV40 ori
/note="SV40 origin of replication"
primer_bind 3329..3348
/label=SV40pro-F
/note="SV40 promoter/origin, forward primer"
CDS 3483..4277
/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(3537..3556)
/label=Neo-R
/note="Neomycin resistance gene, reverse primer"
primer_bind 4147..4166
/label=Neo-F
/note="Neomycin resistance gene, forward primer"
polyA_signal 4451..4572
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
primer_bind complement(4488..4507)
/label=SV40pA-R
/note="SV40 polyA, reverse primer"
primer_bind 4542..4561
/label=EBV-rev
/note="SV40 polyA terminator, reverse primer"
primer_bind complement(4621..4637)
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(4621..4637)
/label=M13 Reverse
/note="In lacZ gene. Also called M13-rev"
primer_bind complement(4634..4656)
/label=M13/pUC Reverse
/note="In lacZ gene"
protein_bind 4645..4661
/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(4669..4699)
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 4714..4735
/label=CAP binding site
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding activates transcription in the presence
of cAMP."
primer_bind complement(4852..4869)
/label=L4440
/note="L4440 vector, forward primer"
rep_origin complement(5023..5611)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind complement(5103..5122)
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
CDS complement(5782..6642)
/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 6405..6424
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
promoter complement(6643..6747)
/gene="bla"
/label=AmpR promoter
ORIGIN
1 cggatcggga gatctcccga tcccctatgg tgcactctca gtacaatctg ctctgatgcc
61 gcatagttaa gccagtatct gctccctgct tgtgtgttgg aggtcgctga gtagtgcgcg
121 agcaaaattt aagctacaac aaggcaaggc ttgaccgaca attgcatgaa gaatctgctt
181 agggttaggc gttttgcgct gcttcgcgat gtacgggcca gatatacgcg ttgacattga
241 ttattgacta gttattaata gtaatcaatt acggggtcat tagttcatag cccatatatg
301 gagttccgcg ttacataact tacggtaaat ggcccgcctg gctgaccgcc caacgacccc
361 cgcccattga cgtcaataat gacgtatgtt cccatagtaa cgccaatagg gactttccat
421 tgacgtcaat gggtggagta tttacggtaa actgcccact tggcagtaca tcaagtgtat
481 catatgccaa gtacgccccc tattgacgtc aatgacggta aatggcccgc ctggcattat
541 gcccagtaca tgaccttatg ggactttcct acttggcagt acatctacgt attagtcatc
601 gctattacca tggtgatgcg gttttggcag tacatcaatg ggcgtggata gcggtttgac
661 tcacggggat ttccaagtct ccaccccatt gacgtcaatg ggagtttgtt ttggcaccaa
721 aatcaacggg actttccaaa atgtcgtaac aactccgccc cattgacgca aatgggcggt
781 aggcgtgtac ggtgggaggt ctatataagc agagctctct ggctaactag agaacccact
841 gcttactggc ttatcgaaat taatacgact cactataggg agacccaagc ttatggtgag
901 caagggcgag gagctgttca ccggggtggt gcccatcctg gtcgagctgg acggcgacgt
961 aaacggccac aagttcagcg tgtccggcga gggcgagggc gatgccacct acggcaagct
1021 gaccctgaag ttcatctgca ccaccggcaa gctgcccgtg ccctggccca ccctcgtgac
1081 caccctgacc tacggcgtgc agtgcttcag ccgctacccc gaccacatga agcagcacga
1141 cttcttcaag tccgccatgc ccgaaggcta cgtccaggag cgcaccatct tcttcaagga
1201 cgacggcaac tacaagaccc gcgccgaggt gaagttcgag ggcgacaccc tggtgaaccg
1261 catcgagctg aagggcatcg acttcaagga ggacggcaac atcctggggc acaagctgga
1321 gtacaactac aacagccaca acgtctatat catggccgac aagcagaaga acggcatcaa
1381 ggtgaacttc aagatccgcc acaacatcga ggacggcagc gtgcagctcg ccgaccacta
1441 ccagcagaac acccccatcg gcgacggccc cgtgctgctg cccgacaacc actacctgag
1501 cacccagtcc gccctgagca aagaccccaa cgagaagcgc gatcacatgg tcctgctgga
1561 gttcgtgacc gccgccggga tcactctcgg catggacgag ctgtacaagg gatccatgta
1621 cccatacgat gttccggatt acgcttaccc atacgatgtt ccggattacg ctggtacctc
1681 taggaagaaa gtgttgctga aggttatcat cctgggagat tctggagtcg ggaagaactc
1741 actcatgaac cagtatgtga ataagaaatt cagcaatcag tacaaagcca caataggagc
1801 tgactttctg accaaggagg tgatggtgga tgacaggcta gtcacaatgc agatatggga
1861 cacagcagga caggaacggt tccagtctct cggtgtggcc ttctacagag gtgcagactg
1921 ctgcgttctg gtatttgatg tgactgcccc caacacattc aaaaccctag atagctggag
1981 agatgagttt ctcatccagg ccagtccccg agatcctgaa aacttcccat ttgttgtgtt
2041 gggaaacaag attgacctcg aaaacagaca agtggccaca aagcgggcac aggcctggtg
2101 ctacagcaaa aacaacattc cctactttga gaccagtgcc aaggaggcca tcaacgtgga
2161 gcaggcgttc cagacgattg cacggaatgc acttaagcag gaaacggagg tggagctgta
2221 caacgaattt cctgaaccta tcaaactgga caagaatgac cgggccaagg cctcggcaga
2281 aagctgcagt tgctaagcgg ccgctcgagc atgcatctag agggccctat tctatagtgt
2341 cacctaaatg ctagagctcg ctgatcagcc tcgactgtgc cttctagttg ccagccatct
2401 gttgtttgcc cctcccccgt gccttccttg accctggaag gtgccactcc cactgtcctt
2461 tcctaataaa atgaggaaat tgcatcgcat tgtctgagta ggtgtcattc tattctgggg
2521 ggtggggtgg ggcaggacag caagggggag gattgggaag acaatagcag gcatgctggg
2581 gatgcggtgg gctctatggc ttctgaggcg gaaagaacca gctggggctc tagggggtat
2641 ccccacgcgc cctgtagcgg cgcattaagc gcggcgggtg tggtggttac gcgcagcgtg
2701 accgctacac ttgccagcgc cctagcgccc gctcctttcg ctttcttccc ttcctttctc
2761 gccacgttcg ccggctttcc ccgtcaagct ctaaatcggg ggctcccttt agggttccga
2821 tttagtgctt tacggcacct cgaccccaaa aaacttgatt agggtgatgg ttcacgtagt
2881 gggccatcgc cctgatagac ggtttttcgc cctttgacgt tggagtccac gttctttaat
2941 agtggactct tgttccaaac tggaacaaca ctcaacccta tctcggtcta ttcttttgat
3001 ttataaggga ttttgccgat ttcggcctat tggttaaaaa atgagctgat ttaacaaaaa
3061 tttaacgcga attaattctg tggaatgtgt gtcagttagg gtgtggaaag tccccaggct
3121 ccccagcagg cagaagtatg caaagcatgc atctcaatta gtcagcaacc aggtgtggaa
3181 agtccccagg ctccccagca ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa
3241 ccatagtccc gcccctaact ccgcccatcc cgcccctaac tccgcccagt tccgcccatt
3301 ctccgcccca tggctgacta atttttttta tttatgcaga ggccgaggcc gcctctgcct
3361 ctgagctatt ccagaagtag tgaggaggct tttttggagg cctaggcttt tgcaaaaagc
3421 tcccgggagc ttgtatatcc attttcggat ctgatcaaga gacaggatga ggatcgtttc
3481 gcatgattga acaagatgga ttgcacgcag gttctccggc cgcttgggtg gagaggctat
3541 tcggctatga ctgggcacaa cagacaatcg gctgctctga tgccgccgtg ttccggctgt
3601 cagcgcaggg gcgcccggtt ctttttgtca agaccgacct gtccggtgcc ctgaatgaac
3661 tgcaggacga ggcagcgcgg ctatcgtggc tggccacgac gggcgttcct tgcgcagctg
3721 tgctcgacgt tgtcactgaa gcgggaaggg actggctgct attgggcgaa gtgccggggc
3781 aggatctcct gtcatctcac cttgctcctg ccgagaaagt atccatcatg gctgatgcaa
3841 tgcggcggct gcatacgctt gatccggcta cctgcccatt cgaccaccaa gcgaaacatc
3901 gcatcgagcg agcacgtact cggatggaag ccggtcttgt cgatcaggat gatctggacg
3961 aagagcatca ggggctcgcg ccagccgaac tgttcgccag gctcaaggcg cgcatgcccg
4021 acggcgagga tctcgtcgtg acccatggcg atgcctgctt gccgaatatc atggtggaaa
4081 atggccgctt ttctggattc atcgactgtg gccggctggg tgtggcggac cgctatcagg
4141 acatagcgtt ggctacccgt gatattgctg aagagcttgg cggcgaatgg gctgaccgct
4201 tcctcgtgct ttacggtatc gccgctcccg attcgcagcg catcgccttc tatcgccttc
4261 ttgacgagtt cttctgagcg ggactctggg gttcgaaatg accgaccaag cgacgcccaa
4321 cctgccatca cgagatttcg attccaccgc cgccttctat gaaaggttgg gcttcggaat
4381 cgttttccgg gacgccggct ggatgatcct ccagcgcggg gatctcatgc tggagttctt
4441 cgcccacccc aacttgttta ttgcagctta taatggttac aaataaagca atagcatcac
4501 aaatttcaca aataaagcat ttttttcact gcattctagt tgtggtttgt ccaaactcat
4561 caatgtatct tatcatgtct gtataccgtc gacctctagc tagagcttgg cgtaatcatg
4621 gtcatagctg tttcctgtgt gaaattgtta tccgctcaca attccacaca acatacgagc
4681 cggaagcata aagtgtaaag cctggggtgc ctaatgagtg agctaactca cattaattgc
4741 gttgcgctca ctgcccgctt tccagtcggg aaacctgtcg tgccagctgc attaatgaat
4801 cggccaacgc gcggggagag gcggtttgcg tattgggcgc tcttccgctt cctcgctcac
4861 tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta tcagctcact caaaggcggt
4921 aatacggtta tccacagaat caggggataa cgcaggaaag aacatgtgag caaaaggcca
4981 gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc
5041 ccctgacgag catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact
5101 ataaagatac caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct
5161 gccgcttacc ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcatag
5221 ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca
5281 cgaacccccc gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa
5341 cccggtaaga cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc
5401 gaggtatgta ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag
5461 aagaacagta tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg
5521 tagctcttga tccggcaaac aaaccaccgc tggtagcggt ggtttttttg tttgcaagca
5581 gcagattacg cgcagaaaaa aaggatctca agaagatcct ttgatctttt ctacggggtc
5641 tgacgctcag tggaacgaaa actcacgtta agggattttg gtcatgagat tatcaaaaag
5701 gatcttcacc tagatccttt taaattaaaa atgaagtttt aaatcaatct aaagtatata
5761 tgagtaaact tggtctgaca gttaccaatg cttaatcagt gaggcaccta tctcagcgat
5821 ctgtctattt cgttcatcca tagttgcctg actccccgtc gtgtagataa ctacgatacg
5881 ggagggctta ccatctggcc ccagtgctgc aatgataccg cgagacccac gctcaccggc
5941 tccagattta tcagcaataa accagccagc cggaagggcc gagcgcagaa gtggtcctgc
6001 aactttatcc gcctccatcc agtctattaa ttgttgccgg gaagctagag taagtagttc
6061 gccagttaat agtttgcgca acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc
6121 gtcgtttggt atggcttcat tcagctccgg ttcccaacga tcaaggcgag ttacatgatc
6181 ccccatgttg tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg tcagaagtaa
6241 gttggccgca gtgttatcac tcatggttat ggcagcactg cataattctc ttactgtcat
6301 gccatccgta agatgctttt ctgtgactgg tgagtactca accaagtcat tctgagaata
6361 gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata cgggataata ccgcgccaca
6421 tagcagaact ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa aactctcaag
6481 gatcttaccg ctgttgagat ccagttcgat gtaacccact cgtgcaccca actgatcttc
6541 agcatctttt actttcacca gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc
6601 aaaaaaggga ataagggcga cacggaaatg ttgaatactc atactcttcc tttttcaata
6661 ttattgaagc atttatcagg gttattgtct catgagcgga tacatatttg aatgtattta
6721 gaaaaataaa caaatagggg ttccgcgcac atttccccga aaagtgccac ctgacgtcga
//
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EGFP-Rab7A T22N质粒载体应用举例
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