| 质粒名称: | EGFP-Rab5A Q79L |
|---|---|
| 出品公司: | 爱迪基因 |
| 目录编号: | 28046 |
| 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 |
| 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: | Rab5A |
| 物种来源: | H. sapiens (human) |
| 插入大小: | 647 bp |
| 基因突变: | Q79L |
| GenBank编号 | -- |
| 启动子 | -- |
| 标签/融合蛋白: | EGFP on N-term |
| 克隆方法: | 限制性内切酶 |
| 5' 克隆位点: | -- |
| 3' 克隆位点: | -- |
| 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-Rab5A Q79L was a gift from Qing Zhong (Addgene plasmid # 28046 ; http://n2t.net/addgene:28046 ; RRID:Addgene_28046) |
| 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
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 EGFP-Rab5A Q79L 6748 bp ds-DNA circular SYN 06-AUG-2018
DEFINITION synthetic circular DNA
ACCESSION .
VERSION .
KEYWORDS EGFP-Rab5A Q79L
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 6748)
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 6748)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Aug 6, 2018 from SnapGene Server 1.1.58
http://www.snapgene.com
FEATURES Location/Qualifiers
source 1..6748
/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 897..1613
/codon_start=1
/product="enhanced GFP"
/label=EGFP
/note="mammalian codon-optimized"
/translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
EFVTAAGITLGMDELYK"
primer_bind complement(942..963)
/label=EGFP-N
/note="EGFP, reverse primer"
primer_bind complement(1203..1222)
/label=EXFP-R
/note="For distinguishing EGFP variants, reverse primer"
primer_bind 1550..1571
/label=EGFP-C
/note="EGFP, forward primer"
promoter complement(2303..2321)
/label=SP6 promoter
/note="promoter for bacteriophage SP6 RNA polymerase"
primer_bind complement(2304..2321)
/label=SP6
/note="SP6 promoter, forward primer"
primer_bind complement(2341..2358)
/label=BGH-rev
/note="Bovine growth hormone terminator, reverse primer.
Also called BGH reverse"
polyA_signal 2347..2571
/label=bGH poly(A) signal
/note="bovine growth hormone polyadenylation signal"
rep_origin 2617..3045
/direction=RIGHT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
primer_bind complement(2704..2723)
/label=F1ori-R
/note="F1 origin, reverse primer"
primer_bind 2914..2935
/label=F1ori-F
/note="F1 origin, forward primer"
primer_bind complement(3054..3074)
/label=pBABE 3'
/note="SV40 enhancer, reverse primer for pBABE vectors"
promoter 3059..3388
/label=SV40 promoter
/note="SV40 enhancer and early promoter"
rep_origin 3239..3374
/label=SV40 ori
/note="SV40 origin of replication"
primer_bind 3301..3320
/label=SV40pro-F
/note="SV40 promoter/origin, forward primer"
CDS 3455..4249
/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(3509..3528)
/label=Neo-R
/note="Neomycin resistance gene, reverse primer"
primer_bind 4119..4138
/label=Neo-F
/note="Neomycin resistance gene, forward primer"
polyA_signal 4423..4544
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
primer_bind complement(4460..4479)
/label=SV40pA-R
/note="SV40 polyA, reverse primer"
primer_bind 4514..4533
/label=EBV-rev
/note="SV40 polyA terminator, reverse primer"
primer_bind complement(4593..4609)
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(4593..4609)
/label=M13 Reverse
/note="In lacZ gene. Also called M13-rev"
primer_bind complement(4606..4628)
/label=M13/pUC Reverse
/note="In lacZ gene"
protein_bind 4617..4633
/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(4641..4671)
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 4686..4707
/label=CAP binding site
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding activates transcription in the presence
of cAMP."
primer_bind complement(4824..4841)
/label=L4440
/note="L4440 vector, forward primer"
rep_origin complement(4995..5583)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind complement(5075..5094)
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
CDS complement(5754..6614)
/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 6377..6396
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
promoter complement(6615..6719)
/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 aagcttatgg
901 tgagcaaggg cgaggagctg ttcaccgggg tggtgcccat cctggtcgag ctggacggcg
961 acgtaaacgg ccacaagttc agcgtgtccg gcgagggcga gggcgatgcc acctacggca
1021 agctgaccct gaagttcatc tgcaccaccg gcaagctgcc cgtgccctgg cccaccctcg
1081 tgaccaccct gacctacggc gtgcagtgct tcagccgcta ccccgaccac atgaagcagc
1141 acgacttctt caagtccgcc atgcccgaag gctacgtcca ggagcgcacc atcttcttca
1201 aggacgacgg caactacaag acccgcgccg aggtgaagtt cgagggcgac accctggtga
1261 accgcatcga gctgaagggc atcgacttca aggaggacgg caacatcctg gggcacaagc
1321 tggagtacaa ctacaacagc cacaacgtct atatcatggc cgacaagcag aagaacggca
1381 tcaaggtgaa cttcaagatc cgccacaaca tcgaggacgg cagcgtgcag ctcgccgacc
1441 actaccagca gaacaccccc atcggcgacg gccccgtgct gctgcccgac aaccactacc
1501 tgagcaccca gtccgccctg agcaaagacc ccaacgagaa gcgcgatcac atggtcctgc
1561 tggagttcgt gaccgccgcc gggatcactc tcggcatgga cgagctgtac aagggatcca
1621 tggctagtcg aggcgcaaca agacccaacg ggccaaatac tggaaataaa atatgccagt
1681 tcaaactagt acttctggga gagtccgctg ttggcaaatc aagcctagtg cttcgttttg
1741 tgaaaggcca atttcatgaa tttcaagaga gtaccattgg ggctgctttt ctaacccaaa
1801 ctgtatgtct tgatgacact acagtaaagt ttgaaatatg ggatacagct ggtctagaac
1861 gataccatag cctagcacca atgtactaca gaggagcaca agcagccata gttgtatatg
1921 atatcacaaa tgaggagtcc tttgcaagag caaaaaattg ggttaaagaa cttcagaggc
1981 aagcaagtcc taacattgta atagctttat cgggaaacaa ggccgaccta gcaaataaaa
2041 gagcagtaga tttccaggaa gcacagtcct atgcagatga caatagttta ttattcatgg
2101 agacatccgc taaaacatca atgaatgtaa atgaaatatt catggcaata gctaaaaaat
2161 tgccaaagaa tgaaccacaa aatccaggag caaattctgc cagaggaaga ggagtagacc
2221 ttaccgaacc cacacaacca accaggaatc agtgttgtag taactgaggc ggccgctcga
2281 gcatgcatct agagggccct attctatagt gtcacctaaa tgctagagct cgctgatcag
2341 cctcgactgt gccttctagt tgccagccat ctgttgtttg cccctccccc gtgccttcct
2401 tgaccctgga aggtgccact cccactgtcc tttcctaata aaatgaggaa attgcatcgc
2461 attgtctgag taggtgtcat tctattctgg ggggtggggt ggggcaggac agcaaggggg
2521 aggattggga agacaatagc aggcatgctg gggatgcggt gggctctatg gcttctgagg
2581 cggaaagaac cagctggggc tctagggggt atccccacgc gccctgtagc ggcgcattaa
2641 gcgcggcggg tgtggtggtt acgcgcagcg tgaccgctac acttgccagc gccctagcgc
2701 ccgctccttt cgctttcttc ccttcctttc tcgccacgtt cgccggcttt ccccgtcaag
2761 ctctaaatcg ggggctccct ttagggttcc gatttagtgc tttacggcac ctcgacccca
2821 aaaaacttga ttagggtgat ggttcacgta gtgggccatc gccctgatag acggtttttc
2881 gccctttgac gttggagtcc acgttcttta atagtggact cttgttccaa actggaacaa
2941 cactcaaccc tatctcggtc tattcttttg atttataagg gattttgccg atttcggcct
3001 attggttaaa aaatgagctg atttaacaaa aatttaacgc gaattaattc tgtggaatgt
3061 gtgtcagtta gggtgtggaa agtccccagg ctccccagca ggcagaagta tgcaaagcat
3121 gcatctcaat tagtcagcaa ccaggtgtgg aaagtcccca ggctccccag caggcagaag
3181 tatgcaaagc atgcatctca attagtcagc aaccatagtc ccgcccctaa ctccgcccat
3241 cccgccccta actccgccca gttccgccca ttctccgccc catggctgac taattttttt
3301 tatttatgca gaggccgagg ccgcctctgc ctctgagcta ttccagaagt agtgaggagg
3361 cttttttgga ggcctaggct tttgcaaaaa gctcccggga gcttgtatat ccattttcgg
3421 atctgatcaa gagacaggat gaggatcgtt tcgcatgatt gaacaagatg gattgcacgc
3481 aggttctccg gccgcttggg tggagaggct attcggctat gactgggcac aacagacaat
3541 cggctgctct gatgccgccg tgttccggct gtcagcgcag gggcgcccgg ttctttttgt
3601 caagaccgac ctgtccggtg ccctgaatga actgcaggac gaggcagcgc ggctatcgtg
3661 gctggccacg acgggcgttc cttgcgcagc tgtgctcgac gttgtcactg aagcgggaag
3721 ggactggctg ctattgggcg aagtgccggg gcaggatctc ctgtcatctc accttgctcc
3781 tgccgagaaa gtatccatca tggctgatgc aatgcggcgg ctgcatacgc ttgatccggc
3841 tacctgccca ttcgaccacc aagcgaaaca tcgcatcgag cgagcacgta ctcggatgga
3901 agccggtctt gtcgatcagg atgatctgga cgaagagcat caggggctcg cgccagccga
3961 actgttcgcc aggctcaagg cgcgcatgcc cgacggcgag gatctcgtcg tgacccatgg
4021 cgatgcctgc ttgccgaata tcatggtgga aaatggccgc ttttctggat tcatcgactg
4081 tggccggctg ggtgtggcgg accgctatca ggacatagcg ttggctaccc gtgatattgc
4141 tgaagagctt ggcggcgaat gggctgaccg cttcctcgtg ctttacggta tcgccgctcc
4201 cgattcgcag cgcatcgcct tctatcgcct tcttgacgag ttcttctgag cgggactctg
4261 gggttcgaaa tgaccgacca agcgacgccc aacctgccat cacgagattt cgattccacc
4321 gccgccttct atgaaaggtt gggcttcgga atcgttttcc gggacgccgg ctggatgatc
4381 ctccagcgcg gggatctcat gctggagttc ttcgcccacc ccaacttgtt tattgcagct
4441 tataatggtt acaaataaag caatagcatc acaaatttca caaataaagc atttttttca
4501 ctgcattcta gttgtggttt gtccaaactc atcaatgtat cttatcatgt ctgtataccg
4561 tcgacctcta gctagagctt ggcgtaatca tggtcatagc tgtttcctgt gtgaaattgt
4621 tatccgctca caattccaca caacatacga gccggaagca taaagtgtaa agcctggggt
4681 gcctaatgag tgagctaact cacattaatt gcgttgcgct cactgcccgc tttccagtcg
4741 ggaaacctgt cgtgccagct gcattaatga atcggccaac gcgcggggag aggcggtttg
4801 cgtattgggc gctcttccgc ttcctcgctc actgactcgc tgcgctcggt cgttcggctg
4861 cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt tatccacaga atcaggggat
4921 aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc
4981 gcgttgctgg cgtttttcca taggctccgc ccccctgacg agcatcacaa aaatcgacgc
5041 tcaagtcaga ggtggcgaaa cccgacagga ctataaagat accaggcgtt tccccctgga
5101 agctccctcg tgcgctctcc tgttccgacc ctgccgctta ccggatacct gtccgccttt
5161 ctcccttcgg gaagcgtggc gctttctcat agctcacgct gtaggtatct cagttcggtg
5221 taggtcgttc gctccaagct gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc
5281 gccttatccg gtaactatcg tcttgagtcc aacccggtaa gacacgactt atcgccactg
5341 gcagcagcca ctggtaacag gattagcaga gcgaggtatg taggcggtgc tacagagttc
5401 ttgaagtggt ggcctaacta cggctacact agaagaacag tatttggtat ctgcgctctg
5461 ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa acaaaccacc
5521 gctggtagcg gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct
5581 caagaagatc ctttgatctt ttctacgggg tctgacgctc agtggaacga aaactcacgt
5641 taagggattt tggtcatgag attatcaaaa aggatcttca cctagatcct tttaaattaa
5701 aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa cttggtctga cagttaccaa
5761 tgcttaatca gtgaggcacc tatctcagcg atctgtctat ttcgttcatc catagttgcc
5821 tgactccccg tcgtgtagat aactacgata cgggagggct taccatctgg ccccagtgct
5881 gcaatgatac cgcgagaccc acgctcaccg gctccagatt tatcagcaat aaaccagcca
5941 gccggaaggg ccgagcgcag aagtggtcct gcaactttat ccgcctccat ccagtctatt
6001 aattgttgcc gggaagctag agtaagtagt tcgccagtta atagtttgcg caacgttgtt
6061 gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc
6121 ggttcccaac gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa agcggttagc
6181 tccttcggtc ctccgatcgt tgtcagaagt aagttggccg cagtgttatc actcatggtt
6241 atggcagcac tgcataattc tcttactgtc atgccatccg taagatgctt ttctgtgact
6301 ggtgagtact caaccaagtc attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc
6361 ccggcgtcaa tacgggataa taccgcgcca catagcagaa ctttaaaagt gctcatcatt
6421 ggaaaacgtt cttcggggcg aaaactctca aggatcttac cgctgttgag atccagttcg
6481 atgtaaccca ctcgtgcacc caactgatct tcagcatctt ttactttcac cagcgtttct
6541 gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa
6601 tgttgaatac tcatactctt cctttttcaa tattattgaa gcatttatca gggttattgt
6661 ctcatgagcg gatacatatt tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc
6721 acatttcccc gaaaagtgcc acctgacg
//
