| 质粒名称: | GFP-Rab27A |
|---|---|
| 出品公司: | addgene |
| 目录编号: | 89237 |
| 存储实验室: | William Gahl |
|
载体骨架基本信息 |
|
| 载体名称: | peGFP-C1 |
| 载体出品公司: | Clontech |
| 空载体大小: | 4700bp |
| 完整质粒大小: | 5360bp |
| 载体类型: | 哺乳细胞表达 |
| 筛选标记: | 新霉素（G418） |
| 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: | human Rab27A |
| 物种来源: | H. sapiens (human) |
| 插入大小: | 660 bp |
| 基因突变: | wild type |
| GenBank编号 | BC132800.1 |
| 启动子: | CMV |
| 标签/融合蛋白: | eGFP (C terminal on backbone) |
| 克隆方法: | 限制性内切酶 |
| 5’克隆位点: | EcoR1(未破坏) |
| 3’克隆位点: | BamHI (未破坏) |
| 5’测序引物 | 5’: CATGGTCCTGCTGGAGTTCGTGACCG |
| 3’测序引物 | 5’: ATAAGCTGCAATAAACAAGTTAACAA |
|
文献引用方法 |
|
| 材料和方法部分: | GFP-Rab27A was a gift from William Gahl (Addgene plasmid # 89237 ; http://n2t.net/addgene:89237 ; RRID:Addgene_89237) |
| 参考文献部分: | A novel missense mutation (G43S) in the switch I region of Rab27A causing Griscelli syndrome. Westbroek W, Tuchman M, Tinloy B, De Wever O, Vilboux T, Hertz JM, Hasle H, Heilmann C, Helip-Wooley A, Kleta R, Gahl WA. Mol Genet Metab. 2008 Jun;94(2):248-54. doi: 10.1016/j.ymgme.2008.02.009. Epub 2008 Apr 7. 10.1016/j.ymgme.2008.02.009 PubMed 18397837 |
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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 GFP-Rab27A 5416 bp ds-DNA circular SYN 12-MAY-2021
DEFINITION Expression of GFP-tagged human Rab27A in mammalian cells.
ACCESSION .
VERSION .
KEYWORDS GFP-Rab27A
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 5416)
AUTHORS Westbroek W, Tuchman M, Tinloy B, De Wever O, Vilboux T, Hertz JM,
Hasle H, Heilmann C, Helip-Wooley A, Kleta R, Gahl WA
TITLE A novel missense mutation (G43S) in the switch I region of Rab27A
causing Griscelli syndrome.
JOURNAL Mol Genet Metab. 2008 Jun;94(2):248-54. doi:
10.1016/j.ymgme.2008.02.009. Epub 2008 Apr 7.
PUBMED 18397837
REFERENCE 2 (bases 1 to 5416)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58
http://www.biofeng.com
FEATURES Location/Qualifiers
source 1..5416
/organism="synthetic DNA construct"
/mol_type="other DNA"
rep_origin 15..603
/direction=RIGHT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind 504..523
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
enhancer 778..1081
/label=CMV enhancer
/note="human cytomegalovirus immediate early enhancer"
promoter 1082..1285
/label=CMV promoter
/note="human cytomegalovirus (CMV) immediate early
promoter"
primer_bind 1236..1256
/label=CMV-F
/note="Human CMV immediate early promoter, forward primer"
regulatory 1324..1333
/regulatory_class="other"
/note="vertebrate consensus sequence for strong initiation
of translation (Kozak, 1987)"
CDS 1330..2046
/codon_start=1
/product="enhanced GFP"
/label=EGFP
/translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
EFVTAAGITLGMDELYK"
CDS 1330..2046
/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(1375..1396)
/label=EGFP-N
/note="EGFP, reverse primer"
primer_bind complement(1636..1655)
/label=EXFP-R
/note="For distinguishing EGFP variants, reverse primer"
primer_bind 1983..2004
/label=EGFP-C
/note="EGFP, forward primer"
polyA_signal 2921..3042
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
primer_bind complement(2958..2977)
/label=SV40pA-R
/note="SV40 polyA, reverse primer"
primer_bind 3012..3031
/label=EBV-rev
/note="SV40 polyA terminator, reverse primer"
rep_origin complement(3049..3504)
/direction=LEFT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
primer_bind complement(3186..3207)
/label=F1ori-F
/note="F1 origin, forward primer"
primer_bind 3398..3417
/label=F1ori-R
/note="F1 origin, reverse primer"
promoter 3531..3635
/gene="bla"
/label=AmpR promoter
promoter 3637..3994
/label=SV40 promoter
/note="SV40 enhancer and early promoter"
primer_bind complement(3660..3680)
/label=pBABE 3'
/note="SV40 enhancer, reverse primer for pBABE vectors"
rep_origin 3845..3980
/label=SV40 ori
/note="SV40 origin of replication"
primer_bind 3907..3926
/label=SV40pro-F
/note="SV40 promoter/origin, forward primer"
CDS 4029..4823
/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
GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
primer_bind complement(4083..4102)
/label=Neo-R
/note="Neomycin resistance gene, reverse primer"
primer_bind 4693..4712
/label=Neo-F
/note="Neomycin resistance gene, forward primer"
primer_bind complement(5011..5030)
/label=TK-pA-R
/note="Thymidine kinase polyA, reverse primer"
polyA_signal 5055..5102
/label=HSV TK poly(A) signal
/note="herpes simplex virus thymidine kinase
polyadenylation signal (Cole and Stacy, 1985)"
ORIGIN
1 atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa
61 aaaccaccgc taccagcggt ggtttgtttg ccggatcaag agctaccaac tctttttccg
121 aaggtaactg gcttcagcag agcgcagata ccaaatactg ttcttctagt gtagccgtag
181 ttaggccacc acttcaagaa ctctgtagca ccgcctacat acctcgctct gctaatcctg
241 ttaccagtgg ctgctgccag tggcgataag tcgtgtctta ccgggttgga ctcaagacga
301 tagttaccgg ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac acagcccagc
361 ttggagcgaa cgacctacac cgaactgaga tacctacagc gtgagctatg agaaagcgcc
421 acgcttcccg aagggagaaa ggcggacagg tatccggtaa gcggcagggt cggaacagga
481 gagcgcacga gggagcttcc agggggaaac gcctggtatc tttatagtcc tgtcgggttt
541 cgccacctct gacttgagcg tcgatttttg tgatgctcgt caggggggcg gagcctatgg
601 aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct tttgctggcc ttttgctcac
661 atgttctttc ctgcgttatc ccctgattct gtggataacc gtattaccgc catgcattag
721 ttattaatag taatcaatta cggggtcatt agttcatagc ccatatatgg agttccgcgt
781 tacataactt acggtaaatg gcccgcctgg ctgaccgccc aacgaccccc gcccattgac
841 gtcaataatg acgtatgttc ccatagtaac gccaataggg actttccatt gacgtcaatg
901 ggtggagtat ttacggtaaa ctgcccactt ggcagtacat caagtgtatc atatgccaag
961 tacgccccct attgacgtca atgacggtaa atggcccgcc tggcattatg cccagtacat
1021 gaccttatgg gactttccta cttggcagta catctacgta ttagtcatcg ctattaccat
1081 ggtgatgcgg ttttggcagt acatcaatgg gcgtggatag cggtttgact cacggggatt
1141 tccaagtctc caccccattg acgtcaatgg gagtttgttt tggcaccaaa atcaacggga
1201 ctttccaaaa tgtcgtaaca actccgcccc attgacgcaa atgggcggta ggcgtgtacg
1261 gtgggaggtc tatataagca gagctggttt agtgaaccgt cagatccgct agcgctaccg
1321 gtcgccacca tggtgagcaa gggcgaggag ctgttcaccg gggtggtgcc catcctggtc
1381 gagctggacg gcgacgtaaa cggccacaag ttcagcgtgt ccggcgaggg cgagggcgat
1441 gccacctacg gcaagctgac cctgaagttc atctgcacca ccggcaagct gcccgtgccc
1501 tggcccaccc tcgtgaccac cctgacctac ggcgtgcagt gcttcagccg ctaccccgac
1561 cacatgaagc agcacgactt cttcaagtcc gccatgcccg aaggctacgt ccaggagcgc
1621 accatcttct tcaaggacga cggcaactac aagacccgcg ccgaggtgaa gttcgagggc
1681 gacaccctgg tgaaccgcat cgagctgaag ggcatcgact tcaaggagga cggcaacatc
1741 ctggggcaca agctggagta caactacaac agccacaacg tctatatcat ggccgacaag
1801 cagaagaacg gcatcaaggt gaacttcaag atccgccaca acatcgagga cggcagcgtg
1861 cagctcgccg accactacca gcagaacacc cccatcggcg acggccccgt gctgctgccc
1921 gacaaccact acctgagcac ccagtccgcc ctgagcaaag accccaacga gaagcgcgat
1981 cacatggtcc tgctggagtt cgtgaccgcc gccgggatca ctctcggcat ggacgagctg
2041 tacaagtccg gactcagatc tcgagctcaa gcttcgaatt ctatgtctga tggagattat
2101 gattacctca tcaagttttt agctttggga gactctggtg tagggaagac cagtgtactt
2161 taccaatata cagatggtaa atttaactcc aaatttatca caacagtggg cattgatttc
2221 agggaaaaaa gagtggtgta cagagccagt gggccggatg gagccactgg cagaggccag
2281 agaatccacc tgcagttatg ggacacagca gggcaggaga ggtttcgtag cttaacgaca
2341 gcgttcttca gagatgctat gggttttctt ctactttttg atctgacaaa tgagcaaagt
2401 ttcctcaatg tcagaaactg gataagccag ctacagatgc atgcatattg tgaaaaccca
2461 gatatagtgc tgtgtggaaa caagagtgat ctggaggacc agagagtagt gaaagaggag
2521 gaagccatag cactcgcaga gaaatatgga atcccctact ttgaaactag tgctgccaat
2581 gggacaaaca taagccaagc aattgagatg cttctggacc tgataatgaa gcgaatggaa
2641 cggtgtgtgg acaagtcctg gattcctgaa ggagtggtgc gatcaaatgg tcatgcctct
2701 acggatcagt taagtgaaga aaaggagaaa ggggcatgtg gctgttgaga agtcaagtaa
2761 gcgacatagt agttcaggtg gcccatgcct gggatccacc ggatctagat aactgatcat
2821 aatcagccat accacatttg tagaggtttt acttgcttta aaaaacctcc cacacctccc
2881 cctgaacctg aaacataaaa tgaatgcaat tgttgttgtt aacttgttta ttgcagctta
2941 taatggttac aaataaagca atagcatcac aaatttcaca aataaagcat ttttttcact
3001 gcattctagt tgtggtttgt ccaaactcat caatgtatct taacgcgtaa attgtaagcg
3061 ttaatatttt gttaaaattc gcgttaaatt tttgttaaat cagctcattt tttaaccaat
3121 aggccgaaat cggcaaaatc ccttataaat caaaagaata gaccgagata gggttgagtg
3181 ttgttccagt ttggaacaag agtccactat taaagaacgt ggactccaac gtcaaagggc
3241 gaaaaaccgt ctatcagggc gatggcccac tacgtgaacc atcaccctaa tcaagttttt
3301 tggggtcgag gtgccgtaaa gcactaaatc ggaaccctaa agggagcccc cgatttagag
3361 cttgacgggg aaagccggcg aacgtggcga gaaaggaagg gaagaaagcg aaaggagcgg
3421 gcgctagggc gctggcaagt gtagcggtca cgctgcgcgt aaccaccaca cccgccgcgc
3481 ttaatgcgcc gctacagggc gcgtcaggtg gcacttttcg gggaaatgtg cgcggaaccc
3541 ctatttgttt atttttctaa atacattcaa atatgtatcc gctcatgaga caataaccct
3601 gataaatgct tcaataatat tgaaaaagga agagtcctga ggcggaaaga accagctgtg
3661 gaatgtgtgt cagttagggt gtggaaagtc cccaggctcc ccagcaggca gaagtatgca
3721 aagcatgcat ctcaattagt cagcaaccag gtgtggaaag tccccaggct ccccagcagg
3781 cagaagtatg caaagcatgc atctcaatta gtcagcaacc atagtcccgc ccctaactcc
3841 gcccatcccg cccctaactc cgcccagttc cgcccattct ccgccccatg gctgactaat
3901 tttttttatt tatgcagagg ccgaggccgc ctcggcctct gagctattcc agaagtagtg
3961 aggaggcttt tttggaggcc taggcttttg caaagatcga tcaagagaca ggatgaggat
4021 cgtttcgcat gattgaacaa gatggattgc acgcaggttc tccggccgct tgggtggaga
4081 ggctattcgg ctatgactgg gcacaacaga caatcggctg ctctgatgcc gccgtgttcc
4141 ggctgtcagc gcaggggcgc ccggttcttt ttgtcaagac cgacctgtcc ggtgccctga
4201 atgaactgca agacgaggca gcgcggctat cgtggctggc cacgacgggc gttccttgcg
4261 cagctgtgct cgacgttgtc actgaagcgg gaagggactg gctgctattg ggcgaagtgc
4321 cggggcagga tctcctgtca tctcaccttg ctcctgccga gaaagtatcc atcatggctg
4381 atgcaatgcg gcggctgcat acgcttgatc cggctacctg cccattcgac caccaagcga
4441 aacatcgcat cgagcgagca cgtactcgga tggaagccgg tcttgtcgat caggatgatc
4501 tggacgaaga gcatcagggg ctcgcgccag ccgaactgtt cgccaggctc aaggcgagca
4561 tgcccgacgg cgaggatctc gtcgtgaccc atggcgatgc ctgcttgccg aatatcatgg
4621 tggaaaatgg ccgcttttct ggattcatcg actgtggccg gctgggtgtg gcggaccgct
4681 atcaggacat agcgttggct acccgtgata ttgctgaaga gcttggcggc gaatgggctg
4741 accgcttcct cgtgctttac ggtatcgccg ctcccgattc gcagcgcatc gccttctatc
4801 gccttcttga cgagttcttc tgagcgggac tctggggttc gaaatgaccg accaagcgac
4861 gcccaacctg ccatcacgag atttcgattc caccgccgcc ttctatgaaa ggttgggctt
4921 cggaatcgtt ttccgggacg ccggctggat gatcctccag cgcggggatc tcatgctgga
4981 gttcttcgcc caccctaggg ggaggctaac tgaaacacgg aaggagacaa taccggaagg
5041 aacccgcgct atgacggcaa taaaaagaca gaataaaacg cacggtgttg ggtcgtttgt
5101 tcataaacgc ggggttcggt cccagggctg gcactctgtc gataccccac cgagacccca
5161 ttggggccaa tacgcccgcg tttcttcctt ttccccaccc caccccccaa gttcgggtga
5221 aggcccaggg ctcgcagcca acgtcggggc ggcaggccct gccatagcct caggttactc
5281 atatatactt tagattgatt taaaacttca tttttaattt aaaaggatct aggtgaagat
5341 cctttttgat aatctcatga ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc
5401 agaccccgta gaaaag
//
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