| 質粒名稱: | mCh-Rab7A |
|---|---|
| 出品公司: | addgene |
| 目錄編號: | 61804 |
| 存儲實驗室: | Gia Voeltz |
|
載體骨架基本信息 |
|
| 載體名稱: | pAcGFP1-C1 |
| 載體出品公司: | Clontech |
| 空載體大小: | 4700bp |
| 完整質粒大小: | 5345bp |
| 載體類型: | 哺乳細胞表達 |
| 篩選標記: | 新霉素（G418） |
| 載體抗性: | 卡那霉素 |
| 生長溫度: | 37 ℃ |
| 宿主菌: | XL1 Blue |
| 拷貝數: | 未知 |
|
插入基因信息 |
|
| 插入基因名稱: | Rab7A |
| 物種來源: | H. sapiens (human) |
| 插入大小: | 624bp |
| GenBank編號 | JNM_004637.5 |
| 啟動子: | CMV |
| 標簽/融合蛋白: | mCh (N terminal on backbone) |
| 克隆方法: | 限制性內切酶 |
| 5’克隆位點: | XhoI(未破壞) |
| 3’克隆位點: | HindIII (未破壞) |
| 5’測序引物 | CCA TCG TGG AAC AGT ACG AACG |
| 3’測序引物 | none |
|
文獻引用方法 |
|
| 材料和方法部分: | mCh-Rab7A was a gift from Gia Voeltz (Addgene plasmid # 61804 ; //n2t.net/addgene:61804 ; RRID:Addgene_61804) |
| 參考文獻部分: | ER Contact Sites Define the Position and Timing of Endosome Fission. Rowland AA, Chitwood PJ, Phillips MJ, Voeltz GK. Cell. 2014 Nov 20;159(5):1027-41. doi: 10.1016/j.cell.2014.10.023. 10.1016/j.cell.2014.10.023 PubMed 25416 |
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mCh-Rab7A載體質粒基本信息
mCh-Rab7A質粒圖譜載體圖譜和mCh-Rab7A載體序列質粒序列多克隆位點信息
mCh-Rab7A質粒載體簡介
mCh-Rab7A質粒序列載體序列
LOCUS mCh-Rab7A 5345 bp ds-DNA circular SYN 12-MAY-2021
DEFINITION Human Rab7A fluorescently tagged with mCh on the N-terminus for
mammalian expression.
ACCESSION .
VERSION .
KEYWORDS mCh-Rab7A
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 5345)
AUTHORS Rowland AA, Chitwood PJ, Phillips MJ, Voeltz GK
TITLE ER Contact Sites Define the Position and Timing of Endosome Fission.
JOURNAL Cell. 2014 Nov 20;159(5):1027-41. doi: 10.1016/j.cell.2014.10.023.
PUBMED 25416943
REFERENCE 2 (bases 1 to 5345)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58
//gt12365.cn
FEATURES Location/Qualifiers
source 1..5345
/organism="synthetic DNA construct"
/mol_type="other DNA"
enhancer 68..371
/label=CMV enhancer
/note="human cytomegalovirus immediate early enhancer"
promoter 372..575
/label=CMV promoter
/note="human cytomegalovirus (CMV) immediate early
promoter"
primer_bind 526..546
/label=CMV-F
/note="Human CMV immediate early promoter, forward primer"
regulatory 614..623
/regulatory_class="other"
/note="vertebrate consensus sequence for strong initiation
of translation (Kozak, 1987)"
CDS 620..1327
/codon_start=1
/product="monomeric derivative of DsRed fluorescent protein
(Shaner et al., 2004)"
/label=mCherry
/note="mammalian codon-optimized"
/translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEG
TQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNF
EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK
GEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERA
EGRHSTGGMDELYK"
primer_bind complement(765..783)
/label=mCherry-R
/note="mCherry, reverse primer"
primer_bind 1033..1052
/label=mCherry-F
/note="mCherry, forward primer"
polyA_signal 2140..2261
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
primer_bind complement(2177..2196)
/label=SV40pA-R
/note="SV40 polyA, reverse primer"
primer_bind 2231..2250
/label=EBV-rev
/note="SV40 polyA terminator, reverse primer"
rep_origin complement(2268..2723)
/direction=LEFT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
primer_bind complement(2405..2426)
/label=F1ori-F
/note="F1 origin, forward primer"
primer_bind 2617..2636
/label=F1ori-R
/note="F1 origin, reverse primer"
promoter 2750..2854
/gene="bla"
/label=AmpR promoter
promoter 2856..3213
/label=SV40 promoter
/note="SV40 enhancer and early promoter"
primer_bind complement(2879..2899)
/label=pBABE 3'
/note="SV40 enhancer, reverse primer for pBABE vectors"
rep_origin 3064..3199
/label=SV40 ori
/note="SV40 origin of replication"
primer_bind 3126..3145
/label=SV40pro-F
/note="SV40 promoter/origin, forward primer"
CDS 3248..4042
/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(3302..3321)
/label=Neo-R
/note="Neomycin resistance gene, reverse primer"
primer_bind 3912..3931
/label=Neo-F
/note="Neomycin resistance gene, forward primer"
primer_bind complement(4230..4249)
/label=TK-pA-R
/note="Thymidine kinase polyA, reverse primer"
polyA_signal 4274..4321
/label=HSV TK poly(A) signal
/note="herpes simplex virus thymidine kinase
polyadenylation signal (Cole and Stacy, 1985)"
rep_origin 4650..5238
/direction=RIGHT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind 5139..5158
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
ORIGIN
1 catgcattag ttattaatag taatcaatta cggggtcatt agttcatagc ccatatatgg
61 agttccgcgt tacataactt acggtaaatg gcccgcctgg ctgaccgccc aacgaccccc
121 gcccattgac gtcaataatg acgtatgttc ccatagtaac gccaataggg actttccatt
181 gacgtcaatg ggtggagtat ttacggtaaa ctgcccactt ggcagtacat caagtgtatc
241 atatgccaag tacgccccct attgacgtca atgacggtaa atggcccgcc tggcattatg
301 cccagtacat gaccttatgg gactttccta cttggcagta catctacgta ttagtcatcg
361 ctattaccat ggtgatgcgg ttttggcagt acatcaatgg gcgtggatag cggtttgact
421 cacggggatt tccaagtctc caccccattg acgtcaatgg gagtttgttt tggcaccaaa
481 atcaacggga ctttccaaaa tgtcgtaaca actccgcccc attgacgcaa atgggcggta
541 ggcgtgtacg gtgggaggtc tatataagca gagctggttt agtgaaccgt cagatccgct
601 agcgctaccg gtcgccacca tggtgagcaa gggcgaggag gataacatgg ccatcatcaa
661 ggagttcatg cgcttcaagg tgcacatgga gggctccgtg aacggccacg agttcgagat
721 cgagggcgag ggcgagggcc gcccctacga gggcacccag accgccaagc tgaaggtgac
781 caagggtggc cccctgccct tcgcctggga catcctgtcc cctcagttca tgtacggctc
841 caaggcctac gtgaagcacc ccgccgacat ccccgactac ttgaagctgt ccttccccga
901 gggcttcaag tgggagcgcg tgatgaactt cgaggacggc ggcgtggtga ccgtgaccca
961 ggactcctcc ctgcaggacg gcgagttcat ctacaaggtg aagctgcgcg gcaccaactt
1021 cccctccgac ggccccgtaa tgcagaagaa gaccatgggc tgggaggcct cctccgagcg
1081 gatgtacccc gaggacggcg ccctgaaggg cgagatcaag cagaggctga agctgaagga
1141 cggcggccac tacgacgctg aggtcaagac cacctacaag gccaagaagc ccgtgcagct
1201 gcccggcgcc tacaacgtca acatcaagtt ggacatcacc tcccacaacg aggactacac
1261 catcgtggaa cagtacgaac gcgccgaggg ccgccactcc accggcggca tggacgagct
1321 gtacaagtcc ggactcagat ctcgagctat gacctctagg aagaaagtgt tgctgaaggt
1381 tatcatcctg ggagattctg gagtcgggaa gacatcactc atgaaccagt atgtgaataa
1441 gaaattcagc aatcagtaca aagccacaat aggagctgac tttctgacca aggaggtgat
1501 ggtggatgac aggctagtca caatgcagat atgggacaca gcaggacagg aacggttcca
1561 gtctctcggt gtggccttct acagaggtgc agactgctgc gttctggtat ttgatgtgac
1621 tgcccccaac acattcaaaa ccctagatag ctggagagat gagtttctca tccaggccag
1681 tccccgagat cctgaaaact tcccatttgt tgtgttggga aacaagattg acctcgaaaa
1741 cagacaagtg gccacaaagc gggcacaggc ctggtgctac agcaaaaaca acattcccta
1801 ctttgagacc agtgccaagg aggccatcaa cgtggagcag gcgttccaga cgattgcacg
1861 gaatgcactt aagcaggaaa cggaggtgga gctgtacaac gaatttcctg aacctatcaa
1921 actggacaag aatgaccggg ccaaggcctc ggcagaaagc tgcagttgct gaaagcttcg
1981 aattctgcag tcgacggtac cgcgggcccg ggatccaccg gatctagata actgatcata
2041 atcagccata ccacatttgt agaggtttta cttgctttaa aaaacctccc acacctcccc
2101 ctgaacctga aacataaaat gaatgcaatt gttgttgtta acttgtttat tgcagcttat
2161 aatggttaca aataaagcaa tagcatcaca aatttcacaa ataaagcatt tttttcactg
2221 cattctagtt gtggtttgtc caaactcatc aatgtatctt aacgcgtaaa ttgtaagcgt
2281 taatattttg ttaaaattcg cgttaaattt ttgttaaatc agctcatttt ttaaccaata
2341 ggccgaaatc ggcaaaatcc cttataaatc aaaagaatag accgagatag ggttgagtgt
2401 tgttccagtt tggaacaaga gtccactatt aaagaacgtg gactccaacg tcaaagggcg
2461 aaaaaccgtc tatcagggcg atggcccact acgtgaacca tcaccctaat caagtttttt
2521 ggggtcgagg tgccgtaaag cactaaatcg gaaccctaaa gggagccccc gatttagagc
2581 ttgacgggga aagccggcga acgtggcgag aaaggaaggg aagaaagcga aaggagcggg
2641 cgctagggcg ctggcaagtg tagcggtcac gctgcgcgta accaccacac ccgccgcgct
2701 taatgcgccg ctacagggcg cgtcaggtgg cacttttcgg ggaaatgtgc gcggaacccc
2761 tatttgttta tttttctaaa tacattcaaa tatgtatccg ctcatgagac aataaccctg
2821 ataaatgctt caataatatt gaaaaaggaa gagtcctgag gcggaaagaa ccagctgtgg
2881 aatgtgtgtc agttagggtg tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa
2941 agcatgcatc tcaattagtc agcaaccagg tgtggaaagt ccccaggctc cccagcaggc
3001 agaagtatgc aaagcatgca tctcaattag tcagcaacca tagtcccgcc cctaactccg
3061 cccatcccgc ccctaactcc gcccagttcc gcccattctc cgccccatgg ctgactaatt
3121 ttttttattt atgcagaggc cgaggccgcc tcggcctctg agctattcca gaagtagtga
3181 ggaggctttt ttggaggcct aggcttttgc aaagatcgat caagagacag gatgaggatc
3241 gtttcgcatg attgaacaag atggattgca cgcaggttct ccggccgctt gggtggagag
3301 gctattcggc tatgactggg cacaacagac aatcggctgc tctgatgccg ccgtgttccg
3361 gctgtcagcg caggggcgcc cggttctttt tgtcaagacc gacctgtccg gtgccctgaa
3421 tgaactgcaa gacgaggcag cgcggctatc gtggctggcc acgacgggcg ttccttgcgc
3481 agctgtgctc gacgttgtca ctgaagcggg aagggactgg ctgctattgg gcgaagtgcc
3541 ggggcaggat ctcctgtcat ctcaccttgc tcctgccgag aaagtatcca tcatggctga
3601 tgcaatgcgg cggctgcata cgcttgatcc ggctacctgc ccattcgacc accaagcgaa
3661 acatcgcatc gagcgagcac gtactcggat ggaagccggt cttgtcgatc aggatgatct
3721 ggacgaagag catcaggggc tcgcgccagc cgaactgttc gccaggctca aggcgagcat
3781 gcccgacggc gaggatctcg tcgtgaccca tggcgatgcc tgcttgccga atatcatggt
3841 ggaaaatggc cgcttttctg gattcatcga ctgtggccgg ctgggtgtgg cggaccgcta
3901 tcaggacata gcgttggcta cccgtgatat tgctgaagag cttggcggcg aatgggctga
3961 ccgcttcctc gtgctttacg gtatcgccgc tcccgattcg cagcgcatcg ccttctatcg
4021 ccttcttgac gagttcttct gagcgggact ctggggttcg aaatgaccga ccaagcgacg
4081 cccaacctgc catcacgaga tttcgattcc accgccgcct tctatgaaag gttgggcttc
4141 ggaatcgttt tccgggacgc cggctggatg atcctccagc gcggggatct catgctggag
4201 ttcttcgccc accctagggg gaggctaact gaaacacgga aggagacaat accggaagga
4261 acccgcgcta tgacggcaat aaaaagacag aataaaacgc acggtgttgg gtcgtttgtt
4321 cataaacgcg gggttcggtc ccagggctgg cactctgtcg ataccccacc gagaccccat
4381 tggggccaat acgcccgcgt ttcttccttt tccccacccc accccccaag ttcgggtgaa
4441 ggcccagggc tcgcagccaa cgtcggggcg gcaggccctg ccatagcctc aggttactca
4501 tatatacttt agattgattt aaaacttcat ttttaattta aaaggatcta ggtgaagatc
4561 ctttttgata atctcatgac caaaatccct taacgtgagt tttcgttcca ctgagcgtca
4621 gaccccgtag aaaagatcaa aggatcttct tgagatcctt tttttctgcg cgtaatctgc
4681 tgcttgcaaa caaaaaaacc accgctacca gcggtggttt gtttgccgga tcaagagcta
4741 ccaactcttt ttccgaaggt aactggcttc agcagagcgc agataccaaa tactgttctt
4801 ctagtgtagc cgtagttagg ccaccacttc aagaactctg tagcaccgcc tacatacctc
4861 gctctgctaa tcctgttacc agtggctgct gccagtggcg ataagtcgtg tcttaccggg
4921 ttggactcaa gacgatagtt accggataag gcgcagcggt cgggctgaac ggggggttcg
4981 tgcacacagc ccagcttgga gcgaacgacc tacaccgaac tgagatacct acagcgtgag
5041 ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc
5101 agggtcggaa caggagagcg cacgagggag cttccagggg gaaacgcctg gtatctttat
5161 agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg
5221 gggcggagcc tatggaaaaa cgccagcaac gcggcctttt tacggttcct ggccttttgc
5281 tggccttttg ctcacatgtt ctttcctgcg ttatcccctg attctgtgga taaccgtatt
5341 accgc
//
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