质粒名称: | mCh-Rab7A |
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出品公司: | addgene |
目录编号: | 61804 |
存储实验室: | Gia Voeltz |
载体骨架基本信息 |
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载体名称: | pAcGFP1-C1 |
载体出品公司: | Clontech |
空载体大小: | 4700bp |
完整质粒大小: | 5345bp |
载体类型: | 哺乳细胞表达 |
筛选标记: | 新霉素(G418) |
载体抗性: | 卡那霉素 |
生长温度: | 37 ℃ |
宿主菌: | XL1 Blue |
拷贝数: | 未知 |
插入基因信息 |
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插入基因名称: | Rab7A |
物种来源: | H. sapiens (human) |
插入大小: | 624bp |
GenBank编号 | JNM_004637.5 |
启动子: | CMV |
标签/融合蛋白: | mCh (N terminal on backbone) |
克隆方法: | 限制性内切酶 |
5’克隆位点: | XhoI(未破坏) |
3’克隆位点: | HindIII (未破坏) |
5’测序引物 | CCA TCG TGG AAC AGT ACG AACG |
3’测序引物 | none |
文献引用方法 |
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材料和方法部分: | mCh-Rab7A was a gift from Gia Voeltz (Addgene plasmid # 61804 ; http://n2t.net/addgene:61804 ; RRID:Addgene_61804) |
参考文献部分: | 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载体质粒基本信息
mCh-Rab7A质粒图谱载体图谱和mCh-Rab7A载体序列质粒序列多克隆位点信息
mCh-Rab7A质粒载体简介
mCh-Rab7A质粒序列载体序列
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 http://www.biofeng.com 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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mCh-Rab7A质粒载体应用举例
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