| 质粒名称: | mCherry-Sec61b-C1 |
|---|---|
| 出品公司: | Addgene |
| 目录编号: | 90994 |
| 存储实验室: | Jennifer Lippincott-Schwartz |
|
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: | 4732bp |
| 完整质粒大小: | 5000bp |
| 载体类型: | 哺乳细胞表达 |
| 筛选标记: | 新霉素（G418） |
| 载体抗性: | Kanamycin, 50 μg/mL |
| 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: | mCherry-Sec61b |
| 物种来源: | H. sapiens (human), Synthetic |
| 插入大小: | 702bp |
| 启动子: | CMV |
| 标签/融合蛋白: | mCherry (red fluorescence) (N terminal on insert) |
| 克隆方法: | 限制性内切酶 |
| 5’克隆位点: | AgeI (not destroyed) |
| 3’克隆位点: | BsrG1 (not destroyed) |
| 5’测序引物 | CMV-F |
| 3’测序引物 | SV40polyA |
|
文献引用方法 |
|
| 材料和方法部分: | mCherry-Sec61b-C1 was a gift from Jennifer Lippincott-Schwartz (Addgene plasmid # 90994 ; http://n2t.net/addgene:90994 ; RRID:Addgene_90994) |
| 参考文献部分: | Increased spatiotemporal resolution reveals highly dynamic dense tubular matrices in the peripheral ER. Nixon-Abell J, Obara CJ, Weigel AV, Li D, Legant WR, Xu CS, Pasolli HA, Harvey K, Hess HF, Betzig E, Blackstone C, Lippincott-Schwartz J. Science. 2016 Oct 28;354(6311). pii: aaf3928. Epub 2016 Oct 27. 10.1126/science.aaf3928 PubMed 27789813 |
买家导航
mCherry-Sec61b-C1载体质粒基本信息
mCherry-Sec61b-C1质粒图谱载体图谱和 mCherry-Sec61b-C1载体序列质粒序列多克隆位点信息
mCherry-Sec61b-C1质粒载体简介
mCherry-Sec61b-C1质粒序列载体序列
LOCUS mCherry-Sec61b-C1 4999 bp DNA circular SYN 14-APR-2025
DEFINITION Expresses mCherry-tagged Sec61b, brightly labels endoplasmic
reticulum in mammalian cells.
ACCESSION .
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 4999)
AUTHORS Nixon-Abell J, Obara CJ, Weigel AV, Li D, Legant WR, Xu CS, Pasolli
HA, Harvey K, Hess HF, Betzig E, Blackstone C, Lippincott-Schwartz J
TITLE Increased spatiotemporal resolution reveals highly dynamic dense
tubular matrices in the peripheral ER.
JOURNAL Science. 2016 Oct 28;354(6311). pii: aaf3928. Epub 2016 Oct 27.
PUBMED 27789813
REFERENCE 2 (bases 1 to 4999)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Apr 14, 2025 from SnapGene Server 8.0.1
https://www.biofeng.com
COMMENT SGRef: number: 1; type: "Journal Article"; journalName: "Science.";
date: "2016-10-28"; volume: "354(6311). pii"; pages: " aaf3928. Epub
2016 Oct 27"
COMMENT Sequence Label: mCherry-Sec61b-C1
FEATURES Location/Qualifiers
source 1..4999
/mol_type="other DNA"
/organism="synthetic DNA construct"
polyA_signal 403..450
/label=HSV TK poly(A) signal
/note="herpes simplex virus thymidine kinase
polyadenylation signal (Cole and Stacy, 1985)"
primer_bind 475..494
/label=TK-pA-R
/note="Thymidine kinase polyA, reverse primer"
CDS complement(682..1476)
/codon_start=1
/product="aminoglycoside phosphotransferase"
/label=NeoR/KanR
/note="confers resistance to neomycin, kanamycin, and G418
(Geneticin(R))"
/translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
CDS complement(682..1476)
/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(793..812)
/label=Neo-F
/note="Neomycin resistance gene, forward primer"
primer_bind 1403..1422
/label=Neo-R
/note="Neomycin resistance gene, reverse primer"
promoter complement(1511..1868)
/label=SV40 promoter
/note="SV40 enhancer and early promoter"
rep_origin 1525..1660
/label=SV40 ori
/note="SV40 origin of replication"
enhancer 1677..1868
/label=SV40 enhancer
/note="enhancer for the SV40 early promoter (Herr, 1993)"
primer_bind 1825..1845
/label=pBABE 3'
/note="SV40 enhancer, reverse primer for pBABE vectors"
promoter complement(1870..1974)
/gene="bla"
/label=AmpR promoter
rep_origin 2001..2456
/direction=RIGHT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
primer_bind complement(2088..2107)
/label=F1ori-R
/note="F1 origin, reverse primer"
primer_bind 2298..2319
/label=F1ori-F
/note="F1 origin, forward primer"
polyA_signal 2463..2584
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
primer_bind complement(2474..2493)
/label=EBV-rev
/note="SV40 polyA terminator, reverse primer"
primer_bind 2528..2547
/label=SV40pA-R
/note="SV40 polyA, reverse primer"
CDS complement(2745..3035)
/codon_start=1
/gene="SEC61B"
/product="human Sec61-beta subunit of the translocon in the
endoplasmic reticulum"
/label=Sec61-beta
/translation="MPGPTPSGTNVGSSGRSPSKAVAARAAGSTVRQRKNASCGTRSAG
RTTSAGTGGMWRFYTEDSPGLKVGPVPVLVMSLLFIASVFMLHIWGKYTRS"
CDS complement(3051..3758)
/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(3326..3345)
/label=mCherry-F
/note="mCherry, forward primer"
primer_bind 3595..3613
/label=mCherry-R
/note="mCherry, reverse primer"
regulatory 3755..3764
/label=Kozak sequence
/note="vertebrate consensus sequence for strong initiation
of translation (Kozak, 1987)"
/regulatory_class="other"
promoter complement(3803..4006)
/label=CMV promoter
/note="human cytomegalovirus (CMV) immediate early
promoter"
primer_bind complement(3832..3852)
/label=CMV-F
/note="Human CMV immediate early promoter, forward primer"
enhancer 4007..4310
/label=CMV enhancer
/note="human cytomegalovirus immediate early enhancer"
rep_origin complement(join(4485..4999,1..74))
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind complement(4565..4584)
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
ORIGIN
1 aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa
61 aaaaaggatc tcaagaagat cctttgatct tttctacggg gtctgacgct cagtggaacg
121 aaaactcacg ttaagggatt ttggtcatga gattatcaaa aaggatcttc acctagatcc
181 ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa cctgaggcta
241 tggcagggcc tgccgccccg acgttggctg cgagccctgg gccttcaccc gaacttgggg
301 ggtggggtgg ggaaaaggaa gaaacgcggg cgtattggcc ccaatggggt ctcggtgggg
361 tatcgacaga gtgccagccc tgggaccgaa ccccgcgttt atgaacaaac gacccaacac
421 cgtgcgtttt attctgtctt tttattgccg tcatagcgcg ggttccttcc ggtattgtct
481 ccttccgtgt ttcagttagc ctccccctag ggtgggcgaa gaactccagc atgagatccc
541 cgcgctggag gatcatccag ccggcgtccc ggaaaacgat tccgaagccc aacctttcat
601 agaaggcggc ggtggaatcg aaatctcgtg atggcaggtt gggcgtcgct tggtcggtca
661 tttcgaaccc cagagtcccg ctcagaagaa ctcgtcaaga aggcgataga aggcgatgcg
721 ctgcgaatcg ggagcggcga taccgtaaag cacgaggaag cggtcagccc attcgccgcc
781 aagctcttca gcaatatcac gggtagccaa cgctatgtcc tgatagcggt ccgccacacc
841 cagccggcca cagtcgatga atccagaaaa gcggccattt tccaccatga tattcggcaa
901 gcaggcatcg ccatgggtca cgacgagatc ctcgccgtcg ggcatgctcg ccttgagcct
961 ggcgaacagt tcggctggcg cgagcccctg atgctcttcg tccagatcat cctgatcgac
1021 aagaccggct tccatccgag tacgtgctcg ctcgatgcga tgtttcgctt ggtggtcgaa
1081 tgggcaggta gccggatcaa gcgtatgcag ccgccgcatt gcatcagcca tgatggatac
1141 tttctcggca ggagcaaggt gagatgacag gagatcctgc cccggcactt cgcccaatag
1201 cagccagtcc cttcccgctt cagtgacaac gtcgagcaca gctgcgcaag gaacgcccgt
1261 cgtggccagc cacgatagcc gcgctgcctc gtcttgcagt tcattcaggg caccggacag
1321 gtcggtcttg acaaaaagaa ccgggcgccc ctgcgctgac agccggaaca cggcggcatc
1381 agagcagccg attgtctgtt gtgcccagtc atagccgaat agcctctcca cccaagcggc
1441 cggagaacct gcgtgcaatc catcttgttc aatcatgcga aacgatcctc atcctgtctc
1501 ttgatcgatc tttgcaaaag cctaggcctc caaaaaagcc tcctcactac ttctggaata
1561 gctcagaggc cgaggcggcc tcggcctctg cataaataaa aaaaattagt cagccatggg
1621 gcggagaatg ggcggaactg ggcggagtta ggggcgggat gggcggagtt aggggcggga
1681 ctatggttgc tgactaattg agatgcatgc tttgcatact tctgcctgct ggggagcctg
1741 gggactttcc acacctggtt gctgactaat tgagatgcat gctttgcata cttctgcctg
1801 ctggggagcc tggggacttt ccacacccta actgacacac attccacagc tggttctttc
1861 cgcctcagga ctcttccttt ttcaatatta ttgaagcatt tatcagggtt attgtctcat
1921 gagcggatac atatttgaat gtatttagaa aaataaacaa ataggggttc cgcgcacatt
1981 tccccgaaaa gtgccacctg acgcgccctg tagcggcgca ttaagcgcgg cgggtgtggt
2041 ggttacgcgc agcgtgaccg ctacacttgc cagcgcccta gcgcccgctc ctttcgcttt
2101 cttcccttcc tttctcgcca cgttcgccgg ctttccccgt caagctctaa atcgggggct
2161 ccctttaggg ttccgattta gtgctttacg gcacctcgac cccaaaaaac ttgattaggg
2221 tgatggttca cgtagtgggc catcgccctg atagacggtt tttcgccctt tgacgttgga
2281 gtccacgttc tttaatagtg gactcttgtt ccaaactgga acaacactca accctatctc
2341 ggtctattct tttgatttat aagggatttt gccgatttcg gcctattggt taaaaaatga
2401 gctgatttaa caaaaattta acgcgaattt taacaaaata ttaacgctta caatttacgc
2461 gttaagatac attgatgagt ttggacaaac cacaactaga atgcagtgaa aaaaatgctt
2521 tatttgtgaa atttgtgatg ctattgcttt atttgtaacc attataagct gcaataaaca
2581 agttaacaac aacaattgca ttcattttat gtttcaggtt cagggggagg tgtgggaggt
2641 tttttaaagc aagtaaaacc tctacaaatg tggtatggct gattatgatc agttatctag
2701 atccggtgga tcccgggccc gcggtaccgt cgactgcaga attcctacga acgagtgtac
2761 ttgccccaaa tgtgcaacat aaatacagaa gcgatgaaca gaagactcat aaccaatact
2821 ggaacagggc caactttgag cccaggtgaa tcttctgtgt agaatcgcca catccccccg
2881 gtgcctgccg aggttgtgcg gcctgcactc cttgtcccac agctggcatt tttcctctgc
2941 cggacagtgg atcccgccgc ccgggcggcc actgctttgc tgggagagcg ccctgaggat
3001 cccacgttag tgccactggg ggtcggacca ggcatagatc tgagtccgga cttgtacagc
3061 tcgtccatgc cgccggtgga gtggcggccc tcggcgcgtt cgtactgttc cacgatggtg
3121 tagtcctcgt tgtgggaggt gatgtccaac ttgatgttga cgttgtaggc gccgggcagc
3181 tgcacgggct tcttggcctt gtaggtggtc ttgacctcag cgtcgtagtg gccgccgtcc
3241 ttcagcttca gcctctgctt gatctcgccc ttcagggcgc cgtcctcggg gtacatccgc
3301 tcggaggagg cctcccagcc catggtcttc ttctgcatta cggggccgtc ggaggggaag
3361 ttggtgccgc gcagcttcac cttgtagatg aactcgccgt cctgcaggga ggagtcctgg
3421 gtcacggtca ccacgccgcc gtcctcgaag ttcatcacgc gctcccactt gaagccctcg
3481 gggaaggaca gcttcaagta gtcggggatg tcggcggggt gcttcacgta ggccttggag
3541 ccgtacatga actgagggga caggatgtcc caggcgaagg gcagggggcc acccttggtc
3601 accttcagct tggcggtctg ggtgccctcg taggggcggc cctcgccctc gccctcgatc
3661 tcgaactcgt ggccgttcac ggagccctcc atgtgcacct tgaagcgcat gaactccttg
3721 atgatggcca tgttatcctc ctcgcccttg ctcaccatgg tggcgaccgg tagcgctagc
3781 ggatctgacg gttcactaaa ccagctctgc ttatatagac ctcccaccgt acacgcctac
3841 cgcccatttg cgtcaatggg gcggagttgt tacgacattt tggaaagtcc cgttgatttt
3901 ggtgccaaaa caaactccca ttgacgtcaa tggggtggag acttggaaat ccccgtgagt
3961 caaaccgcta tccacgccca ttgatgtact gccaaaaccg catcaccatg gtaatagcga
4021 tgactaatac gtagatgtac tgccaagtag gaaagtccca taaggtcatg tactgggcat
4081 aatgccaggc gggccattta ccgtcattga cgtcaatagg gggcgtactt ggcatatgat
4141 acacttgatg tactgccaag tgggcagttt accgtaaata ctccacccat tgacgtcaat
4201 ggaaagtccc tattggcgtt actatgggaa catacgtcat tattgacgtc aatgggcggg
4261 ggtcgttggg cggtcagcca ggcgggccat ttaccgtaag ttatgtaacg cggaactcca
4321 tatatgggct atgaactaat gaccccgtaa ttgattacta ttaataacta atgcatggcg
4381 gtaatacggt tatccacaga atcaggggat aacgcaggaa agaacatgtg agcaaaaggc
4441 cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca taggctccgc
4501 ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga
4561 ctataaagat accaggcgtt tccccctgga agctccctcg tgcgctctcc tgttccgacc
4621 ctgccgctta ccggatacct gtccgccttt ctcccttcgg gaagcgtggc gctttctcat
4681 agctcacgct gtaggtatct cagttcggtg taggtcgttc gctccaagct gggctgtgtg
4741 cacgaacccc ccgttcagcc cgaccgctgc gccttatccg gtaactatcg tcttgagtcc
4801 aacccggtaa gacacgactt atcgccactg gcagcagcca ctggtaacag gattagcaga
4861 gcgaggtatg taggcggtgc tacagagttc ttgaagtggt ggcctaacta cggctacact
4921 agaagaacag tatttggtat ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt
4981 ggtagctctt gatccggca
//
