| 质粒名称: | mApple-Sec61b-C1 |
|---|---|
| 出品公司: | addgene |
| 目录编号: | 90993 |
| 存储实验室: | Jennifer Lippincott-Schwartz |
|
载体骨架基本信息 |
|
| 载体名称: | pN1 |
| 载体出品公司: | Clontech |
| 空载体大小: | 4732 bp |
| 载体修饰: | 人细胞色素c氧化酶亚基VIII（human cytochrome C oxidase subunit VIII） |
| 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: | mApple-Sec61b |
| 物种来源: | H. sapiens (human), Synthetic |
| 插入大小: | 1014 bp |
| 启动子: | CMV |
| 标签/融合蛋白: | mApple (red fluorescence) (N terminal on insert) |
| 克隆方法: | 限制性内切酶 |
| 5’克隆位点: | AgeI(未破坏) |
| 3’克隆位点: | BsrG1 (未破坏) |
| 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: | mApple-Sec61b-C1 was a gift from Jennifer Lippincott-Schwartz (Addgene plasmid # 90993 ; http://n2t.net/addgene:90993 ; RRID:Addgene_90993) |
| 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
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 mApple-Sec61b-C1 4999 bp ds-DNA circular SYN 12-MAY-2021
DEFINITION Expresses mApple-tagged Sec61b, brightly labels endoplasmic
reticulum in mammalian cells.
ACCESSION .
VERSION .
KEYWORDS mApple-Sec61b-C1
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 May 12, 2021 from SnapGene Server 1.1.58
http://www.biofeng.com
FEATURES Location/Qualifiers
source 1..4999
/organism="synthetic DNA construct"
/mol_type="other DNA"
primer_bind 54..73
/label=mCherry-F
/note="mCherry, forward primer"
CDS 364..654
/codon_start=1
/gene="SEC61B"
/product="human Sec61-beta subunit of the translocon in the
endoplasmic reticulum"
/label=Sec61-beta
/translation="MPGPTPSGTNVGSSGRSPSKAVAARAAGSTVRQRKNASCGTRSAG
RTTSAGTGGMWRFYTEDSPGLKVGPVPVLVMSLLFIASVFMLHIWGKYTRS"
polyA_signal 815..936
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
primer_bind complement(852..871)
/label=SV40pA-R
/note="SV40 polyA, reverse primer"
primer_bind 906..925
/label=EBV-rev
/note="SV40 polyA terminator, reverse primer"
rep_origin complement(943..1398)
/direction=LEFT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
primer_bind complement(1080..1101)
/label=F1ori-F
/note="F1 origin, forward primer"
primer_bind 1292..1311
/label=F1ori-R
/note="F1 origin, reverse primer"
promoter 1425..1529
/gene="bla"
/label=AmpR promoter
promoter 1531..1888
/label=SV40 promoter
/note="SV40 enhancer and early promoter"
primer_bind complement(1554..1574)
/label=pBABE 3'
/note="SV40 enhancer, reverse primer for pBABE vectors"
rep_origin 1739..1874
/label=SV40 ori
/note="SV40 origin of replication"
primer_bind 1801..1820
/label=SV40pro-F
/note="SV40 promoter/origin, forward primer"
CDS 1923..2717
/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(1977..1996)
/label=Neo-R
/note="Neomycin resistance gene, reverse primer"
primer_bind 2587..2606
/label=Neo-F
/note="Neomycin resistance gene, forward primer"
primer_bind complement(2905..2924)
/label=TK-pA-R
/note="Thymidine kinase polyA, reverse primer"
polyA_signal 2949..2996
/label=HSV TK poly(A) signal
/note="herpes simplex virus thymidine kinase
polyadenylation signal (Cole and Stacy, 1985)"
rep_origin 3325..3913
/direction=RIGHT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind 3814..3833
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
enhancer 4088..4391
/label=CMV enhancer
/note="human cytomegalovirus immediate early enhancer"
promoter 4392..4595
/label=CMV promoter
/note="human cytomegalovirus (CMV) immediate early
promoter"
primer_bind 4546..4566
/label=CMV-F
/note="Human CMV immediate early promoter, forward primer"
regulatory 4634..4643
/regulatory_class="other"
/note="vertebrate consensus sequence for strong initiation
of translation (Kozak, 1987)"
CDS 4640..348
/codon_start=1
/product="photostable monomeric derivative of DsRed (Shaner
et al., 2008)"
/label=mApple
/note="mammalian codon-optimized"
/translation="MVSKGEENNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEA
FQTAKLKVTKGGPLPFAWDILSPQFMYGSKVYIKHPADIPDYFKLSFPEGFRWERVMNF
EDGGIIHVNQDSSLQDGVFIYKVKLRGTNFPSDGPVMQKKTMGWEASEERMYPEDGALK
SEIKKRLKLKDGGHYAAEVKTTYKAKKPVQLPGAYIVDIKLDIVSHNEDYTIVEQYERA
EGRHSTGGMDELYK"
ORIGIN
1 ggcgtgttca tctacaaggt gaagctgcgc ggcaccaact tcccctccga cggccccgta
61 atgcagaaga agaccatggg ctgggaggcc tccgaggagc ggatgtaccc cgaggacggc
121 gccctgaaga gcgagatcaa gaagaggctg aagctgaagg acggcggcca ctacgccgcc
181 gaggtcaaga ccacctacaa ggccaagaag cccgtgcagc tgcccggcgc ctacatcgtc
241 gacatcaagt tggacatcgt gtcccacaac gaggactaca ccatcgtgga acagtacgaa
301 cgcgccgagg gccgccactc caccggcggc atggacgagc tgtacaagtc cggactcaga
361 tctatgcctg gtccgacccc cagtggcact aacgtgggat cctcagggcg ctctcccagc
421 aaagcagtgg ccgcccgggc ggcgggatcc actgtccggc agaggaaaaa tgccagctgt
481 gggacaagga gtgcaggccg cacaacctcg gcaggcaccg gggggatgtg gcgattctac
541 acagaagatt cacctgggct caaagttggc cctgttccag tattggttat gagtcttctg
601 ttcatcgctt ctgtatttat gttgcacatt tggggcaagt acactcgttc gtaggaattc
661 tgcagtcgac ggtaccgcgg gcccgggatc caccggatct agataactga tcataatcag
721 ccataccaca tttgtagagg ttttacttgc tttaaaaaac ctcccacacc tccccctgaa
781 cctgaaacat aaaatgaatg caattgttgt tgttaacttg tttattgcag cttataatgg
841 ttacaaataa agcaatagca tcacaaattt cacaaataaa gcattttttt cactgcattc
901 tagttgtggt ttgtccaaac tcatcaatgt atcttaacgc gtaaattgta agcgttaata
961 ttttgttaaa attcgcgtta aatttttgtt aaatcagctc attttttaac caataggccg
1021 aaatcggcaa aatcccttat aaatcaaaag aatagaccga gatagggttg agtgttgttc
1081 cagtttggaa caagagtcca ctattaaaga acgtggactc caacgtcaaa gggcgaaaaa
1141 ccgtctatca gggcgatggc ccactacgtg aaccatcacc ctaatcaagt tttttggggt
1201 cgaggtgccg taaagcacta aatcggaacc ctaaagggag cccccgattt agagcttgac
1261 ggggaaagcc ggcgaacgtg gcgagaaagg aagggaagaa agcgaaagga gcgggcgcta
1321 gggcgctggc aagtgtagcg gtcacgctgc gcgtaaccac cacacccgcc gcgcttaatg
1381 cgccgctaca gggcgcgtca ggtggcactt ttcggggaaa tgtgcgcgga acccctattt
1441 gtttattttt ctaaatacat tcaaatatgt atccgctcat gagacaataa ccctgataaa
1501 tgcttcaata atattgaaaa aggaagagtc ctgaggcgga aagaaccagc tgtggaatgt
1561 gtgtcagtta gggtgtggaa agtccccagg ctccccagca ggcagaagta tgcaaagcat
1621 gcatctcaat tagtcagcaa ccaggtgtgg aaagtcccca ggctccccag caggcagaag
1681 tatgcaaagc atgcatctca attagtcagc aaccatagtc ccgcccctaa ctccgcccat
1741 cccgccccta actccgccca gttccgccca ttctccgccc catggctgac taattttttt
1801 tatttatgca gaggccgagg ccgcctcggc ctctgagcta ttccagaagt agtgaggagg
1861 cttttttgga ggcctaggct tttgcaaaga tcgatcaaga gacaggatga ggatcgtttc
1921 gcatgattga acaagatgga ttgcacgcag gttctccggc cgcttgggtg gagaggctat
1981 tcggctatga ctgggcacaa cagacaatcg gctgctctga tgccgccgtg ttccggctgt
2041 cagcgcaggg gcgcccggtt ctttttgtca agaccgacct gtccggtgcc ctgaatgaac
2101 tgcaagacga ggcagcgcgg ctatcgtggc tggccacgac gggcgttcct tgcgcagctg
2161 tgctcgacgt tgtcactgaa gcgggaaggg actggctgct attgggcgaa gtgccggggc
2221 aggatctcct gtcatctcac cttgctcctg ccgagaaagt atccatcatg gctgatgcaa
2281 tgcggcggct gcatacgctt gatccggcta cctgcccatt cgaccaccaa gcgaaacatc
2341 gcatcgagcg agcacgtact cggatggaag ccggtcttgt cgatcaggat gatctggacg
2401 aagagcatca ggggctcgcg ccagccgaac tgttcgccag gctcaaggcg agcatgcccg
2461 acggcgagga tctcgtcgtg acccatggcg atgcctgctt gccgaatatc atggtggaaa
2521 atggccgctt ttctggattc atcgactgtg gccggctggg tgtggcggac cgctatcagg
2581 acatagcgtt ggctacccgt gatattgctg aagagcttgg cggcgaatgg gctgaccgct
2641 tcctcgtgct ttacggtatc gccgctcccg attcgcagcg catcgccttc tatcgccttc
2701 ttgacgagtt cttctgagcg ggactctggg gttcgaaatg accgaccaag cgacgcccaa
2761 cctgccatca cgagatttcg attccaccgc cgccttctat gaaaggttgg gcttcggaat
2821 cgttttccgg gacgccggct ggatgatcct ccagcgcggg gatctcatgc tggagttctt
2881 cgcccaccct agggggaggc taactgaaac acggaaggag acaataccgg aaggaacccg
2941 cgctatgacg gcaataaaaa gacagaataa aacgcacggt gttgggtcgt ttgttcataa
3001 acgcggggtt cggtcccagg gctggcactc tgtcgatacc ccaccgagac cccattgggg
3061 ccaatacgcc cgcgtttctt ccttttcccc accccacccc ccaagttcgg gtgaaggccc
3121 agggctcgca gccaacgtcg gggcggcagg ccctgccata gcctcaggtt actcatatat
3181 actttagatt gatttaaaac ttcattttta atttaaaagg atctaggtga agatcctttt
3241 tgataatctc atgaccaaaa tcccttaacg tgagttttcg ttccactgag cgtcagaccc
3301 cgtagaaaag atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt
3361 gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg ccggatcaag agctaccaac
3421 tctttttccg aaggtaactg gcttcagcag agcgcagata ccaaatactg ttcttctagt
3481 gtagccgtag ttaggccacc acttcaagaa ctctgtagca ccgcctacat acctcgctct
3541 gctaatcctg ttaccagtgg ctgctgccag tggcgataag tcgtgtctta ccgggttgga
3601 ctcaagacga tagttaccgg ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac
3661 acagcccagc ttggagcgaa cgacctacac cgaactgaga tacctacagc gtgagctatg
3721 agaaagcgcc acgcttcccg aagggagaaa ggcggacagg tatccggtaa gcggcagggt
3781 cggaacagga gagcgcacga gggagcttcc agggggaaac gcctggtatc tttatagtcc
3841 tgtcgggttt cgccacctct gacttgagcg tcgatttttg tgatgctcgt caggggggcg
3901 gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct tttgctggcc
3961 ttttgctcac atgttctttc ctgcgttatc ccctgattct gtggataacc gtattaccgc
4021 catgcattag ttattaatag taatcaatta cggggtcatt agttcatagc ccatatatgg
4081 agttccgcgt tacataactt acggtaaatg gcccgcctgg ctgaccgccc aacgaccccc
4141 gcccattgac gtcaataatg acgtatgttc ccatagtaac gccaataggg actttccatt
4201 gacgtcaatg ggtggagtat ttacggtaaa ctgcccactt ggcagtacat caagtgtatc
4261 atatgccaag tacgccccct attgacgtca atgacggtaa atggcccgcc tggcattatg
4321 cccagtacat gaccttatgg gactttccta cttggcagta catctacgta ttagtcatcg
4381 ctattaccat ggtgatgcgg ttttggcagt acatcaatgg gcgtggatag cggtttgact
4441 cacggggatt tccaagtctc caccccattg acgtcaatgg gagtttgttt tggcaccaaa
4501 atcaacggga ctttccaaaa tgtcgtaaca actccgcccc attgacgcaa atgggcggta
4561 ggcgtgtacg gtgggaggtc tatataagca gagctggttt agtgaaccgt cagatccgct
4621 agcgctaccg gtcgccacca tggtgagcaa gggcgaggag aataacatgg ccatcatcaa
4681 ggagttcatg cgcttcaagg tgcacatgga gggctccgtg aacggccacg agttcgagat
4741 cgagggcgag ggcgagggcc gcccctacga ggcctttcag accgctaagc tgaaggtgac
4801 caagggtggc cccctgccct tcgcctggga catcctgtcc cctcagttca tgtacggctc
4861 caaggtctac attaagcacc cagccgacat ccccgactac ttcaagctgt ccttccccga
4921 gggcttcagg tgggagcgcg tgatgaactt cgaggacggc ggcattattc acgttaacca
4981 ggactcctcc ctgcaggac
//
