| 质粒名称: | pLV-EF1a-V5-Sec61B |
|---|---|
| 出品公司: | addgene |
| 目录编号: | 111135 |
| 存储实验室: | Karin Busch |
|
载体骨架基本信息 |
|
| 载体名称: | pSEMS(26m) |
| 载体出品公司: | Covalys, NEB |
| 空载体大小: | 5325bp |
| 完整质粒大小: | 6570 bp |
| 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: | Receptor of tranlocase of outer mitochondrial membrane Tom20/Tom20 |
| 物种来源: | H. sapiens (human) |
| 插入大小: | 435 bp |
| 启动子: | CMV |
| 标签/融合蛋白: | Halo7 Tag (C terminal on insert) |
| 克隆方法: | 限制性内切酶 |
| 5’克隆位点: | EcoRV(未破坏) |
| 3’克隆位点: | EcoRI(未破坏) |
| 5’测序引物 | ATGGTGGGTCGGAACAGCGCCAT |
| 3’测序引物 | TTCCACATCATCTTCAGCCAAG |
|
文献引用方法 |
|
| 材料和方法部分: | pSEMS-Tom20-Halo7Tag was a gift from Karin Busch (Addgene plasmid # 111135 ; http://n2t.net/addgene:111135 ; RRID:Addgene_111135) |
| 参考文献部分: | Nanoscale organization of mitochondrial microcompartments revealed by combining tracking and localization microscopy. Appelhans T, Richter CP, Wilkens V, Hess ST, Piehler J, Busch KB. Nano Lett. 2012 Feb 8;12(2):610-6. doi: 10.1021/nl203343a. Epub 2012 Jan 13. 10.1021/nl203343a PubMed 22201267 |
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 pSEMS-Tom20-Halo7Tag 6570 bp ds-DNA circular SYN 12-MAY-2021
DEFINITION Tom20 with selflabeling tag, e.g. for fluorescence microscopy.
ACCESSION .
VERSION .
KEYWORDS pSEMS-Tom20-Halo7Tag
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 6570)
AUTHORS Appelhans T, Richter CP, Wilkens V, Hess ST, Piehler J, Busch KB
TITLE Nanoscale organization of mitochondrial microcompartments revealed
by combining tracking and localization microscopy.
JOURNAL Nano Lett. 2012 Feb 8;12(2):610-6. doi: 10.1021/nl203343a. Epub 2012
Jan 13.
PUBMED 22201267
REFERENCE 2 (bases 1 to 6570)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58
http://www.biofeng.com
FEATURES Location/Qualifiers
source 1..6570
/organism="synthetic DNA construct"
/mol_type="other DNA"
primer_bind complement(44..63)
/label=pRS-marker
/note="pRS vectors, use to sequence yeast selectable
marker"
enhancer 235..614
/label=CMV enhancer
/note="human cytomegalovirus immediate early enhancer"
promoter 615..818
/label=CMV promoter
/note="human cytomegalovirus (CMV) immediate early
promoter"
primer_bind 769..789
/label=CMV-F
/note="Human CMV immediate early promoter, forward primer"
primer_bind 863..882
/label=T7
/note="T7 promoter, forward primer"
promoter 863..881
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
CDS 1371..2255
/codon_start=1
/product="modified bacterial dehalogenase that forms
covalent bonds with chloroalkane derivatives"
/label=HaloTag(R)
/translation="EIGTGFPFDPHYVEVLGERMHYVDVGPRDGTPVLFLHGNPTSSYV
WRNIIPHVAPTHRCIAPDLIGMGKSDKPDLGYFFDDHVRFMDAFIEALGLEEVVLVIHD
WGSALGFHWAKRNPERVKGIAFMEFIRPIPTWDEWPEFARETFQAFRTTDVGRKLIIDQ
NVFIEGTLPMGVVRPLTEVEMDHYREPFLNPVDREPLWRFPNELPIAGEPANIVALVEE
YMDWLHQSPVPKLLFWGTPGVLIPPAEAARLAKSLPNCKAVDIGPGLNLLQEDNPDLIG
SEIARWLSTLEISG"
intron 2361..2590
/label=chimeric intron
/note="chimera between introns from adenovirus and
immunoglobulin heavy chain genes"
primer_bind 2573..2592
/label=pMT2-F
/note="Synthetic intron, forward primer"
misc_feature 2648..3221
/label=IRES
/note="internal ribosome entry site (IRES) of the
encephalomyocarditis virus (EMCV)"
primer_bind complement(2815..2832)
/label=IRES reverse
/note="IRES internal ribosome entry site, reverse primer.
Also called pCDH-rev"
primer_bind 3042..3061
/label=IRES-F
/note="IRES internal ribosome entry site, forward primer"
CDS 3257..4060
/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="MGSAIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQ
GRPVLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQD
LLSSHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDE
EHQGLAPAELFARLKARMPDGDDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQ
DIALATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
primer_bind complement(3320..3339)
/label=Neo-R
/note="Neomycin resistance gene, reverse primer"
primer_bind 3930..3949
/label=Neo-F
/note="Neomycin resistance gene, forward primer"
polyA_signal 4190..4311
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
primer_bind complement(4227..4246)
/label=SV40pA-R
/note="SV40 polyA, reverse primer"
primer_bind complement(4413..4429)
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(4413..4429)
/label=M13 Reverse
/note="In lacZ gene. Also called M13-rev"
primer_bind complement(4426..4448)
/label=M13/pUC Reverse
/note="In lacZ gene"
protein_bind 4437..4453
/label=lac operator
/bound_moiety="lac repressor encoded by lacI"
/note="The lac repressor binds to the lac operator to
inhibit transcription in E. coli. This inhibition can be
relieved by adding lactose or
isopropyl-beta-D-thiogalactopyranoside (IPTG)."
promoter complement(4461..4491)
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 4506..4527
/label=CAP binding site
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding activates transcription in the presence
of cAMP."
primer_bind complement(4644..4661)
/label=L4440
/note="L4440 vector, forward primer"
rep_origin complement(4815..5403)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind complement(4895..4914)
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
CDS complement(5574..6434)
/codon_start=1
/gene="bla"
/product="beta-lactamase"
/label=AmpR
/note="confers resistance to ampicillin, carbenicillin, and
related antibiotics"
/translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
LIKHW"
primer_bind 6197..6216
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
promoter complement(6435..6539)
/gene="bla"
/label=AmpR promoter
ORIGIN
1 gacggatcgg gagatctccc gatcccctat ggtgcactct cagtacaatc tgctctgatg
61 ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg
121 cgagcaaaat ttaagctaca acaaggcaag gcttgaccga caattgcatg aagaatctgc
181 ttagggttag gcgttttgcg ctgcttcgcg atgtacgggc cagatatacg cgttgacatt
241 gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata
301 tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc
361 cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc
421 attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt
481 atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt
541 atgcccagta catgacctta tgggactttc ctacttggca gtacatctac gtattagtca
601 tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg
661 actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc
721 aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg
781 gtaggcgtgt acggtgggag gtctatataa gcagagctct ctggctaact agagaaccca
841 ctgcttactg gcttatcgaa attaatacga ctcactatag ggagacccaa gcttggtacc
901 gagctcggat cgatatcatg gtgggtcgga acagcgccat cgccgccggt gtatgcgggg
961 cccttttcat tgggtactgc atctacttcg accgcaaaag acgaagtgac cccaacttca
1021 agaacaggct tcgagaacga agaaagaaac agaagcttgc caaggagaga gctgggcttt
1081 ccaagttacc tgaccttaaa gatgctgaag ctgttcagaa gttcttcctt gaagaaatac
1141 agcttggtga agagttacta gctcaaggtg aatatgagaa gggcgtagac catctgacaa
1201 atgcaattgc tgtgtgtgga cagccacagc agttactgca ggtcttacag caaactcttc
1261 caccaccagt gttccagatg cttctgacta agctcccaac aattagtcag agaattgtaa
1321 gtgctcagag cttggctgaa gatgatgtgg aagaattcca tatgggatcc gaaatcggta
1381 ctggctttcc attcgacccc cattatgtgg aagtcctggg cgagcgcatg cactacgtcg
1441 atgttggtcc gcgcgatggc acccctgtgc tgttcctgca cggtaacccg acctcctcct
1501 acgtgtggcg caacatcatc ccgcatgttg caccgaccca tcgctgcatt gctccagacc
1561 tgatcggtat gggcaaatcc gacaaaccag acctgggtta tttcttcgac gaccacgtcc
1621 gcttcatgga tgccttcatc gaagccctgg gtctggaaga ggtcgtcctg gtcattcacg
1681 actggggctc cgctctgggt ttccactggg ccaagcgcaa tccagagcgc gtcaaaggta
1741 ttgcatttat ggagttcatc cgccctatcc cgacctggga cgaatggcca gaatttgccc
1801 gcgagacctt ccaggccttc cgcaccaccg acgtcggccg caagctgatc atcgatcaga
1861 acgtttttat cgagggtacg ctgccgatgg gtgtcgtccg cccgctgact gaagtcgaga
1921 tggaccatta ccgcgagccg ttcctgaatc ctgttgaccg cgagccactg tggcgcttcc
1981 caaacgagct gccaatcgcc ggtgagccag cgaacatcgt cgcgctggtc gaagaataca
2041 tggactggct gcaccagtcc cctgtcccga agctgctgtt ctggggcacc ccaggcgttc
2101 tgatcccacc ggccgaagcc gctcgcctgg ccaaaagcct gcctaactgc aaggctgtgg
2161 acatcggccc gggtctgaat ctgctgcaag aagacaaccc ggacctgatc ggcagcgaga
2221 tcgcgcgctg gctgtctact ctggagattt ccggtcctgc aggtataggc gcgccaggat
2281 ccctcgagtg aggcggccgc atagataact gatccagtgt gctggaatta attcgctgtc
2341 tgcgagggcc agctgttggg gtgagtactc cctctcaaaa gcgggcatga cttctgcgct
2401 aagattgtca gtttccaaaa acgaggagga tttgatattc acctggcccg cggtgatgcc
2461 tttgagggtg gccgcgtcca tctggtcaga aaagacaatc tttttgttgt caagcttgag
2521 gtgtggcagg cttgagatct ggccatacac ttgagtgaca atgacatcca ctttgccttt
2581 ctctccacag gtgtccactc ccaggtccaa ctgcaggtcg agcatgcatc tagggcggcc
2641 aattccgccc ctctcccycc ccccccccta acgttactgg ccgaagccgc ttggaataag
2701 gccggtgtgc gtttgtctat atgttatttt ccaccatatt gccgtctttt ggcaatgtga
2761 gggcccggaa acctggccct gtcttcttga cgagcattcc taggggtctt tcccctctcg
2821 ccaaaggaat gcaaggtctg ttgaatgtcg tgaaggaagc agttcctctg gaagcttctt
2881 gaagacaaac aacgtctgta gcgacccttt gcaggcagcg gaacccccca cctggcgaca
2941 ggtgcctctg cggccaaaag ccacgtgtat aagatacacc tgcaaaggcg gcacaacccc
3001 agtgccacgt tgtgagttgg atagttgtgg aaagagtcaa atggctctcc tcaagcgtat
3061 tcaacaaggg gctgaaggat gcccagaagg taccccattg tatgggatct gatctggggc
3121 ctcggtgcac atgctttaca tgtgtttagt cgaggttaaa aaaacgtcta ggccccccga
3181 accacgggga cgtggttttc ctttgaaaaa cacgatgata agcttgccac aacccgggat
3241 aattcctgca gccaatatgg gatcggccat tgaacaagat ggattgcacg caggttctcc
3301 ggccgcttgg gtggagaggc tattcggcta tgactgggca caacagacaa tcggctgctc
3361 tgatgccgcc gtgttccggc tgtcagcgca ggggcgcccg gttctttttg tcaagaccga
3421 cctgtccggt gccctgaatg aactgcagga cgaggcagcg cggctatcgt ggctggccac
3481 gacgggcgtt ccttgcgcag ctgtgctcga cgttgtcact gaagcgggaa gggactggct
3541 gctattgggc gaagtgccgg ggcaggatct cctgtcatct caccttgctc ctgccgagaa
3601 agtatccatc atggctgatg caatgcggcg gctgcatacg cttgatccgg ctacctgccc
3661 attcgaccac caagcgaaac atcgcatcga gcgagcacgt actcggatgg aagccggtct
3721 tgtcgatcag gatgatctgg acgaagagca tcaggggctc gcgccagccg aactgttcgc
3781 caggctcaag gcgcgcatgc ccgacggcga tgatctcgtc gtgacccatg gcgatgcctg
3841 cttgccgaat atcatggtgg aaaatggccg cttttctgga ttcatcgact gtggccggct
3901 gggtgtggcg gaccgctatc aggacatagc gttggctacc cgtgatattg ctgaagagct
3961 tggcggcgaa tgggctgacc gcttcctcgt gctttacggt atcgccgctc ccgattcgca
4021 gcgcatcgcc ttctatcgcc ttcttgacga gttcttctga ggggatcaat tctctagata
4081 actgatcata atcagccata ccacatttgt agaggtttta cttgctttaa aaaacctccc
4141 acacctcccc ctgaacctga aacataaaat gaatgcaatt gttgttgtta acttgtttat
4201 tgcagcttat aatggttaca aataaagcaa tagcatcaca aatttcacaa ataaagcatt
4261 tttttcactg cattctagtt gtggtttgtc caaactcatc aatgtatctt aacgcgtcga
4321 gtgcattcta gttgtggttt gtccaaactc atcaatgtat cttatcatgt ctgtataccg
4381 tcgacctcta gctagagctt ggcgtaatca tggtcatagc tgtttcctgt gtgaaattgt
4441 tatccgctca caattccaca caacatacga gccggaagca taaagtgtaa agcctggggt
4501 gcctaatgag tgagctaact cacattaatt gcgttgcgct cactgcccgc tttccagtcg
4561 ggaaacctgt cgtgccagct gcattaatga atcggccaac gcgcggggag aggcggtttg
4621 cgtattgggc gctcttccgc ttcctcgctc actgactcgc tgcgctcggt cgttcggctg
4681 cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt tatccacaga atcaggggat
4741 aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc
4801 gcgttgctgg cgtttttcca taggctccgc ccccctgacg agcatcacaa aaatcgacgc
4861 tcaagtcaga ggtggcgaaa cccgacagga ctataaagat accaggcgtt tccccctgga
4921 agctccctcg tgcgctctcc tgttccgacc ctgccgctta ccggatacct gtccgccttt
4981 ctcccttcgg gaagcgtggc gctttctcat agctcacgct gtaggtatct cagttcggtg
5041 taggtcgttc gctccaagct gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc
5101 gccttatccg gtaactatcg tcttgagtcc aacccggtaa gacacgactt atcgccactg
5161 gcagcagcca ctggtaacag gattagcaga gcgaggtatg taggcggtgc tacagagttc
5221 ttgaagtggt ggcctaacta cggctacact agaagaacag tatttggtat ctgcgctctg
5281 ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa acaaaccacc
5341 gctggtagcg gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct
5401 caagaagatc ctttgatctt ttctacgggg tctgacgctc agtggaacga aaactcacgt
5461 taagggattt tggtcatgag attatcaaaa aggatcttca cctagatcct tttaaattaa
5521 aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa cttggtctga cagttaccaa
5581 tgcttaatca gtgaggcacc tatctcagcg atctgtctat ttcgttcatc catagttgcc
5641 tgactccccg tcgtgtagat aactacgata cgggagggct taccatctgg ccccagtgct
5701 gcaatgatac cgcgagaccc acgctcaccg gctccagatt tatcagcaat aaaccagcca
5761 gccggaaggg ccgagcgcag aagtggtcct gcaactttat ccgcctccat ccagtctatt
5821 aattgttgcc gggaagctag agtaagtagt tcgccagtta atagtttgcg caacgttgtt
5881 gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc
5941 ggttcccaac gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa agcggttagc
6001 tccttcggtc ctccgatcgt tgtcagaagt aagttggccg cagtgttatc actcatggtt
6061 atggcagcac tgcataattc tcttactgtc atgccatccg taagatgctt ttctgtgact
6121 ggtgagtact caaccaagtc attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc
6181 ccggcgtcaa tacgggataa taccgcgcca catagcagaa ctttaaaagt gctcatcatt
6241 ggaaaacgtt cttcggggcg aaaactctca aggatcttac cgctgttgag atccagttcg
6301 atgtaaccca ctcgtgcacc caactgatct tcagcatctt ttactttcac cagcgtttct
6361 gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa
6421 tgttgaatac tcatactctt cctttttcaa tattattgaa gcatttatca gggttattgt
6481 ctcatgagcg gatacatatt tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc
6541 acatttcccc gaaaagtgcc acctgacgtc
//
