| 质粒名称: | pnEK-NvS_M |
|---|---|
| 出品公司: | Addgene |
| 目录编号: | 146926 |
| 存储实验室: | Elisa Izaurralde |
|
载体骨架基本信息 |
|
| 载体名称: | pnEK |
| 载体出品公司: | -- |
| 空载体大小: | -- |
| 完整质粒大小: | -- |
| 载体修饰: | -- |
| 载体类型: | 细菌表达 |
| 筛选标记: | -- |
| 载体抗性: | 卡那霉素50 μg/mL |
| 生长温度: | 37 ℃ |
| 宿主菌: | DH5a |
| 拷贝数: | 低拷贝 |
|
插入基因信息 |
|
| 插入基因名称: | -- |
| 物种来源: | -- |
| 插入大小: | -- |
| 基因突变: | -- |
| GenBank编号 | -- |
| 启动子: | -- |
| 标签/融合蛋白: | -- |
| 克隆方法: | -- |
| 5’克隆位点: | -- |
| 3’克隆位点: | -- |
| 5’测序引物 | -- |
| 3’测序引物 | -- |
|
文献引用方法 |
|
| 材料和方法部分: | pnEK-NvS_M was a gift from Elisa Izaurralde (Addgene plasmid # 146926 ; http://n2t.net/addgene:146926 ; RRID:Addgene_146926) |
| 参考文献部分: | Deciphering correct strategies for multiprotein complex assembly by co-expression: application to complexes as large as the histone octamer. Diebold ML, Fribourg S, Koch M, Metzger T, Romier C. J Struct Biol. 2011 Aug;175(2):178-88. doi: 10.1016/j.jsb.2011.02.001. Epub 2011 Feb 12. 10.1016/j.jsb.2011.02.001 PubMed 21320604 |
买家导航
pnEK-NvS_M载体质粒基本信息
pnEK-NvS_M质粒图谱载体图谱和pnEK-NvS_M载体序列质粒序列多克隆位点信息
pnEK-NvS_M质粒载体简介
pnEK-NvS_M质粒序列载体序列
LOCUS pnEK-NvS_M 5335 bp ds-DNA circular SYN 24-FEB-2022
DEFINITION Bacterial Expression vector.
ACCESSION .
VERSION .
KEYWORDS pnEK-NvS_M
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 5335)
AUTHORS Diebold ML, Fribourg S, Koch M, Metzger T, Romier C
TITLE Deciphering correct strategies for multiprotein complex assembly by
co-expression: application to complexes as large as the histone
octamer.
JOURNAL J Struct Biol. 2011 Aug;175(2):178-88. doi:
10.1016/j.jsb.2011.02.001. Epub 2011 Feb 12.
PUBMED 21320604
REFERENCE 2 (bases 1 to 5335)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Feb 24, 2022 from SnapGene Server 1.1.58
http://www.snapgene.com
FEATURES Location/Qualifiers
source 1..5335
/organism="synthetic DNA construct"
/mol_type="other DNA"
rep_origin 12..467
/direction=RIGHT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
primer_bind complement(99..118)
/label=F1ori-R
/note="F1 origin, reverse primer"
primer_bind 309..330
/label=F1ori-F
/note="F1 origin, forward primer"
CDS complement(559..1374)
/codon_start=1
/gene="aph(3')-Ia"
/product="aminoglycoside phosphotransferase"
/label=KanR
/note="confers resistance to kanamycin in bacteria or G418
(Geneticin(R)) in eukaryotes"
/translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
primer_bind 1282..1301
/label=Kan-R
/note="Kanamycin resistance gene, reverse primer"
primer_bind 1366..1385
/label=pENTR-R
/note="pENTR vectors, reverse primer"
rep_origin 1496..2084
/direction=RIGHT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind 1985..2004
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
primer_bind 2238..2255
/label=L4440
/note="L4440 vector, forward primer"
misc_feature 2270..2409
/label=bom
/note="basis of mobility region from pBR322"
primer_bind complement(2335..2354)
/label=pRS-marker
/note="pRS vectors, use to sequence yeast selectable
marker"
primer_bind 2495..2517
/label=pGEX 3'
/note="pGEX vectors, reverse primer"
CDS complement(2511..2702)
/codon_start=1
/gene="rop"
/product="Rop protein, which maintains plasmids at low copy
number"
/label=rop
/translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
DELYRSCLARFGDDGENL"
CDS complement(3511..4593)
/codon_start=1
/gene="lacI"
/product="lac repressor"
/label=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)."
/translation="MKPVTLYDVAEYAGVSYQTVSRVVNQASHVSAKTREKVEAAMAEL
NYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSRADQLGASVVVSMVERSGV
EACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSIIFSH
EDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREGDWSA
MSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTEDSSC
YIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR
ALADSLMQLARQVSRLESGQ"
primer_bind 4555..4574
/label=LacI-R
/note="LacI, reverse primer"
promoter complement(4594..4671)
/gene="lacI"
/label=lacI promoter
primer_bind 4877..4896
/label=pBRrevBam
/note="pBR322 vectors, tet region, downstream of BamHI,
reverse primer"
primer_bind 4980..4999
/label=T7
/note="T7 promoter, forward primer"
promoter 4980..4998
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
protein_bind 4999..5023
/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)."
RBS 5038..5060
/note="efficient ribosome binding site from bacteriophage
T7 gene 10 (Olins and Rangwala, 1989)"
CDS 5079..5102
/codon_start=1
/product="peptide that binds Strep-Tactin(R), an engineered
form of streptavidin"
/label=Strep-Tag II
/translation="WSHPQFEK"
CDS 5124..5144
/codon_start=1
/product="tobacco etch virus (TEV) protease recognition and
cleavage site"
/label=TEV site
/translation="ENLYFQG"
primer_bind complement(5249..5267)
/label=T7 Term
/note="T7 terminator, reverse primer"
terminator 5263..5310
/label=T7 terminator
/note="transcription terminator for bacteriophage T7 RNA
polymerase"
ORIGIN
1 tggcgaatgg gacgcgccct gtagcggcgc attaagcgcg gcgggtgtgg tggttacgcg
61 cagcgtgacc gctacacttg ccagcgccct agcgcccgct cctttcgctt tcttcccttc
121 ctttctcgcc acgttcgccg gctttccccg tcaagctcta aatcgggggc tccctttagg
181 gttccgattt agtgctttac ggcacctcga ccccaaaaaa cttgattagg gtgatggttc
241 acgtagtggg ccatcgccct gatagacggt ttttcgccct ttgacgttgg agtccacgtt
301 ctttaatagt ggactcttgt tccaaactgg aacaacactc aaccctatct cggtctattc
361 ttttgattta taagggattt tgccgatttc ggcctattgg ttaaaaaatg agctgattta
421 acaaaaattt aacgcgaatt ttaacaaaat attaacgctt acaatttagg tggcactttt
481 cggggaaatg tgcgcggaac ccctatttgt ttatttttct aaatacattc aaatatgtat
541 ccgctcatga attaattctt agaaaaactc atcgagcatc aaatgaaact gcaatttatt
601 catatcagga ttatcaatac catatttttg aaaaagccgt ttctgtaatg aaggagaaaa
661 ctcaccgagg cagttccata ggatggcaag atcctggtat cggtctgcga ttccgactcg
721 tccaacatca atacaaccta ttaatttccc ctcgtcaaaa ataaggttat caagtgagaa
781 atcaccatga gtgacgactg aatccggtga gaatggcaaa agtttatgca tttctttcca
841 gacttgttca acaggccagc cattacgctc gtcatcaaaa tcactcgcat caaccaaacc
901 gttattcatt cgtgattgcg cctgagcgag acgaaatacg cgatcgctgt taaaaggaca
961 attacaaaca ggaatcgaat gcaaccggcg caggaacact gccagcgcat caacaatatt
1021 ttcacctgaa tcaggatatt cttctaatac ctggaatgct gttttcccgg ggatcgcagt
1081 ggtgagtaac catgcatcat caggagtacg gataaaatgc ttgatggtcg gaagaggcat
1141 aaattccgtc agccagttta gtctgaccat ctcatctgta acatcattgg caacgctacc
1201 tttgccatgt ttcagaaaca actctggcgc atcgggcttc ccatacaatc gatagattgt
1261 cgcacctgat tgcccgacat tatcgcgagc ccatttatac ccatataaat cagcatccat
1321 gttggaattt aatcgcggcc tagagcaaga cgtttcccgt tgaatatggc tcataacacc
1381 ccttgtatta ctgtttatgt aagcagacag ttttattgtt catgaccaaa atcccttaac
1441 gtgagttttc gttccactga gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag
1501 atcctttttt tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg
1561 tggtttgttt gccggatcaa gagctaccaa ctctttttcc gaaggtaact ggcttcagca
1621 gagcgcagat accaaatact gtccttctag tgtagccgta gttaggccac cacttcaaga
1681 actctgtagc accgcctaca tacctcgctc tgctaatcct gttaccagtg gctgctgcca
1741 gtggcgataa gtcgtgtctt accgggttgg actcaagacg atagttaccg gataaggcgc
1801 agcggtcggg ctgaacgggg ggttcgtgca cacagcccag cttggagcga acgacctaca
1861 ccgaactgag atacctacag cgtgagctat gagaaagcgc cacgcttccc gaagggagaa
1921 aggcggacag gtatccggta agcggcaggg tcggaacagg agagcgcacg agggagcttc
1981 cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc
2041 gtcgattttt gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg
2101 cctttttacg gttcctggcc ttttgctggc cttttgctca catgttcttt cctgcgttat
2161 cccctgattc tgtggataac cgtattaccg cctttgagtg agctgatacc gctcgccgca
2221 gccgaacgac cgagcgcagc gagtcagtga gcgaggaagc ggaagagcgc ctgatgcggt
2281 attttctcct tacgcatctg tgcggtattt cacaccgcaa tggtgcactc tcagtacaat
2341 ctgctctgat gccgcatagt taagccagta tacactccgc tatcgctacg tgactgggtc
2401 atggctgcgc cccgacaccc gccaacaccc gctgacgcgc cctgacgggc ttgtctgctc
2461 ccggcatccg cttacagaca agctgtgacc gtctccggga gctgcatgtg tcagaggttt
2521 tcaccgtcat caccgaaacg cgcgaggcag ctgcggtaaa gctcatcagc gtggtcgtga
2581 agcgattcac agatgtctgc ctgttcatcc gcgtccagct cgttgagttt ctccagaagc
2641 gttaatgtct ggcttctgat aaagcgggcc atgttaaggg cggttttttc ctgtttggtc
2701 actgatgcct ccgtgtaagg gggatttctg ttcatggggg taatgatacc gatgaaacga
2761 gagaggatgc tcacgatacg ggttactgat gatgaacatg cccggttact ggaacgttgt
2821 gagggtaaac aactggcggt atggatgcgg cgggaccaga gaaaaatcac tcagggtcaa
2881 tgccagcgct tcgttaatac agatgtaggt gttccacagg gtagccagca gcatcctgcg
2941 atgcagatcc ggaacataat ggtgcagggc gctgacttcc gcgtttccag actttacgaa
3001 acacggaaac cgaagaccat tcatgttgtt gctcaggtcg cagacgtttt gcagcagcag
3061 tcgcttcacg ttcgctcgcg tatcggtgat tcattctgct aaccagtaag gcaaccccgc
3121 cagcctagcc gggtcctcaa cgacaggagc acgatcatgc gcacccgtgg ggccgccatg
3181 ccggcgataa tggcctgctt ctcgccgaaa cgtttggtgg cgggaccagt gacgaaggct
3241 tgagcgaggg cgtgcaagat tccgaatacc gcaagcgaca ggccgatcat cgtcgcgctc
3301 cagcgaaagc ggtcctcgcc gaaaatgacc cagagcgctg ccggcacctg tcctacgagt
3361 tgcatgataa agaagacagt cataagtgcg gcgacgatag tcatgccccg cgcccaccgg
3421 aaggagctga ctgggttgaa ggctctcaag ggcatcggtc gagatcccgg tgcctaatga
3481 gtgagctaac ttacattaat tgcgttgcgc tcactgcccg ctttccagtc gggaaacctg
3541 tcgtgccagc tgcattaatg aatcggccaa cgcgcgggga gaggcggttt gcgtattggg
3601 cgccagggtg gtttttcttt tcaccagtga gacgggcaac agctgattgc ccttcaccgc
3661 ctggccctga gagagttgca gcaagcggtc cacgctggtt tgccccagca ggcgaaaatc
3721 ctgtttgatg gtggttaacg gcgggatata acatgagctg tcttcggtat cgtcgtatcc
3781 cactaccgag atatccgcac caacgcgcag cccggactcg gtaatggcgc gcattgcgcc
3841 cagcgccatc tgatcgttgg caaccagcat cgcagtggga acgatgccct cattcagcat
3901 ttgcatggtt tgttgaaaac cggacatggc actccagtcg ccttcccgtt ccgctatcgg
3961 ctgaatttga ttgcgagtga gatatttatg ccagccagcc agacgcagac gcgccgagac
4021 agaacttaat gggcccgcta acagcgcgat ttgctggtga cccaatgcga ccagatgctc
4081 cacgcccagt cgcgtaccgt cttcatggga gaaaataata ctgttgatgg gtgtctggtc
4141 agagacatca agaaataacg ccggaacatt agtgcaggca gcttccacag caatggcatc
4201 ctggtcatcc agcggatagt taatgatcag cccactgacg cgttgcgcga gaagattgtg
4261 caccgccgct ttacaggctt cgacgccgct tcgttctacc atcgacacca ccacgctggc
4321 acccagttga tcggcgcgag atttaatcgc cgcgacaatt tgcgacggcg cgtgcagggc
4381 cagactggag gtggcaacgc caatcagcaa cgactgtttg cccgccagtt gttgtgccac
4441 gcggttggga atgtaattca gctccgccat cgccgcttcc actttttccc gcgttttcgc
4501 agaaacgtgg ctggcctggt tcaccacgcg ggaaacggtc tgataagaga caccggcata
4561 ctctgcgaca tcgtataacg ttactggttt cacattcacc accctgaatt gactctcttc
4621 cgggcgctat catgccatac cgcgaaaggt tttgcgccat tcgatggtgt ccgggatctc
4681 gacgctctcc cttatgcgac tcctgcatta ggaagcagcc cagtagtagg ttgaggccgt
4741 tgagcaccgc cgccgcaagg aatggtgcat gcaaggagat ggcgcccaac agtcccccgg
4801 ccacggggcc tgccaccata cccacgccga aacaagcgct catgagcccg aagtggcgag
4861 cccgatcttc cccatcggtg atgtcggcga tataggcgcc agcaaccgca cctgtggcgc
4921 cggtgatgcc ggccacgatg cgtccggcgt agaggatcga gatctcgatc ccgcgaaatt
4981 aatacgactc actatagggg aattgtgagc ggataacaat tccccactag taataatttt
5041 gtttaacttt aagaaggaga tataccatgg gcagcagctg gagccacccg cagttcgaaa
5101 aaagcagcgg taccggcagc ggcgaaaacc tttacttcca gggccatatg ctcgagctta
5161 agcaattggg atcctaatag tctagagcta gccctaggag atccggctgc taacaaagcc
5221 cgaaaggaag ctgagttggc tgctgccacc gctgagcaat aactagcata accccttggg
5281 gcctctaaac gggtcttgag gggttttttg ctgaaaggag gaactatatc cggat
//
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