pET100/D-TOPO

保存条件 ：-20℃

保质期 ：6个月

英文名 ：pET100/D-TOPO

库存 ：大量

供应商 ：钰博生物

规格 ：1ug/5ug

pET100/D-TOPO

载体基本信息

出品公司:	Ybscience
载体名称:	pET100 ； pET100/D-TOPO
质粒类型:	大肠杆菌表达载体；细菌表达载体；TOPO载体
高拷贝/低拷贝:	低拷贝
克隆方法:	TOPO
启动子:	T7
载体大小:	5764 bp
5' 测序引物及序列:	T7 promoter 5’TAATACGACTCACTATAGGG 3’
3' 测序引物及序列:	T7 terminator 5’GCTAGTTATTGCTCAGCGG 3’
载体标签:	6xHis tag、Xpress tag（N-term）；T7 tag （C-term）
载体抗性:	氨苄青霉素
克隆菌株:	TOP10
表达菌株:	BL21-Star(DE3) 、BL21-Star(DE3) pLysS
备注:	pET100含有T7聚合酶结合位点和RBS核糖体结合位点，可以做体外转录和翻译。
组成型/诱导型:	诱导型
病毒/非病毒:	非病毒

 

pET100载体质粒图谱和多克隆位点信息

 

pET100载体简介

 

pET100载体序列

LOCUS       pET100/D-TOPO               5764 bp ds-DNA     circular SYN 15-AUG-2016DEFINITION  synthetic circular DNA  ORGANISM  synthetic DNA constructREFERENCE   1  (bases 1 to 5764)  AUTHORS   Biofeng Lab.  TITLE     Direct SubmissionFEATURES             Location/Qualifiers     source          1..5764                     /organism="synthetic DNA construct"                     /mol_type="other DNA"     promoter        209..227                     /note="T7 promoter"                     /note="promoter for bacteriophage T7 RNA polymerase"     protein_bind    228..252                     /bound_moiety="lac repressor encoded by lacI"                     /note="lac operator"                     /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             282..288                     /note="efficient ribosome binding site from bacteriophage                      T7 gene 10 (Olins and Rangwala, 1989)"     misc_feature    297..299                     /note="ATG"                     /note="Initiation ATG:"     CDS             309..326                     /codon_start=1                     /product="6xHis affinity tag"                     /note="6xHis"                     /translation="HHHHHH"     CDS             330..362                     /codon_start=1                     /product="leader peptide from bacteriophage T7 gene 10                     T7 tag (gene 10 leader)"                     /note="T7 tag"                     /note="promotes efficient translation in E. coli"                     /translation="MASMTGGQQMG"     CDS             366..389                     /codon_start=1                     /product="Xpress(TM) epitope tag, including an enterokinase                     recognition and cleavage site"                     /note="Xpress(TM) tag"                     /translation="DLYDDDDK"     misc_feature    396..400                     /note="TOPO site1"                     /note="TOPO recognition site1"     misc_feature    405..409                     /note="TOPO site 2"                     /note="TOPO recognition site 2"     terminator      427..555                     /note="T7 terminator"                     /note="transcription terminator for bacteriophage T7 RNA                      polymerase"     promoter        complement(705..733)                     /note="tet promoter"                     /note="E. coli promoter for tetracycline efflux protein                      gene"     promoter        856..954                     /gene="bla"                     /note="bla promoter"     CDS             955..1815                     /codon_start=1                     /gene="bla"                     /product="beta-lactamase"                     /note="AmpR"                     /note="confers resistance to ampicillin, carbenicillin, and                     related antibiotics"                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW                     EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS                     LIKHW"     rep_origin      2022..2757                     /direction=RIGHT                     /note="pBR322"                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of                      replication"     misc_feature    2760..2899                     /note="bom"                     /note="basis of mobility region from pBR322"     CDS             complement(3001..3192)                     /codon_start=1                     /gene="rop"                     /product="Rop protein, which maintains plasmids at low copy                     number"                     /note="rop"                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA                     DELYRSCLARFGDDGENL"     CDS             complement(4507..5595)                     /codon_start=1                     /gene="lacI"                     /product="lac repressor"                     /note="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="MVNVKPVTLYDVAEYAGVSYQTVSRVVNQASHVSAKTREKVEAAM                     AELNYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSRADQLGASVVVSMVER                     SGVEACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSII                     FSHEDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREGD                     WSAMSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTED                     SSCYIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTA                     SPRALADSLMQLARQVSRLESGQ"     promoter        complement(5599..5664)                     /gene="lacI"                     /note="lacI promoter"ORIGIN        1 caaggagatg gcgcccaaca gtcccccggc cacggggcct gccaccatac ccacgccgaa       61 acaagcgctc atgagcccga agtggcgagc ccgatcttcc ccatcggtga tgtcggcgat      121 ataggcgcca gcaaccgcac ctgtggcgcc ggtgatgccg gccacgatgc gtccggcgta      181 gaggatcgag atctcgatcc cgcgaaatta atacgactca ctatagggga attgtgagcg      241 gataacaatt cccctctaga aataattttg tttaacttta agaaggagat atacatatgc      301 ggggttctca tcatcatcat catcatggta tggctagcat gactggtgga cagcaaatgg      361 gtcgggatct gtacgacgat gacgataagg atcatccctt caccaagggc gagctcaacg      421 atccggctgc taacaaagcc cgaaaggaag ctgagttggc tgctgccacc gctgagcaat      481 aactagcata accccttggg gcctctaaac gggtcttgag gagttttttg ctgaaaggag      541 gaactatatc cggatatccc gcaagaggcc cggcagtacc ggcataacca agcctatgcc      601 tacagcatcc agggtgacgg tgccgaggat gacgatgagc gcattgttag atttcataca      661 cggtgcctga ctgcgttagc aatttaactg tgataaacta ccgcattaaa gcttatcgat      721 gataagctgt caaacatgag aattaattct tgaagacgaa agggcctcgt gatacgccta      781 tttttatagg ttaatgtcat gataataatg gtttcttaga cgtcaggtgg cacttttcgg      841 ggaaatgtgc gcggaacccc tatttgttta tttttctaaa tacattcaaa tatgtatccg      901 ctcatgagac aataaccctg ataaatgctt caataatatt gaaaaaggaa gagtatgagt      961 attcaacatt tccgtgtcgc ccttattccc ttttttgcgg cattttgcct tcctgttttt     1021 gctcacccag aaacgctggt gaaagtaaaa gatgctgaag atcagttggg tgcacgagtg     1081 ggttacatcg aactggatct caacagcggt aagatccttg agagttttcg ccccgaagaa     1141 cgttttccaa tgatgagcac ttttaaagtt ctgctatgtg gcgcggtatt atcccgtgtt     1201 gacgccgggc aagagcaact cggtcgccgc atacactatt ctcagaatga cttggttgag     1261 tactcaccag tcacagaaaa gcatcttacg gatggcatga cagtaagaga attatgcagt     1321 gctgccataa ccatgagtga taacactgcg gccaacttac ttctgacaac gatcggagga     1381 ccgaaggagc taaccgcttt tttgcacaac atgggggatc atgtaactcg ccttgatcgt     1441 tgggaaccgg agctgaatga agccatacca aacgacgagc gtgacaccac gatgcctgca     1501 gcaatggcaa caacgttgcg caaactatta actggcgaac tacttactct agcttcccgg     1561 caacaattaa tagactggat ggaggcggat aaagttgcag gaccacttct gcgctcggcc     1621 cttccggctg gctggtttat tgctgataaa tctggagccg gtgagcgtgg gtctcgcggt     1681 atcattgcag cactggggcc agatggtaag ccctcccgta tcgtagttat ctacacgacg     1741 gggagtcagg caactatgga tgaacgaaat agacagatcg ctgagatagg tgcctcactg     1801 attaagcatt ggtaactgtc agaccaagtt tactcatata tactttagat tgatttaaaa     1861 cttcattttt aatttaaaag gatctaggtg aagatccttt ttgataatct catgaccaaa     1921 atcccttaac gtgagttttc gttccactga gcgtcagacc ccgtagaaaa gatcaaagga     1981 tcttcttgag atcctttttt tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg     2041 ctaccagcgg tggtttgttt gccggatcaa gagctaccaa ctctttttcc gaaggtaact     2101 ggcttcagca gagcgcagat accaaatact gtccttctag tgtagccgta gttaggccac     2161 cacttcaaga actctgtagc accgcctaca tacctcgctc tgctaatcct gttaccagtg     2221 gctgctgcca gtggcgataa gtcgtgtctt accgggttgg actcaagacg atagttaccg     2281 gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca cacagcccag cttggagcga     2341 acgacctaca ccgaactgag atacctacag cgtgagctat gagaaagcgc cacgcttccc     2401 gaagggagaa aggcggacag gtatccggta agcggcaggg tcggaacagg agagcgcacg     2461 agggagcttc cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc     2521 tgacttgagc gtcgattttt gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc     2581 agcaacgcgg cctttttacg gttcctggcc ttttgctggc cttttgctca catgttcttt     2641 cctgcgttat cccctgattc tgtggataac cgtattaccg cctttgagtg agctgatacc     2701 gctcgccgca gccgaacgac cgagcgcagc gagtcagtga gcgaggaagc ggaagagcgc     2761 ctgatgcggt attttctcct tacgcatctg tgcggtattt cacaccgcaa tggtgcactc     2821 tcagtacaat ctgctctgat gccgcatagt taagccagta tacactccgc tatcgctacg     2881 tgactgggtc atggctgcgc cccgacaccc gccaacaccc gctgacgcgc cctgacgggc     2941 ttgtctgctc ccggcatccg cttacagaca agctgtgacc gtctccggga gctgcatgtg     3001 tcagaggttt tcaccgtcat caccgaaacg cgcgaggcag ctgcggtaaa gctcatcagc     3061 gtggtcgtga agcgattcac agatgtctgc ctgttcatcc gcgtccagct cgttgagttt     3121 ctccagaagc gttaatgtct ggcttctgat aaagcgggcc atgttaaggg cggttttttc     3181 ctgtttggtc actgatgcct ccgtgtaagg gggatttctg ttcatggggg taatgatacc     3241 gatgaaacga gagaggatgc tcacgatacg ggttactgat gatgaacatg cccggttact     3301 ggaacgttgt gagggtaaac aactggcggt atggatgcgg cgggaccaga gaaaaatcac     3361 tcagggtcaa tgccagcgct tcgttaatac agatgtaggt gttccacagg gtagccagca     3421 gcatcctgcg atgcagatcc ggaacataat ggtgcagggc gctgacttcc gcgtttccag     3481 actttacgaa acacggaaac cgaagaccat tcatgttgtt gctcaggtcg cagacgtttt     3541 gcagcagcag tcgcttcacg ttcgctcgcg tatcggtgat tcattctgct aaccagtaag     3601 gcaaccccgc cagcctagcc gggtcctcaa cgacaggagc acgatcatgc gcacccgtgg     3661 ccaggaccca acgctgcccg agatgcgccg cgtgcggctg ctggagatgg cggacgcgat     3721 ggatatgttc tgccaagggt tggtttgcgc attcacagtt ctccgcaaga attgattggc     3781 tccaattctt ggagtggtga atccgttagc gaggtgccgc cggcttccat tcaggtcgag     3841 gtggcccggc tccatgcacc gcgacgcaac gcggggaggc agacaaggta tagggcggcg     3901 cctacaatcc atgccaaccc gttccatgtg ctcgccgagg cggcataaat cgccgtgacg     3961 atcagcggtc caatgatcga agttaggctg gtaagagccg cgagcgatcc ttgaagctgt     4021 ccctgatggt cgtcatctac ctgcctggac agcatggcct gcaacgcggg catcccgatg     4081 ccgccggaag cgagaagaat cataatgggg aaggccatcc agcctcgcgt cgcgaacgcc     4141 agcaagacgt agcccagcgc gtcggccgcc atgccggcga taatggcctg cttctcgccg     4201 aaacgtttgg tggcgggacc agtgacgaag gcttgagcga gggcgtgcaa gattccgaat     4261 accgcaagcg acaggccgat catcgtcgcg ctccagcgaa agcggtcctc gccgaaaatg     4321 acccagagcg ctgccggcac ctgtcctacg agttgcatga taaagaagac agtcataagt     4381 gcggcgacga tagtcatgcc ccgcgcccac cggaaggagc tgactgggtt gaaggctctc     4441 aagggcatcg gtcgagatcc cggtgcctaa tgagtgagct aacttacatt aattgcgttg     4501 cgctcactgc ccgctttcca gtcgggaaac ctgtcgtgcc agctgcatta atgaatcggc     4561 caacgcgcgg ggagaggcgg tttgcgtatt gggcgccagg gtggtttttc ttttcaccag     4621 tgagacgggc aacagctgat tgcccttcac cgcctggccc tgagagagtt gcagcaagcg     4681 gtccacgctg gtttgcccca gcaggcgaaa atcctgtttg atggtggtta acggcgggat     4741 ataacatgag ctgtcttcgg tatcgtcgta tcccactacc gagatatccg caccaacgcg     4801 cagcccggac tcggtaatgg cgcgcattgc gcccagcgcc atctgatcgt tggcaaccag     4861 catcgcagtg ggaacgatgc cctcattcag catttgcatg gtttgttgaa aaccggacat     4921 ggcactccag tcgccttccc gttccgctat cggctgaatt tgattgcgag tgagatattt     4981 atgccagcca gccagacgca gacgcgccga gacagaactt aatgggcccg ctaacagcgc     5041 gatttgctgg tgacccaatg cgaccagatg ctccacgccc agtcgcgtac cgtcttcatg     5101 ggagaaaata atactgttga tgggtgtctg gtcagagaca tcaagaaata acgccggaac     5161 attagtgcag gcagcttcca cagcaatggc atcctggtca tccagcggat agttaatgat     5221 cagcccactg acgcgttgcg cgagaagatt gtgcaccgcc gctttacagg cttcgacgcc     5281 gcttcgttct accatcgaca ccaccacgct ggcacccagt tgatcggcgc gagatttaat     5341 cgccgcgaca atttgcgacg gcgcgtgcag ggccagactg gaggtggcaa cgccaatcag     5401 caacgactgt ttgcccgcca gttgttgtgc cacgcggttg ggaatgtaat tcagctccgc     5461 catcgccgct tccacttttt cccgcgtttt cgcagaaacg tggctggcct ggttcaccac     5521 gcgggaaacg gtctgataag agacaccggc atactctgcg acatcgtata acgttactgg     5581 tttcacattc accaccctga attgactctc ttccgggcgc tatcatgcca taccgcgaaa     5641 ggttttgcgc cattcgatgg tgtccgggat ctcgacgctc tcccttatgc gactcctgca     5701 ttaggaagca gcccagtagt aggttgaggc cgttgagcac cgccgccgca aggaatggtg     5761 catg//