pCold TF - 91蜜桃视频

产物信息
基本信息
质粒介绍
质粒图谱
质粒序列

产物信息

产物货号	产物名称	产物规格	优惠价
XY0076	pCold TF	5μ驳	?请询价

使用说明

信裕质粒平台的各批次质粒菌株发货前均经过严格的多重验证，如存在质量问题，请在收到产物的三个月内通知我司。收到质粒后请短暂离心，加入20μl ddH2O溶解质粒，取2μl转化至对应感受态中，挑取单克隆重新提取质粒后使用。如需获取其他详细信息请登录我司官网查询。

基本信息

启动子:	CspA promoter
复制子:	ColE1 ori，f1 ori
质粒分类:	大肠系列质粒；大肠表达质粒；辫颁辞濒诲系列质粒
质粒大小:	5769bp
质粒标签:	N-His, N-Trigger factor, N-HRV 3C, N-Thrombin, N-Factor Xa
原核抗性:	氨苄青霉素础尘辫（100μ驳/尘濒）
克隆菌株:	顿贬5α等大肠杆菌
培养条件:	37℃，有氧，尝叠
表达宿主:	叠尝21等大肠杆菌
培养条件:	37℃，有氧，尝叠
诱导方式:	滨笔罢骋或乳糖及其类似物
5'测序引物:	辫颁辞濒诲-罢贵-贵1（颁颁础颁罢罢罢颁础础颁骋础骋颁罢骋础罢骋）
3'测序引物:	辫颁辞濒诲-搁（骋骋颁础骋骋骋础罢颁罢罢础骋础罢罢颁罢骋）
备注:	冷启动表达质粒

质粒介绍

Takara的pCold TF载体是一种融合表达可溶性的标签"触发因子（Trigger Factor,TF）伴侣"的冷休克载体。"触发因子"（48KD）是原核核糖体相关的分子伴侣蛋白（48 kDa），有利于新表达的多肽的共翻译折叠。正因为它来源于大肠杆菌，保证了触发因子能够在大肠杆菌表达系统中高表达。 pCold TF载体由CSPA启动子及其下游的序列组成，其下游序列主要包括包括5'非翻译区（5'非编码区），翻译增强元件（TEE），His标签序列，多克隆位点（MCS）。Lac操纵子插入在CSPA启动子下游，以确保严格的表达调控。此外，HRV 3C蛋白酶，凝血酶，和Factor Xa识别位点位于“Trigger Factor”和多克隆位点（MCS）之间，以便表达的融合蛋白上将标签去除。大多数大肠杆菌菌株都可以可以作为本载体的表达宿主。pCold TF载体提供冷休克诱导的方法高水平表达目的蛋白，再加上触发因子（伴侣）的表达来提高正确的蛋白质折叠，从而实现高效可溶性表达其他系统中难以对付的蛋白。
pColdTF质粒是利用冷休克基因CSPA启动子设计高效率蛋白质表达载体。在E.COLI启动子下游处，插入了lac操纵子，以便严格控制表达。此外，5’非翻译区（5’UTR），翻译增强元件（TEE）,His标签序列，Xa因子切割位点和多克隆位点（MCS）位于CAPS启动子的下游。当培养温度切换到低温时，大肠杆菌暂时停止生长，大部分的大肠杆菌蛋白质表达减少，但是一类称为冷休克蛋白的蛋白质被特异性诱导表达。

质粒图谱

质粒序列

LOCUS       Exported                5769 bp ds-DNA     circular SYN 13-SEP-2016
DEFINITION synthetic circular DNA
KEYWORDS    Untitled 9
SOURCE      synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE   1 (bases 1 to 5769)
TITLE     Direct Submission
JOURNAL   Exported Tuesday, September 13, 2016 from MiaoLingPlasmid
            http://www.miaolingbio.com
FEATURES             Location/Qualifiers
     source          1..5769
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     protein_bind    84..100
                     /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)."
     CDS             271..288
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /note="6xHis"
                     /translation="HHHHHH"
     CDS             1594..1617
                     /codon_start=1
                     /product="recognition and cleavage site for human
                     rhinovirus 3C and PreScission proteases"
                     /note="HRV 3C site"
                     /translation="LEVLFQGP"
     CDS             1627..1644
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /note="thrombin site"
                     /translation="LVPRGS"
     CDS             1651..1662
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /note="Factor Xa site"
                     /translation="IEGR"
     rep_origin      complement(2073..2528)
                     /direction=LEFT
                     /note="f1 ori"
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     promoter        2628..2732
                     /gene="bla"
                     /note="AmpR promoter"
     CDS             2733..3593
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /note="AmpR"
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     rep_origin      3764..4352
                     /direction=RIGHT
                     /note="ori"
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
     CDS             complement(4524..5606)
                     /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="MKPVTLYDVAEYAGVSYQTVSRVVNQASHVSAKTREKVEAAMAEL
                     NYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSRADQLGASVVVSMVERSGV
                     EACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSIIFSH
                     EDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREGDWSA
                     MSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTEDSSC
                     YIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR
                     ALADSLMQLARQVSRLESGQ"
     promoter        complement(5607..5684)
                     /gene="lacI"
                     /note="lacI promoter"
ORIGIN
        1 aaggaatggt gtggccgatt aatcataaat atgaaaaata attgttgcat cacccgccaa
       61 tgcgtggctt aatgcacatc aaattgtgag cggataacaa tttgatgtgc tagcgcatat
      121 ccagtgtagt aaggcaagtc ccttcaagag ttatcgttga tacccctcgt agtgcacatt
      181 cctttaacgc ttcaaaatct gtaaagcacg ccatatcgcc gaaaggcaca cttaattatt
      241 aagaggtaat acaccatgaa tcacaaagtg catcatcatc atcatcacat gcaagtttca
      301 gttgaaacca ctcaaggcct tggccgccgt gtaacgatta ctatcgctgc tgacagcatc
      361 gagaccgctg ttaaaagcga gctggtcaac gttgcgaaaa aagtacgtat tgacggcttc
      421 cgcaagggca aagtgccaat gaatatcgtt gctcagcgtt atggcgcgtc tgtacgccag
      481 gacgttctgg gtgacctgat gagccgtaac ttcattgacg ccatcattaa agaaaaaatc
      541 aatccggctg gcgcaccgac ttatgttccg ggcgaataca agctgggtga agacttcact
      601 tactctgtag agtttgaagt ttatccggaa gttgaactgc aaggtctgga agcgatcgaa
      661 gttgaaaaac cgatcgttga agtgaccgac gctgacgttg acggcatgct ggatactctg
      721 cgtaaacagc aggcgacctg gaaagaaaaa gacggcgctg ttgaagcaga agaccgcgtg
      781 accatcgact tcaccggttc tgtagacggc gaagagttcg aaggcggtaa agcgtctgat
      841 ttcgtactgg cgatgggcca gggtcgtatg atcccgggct ttgaagacgg tatcaaaggc
      901 cacaaagctg gcgaagagtt caccatcgac gtgaccttcc cggaagaata ccacgcagaa
      961 aacctgaaag gtaaagcagc gaaattcgct atcaacctga agaaagttga agagcgtgaa
     1021 ctgccggaac tgaccgcaga gttcatcaaa cgtttcggcg ttgaagatgg ttccgtagaa
     1081 ggtctgcgcg ctgaagtgcg taaaaacatg gagcgcgagc tgaagagcgc catccgtaac
     1141 cgcgttaagt ctcaggcgat cgaaggtctg gtaaaagcta acgacatcga cgtaccggct
     1201 gcgctgatcg acagcgaaat cgacgttctg cgtcgccagg ctgcacagcg tttcggtggc
     1261 aacgaaaaac aagctctgga actgccgcgc gaactgttcg aagaacaggc taaacgccgc
     1321 gtagttgttg gcctgctgct gggcgaagtt atccgcacca acgagctgaa agctgacgaa
     1381 gagcgcgtga aaggcctgat cgaagagatg gcttctgcgt acgaagatcc gaaagaagtt
     1441 atcgagttct acagcaaaaa caaagaactg atggacaaca tgcgcaatgt tgctctggaa
     1501 gaacaggctg ttgaagctgt actggcgaaa gcgaaagtga ctgaaaaaga aaccactttc
     1561 aacgagctga tgaaccagca ggcgtccgcg ggtctggaag ttctgttcca ggggccctcc
     1621 gcgggtctgg tgccacgcgg tagtggtggt atcgaaggta ggcatatgga gctcggtacc
     1681 ctcgagggat ccgaattcaa gcttgtcgac ctgcagtcta gataggtaat ctctgcttaa
     1741 aagcacagaa tctaagatcc ctgccatttg gcggggattt ttttatttgt tttcaggaaa
     1801 taaataatcg atcgcgtaat aaaatctatt attatttttg tgaagaataa atttgggtgc
     1861 aatgagaatg cgcaggccct ttcgtctcgc gcgtttcggt gatgacggtg aaaacctctg
     1921 acacatgcag ctcccggaga cggtcacagc ttgtctgtaa gcggatgccg ggagcagaca
     1981 agcccgtcag ggcgcgtcag cgggtgttgg cgggtgtcgg ggctggctta actatgcggc
     2041 atcagagcag attgtactga gagtgcacca taaaattgta aacgttaata ttttgttaaa
     2101 attcgcgtta aatttttgtt aaatcagctc attttttaac caataggccg aaatcggcaa
     2161 aatcccttat aaatcaaaag aatagcccga gatagggttg agtgttgttc cagtttggaa
     2221 caagagtcca ctattaaaga acgtggactc caacgtcaaa gggcgaaaaa ccgtctatca
     2281 gggcgatggc ccactacgtg aaccatcacc caaatcaagt tttttggggt cgaggtgccg
     2341 taaagcacta aatcggaacc ctaaagggag cccccgattt agagcttgac ggggaaagcc
     2401 ggcgaacgtg gcgagaaagg aagggaagaa agcgaaagga gcgggcgcta gggcgctggc
     2461 aagtgtagcg gtcacgctgc gcgtaaccac cacacccgcc gcgcttaatg cgccgctaca
     2521 gggcgcgtac tatggttgct ttgacgtatg cggtgtgaaa taccgcacag atgcgtaagg
     2581 agaaaatacc gcatcaggcg tcaggtggca cttttcgggg aaatgtgcgc ggaaccccta
     2641 tttgtttatt tttctaaata cattcaaata tgtatccgct catgagacaa taaccctgat
     2701 aaatgcttca ataatattga aaaaggaaga gtatgagtat tcaacatttc cgtgtcgccc
     2761 ttattccctt ttttgcggca ttttgccttc ctgtttttgc tcacccagaa acgctggtga
     2821 aagtaaaaga tgctgaagat cagttgggtg cacgagtggg ttacatcgaa ctggatctca
     2881 acagcggtaa gatccttgag agttttcgcc ccgaagaacg ttttccaatg atgagcactt
     2941 ttaaagttct gctatgtggc gcggtattat cccgtattga cgccgggcaa gagcaactcg
     3001 gtcgccgcat acactattct cagaatgact tggttgagta ctcaccagtc acagaaaagc
     3061 atcttacgga tggcatgaca gtaagagaat tatgcagtgc tgccataacc atgagtgata
     3121 acactgcggc caacttactt ctgacaacga tcggaggacc gaaggagcta accgcttttt
     3181 tgcacaacat gggggatcat gtaactcgcc ttgatcgttg ggaaccggag ctgaatgaag
     3241 ccataccaaa cgacgagcgt gacaccacga tgcctgtagc aatggcaaca acgttgcgca
     3301 aactattaac tggcgaacta cttactctag cttcccggca acaattaata gactggatgg
     3361 aggcggataa agttgcagga ccacttctgc gctcggccct tccggctggc tggtttattg
     3421 ctgataaatc tggagccggt gagcgtgggt ctcgcggtat cattgcagca ctggggccag
     3481 atggtaagcc ctcccgtatc gtagttatct acacgacggg gagtcaggca actatggatg
     3541 aacgaaatag acagatcgct gagataggtg cctcactgat taagcattgg taactgtcag
     3601 accaagttta ctcatatata ctttagattg atttaaaact tcatttttaa tttaaaagga
     3661 tctaggtgaa gatccttttt gataatctca tgaccaaaat cccttaacgt gagttttcgt
     3721 tccactgagc gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat cctttttttc
     3781 tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct accagcggtg gtttgtttgc
     3841 cggatcaaga gctaccaact ctttttccga aggtaactgg cttcagcaga gcgcagatac
     3901 caaatactgt tcttctagtg tagccgtagt taggccacca cttcaagaac tctgtagcac
     3961 cgcctacata cctcgctctg ctaatcctgt taccagtggc tgctgccagt ggcgataagt
     4021 cgtgtcttac cgggttggac tcaagacgat agttaccgga taaggcgcag cggtcgggct
     4081 gaacgggggg ttcgtgcaca cagcccagct tggagcgaac gacctacacc gaactgagat
     4141 acctacagcg tgagctatga gaaagcgcca cgcttcccga agggagaaag gcggacaggt
     4201 atccggtaag cggcagggtc ggaacaggag agcgcacgag ggagcttcca gggggaaacg
     4261 cctggtatct ttatagtcct gtcgggtttc gccacctctg acttgagcgt cgatttttgt
     4321 gatgctcgtc aggggggcgg agcctatgga aaaacgccag caacgcggcc tttttacggt
     4381 tcctggcctt ttgctggcct tttgctcaca tagtcatgcc ccgcgcccac cggaaggagc
     4441 tgactgggtt gaaggctctc aagggcatcg gtcgagatcc cggtgcctaa tgagtgagct
     4501 aacttacatt aattgcgttg cgctcactgc ccgctttcca gtcgggaaac ctgtcgtgcc
     4561 agctgcatta atgaatcggc caacgcgcgg ggagaggcgg tttgcgtatt gggcgccagg
     4621 gtggtttttc ttttcaccag tgagacgggc aacagctgat tgcccttcac cgcctggccc
     4681 tgagagagtt gcagcaagcg gtccacgctg gtttgcccca gcaggcgaaa atcctgtttg
     4741 atggtggtta acggcgggat ataacatgag ctgtcttcgg tatcgtcgta tcccactacc
     4801 gagatatccg caccaacgcg cagcccggac tcggtaatgg cgcgcattgc gcccagcgcc
     4861 atctgatcgt tggcaaccag catcgcagtg ggaacgatgc cctcattcag catttgcatg
     4921 gtttgttgaa aaccggacat ggcactccag tcgccttccc gttccgctat cggctgaatt
     4981 tgattgcgag tgagatattt atgccagcca gccagacgca gacgcgccga gacagaactt
     5041 aatgggcccg ctaacagcgc gatttgctgg tgacccaatg cgaccagatg ctccacgccc
     5101 agtcgcgtac cgtcttcatg ggagaaaata atactgttga tgggtgtctg gtcagagaca
     5161 tcaagaaata acgccggaac attagtgcag gcagcttcca cagcaatggc atcctggtca
     5221 tccagcggat agttaatgat cagcccactg acgcgttgcg cgagaagatt gtgcaccgcc
     5281 gctttacagg cttcgacgcc gcttcgttct accatcgaca ccaccacgct ggcacccagt
     5341 tgatcggcgc gagatttaat cgccgcgaca atttgcgacg gcgcgtgcag ggccagactg
     5401 gaggtggcaa cgccaatcag caacgactgt ttgcccgcca gttgttgtgc cacgcggttg
     5461 ggaatgtaat tcagctccgc catcgccgct tccacttttt cccgcgtttt cgcagaaacg
     5521 tggctggcct ggttcaccac gcgggaaacg gtctgataag agacaccggc atactctgcg
     5581 acatcgtata acgttactgg tttcacattc accaccctga attgactctc ttccgggcgc
     5641 tatcatgcca taccgcgaaa ggttttgcgc cattcgatgg tgtccgggat ctcgacgctc
     5701 tcccttatgc gactcctgca ttaggaagca gcccagtagt aggttgaggc cgttgagcac
     5761 cgccgccgc
//