pCold II

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pColdII载体基本信息

出品公司: Takara
载体名称: pCold II, pColdII, pCold-II
质粒类型: 大肠杆菌蛋白表达载体
表达水平:
启动子: CSPA
克隆方法: 多克隆位点,限制性内切酶
载体大小: 4392 bp
5' 测序引物及序列: pCold-F: 5'-ACGCCATATCGCCGAAAGG-3'
3' 测序引物及序列: pCold-R: 5'-GGCAGGGATCTTAGATTCTG-3'
载体标签: N-His,
载体抗性: Ampicillin 氨苄
备注:
温度诱导,冷休克蛋白表达载体
产品目录号: 3362
稳定性: 瞬时表达 Transient
组成型: 诱导表达
病毒/非病毒: 非病毒

pColdII载体质粒图谱和多克隆位点信息

pCold II载体图谱

pCold II多克隆位点

pColdII载体简介

阐明蛋白质的结构和功能是后基因组研究的一个重要课题。高效率的蛋白质表达系统是研究这些课题的不可缺少的基本技术。大肠杆菌作为宿主的表达系统中被广泛 使用于生产重组蛋白。大肠杆菌表达系统具有良好的优势,方法简单,成本低。然而,对于某些基因,蛋白表达很困难,表达的蛋白质不溶。

为了解决这些问题,TAKARA与Masayori Inouye教授(美国新泽西医学和牙科大学)进行联合研究,以制定一个高效基于低温冷休克蛋白基因的大肠杆菌表达载体。pCold DNA系列产品具有上述的优点,并提供了一个重要功能和结构的分析以及其他蛋白质研究领域的工具。

<产品概况和特点>
当培养的大肠杆菌的温度充分降低,几乎暂时停止生长大部分的蛋白表达减少,而此时一组蛋白质称为“冷休克蛋白”被专门诱导表达。

冷休克表达载体,pCold I-IV,是利用冷休克基因CSPA启动子设计高效率蛋白质表达载体。 CSPA的启动子的下游处,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)位于在CSPA启动子的下游的。由于该产品采用了大肠杆菌的启动子,大多数大肠杆菌菌株可被用作表达宿主。有四种类型的 pCold载体,他们的下列原件的位置稍有不同,TEE,His标签序列和Factor XA酶切位点。

pColdII载体序列

LOCUS       pCold\II                4392 bp    DNA     circular SYN 10-AUG-2004
DEFINITION  Expression vector pColdII DNA, complete sequence.
ACCESSION   AB186389
VERSION     AB186389.1  GI:51090250
KEYWORDS    .
SOURCE      Expression vector pColdII
  ORGANISM  Expression vector pColdII
            other sequences; artificial sequences; vectors.
COMMENT     This file is created by Vector NTI
            http://www.biofeng.com/
COMMENT     VNTAUTHORNAME|biofeng.com|
FEATURES             Location/Qualifiers
     source          1..4392
                     /organism="Expression vector pColdII"
                     /mol_type="other DNA"
                     /db_xref="taxon:289230"
                     /lab_host="Escherichia coli"
                     /note="expresses an open reading frame as a fusion to a
                     His6 containing translation enhancing element. This
                     plasmid is derived from pUC118."
     gene            15..497
                     /gene="cspA"
     promoter        15..81
                     /gene="cspA"
     misc_feature    82..121
                     /gene="cspA"
                     /note="synthetic lac operator sequence"
     5'UTR           122..255
                     /gene="cspA"
                     /note="modified cspA 5'UTR"
     misc_feature    256..270
                     /gene="cspA"
                     /note="TEE (translation enhancing element)"
     misc_feature    271..288
                     /gene="cspA"
                     /note="His6 tag"
     misc_feature    286..345
                     /gene="cspA"
                     /note="MCS = Multiple cloning sites contain the follow
                     restriction sites: NdeI, SacI, KpnI, XhoI, BamHI, EcoRI,
                     HindIII, SalI, PstI, XbaI"
     3'UTR           353..497
                     /gene="cspA"
                     /note="cspA 3'UTR"
     misc_feature    complement(696..1169)
                     /note="M13 intergenic region"
     gene            1356..2216
                     /gene="Amp"
     CDS             1356..2216
                     /gene="Amp"
                     /codon_start=1
                     /transl_table=11
                     /product="beta-lactamase"
                     /protein_id="BAD35136.1"
                     /db_xref="GI:51090251"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGY
                     IELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVE
                     YSPVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRL
                     DRWEPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPL
                     LRSALPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIA
                     EIGASLIKHW"
     rep_origin      2376..2990
                     /note="ColE1 origin"
     gene            complement(3147..4229)
                     /gene="lacI"
     CDS             complement(3147..4229)
                     /gene="lacI"
                     /codon_start=1
                     /transl_table=11
                     /product="lactose operon repressor"
                     /protein_id="BAD35137.1"
                     /db_xref="GI:51090252"
                     /translation="MKPVTLYDVAEYAGVSYQTVSRVVNQASHVSAKTREKVEAAMAE
                     LNYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSRADQLGASVVVSMVERS
                     GVEACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSII
                     FSHEDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREG
                     DWSAMSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDT
                     EDSSCYIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNT
                     QTASPRALADSLMQLARQVSRLESGQ"
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 atcatcatat ggagctcggt
      301 accctcgagg gatccgaatt caagcttgtc gacctgcagt ctagataggt aatctctgct
      361 taaaagcaca gaatctaaga tccctgccat ttggcgggga tttttttatt tgttttcagg
      421 aaataaataa tcgatcgcgt aataaaatct attattattt ttgtgaagaa taaatttggg
      481 tgcaatgaga atgcgcaggc cctttcgtct cgcgcgtttc ggtgatgacg gtgaaaacct
      541 ctgacacatg cagctcccgg agacggtcac agcttgtctg taagcggatg ccgggagcag
      601 acaagcccgt cagggcgcgt cagcgggtgt tggcgggtgt cggggctggc ttaactatgc
      661 ggcatcagag cagattgtac tgagagtgca ccataaaatt gtaaacgtta atattttgtt
      721 aaaattcgcg ttaaattttt gttaaatcag ctcatttttt aaccaatagg ccgaaatcgg
      781 caaaatccct tataaatcaa aagaatagcc cgagataggg ttgagtgttg ttccagtttg
      841 gaacaagagt ccactattaa agaacgtgga ctccaacgtc aaagggcgaa aaaccgtcta
      901 tcagggcgat ggcccactac gtgaaccatc acccaaatca agttttttgg ggtcgaggtg
      961 ccgtaaagca ctaaatcgga accctaaagg gagcccccga tttagagctt gacggggaaa
     1021 gccggcgaac gtggcgagaa aggaagggaa gaaagcgaaa ggagcgggcg ctagggcgct
     1081 ggcaagtgta gcggtcacgc tgcgcgtaac caccacaccc gccgcgctta atgcgccgct
     1141 acagggcgcg tactatggtt gctttgacgt atgcggtgtg aaataccgca cagatgcgta
     1201 aggagaaaat accgcatcag gcgtcaggtg gcacttttcg gggaaatgtg cgcggaaccc
     1261 ctatttgttt atttttctaa atacattcaa atatgtatcc gctcatgaga caataaccct
     1321 gataaatgct tcaataatat tgaaaaagga agagtatgag tattcaacat ttccgtgtcg
     1381 cccttattcc cttttttgcg gcattttgcc ttcctgtttt tgctcaccca gaaacgctgg
     1441 tgaaagtaaa agatgctgaa gatcagttgg gtgcacgagt gggttacatc gaactggatc
     1501 tcaacagcgg taagatcctt gagagttttc gccccgaaga acgttttcca atgatgagca
     1561 cttttaaagt tctgctatgt ggcgcggtat tatcccgtat tgacgccggg caagagcaac
     1621 tcggtcgccg catacactat tctcagaatg acttggttga gtactcacca gtcacagaaa
     1681 agcatcttac ggatggcatg acagtaagag aattatgcag tgctgccata accatgagtg
     1741 ataacactgc ggccaactta cttctgacaa cgatcggagg accgaaggag ctaaccgctt
     1801 ttttgcacaa catgggggat catgtaactc gccttgatcg ttgggaaccg gagctgaatg
     1861 aagccatacc aaacgacgag cgtgacacca cgatgcctgt agcaatggca acaacgttgc
     1921 gcaaactatt aactggcgaa ctacttactc tagcttcccg gcaacaatta atagactgga
     1981 tggaggcgga taaagttgca ggaccacttc tgcgctcggc ccttccggct ggctggttta
     2041 ttgctgataa atctggagcc ggtgagcgtg ggtctcgcgg tatcattgca gcactggggc
     2101 cagatggtaa gccctcccgt atcgtagtta tctacacgac ggggagtcag gcaactatgg
     2161 atgaacgaaa tagacagatc gctgagatag gtgcctcact gattaagcat tggtaactgt
     2221 cagaccaagt ttactcatat atactttaga ttgatttaaa acttcatttt taatttaaaa
     2281 ggatctaggt gaagatcctt tttgataatc tcatgaccaa aatcccttaa cgtgagtttt
     2341 cgttccactg agcgtcagac cccgtagaaa agatcaaagg atcttcttga gatccttttt
     2401 ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt
     2461 tgccggatca agagctacca actctttttc cgaaggtaac tggcttcagc agagcgcaga
     2521 taccaaatac tgttcttcta gtgtagccgt agttaggcca ccacttcaag aactctgtag
     2581 caccgcctac atacctcgct ctgctaatcc tgttaccagt ggctgctgcc agtggcgata
     2641 agtcgtgtct taccgggttg gactcaagac gatagttacc ggataaggcg cagcggtcgg
     2701 gctgaacggg gggttcgtgc acacagccca gcttggagcg aacgacctac accgaactga
     2761 gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca
     2821 ggtatccggt aagcggcagg gtcggaacag gagagcgcac gagggagctt ccagggggaa
     2881 acgcctggta tctttatagt cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt
     2941 tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg gcctttttac
     3001 ggttcctggc cttttgctgg ccttttgctc acatagtcat gccccgcgcc caccggaagg
     3061 agctgactgg gttgaaggct ctcaagggca tcggtcgaga tcccggtgcc taatgagtga
     3121 gctaacttac attaattgcg ttgcgctcac tgcccgcttt ccagtcggga aacctgtcgt
     3181 gccagctgca ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt attgggcgcc
     3241 agggtggttt ttcttttcac cagtgagacg ggcaacagct gattgccctt caccgcctgg
     3301 ccctgagaga gttgcagcaa gcggtccacg ctggtttgcc ccagcaggcg aaaatcctgt
     3361 ttgatggtgg ttaacggcgg gatataacat gagctgtctt cggtatcgtc gtatcccact
     3421 accgagatat ccgcaccaac gcgcagcccg gactcggtaa tggcgcgcat tgcgcccagc
     3481 gccatctgat cgttggcaac cagcatcgca gtgggaacga tgccctcatt cagcatttgc
     3541 atggtttgtt gaaaaccgga catggcactc cagtcgcctt cccgttccgc tatcggctga
     3601 atttgattgc gagtgagata tttatgccag ccagccagac gcagacgcgc cgagacagaa
     3661 cttaatgggc ccgctaacag cgcgatttgc tggtgaccca atgcgaccag atgctccacg
     3721 cccagtcgcg taccgtcttc atgggagaaa ataatactgt tgatgggtgt ctggtcagag
     3781 acatcaagaa ataacgccgg aacattagtg caggcagctt ccacagcaat ggcatcctgg
     3841 tcatccagcg gatagttaat gatcagccca ctgacgcgtt gcgcgagaag attgtgcacc
     3901 gccgctttac aggcttcgac gccgcttcgt tctaccatcg acaccaccac gctggcaccc
     3961 agttgatcgg cgcgagattt aatcgccgcg acaatttgcg acggcgcgtg cagggccaga
     4021 ctggaggtgg caacgccaat cagcaacgac tgtttgcccg ccagttgttg tgccacgcgg
     4081 ttgggaatgt aattcagctc cgccatcgcc gcttccactt tttcccgcgt tttcgcagaa
     4141 acgtggctgg cctggttcac cacgcgggaa acggtctgat aagagacacc ggcatactct
     4201 gcgacatcgt ataacgttac tggtttcaca ttcaccaccc tgaattgact ctcttccggg
     4261 cgctatcatg ccataccgcg aaaggttttg cgccattcga tggtgtccgg gatctcgacg
     4321 ctctccctta tgcgactcct gcattaggaa gcagcccagt agtaggttga ggccgttgag
     4381 caccgccgcc gc
//

pColdII载体质粒图谱pdf版和相关资料下载

pColdII载体应用举例

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