pCold I

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pColdI pCold1载体基本信息

出品公司: Takara
载体名称: pCold I, pColdI, pCold1, pCold-I
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: 4407 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, N-Factor Xa
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: 3361
稳定性: 瞬时表达 Transient
组成型: 诱导表达
病毒/非病毒: 非病毒

pColdI pCold1载体质粒图谱和多克隆位点信息

pCold I载体图谱

pCold I多克隆位点

pColdI pCold1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。

pColdI pCold1载体序列

LOCUS       pCold\I                4407 bp    DNA     circular SYN 10-AUG-2004
DEFINITION  Expression vector pColdI DNA, complete sequence.
ACCESSION   AB186388
VERSION     AB186388.1  GI:51090247
KEYWORDS    .
SOURCE      Expression vector pColdI
  ORGANISM  Expression vector pColdI
            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..4407
                     /organism="Expression vector pColdI"
                     /mol_type="other DNA"
                     /db_xref="taxon:289229"
                     /lab_host="Escherichia coli"
                     /note="expresses an open reading frame as a factor Xa
                     cleavable fusion to a His6 containing translation
                     enhancing element. This plasmid is derived from pUC118."
     gene            15..512
                     /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    289..300
                     /gene="cspA"
                     /note="Factor Xa site"
     misc_feature    301..360
                     /gene="cspA"
                     /note="MCS = Multiple cloning sites contain the follow
                     restriction sites: NdeI, SacI, KpnI, XhoI, BamHI, EcoRI,
                     HindIII, SalI, PstI, XbaI"
     3'UTR           368..512
                     /gene="cspA"
                     /note="cspA 3'UTR"
     misc_feature    complement(711..1184)
                     /note="M13 intergenic region"
     gene            1371..2231
                     /gene="Amp"
     CDS             1371..2231
                     /gene="Amp"
                     /codon_start=1
                     /transl_table=11
                     /product="beta-lactamase"
                     /protein_id="BAD35134.1"
                     /db_xref="GI:51090248"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGY
                     IELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVE
                     YSPVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRL
                     DRWEPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPL
                     LRSALPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIA
                     EIGASLIKHW"
     rep_origin      2391..3005
                     /note="ColE1 origin"
     gene            complement(3162..4244)
                     /gene="lacI"
     CDS             complement(3162..4244)
                     /gene="lacI"
                     /codon_start=1
                     /transl_table=11
                     /product="lactose operon repressor"
                     /protein_id="BAD35135.1"
                     /db_xref="GI:51090249"
                     /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 cgaaggtagg
      301 catatggagc tcggtaccct cgagggatcc gaattcaagc ttgtcgacct gcagtctaga
      361 taggtaatct ctgcttaaaa gcacagaatc taagatccct gccatttggc ggggattttt
      421 ttatttgttt tcaggaaata aataatcgat cgcgtaataa aatctattat tatttttgtg
      481 aagaataaat ttgggtgcaa tgagaatgcg caggcccttt cgtctcgcgc gtttcggtga
      541 tgacggtgaa aacctctgac acatgcagct cccggagacg gtcacagctt gtctgtaagc
      601 ggatgccggg agcagacaag cccgtcaggg cgcgtcagcg ggtgttggcg ggtgtcgggg
      661 ctggcttaac tatgcggcat cagagcagat tgtactgaga gtgcaccata aaattgtaaa
      721 cgttaatatt ttgttaaaat tcgcgttaaa tttttgttaa atcagctcat tttttaacca
      781 ataggccgaa atcggcaaaa tcccttataa atcaaaagaa tagcccgaga tagggttgag
      841 tgttgttcca gtttggaaca agagtccact attaaagaac gtggactcca acgtcaaagg
      901 gcgaaaaacc gtctatcagg gcgatggccc actacgtgaa ccatcaccca aatcaagttt
      961 tttggggtcg aggtgccgta aagcactaaa tcggaaccct aaagggagcc cccgatttag
     1021 agcttgacgg ggaaagccgg cgaacgtggc gagaaaggaa gggaagaaag cgaaaggagc
     1081 gggcgctagg gcgctggcaa gtgtagcggt cacgctgcgc gtaaccacca cacccgccgc
     1141 gcttaatgcg ccgctacagg gcgcgtacta tggttgcttt gacgtatgcg gtgtgaaata
     1201 ccgcacagat gcgtaaggag aaaataccgc atcaggcgtc aggtggcact tttcggggaa
     1261 atgtgcgcgg aacccctatt tgtttatttt tctaaataca ttcaaatatg tatccgctca
     1321 tgagacaata accctgataa atgcttcaat aatattgaaa aaggaagagt atgagtattc
     1381 aacatttccg tgtcgccctt attccctttt ttgcggcatt ttgccttcct gtttttgctc
     1441 acccagaaac gctggtgaaa gtaaaagatg ctgaagatca gttgggtgca cgagtgggtt
     1501 acatcgaact ggatctcaac agcggtaaga tccttgagag ttttcgcccc gaagaacgtt
     1561 ttccaatgat gagcactttt aaagttctgc tatgtggcgc ggtattatcc cgtattgacg
     1621 ccgggcaaga gcaactcggt cgccgcatac actattctca gaatgacttg gttgagtact
     1681 caccagtcac agaaaagcat cttacggatg gcatgacagt aagagaatta tgcagtgctg
     1741 ccataaccat gagtgataac actgcggcca acttacttct gacaacgatc ggaggaccga
     1801 aggagctaac cgcttttttg cacaacatgg gggatcatgt aactcgcctt gatcgttggg
     1861 aaccggagct gaatgaagcc ataccaaacg acgagcgtga caccacgatg cctgtagcaa
     1921 tggcaacaac gttgcgcaaa ctattaactg gcgaactact tactctagct tcccggcaac
     1981 aattaataga ctggatggag gcggataaag ttgcaggacc acttctgcgc tcggcccttc
     2041 cggctggctg gtttattgct gataaatctg gagccggtga gcgtgggtct cgcggtatca
     2101 ttgcagcact ggggccagat ggtaagccct cccgtatcgt agttatctac acgacgggga
     2161 gtcaggcaac tatggatgaa cgaaatagac agatcgctga gataggtgcc tcactgatta
     2221 agcattggta actgtcagac caagtttact catatatact ttagattgat ttaaaacttc
     2281 atttttaatt taaaaggatc taggtgaaga tcctttttga taatctcatg accaaaatcc
     2341 cttaacgtga gttttcgttc cactgagcgt cagaccccgt agaaaagatc aaaggatctt
     2401 cttgagatcc tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac
     2461 cagcggtggt ttgtttgccg gatcaagagc taccaactct ttttccgaag gtaactggct
     2521 tcagcagagc gcagatacca aatactgttc ttctagtgta gccgtagtta ggccaccact
     2581 tcaagaactc tgtagcaccg cctacatacc tcgctctgct aatcctgtta ccagtggctg
     2641 ctgccagtgg cgataagtcg tgtcttaccg ggttggactc aagacgatag ttaccggata
     2701 aggcgcagcg gtcgggctga acggggggtt cgtgcacaca gcccagcttg gagcgaacga
     2761 cctacaccga actgagatac ctacagcgtg agctatgaga aagcgccacg cttcccgaag
     2821 ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg
     2881 agcttccagg gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc cacctctgac
     2941 ttgagcgtcg atttttgtga tgctcgtcag gggggcggag cctatggaaa aacgccagca
     3001 acgcggcctt tttacggttc ctggcctttt gctggccttt tgctcacata gtcatgcccc
     3061 gcgcccaccg gaaggagctg actgggttga aggctctcaa gggcatcggt cgagatcccg
     3121 gtgcctaatg agtgagctaa cttacattaa ttgcgttgcg ctcactgccc gctttccagt
     3181 cgggaaacct gtcgtgccag ctgcattaat gaatcggcca acgcgcgggg agaggcggtt
     3241 tgcgtattgg gcgccagggt ggtttttctt ttcaccagtg agacgggcaa cagctgattg
     3301 cccttcaccg cctggccctg agagagttgc agcaagcggt ccacgctggt ttgccccagc
     3361 aggcgaaaat cctgtttgat ggtggttaac ggcgggatat aacatgagct gtcttcggta
     3421 tcgtcgtatc ccactaccga gatatccgca ccaacgcgca gcccggactc ggtaatggcg
     3481 cgcattgcgc ccagcgccat ctgatcgttg gcaaccagca tcgcagtggg aacgatgccc
     3541 tcattcagca tttgcatggt ttgttgaaaa ccggacatgg cactccagtc gccttcccgt
     3601 tccgctatcg gctgaatttg attgcgagtg agatatttat gccagccagc cagacgcaga
     3661 cgcgccgaga cagaacttaa tgggcccgct aacagcgcga tttgctggtg acccaatgcg
     3721 accagatgct ccacgcccag tcgcgtaccg tcttcatggg agaaaataat actgttgatg
     3781 ggtgtctggt cagagacatc aagaaataac gccggaacat tagtgcaggc agcttccaca
     3841 gcaatggcat cctggtcatc cagcggatag ttaatgatca gcccactgac gcgttgcgcg
     3901 agaagattgt gcaccgccgc tttacaggct tcgacgccgc ttcgttctac catcgacacc
     3961 accacgctgg cacccagttg atcggcgcga gatttaatcg ccgcgacaat ttgcgacggc
     4021 gcgtgcaggg ccagactgga ggtggcaacg ccaatcagca acgactgttt gcccgccagt
     4081 tgttgtgcca cgcggttggg aatgtaattc agctccgcca tcgccgcttc cactttttcc
     4141 cgcgttttcg cagaaacgtg gctggcctgg ttcaccacgc gggaaacggt ctgataagag
     4201 acaccggcat actctgcgac atcgtataac gttactggtt tcacattcac caccctgaat
     4261 tgactctctt ccgggcgcta tcatgccata ccgcgaaagg ttttgcgcca ttcgatggtg
     4321 tccgggatct cgacgctctc ccttatgcga ctcctgcatt aggaagcagc ccagtagtag
     4381 gttgaggccg ttgagcaccg ccgccgc
//

pColdI pCold1载体质粒图谱pdf版和相关资料下载

pColdI pCold1载体应用举例

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