pCold IV

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
XY0075	pCold IV	20μ濒	?请加蚕蚕询价

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20μ濒 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
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:	4359bp
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:	pCold-F: 5-ACGCCATATCGCCGAAAGG-3
3'测序引物:	pCold-R: 5-GGCAGGGATCTTAGATTCTG-3
备注:	冷休克蛋白表达载体

质粒介绍

pCold IV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                4359 bp ds-DNA     circular SYN 13-SEP-2016
DEFINITION synthetic circular DNA
KEYWORDS    Untitled 8
SOURCE      synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE   1 (bases 1 to 4359)
TITLE     Direct Submission
JOURNAL   Exported Tuesday, September 13, 2016 from MiaoLingPlasmid
            http://www.miaolingbio.com
FEATURES             Location/Qualifiers
     source          1..4359
                     /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)."
     rep_origin      complement(663..1118)
                     /direction=LEFT
                     /note="f1 ori"
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     promoter        1218..1322
                     /gene="bla"
                     /note="AmpR promoter"
     CDS             1323..2183
                     /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      2354..2942
                     /direction=RIGHT
                     /note="ori"
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
     CDS             complement(3114..4196)
                     /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(4197..4274)
                     /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 accatatgga gctcggtacc ctcgagggat ccgaattcaa gcttgtcgac
      301 ctgcagtcta gataggtaat ctctgcttaa aagcacagaa tctaagatcc ctgccatttg
      361 gcggggattt ttttatttgt tttcaggaaa taaataatcg atcgcgtaat aaaatctatt
      421 attatttttg tgaagaataa atttgggtgc aatgagaatg cgcaggccct ttcgtctcgc
      481 gcgtttcggt gatgacggtg aaaacctctg acacatgcag ctcccggaga cggtcacagc
      541 ttgtctgtaa gcggatgccg ggagcagaca agcccgtcag ggcgcgtcag cgggtgttgg
      601 cgggtgtcgg ggctggctta actatgcggc atcagagcag attgtactga gagtgcacca
      661 taaaattgta aacgttaata ttttgttaaa attcgcgtta aatttttgtt aaatcagctc
      721 attttttaac caataggccg aaatcggcaa aatcccttat aaatcaaaag aatagcccga
      781 gatagggttg agtgttgttc cagtttggaa caagagtcca ctattaaaga acgtggactc
      841 caacgtcaaa gggcgaaaaa ccgtctatca gggcgatggc ccactacgtg aaccatcacc
      901 caaatcaagt tttttggggt cgaggtgccg taaagcacta aatcggaacc ctaaagggag
      961 cccccgattt agagcttgac ggggaaagcc ggcgaacgtg gcgagaaagg aagggaagaa
     1021 agcgaaagga gcgggcgcta gggcgctggc aagtgtagcg gtcacgctgc gcgtaaccac
     1081 cacacccgcc gcgcttaatg cgccgctaca gggcgcgtac tatggttgct ttgacgtatg
     1141 cggtgtgaaa taccgcacag atgcgtaagg agaaaatacc gcatcaggcg tcaggtggca
     1201 cttttcgggg aaatgtgcgc ggaaccccta tttgtttatt tttctaaata cattcaaata
     1261 tgtatccgct catgagacaa taaccctgat aaatgcttca ataatattga aaaaggaaga
     1321 gtatgagtat tcaacatttc cgtgtcgccc ttattccctt ttttgcggca ttttgccttc
     1381 ctgtttttgc tcacccagaa acgctggtga aagtaaaaga tgctgaagat cagttgggtg
     1441 cacgagtggg ttacatcgaa ctggatctca acagcggtaa gatccttgag agttttcgcc
     1501 ccgaagaacg ttttccaatg atgagcactt ttaaagttct gctatgtggc gcggtattat
     1561 cccgtattga cgccgggcaa gagcaactcg gtcgccgcat acactattct cagaatgact
     1621 tggttgagta ctcaccagtc acagaaaagc atcttacgga tggcatgaca gtaagagaat
     1681 tatgcagtgc tgccataacc atgagtgata acactgcggc caacttactt ctgacaacga
     1741 tcggaggacc gaaggagcta accgcttttt tgcacaacat gggggatcat gtaactcgcc
     1801 ttgatcgttg ggaaccggag ctgaatgaag ccataccaaa cgacgagcgt gacaccacga
     1861 tgcctgtagc aatggcaaca acgttgcgca aactattaac tggcgaacta cttactctag
     1921 cttcccggca acaattaata gactggatgg aggcggataa agttgcagga ccacttctgc
     1981 gctcggccct tccggctggc tggtttattg ctgataaatc tggagccggt gagcgtgggt
     2041 ctcgcggtat cattgcagca ctggggccag atggtaagcc ctcccgtatc gtagttatct
     2101 acacgacggg gagtcaggca actatggatg aacgaaatag acagatcgct gagataggtg
     2161 cctcactgat taagcattgg taactgtcag accaagttta ctcatatata ctttagattg
     2221 atttaaaact tcatttttaa tttaaaagga tctaggtgaa gatccttttt gataatctca
     2281 tgaccaaaat cccttaacgt gagttttcgt tccactgagc gtcagacccc gtagaaaaga
     2341 tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa
     2401 aaccaccgct accagcggtg gtttgtttgc cggatcaaga gctaccaact ctttttccga
     2461 aggtaactgg cttcagcaga gcgcagatac caaatactgt tcttctagtg tagccgtagt
     2521 taggccacca cttcaagaac tctgtagcac cgcctacata cctcgctctg ctaatcctgt
     2581 taccagtggc tgctgccagt ggcgataagt cgtgtcttac cgggttggac tcaagacgat
     2641 agttaccgga taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca cagcccagct
     2701 tggagcgaac gacctacacc gaactgagat acctacagcg tgagctatga gaaagcgcca
     2761 cgcttcccga agggagaaag gcggacaggt atccggtaag cggcagggtc ggaacaggag
     2821 agcgcacgag ggagcttcca gggggaaacg cctggtatct ttatagtcct gtcgggtttc
     2881 gccacctctg acttgagcgt cgatttttgt gatgctcgtc aggggggcgg agcctatgga
     2941 aaaacgccag caacgcggcc tttttacggt tcctggcctt ttgctggcct tttgctcaca
     3001 tagtcatgcc ccgcgcccac cggaaggagc tgactgggtt gaaggctctc aagggcatcg
     3061 gtcgagatcc cggtgcctaa tgagtgagct aacttacatt aattgcgttg cgctcactgc
     3121 ccgctttcca gtcgggaaac ctgtcgtgcc agctgcatta atgaatcggc caacgcgcgg
     3181 ggagaggcgg tttgcgtatt gggcgccagg gtggtttttc ttttcaccag tgagacgggc
     3241 aacagctgat tgcccttcac cgcctggccc tgagagagtt gcagcaagcg gtccacgctg
     3301 gtttgcccca gcaggcgaaa atcctgtttg atggtggtta acggcgggat ataacatgag
     3361 ctgtcttcgg tatcgtcgta tcccactacc gagatatccg caccaacgcg cagcccggac
     3421 tcggtaatgg cgcgcattgc gcccagcgcc atctgatcgt tggcaaccag catcgcagtg
     3481 ggaacgatgc cctcattcag catttgcatg gtttgttgaa aaccggacat ggcactccag
     3541 tcgccttccc gttccgctat cggctgaatt tgattgcgag tgagatattt atgccagcca
     3601 gccagacgca gacgcgccga gacagaactt aatgggcccg ctaacagcgc gatttgctgg
     3661 tgacccaatg cgaccagatg ctccacgccc agtcgcgtac cgtcttcatg ggagaaaata
     3721 atactgttga tgggtgtctg gtcagagaca tcaagaaata acgccggaac attagtgcag
     3781 gcagcttcca cagcaatggc atcctggtca tccagcggat agttaatgat cagcccactg
     3841 acgcgttgcg cgagaagatt gtgcaccgcc gctttacagg cttcgacgcc gcttcgttct
     3901 accatcgaca ccaccacgct ggcacccagt tgatcggcgc gagatttaat cgccgcgaca
     3961 atttgcgacg gcgcgtgcag ggccagactg gaggtggcaa cgccaatcag caacgactgt
     4021 ttgcccgcca gttgttgtgc cacgcggttg ggaatgtaat tcagctccgc catcgccgct
     4081 tccacttttt cccgcgtttt cgcagaaacg tggctggcct ggttcaccac gcgggaaacg
     4141 gtctgataag agacaccggc atactctgcg acatcgtata acgttactgg tttcacattc
     4201 accaccctga attgactctc ttccgggcgc tatcatgcca taccgcgaaa ggttttgcgc
     4261 cattcgatgg tgtccgggat ctcgacgctc tcccttatgc gactcctgca ttaggaagca
     4321 gcccagtagt aggttgaggc cgttgagcac cgccgccgc
//

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