pCold-SUMO - 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
XY0077	pCold-SUMO	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:	4740bp
质粒标签:	N-His, N-SUMO
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-贵（础颁骋颁颁础罢础罢颁骋颁颁骋础础础骋骋）
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:	冷休克表达载体

质粒介绍

原核表达载体(pCold-SUMO)是在 pCold 载体基础上改造而成，该载体启动子(CS Promoter)来源于南极嗜冷**，在低温下(15 度)才能启动蛋白的表达。低温下**生长缓慢，使得蛋白合成速度减慢，从而*大限度的提高了蛋白正确折叠的几率，提高了蛋白可溶性表达，增强了活性蛋白的表达比率。该表达系统所包含的 SUMO tag可以极大地提高小分子量蛋白的表达量，而且进一步提高了蛋白的可溶表达几率。同时，试剂盒配备的SUMO Protease(含6×His标签)可特异性去除 SUMO tag，从而得到不含任何标签的重组蛋白。TEE信号肽可增强冷启动子调控下目的蛋白的高表达。BL21(DE3)Chaprone E.coli **中含有分子伴侣蛋白质粒，其表达产物可协助重组蛋白正确折叠形成可溶活性蛋白。分子伴侣蛋白质粒为氯霉素抗性（chloramphenicol，Cmr）的表达受控于四环素（tetR）操纵子。
pCold-SUMO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                4740 bp ds-DNA     circular SYN 22-JUN-2017
DEFINITION synthetic circular DNA
KEYWORDS    pCold-SUMO
SOURCE      synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE   1 (bases 1 to 4740)
TITLE     Direct Submission
JOURNAL   Exported Thursday, June 22, 2017 from MiaoLingPlasmid
            http://www.miaolingbio.com
FEATURES             Location/Qualifiers
     source          1..4740
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     misc_feature    15..81
                     /note="cspA promoter"
     misc_feature    82..121
                     /note="lac operator"
     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)."
     misc_feature    122..255
                     /note="cspA 5'UTR"
     misc_feature    243..246
                     /note="SD"
     misc_feature    256..270
                     /note="TEE"
     CDS             271..288
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /note="6xHis"
                     /translation="HHHHHH"
     CDS             289..300
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /note="Factor Xa site"
                     /translation="IEGR"
     CDS             340..633
                     /codon_start=1
                     /gene="S. cerevisiae SMT3 (truncated)"
                     /product="cleavable ubiquitin-like protein tag"
                     /note="SUMO"
                     /translation="MSDSEVNQEAKPEVKPEVKPETHINLKVSDGSSEIFFKIKKTTPL
                     RRLMEAFAKRQGKEMDSLRFLYDGIRIQADQTPEDLDMEDNDIIEAHREQIGG"
     misc_feature    634..693
                     /note="mcs"
     misc_feature    701..845
                     /note="cspA 3'UTR"
     misc_feature    729..746
                     /note="transcription terminator"
     rep_origin      complement(1044..1499)
                     /direction=LEFT
                     /note="f1 ori"
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     promoter        1599..1703
                     /gene="bla"
                     /note="AmpR promoter"
     CDS             1704..2564
                     /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      2735..3323
                     /direction=RIGHT
                     /note="ColE1 ori"
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
     CDS             complement(3495..4577)
                     /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(4578..4655)
                     /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 atcatcatat cgaaggtagg
      301 catatgggcg gtggcggtag cggcggtggc ggtagcggta tgtcggactc agaagtcaat
      361 caagaagcta agccagaggt caagccagaa gtcaagcctg agactcacat caatttaaag
      421 gtgtccgatg gatcttcaga gatcttcttc aagatcaaaa agaccactcc tttaagaagg
      481 ctgatggaag cgttcgctaa aagacagggt aaggaaatgg actccttaag attcttgtac
      541 gacggtatta gaattcaagc tgatcagacc cctgaagatt tggacatgga ggataacgat
      601 attattgagg ctcacagaga acagattggt ggtactagtg agctcggtac cctcgaggga
      661 tccgaattca agcttgtcga cctgcagtct agataggtaa tctctgctta aaagcacaga
      721 atctaagatc cctgccattt ggcggggatt tttttatttg ttttcaggaa ataaataatc
      781 gatcgcgtaa taaaatctat tattattttt gtgaagaata aatttgggtg caatgagaat
      841 gcgcaggccc tttcgtctcg cgcgtttcgg tgatgacggt gaaaacctct gacacatgca
      901 gctcccggag acggtcacag cttgtctgta agcggatgcc gggagcagac aagcccgtca
      961 gggcgcgtca gcgggtgttg gcgggtgtcg gggctggctt aactatgcgg catcagagca
     1021 gattgtactg agagtgcacc ataaaattgt aaacgttaat attttgttaa aattcgcgtt
     1081 aaatttttgt taaatcagct cattttttaa ccaataggcc gaaatcggca aaatccctta
     1141 taaatcaaaa gaatagcccg agatagggtt gagtgttgtt ccagtttgga acaagagtcc
     1201 actattaaag aacgtggact ccaacgtcaa agggcgaaaa accgtctatc agggcgatgg
     1261 cccactacgt gaaccatcac ccaaatcaag ttttttgggg tcgaggtgcc gtaaagcact
    1321 aaatcggaac cctaaaggga gcccccgatt tagagcttga cggggaaagc cggcgaacgt
     1381 ggcgagaaag gaagggaaga aagcgaaagg agcgggcgct agggcgctgg caagtgtagc
     1441 ggtcacgctg cgcgtaacca ccacacccgc cgcgcttaat gcgccgctac agggcgcgta
     1501 ctatggttgc tttgacgtat gcggtgtgaa ataccgcaca gatgcgtaag gagaaaatac
     1561 cgcatcaggc gtcaggtggc acttttcggg gaaatgtgcg cggaacccct atttgtttat
     1621 ttttctaaat acattcaaat atgtatccgc tcatgagaca ataaccctga taaatgcttc
     1681 aataatattg aaaaaggaag agtatgagta ttcaacattt ccgtgtcgcc cttattccct
     1741 tttttgcggc attttgcctt cctgtttttg ctcacccaga aacgctggtg aaagtaaaag
     1801 atgctgaaga tcagttgggt gcacgagtgg gttacatcga actggatctc aacagcggta
     1861 agatccttga gagttttcgc cccgaagaac gttttccaat gatgagcact tttaaagttc
     1921 tgctatgtgg cgcggtatta tcccgtattg acgccgggca agagcaactc ggtcgccgca
     1981 tacactattc tcagaatgac ttggttgagt actcaccagt cacagaaaag catcttacgg
     2041 atggcatgac agtaagagaa ttatgcagtg ctgccataac catgagtgat aacactgcgg
     2101 ccaacttact tctgacaacg atcggaggac cgaaggagct aaccgctttt ttgcacaaca
     2161 tgggggatca tgtaactcgc cttgatcgtt gggaaccgga gctgaatgaa gccataccaa
     2221 acgacgagcg tgacaccacg atgcctgtag caatggcaac aacgttgcgc aaactattaa
     2281 ctggcgaact acttactcta gcttcccggc aacaattaat agactggatg gaggcggata
     2341 aagttgcagg accacttctg cgctcggccc ttccggctgg ctggtttatt gctgataaat
     2401 ctggagccgg tgagcgtggg tctcgcggta tcattgcagc actggggcca gatggtaagc
     2461 cctcccgtat cgtagttatc tacacgacgg ggagtcaggc aactatggat gaacgaaata
     2521 gacagatcgc tgagataggt gcctcactga ttaagcattg gtaactgtca gaccaagttt
     2581 actcatatat actttagatt gatttaaaac ttcattttta atttaaaagg atctaggtga
     2641 agatcctttt tgataatctc atgaccaaaa tcccttaacg tgagttttcg ttccactgag
     2701 cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa
     2761 tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg ccggatcaag
     2821 agctaccaac tctttttccg aaggtaactg gcttcagcag agcgcagata ccaaatactg
     2881 ttcttctagt gtagccgtag ttaggccacc acttcaagaa ctctgtagca ccgcctacat
     2941 acctcgctct gctaatcctg ttaccagtgg ctgctgccag tggcgataag tcgtgtctta
     3001 ccgggttgga ctcaagacga tagttaccgg ataaggcgca gcggtcgggc tgaacggggg
     3061 gttcgtgcac acagcccagc ttggagcgaa cgacctacac cgaactgaga tacctacagc
     3121 gtgagctatg agaaagcgcc acgcttcccg aagggagaaa ggcggacagg tatccggtaa
     3181 gcggcagggt cggaacagga gagcgcacga gggagcttcc agggggaaac gcctggtatc
     3241 tttatagtcc tgtcgggttt cgccacctct gacttgagcg tcgatttttg tgatgctcgt
     3301 caggggggcg gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct
     3361 tttgctggcc ttttgctcac atagtcatgc cccgcgccca ccggaaggag ctgactgggt
     3421 tgaaggctct caagggcatc ggtcgagatc ccggtgccta atgagtgagc taacttacat
     3481 taattgcgtt gcgctcactg cccgctttcc agtcgggaaa cctgtcgtgc cagctgcatt
     3541 aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat tgggcgccag ggtggttttt
     3601 cttttcacca gtgagacggg caacagctga ttgcccttca ccgcctggcc ctgagagagt
     3661 tgcagcaagc ggtccacgct ggtttgcccc agcaggcgaa aatcctgttt gatggtggtt
     3721 aacggcggga tataacatga gctgtcttcg gtatcgtcgt atcccactac cgagatatcc
     3781 gcaccaacgc gcagcccgga ctcggtaatg gcgcgcattg cgcccagcgc catctgatcg
     3841 ttggcaacca gcatcgcagt gggaacgatg ccctcattca gcatttgcat ggtttgttga
     3901 aaaccggaca tggcactcca gtcgccttcc cgttccgcta tcggctgaat ttgattgcga
     3961 gtgagatatt tatgccagcc agccagacgc agacgcgccg agacagaact taatgggccc
     4021 gctaacagcg cgatttgctg gtgacccaat gcgaccagat gctccacgcc cagtcgcgta
     4081 ccgtcttcat gggagaaaat aatactgttg atgggtgtct ggtcagagac atcaagaaat
     4141 aacgccggaa cattagtgca ggcagcttcc acagcaatgg catcctggtc atccagcgga
     4201 tagttaatga tcagcccact gacgcgttgc gcgagaagat tgtgcaccgc cgctttacag
     4261 gcttcgacgc cgcttcgttc taccatcgac accaccacgc tggcacccag ttgatcggcg
     4321 cgagatttaa tcgccgcgac aatttgcgac ggcgcgtgca gggccagact ggaggtggca
     4381 acgccaatca gcaacgactg tttgcccgcc agttgttgtg ccacgcggtt gggaatgtaa
     4441 ttcagctccg ccatcgccgc ttccactttt tcccgcgttt tcgcagaaac gtggctggcc
     4501 tggttcacca cgcgggaaac ggtctgataa gagacaccgg catactctgc gacatcgtat
     4561 aacgttactg gtttcacatt caccaccctg aattgactct cttccgggcg ctatcatgcc
     4621 ataccgcgaa aggttttgcg ccattcgatg gtgtccggga tctcgacgct ctcccttatg
     4681 cgactcctgc attaggaagc agcccagtag taggttgagg ccgttgagca ccgccgccgc
//