| 出品公司: | Takara |
|---|---|
| 载体名称: | pCold TF, pColdTF, pCold-TF |
| 质粒类型: | 大肠杆菌蛋白表达载体 |
| 表达水平: | 高 |
| 启动子: | CSPA |
| 克隆方法: | 多克隆位点，限制性内切酶 |
| 载体大小: | 5769 bp |
| 5' 测序引物及序列: |
pCold-TF-F1: 5'-CCACTTTCAACGAGCTGATG-3'; pCold-TF-F2: 5'-GCGAAAGTGACTGAAAAAG-3' |
| 3' 测序引物及序列: | pCold-R: 5'-GGCAGGGATCTTAGATTCTG-3' |
| 载体标签: | N-His, N-Trigger factor, N-HRV 3C, N-Thrombin, 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: | 3365 |
| 稳定性: | 瞬时表达 Transient |
| 组成型: | 诱导表达 |
| 病毒/非病毒: | 非病毒 |
买家导航
pCold TF载体 pColdTF质粒基本信息
pCold TF质粒图谱 pColdTF载体图谱和pCold TF载体序列 pColdTF质粒序列多克隆位点信息
pCold TF质粒 pColdTF载体简介
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。
pCold TF质粒序列 pColdTF载体序列
LOCUS pCold\TF 5769 bp DNA circular SYN 27-DEC-2006
DEFINITION Expression vector pColdTF DNA, complete sequence.
ACCESSION AB213654
VERSION AB213654.1 GI:66472063
KEYWORDS .
SOURCE Expression vector pColdTF
ORGANISM Expression vector pColdTF
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..5769
/organism="Expression vector pColdTF"
/mol_type="other DNA"
/db_xref="taxon:329577"
/lab_host="Escherichia coli"
/note="expresses an open reading frame as a trigger factor
fusion, containing the translation enhancing element,
His6, protease cleavage site for HRV 3C protease, thrombin
and factor Xa"
gene join(15..255,256..270)
/gene="cspA"
promoter 15..81
/gene="cspA"
misc_feature 82..121
/gene="cspA"
/note="synthetic lac operator sequence"
misc_feature 122..255
/gene="cspA"
/note="modified cspA 5'UTR"
gene 256..1725
/gene="tig"
CDS 256..1725
/gene="tig"
/codon_start=1
/transl_table=11
/product="trigger factor tag"
/protein_id="BAD98926.1"
/db_xref="GI:66472064"
/translation="MNHKVHHHHHHMQVSVETTQGLGRRVTITIAADSIETAVKSELV
NVAKKVRIDGFRKGKVPMNIVAQRYGASVRQDVLGDLMSRNFIDAIIKEKINPAGAPT
YVPGEYKLGEDFTYSVEFEVYPEVELQGLEAIEVEKPIVEVTDADVDGMLDTLRKQQA
TWKEKDGAVEAEDRVTIDFTGSVDGEEFEGGKASDFVLAMGQGRMIPGFEDGIKGHKA
GEEFTIDVTFPEEYHAENLKGKAAKFAINLKKVEERELPELTAEFIKRFGVEDGSVEG
LRAEVRKNMERELKSAIRNRVKSQAIEGLVKANDIDVPAALIDSEIDVLRRQAAQRFG
GNEKQALELPRELFEEQAKRRVVVGLLLGEVIRTNELKADEERVKGLIEEMASAYEDP
KEVIEFYSKNKELMDNMRNVALEEQAVEAVLAKAKVTEKETTFNELMNQQASAGLEVL
FQGPSAGLVPRGSGGIEGRHMELGTLEGSEFKLVDLQSR"
misc_feature 256..270
/gene="cspA"
/note="TEE (translation enhancing element)"
misc_feature 271..288
/gene="tig"
/note="His6 tag"
misc_feature 1594..1617
/gene="tig"
/note="HRV 3C Protease site"
misc_feature 1627..1644
/gene="tig"
/note="Thrombin site"
misc_feature 1651..1662
/gene="tig"
/note="Factor Xa site"
misc_feature 1663..1722
/gene="tig"
/note="MCS = Multiple cloning sites contain the follow
restriction sites: NdeI, SacI, KpnI, XhoI, BamHI, EcoRI,
HindIII, SalI, PstI, XbaI"
gene 1730..1874
/gene="cspA"
misc_feature 1730..1874
/gene="cspA"
/note="cspA 3'UTR"
misc_feature complement(2073..2546)
/note="M13 intergenic region"
gene 2733..3593
/gene="Amp"
CDS 2733..3593
/gene="Amp"
/codon_start=1
/transl_table=11
/product="beta-lactamase"
/protein_id="BAD98927.1"
/db_xref="GI:66472065"
/translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGY
IELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVE
YSPVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRL
DRWEPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPL
LRSALPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIA
EIGASLIKHW"
rep_origin 3753..4367
/note="ColE1 origin"
gene complement(4524..5606)
/gene="lacI"
CDS complement(4524..5606)
/gene="lacI"
/codon_start=1
/transl_table=11
/product="lactose operon repressor"
/protein_id="BAD98928.1"
/db_xref="GI:66472066"
/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 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
//
