| 出品公司: | |
|---|---|
| 别名: | pTrcHis2C、 pTrcHis2 C |
| 质粒类型: | 大肠杆菌表达载体 |
| 表达水平: | 高拷贝 |
| 启动子: | Trc |
| 克隆方法: | 多克隆位点，限制性内切酶 |
| 载体大小: | 4405 bp |
| 5' 测序引物及序列: | pTrcHis-F: gaggtatatattaatgtatcg |
| 3' 测序引物及序列: | pTrcHis-R:GATTTAATCTGTATCAGG |
| 载体标签: | C-His, C-Myc |
| 载体抗性: | Ampicillin |
| 备注: |
Hosts: E.coli NM522, E.coli INValphaF'. Related vectors: pSE420.
|
| 产品目录号: | V365-20 |
| 稳定性: | 瞬时表达 Transient |
| 组成型: | 组成型 Constitutive |
| 病毒/非病毒: | 非病毒 |
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- 详细信息
- 文献和实验
- 技术资料
- 保存条件：
-20℃低温保存
- 保质期：
三年
- 英文名：
pTrcHis2 C
- 库存：
20
- 供应商：
泽叶生物
pTrcHis2C载体基本信息
pTrcHis2C载体质粒图谱和多克隆位点信息
pTrcHis2C载体简介
The pTrcHis2 plasmids are pBR322-derived expression vectors designed for efficient recombinant protein expression and purification in E. coli. High levels of expression are possible using the trc (trp-lac) promoter (Egon et al., 1983) and the rrnB anti-termination region (Li et al., 1984). The trc promoter contains the –35 region of the trp promoter together with the –10 region of the lac promoter (Brosius et al., 1985; Egon et al., 1983; Mulligan et al., 1985). To regulate expression, the gene encoding Lac repressor (lacIq) is provided in the pTrcHis2 vectors, allowing regulation of the trc promoter regardless of whether the host strain contains a gene encoding the Lac repressor.
Isopropyl-Beta-D-thiogalactopyranoside (IPTG) is used to induce expression of your gene. Translation is enhanced by the bacteriophage T7 gene 10 translation enhancer and a minicistron that provides highly efficient translational restart into the open reading frame of the multiple cloning site. DNA inserts are positioned downstream and in frame with the initiation ATG and a C-terminal fusion peptide. The C-terminal peptide encodes the myc epitope and six histidine residues that function as a metal binding site in the expressed protein.
pTrcHis2C载体序列
LOCUS pTrcHis2\C 4405 bp DNA circular SYN 7-JUL-2013
DEFINITION pTrcHis2 C.
SOURCE
ORGANISM other sequences; artificial sequences; vectors.
COMMENT http://www.biofeng.com/
COMMENT This file is created by Vector NTI
COMMENT VNTOAUTHORNAME|biofeng.com|
FEATURES Location/Qualifiers
source 1..4405
/organism="pTrcHis2 A"
/mol_type="other DNA"
/vntifkey="98"
misc_feature 235..257
/vntifkey="21"
/label=M13_pUC_rev_primer
misc_feature 264..333
/vntifkey="21"
/label=rrnB_ATS
misc_feature 370..390
/vntifkey="21"
/label=pTrcHis_fwd_primer
misc_feature 383..409
/vntifkey="21"
/label=minicis
misc_feature complement(588..605)
/vntifkey="21"
/label=pBAD_rev_primer
misc_feature complement(588..605)
/vntifkey="21"
/label=pTrcHis_rev_primer
terminator 638..795
/vntifkey="43"
/label=rrnB_terminator
terminator 761..804
/vntifkey="43"
/label=rrnB_T1_terminator
terminator 936..963
/vntifkey="43"
/label=rrnB_T2_terminator
promoter 1005..1033
/vntifkey="30"
/label=AmpR_promoter
gene 1075..1935
/gene="Ampicillin"
/vntifkey="60"
CDS 1075..1935
/vntifkey="4"
/label=ORF\frame\2
rep_origin 2090..2709
/vntifkey="33"
/label=pBR322_origin
misc_feature 3106..3128
/vntifkey="21"
/label=pGEX_3_primer
misc_feature 3275..4366
/vntifkey="21"
/label=lacI
CDS 3407..4366
/vntifkey="4"
/label=ORF\frame\3
BASE COUNT 1092 a 1098 c 1161 g 1054 t
ORIGIN
1 gtttgacagc ttatcatcga ctgcacggtg caccaatgct tctggcgtca ggcagccatc
61 ggaagctgtg gtatggctgt gcaggtcgta aatcactgca taattcgtgt cgctcaaggc
121 gcactcccgt tctggataat gttttttgcg ccgacatcat aacggttctg gcaaatattc
181 tgaaatgagc tgttgacaat taatcatccg gctcgtataa tgtgtggaat tgtgagcgga
241 taacaatttc acacaggaaa cagcgccgct gagaaaaagc gaagcggcac tgctctttaa
301 caatttatca gacaatctgt gtgggcactc gaccggaatt atcgattaac tttattatta
361 aaaattaaag aggtatatat taatgtatcg attaaataag gaggaataaa ccatggatcc
421 gagctcgaga tctgcagctg gtaccatatg ggaattcgaa gcttacgtag aacaaaaact
481 catctcagaa gaggatctga atagcgccgt cgaccatcat catcatcatc attgagttta
541 aacggtctcc agcttggctg ttttggcgga tgagagaaga ttttcagcct gatacagatt
601 aaatcagaac gcagaagcgg tctgataaaa cagaatttgc ctggcggcag tagcgcggtg
661 gtcccacctg accccatgcc gaactcagaa gtgaaacgcc gtagcgccga tggtagtgtg
721 gggtctcccc atgcgagagt agggaactgc caggcatcaa ataaaacgaa aggctcagtc
781 gaaagactgg gcctttcgtt ttatctgttg tttgtcggtg aacgctctcc tgagtaggac
841 aaatccgccg ggagcggatt tgaacgttgc gaagcaacgg cccggagggt ggcgggcagg
901 acgcccgcca taaactgcca ggcatcaaat taagcagaag gccatcctga cggatggcct
961 ttttgcgttt ctacaaactc tttttgttta tttttctaaa tacattcaaa tatgtatccg
1021 ctcatgagac aataaccctg ataaatgctt caataatatt gaaaaaggaa gagtatgagt
1081 attcaacatt tccgtgtcgc ccttattccc ttttttgcgg cattttgcct tcctgttttt
1141 gctcacccag aaacgctggt gaaagtaaaa gatgctgaag atcagttggg tgcacgagtg
1201 ggttacatcg aactggatct caacagcggt aagatccttg agagttttcg ccccgaagaa
1261 cgttttccaa tgatgagcac ttttaaagtt ctgctatgtg gcgcggtatt atcccgtgtt
1321 gacgccgggc aagagcaact cggtcgccgc atacactatt ctcagaatga cttggttgag
1381 tactcaccag tcacagaaaa gcatcttacg gatggcatga cagtaagaga attatgcagt
1441 gctgccataa ccatgagtga taacactgcg gccaacttac ttctgacaac gatcggagga
1501 ccgaaggagc taaccgcttt tttgcacaac atgggggatc atgtaactcg ccttgatcgt
1561 tgggaaccgg agctgaatga agccatacca aacgacgagc gtgacaccac gatgcctgta
1621 gcaatggcaa caacgttgcg caaactatta actggcgaac tacttactct agcttcccgg
1681 caacaattaa tagactggat ggaggcggat aaagttgcag gaccacttct gcgctcggcc
1741 cttccggctg gctggtttat tgctgataaa tctggagccg gtgagcgtgg gtctcgcggt
1801 atcattgcag cactggggcc agatggtaag ccctcccgta tcgtagttat ctacacgacg
1861 gggagtcagg caactatgga tgaacgaaat agacagatcg ctgagatagg tgcctcactg
1921 attaagcatt ggtaactgtc agaccaagtt tactcatata tactttagat tgatttaaaa
1981 cttcattttt aatttaaaag gatctaggtg aagatccttt ttgataatct catgaccaaa
2041 atcccttaac gtgagttttc gttccactga gcgtcagacc ccgtagaaaa gatcaaagga
2101 tcttcttgag atcctttttt tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg
2161 ctaccagcgg tggtttgttt gccggatcaa gagctaccaa ctctttttcc gaaggtaact
2221 ggcttcagca gagcgcagat accaaatact gtccttctag tgtagccgta gttaggccac
2281 cacttcaaga actctgtagc accgcctaca tacctcgctc tgctaatcct gttaccagtg
2341 gctgctgcca gtggcgataa gtcgtgtctt accgggttgg actcaagacg atagttaccg
2401 gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca cacagcccag cttggagcga
2461 acgacctaca ccgaactgag atacctacag cgtgagctat gagaaagcgc cacgcttccc
2521 gaagggagaa aggcggacag gtatccggta agcggcaggg tcggaacagg agagcgcacg
2581 agggagcttc cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc
2641 tgacttgagc gtcgattttt gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc
2701 agcaacgcgg cctttttacg gttcctggcc ttttgctggc cttttgctca catgttcttt
2761 cctgcgttat cccctgattc tgtggataac cgtattaccg cctttgagtg agctgatacc
2821 gctcgccgca gccgaacgac cgagcgcagc gagtcagtga gcgaggaagc ggaagagcgc
2881 ctgatgcggt attttctcct tacgcatctg tgcggtattt cacaccgcat atggtgcact
2941 ctcagtacaa tctgctctga tgccgcatag ttaagccagt atacactccg ctatcgctac
3001 gtgactgggt catggctgcg ccccgacacc cgccaacacc cgctgacgcg ccctgacggg
3061 cttgtctgct cccggcatcc gcttacagac aagctgtgac cgtctccggg agctgcatgt
3121 gtcagaggtt ttcaccgtca tcaccgaaac gcgcgaggca gcagatcaat tcgcgcgcga
3181 aggcgaagcg gcatgcattt acgttgacac catcgaatgg tgcaaaacct ttcgcggtat
3241 ggcatgatag cgcccggaag agagtcaatt cagggtggtg aatgtgaaac cagtaacgtt
3301 atacgatgtc gcagagtatg ccggtgtctc ttatcagacc gtttcccgcg tggtgaacca
3361 ggccagccac gtttctgcga aaacgcggga aaaagtggaa gcggcgatgg cggagctgaa
3421 ttacattccc aaccgcgtgg cacaacaact ggcgggcaaa cagtcgttgc tgattggcgt
3481 tgccacctcc agtctggccc tgcacgcgcc gtcgcaaatt gtcgcggcga ttaaatctcg
3541 cgccgatcaa ctgggtgcca gcgtggtggt gtcgatggta gaacgaagcg gcgtcgaagc
3601 ctgtaaagcg gcggtgcaca atcttctcgc gcaacgcgtc agtgggctga tcattaacta
3661 tccgctggat gaccaggatg ccattgctgt ggaagctgcc tgcactaatg ttccggcgtt
3721 atttcttgat gtctctgacc agacacccat caacagtatt attttctccc atgaagacgg
3781 tacgcgactg ggcgtggagc atctggtcgc attgggtcac cagcaaatcg cgctgttagc
3841 gggcccatta agttctgtct cggcgcgtct gcgtctggct ggctggcata aatatctcac
3901 tcgcaatcaa attcagccga tagcggaacg ggaaggcgac tggagtgcca tgtccggttt
3961 tcaacaaacc atgcaaatgc tgaatgaggg catcgttccc actgcgatgc tggttgccaa
4021 cgatcagatg gcgctgggcg caatgcgcgc cattaccgag tccgggctgc gcgttggtgc
4081 ggatatctcg gtagtgggat acgacgatac cgaagacagc tcatgttata tcccgccgtc
4141 aaccaccatc aaacaggatt ttcgcctgct ggggcaaacc agcgtggacc gcttgctgca
4201 actctctcag ggccaggcgg tgaagggcaa tcagctgttg cccgtctcac tggtgaaaag
4261 aaaaaccacc ctggcgccca atacgcaaac cgcctctccc cgcgcgttgg ccgattcatt
4321 aatgcagctg gcacgacagg tttcccgact ggaaagcggg cagtgagcgc aacgcaatta
4381 atgtgagtta gcgcgaattg atctg
//
DEFINITION pTrcHis2 C.
SOURCE
ORGANISM other sequences; artificial sequences; vectors.
COMMENT http://www.biofeng.com/
COMMENT This file is created by Vector NTI
COMMENT VNTOAUTHORNAME|biofeng.com|
FEATURES Location/Qualifiers
source 1..4405
/organism="pTrcHis2 A"
/mol_type="other DNA"
/vntifkey="98"
misc_feature 235..257
/vntifkey="21"
/label=M13_pUC_rev_primer
misc_feature 264..333
/vntifkey="21"
/label=rrnB_ATS
misc_feature 370..390
/vntifkey="21"
/label=pTrcHis_fwd_primer
misc_feature 383..409
/vntifkey="21"
/label=minicis
misc_feature complement(588..605)
/vntifkey="21"
/label=pBAD_rev_primer
misc_feature complement(588..605)
/vntifkey="21"
/label=pTrcHis_rev_primer
terminator 638..795
/vntifkey="43"
/label=rrnB_terminator
terminator 761..804
/vntifkey="43"
/label=rrnB_T1_terminator
terminator 936..963
/vntifkey="43"
/label=rrnB_T2_terminator
promoter 1005..1033
/vntifkey="30"
/label=AmpR_promoter
gene 1075..1935
/gene="Ampicillin"
/vntifkey="60"
CDS 1075..1935
/vntifkey="4"
/label=ORF\frame\2
rep_origin 2090..2709
/vntifkey="33"
/label=pBR322_origin
misc_feature 3106..3128
/vntifkey="21"
/label=pGEX_3_primer
misc_feature 3275..4366
/vntifkey="21"
/label=lacI
CDS 3407..4366
/vntifkey="4"
/label=ORF\frame\3
BASE COUNT 1092 a 1098 c 1161 g 1054 t
ORIGIN
1 gtttgacagc ttatcatcga ctgcacggtg caccaatgct tctggcgtca ggcagccatc
61 ggaagctgtg gtatggctgt gcaggtcgta aatcactgca taattcgtgt cgctcaaggc
121 gcactcccgt tctggataat gttttttgcg ccgacatcat aacggttctg gcaaatattc
181 tgaaatgagc tgttgacaat taatcatccg gctcgtataa tgtgtggaat tgtgagcgga
241 taacaatttc acacaggaaa cagcgccgct gagaaaaagc gaagcggcac tgctctttaa
301 caatttatca gacaatctgt gtgggcactc gaccggaatt atcgattaac tttattatta
361 aaaattaaag aggtatatat taatgtatcg attaaataag gaggaataaa ccatggatcc
421 gagctcgaga tctgcagctg gtaccatatg ggaattcgaa gcttacgtag aacaaaaact
481 catctcagaa gaggatctga atagcgccgt cgaccatcat catcatcatc attgagttta
541 aacggtctcc agcttggctg ttttggcgga tgagagaaga ttttcagcct gatacagatt
601 aaatcagaac gcagaagcgg tctgataaaa cagaatttgc ctggcggcag tagcgcggtg
661 gtcccacctg accccatgcc gaactcagaa gtgaaacgcc gtagcgccga tggtagtgtg
721 gggtctcccc atgcgagagt agggaactgc caggcatcaa ataaaacgaa aggctcagtc
781 gaaagactgg gcctttcgtt ttatctgttg tttgtcggtg aacgctctcc tgagtaggac
841 aaatccgccg ggagcggatt tgaacgttgc gaagcaacgg cccggagggt ggcgggcagg
901 acgcccgcca taaactgcca ggcatcaaat taagcagaag gccatcctga cggatggcct
961 ttttgcgttt ctacaaactc tttttgttta tttttctaaa tacattcaaa tatgtatccg
1021 ctcatgagac aataaccctg ataaatgctt caataatatt gaaaaaggaa gagtatgagt
1081 attcaacatt tccgtgtcgc ccttattccc ttttttgcgg cattttgcct tcctgttttt
1141 gctcacccag aaacgctggt gaaagtaaaa gatgctgaag atcagttggg tgcacgagtg
1201 ggttacatcg aactggatct caacagcggt aagatccttg agagttttcg ccccgaagaa
1261 cgttttccaa tgatgagcac ttttaaagtt ctgctatgtg gcgcggtatt atcccgtgtt
1321 gacgccgggc aagagcaact cggtcgccgc atacactatt ctcagaatga cttggttgag
1381 tactcaccag tcacagaaaa gcatcttacg gatggcatga cagtaagaga attatgcagt
1441 gctgccataa ccatgagtga taacactgcg gccaacttac ttctgacaac gatcggagga
1501 ccgaaggagc taaccgcttt tttgcacaac atgggggatc atgtaactcg ccttgatcgt
1561 tgggaaccgg agctgaatga agccatacca aacgacgagc gtgacaccac gatgcctgta
1621 gcaatggcaa caacgttgcg caaactatta actggcgaac tacttactct agcttcccgg
1681 caacaattaa tagactggat ggaggcggat aaagttgcag gaccacttct gcgctcggcc
1741 cttccggctg gctggtttat tgctgataaa tctggagccg gtgagcgtgg gtctcgcggt
1801 atcattgcag cactggggcc agatggtaag ccctcccgta tcgtagttat ctacacgacg
1861 gggagtcagg caactatgga tgaacgaaat agacagatcg ctgagatagg tgcctcactg
1921 attaagcatt ggtaactgtc agaccaagtt tactcatata tactttagat tgatttaaaa
1981 cttcattttt aatttaaaag gatctaggtg aagatccttt ttgataatct catgaccaaa
2041 atcccttaac gtgagttttc gttccactga gcgtcagacc ccgtagaaaa gatcaaagga
2101 tcttcttgag atcctttttt tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg
2161 ctaccagcgg tggtttgttt gccggatcaa gagctaccaa ctctttttcc gaaggtaact
2221 ggcttcagca gagcgcagat accaaatact gtccttctag tgtagccgta gttaggccac
2281 cacttcaaga actctgtagc accgcctaca tacctcgctc tgctaatcct gttaccagtg
2341 gctgctgcca gtggcgataa gtcgtgtctt accgggttgg actcaagacg atagttaccg
2401 gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca cacagcccag cttggagcga
2461 acgacctaca ccgaactgag atacctacag cgtgagctat gagaaagcgc cacgcttccc
2521 gaagggagaa aggcggacag gtatccggta agcggcaggg tcggaacagg agagcgcacg
2581 agggagcttc cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc
2641 tgacttgagc gtcgattttt gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc
2701 agcaacgcgg cctttttacg gttcctggcc ttttgctggc cttttgctca catgttcttt
2761 cctgcgttat cccctgattc tgtggataac cgtattaccg cctttgagtg agctgatacc
2821 gctcgccgca gccgaacgac cgagcgcagc gagtcagtga gcgaggaagc ggaagagcgc
2881 ctgatgcggt attttctcct tacgcatctg tgcggtattt cacaccgcat atggtgcact
2941 ctcagtacaa tctgctctga tgccgcatag ttaagccagt atacactccg ctatcgctac
3001 gtgactgggt catggctgcg ccccgacacc cgccaacacc cgctgacgcg ccctgacggg
3061 cttgtctgct cccggcatcc gcttacagac aagctgtgac cgtctccggg agctgcatgt
3121 gtcagaggtt ttcaccgtca tcaccgaaac gcgcgaggca gcagatcaat tcgcgcgcga
3181 aggcgaagcg gcatgcattt acgttgacac catcgaatgg tgcaaaacct ttcgcggtat
3241 ggcatgatag cgcccggaag agagtcaatt cagggtggtg aatgtgaaac cagtaacgtt
3301 atacgatgtc gcagagtatg ccggtgtctc ttatcagacc gtttcccgcg tggtgaacca
3361 ggccagccac gtttctgcga aaacgcggga aaaagtggaa gcggcgatgg cggagctgaa
3421 ttacattccc aaccgcgtgg cacaacaact ggcgggcaaa cagtcgttgc tgattggcgt
3481 tgccacctcc agtctggccc tgcacgcgcc gtcgcaaatt gtcgcggcga ttaaatctcg
3541 cgccgatcaa ctgggtgcca gcgtggtggt gtcgatggta gaacgaagcg gcgtcgaagc
3601 ctgtaaagcg gcggtgcaca atcttctcgc gcaacgcgtc agtgggctga tcattaacta
3661 tccgctggat gaccaggatg ccattgctgt ggaagctgcc tgcactaatg ttccggcgtt
3721 atttcttgat gtctctgacc agacacccat caacagtatt attttctccc atgaagacgg
3781 tacgcgactg ggcgtggagc atctggtcgc attgggtcac cagcaaatcg cgctgttagc
3841 gggcccatta agttctgtct cggcgcgtct gcgtctggct ggctggcata aatatctcac
3901 tcgcaatcaa attcagccga tagcggaacg ggaaggcgac tggagtgcca tgtccggttt
3961 tcaacaaacc atgcaaatgc tgaatgaggg catcgttccc actgcgatgc tggttgccaa
4021 cgatcagatg gcgctgggcg caatgcgcgc cattaccgag tccgggctgc gcgttggtgc
4081 ggatatctcg gtagtgggat acgacgatac cgaagacagc tcatgttata tcccgccgtc
4141 aaccaccatc aaacaggatt ttcgcctgct ggggcaaacc agcgtggacc gcttgctgca
4201 actctctcag ggccaggcgg tgaagggcaa tcagctgttg cccgtctcac tggtgaaaag
4261 aaaaaccacc ctggcgccca atacgcaaac cgcctctccc cgcgcgttgg ccgattcatt
4321 aatgcagctg gcacgacagg tttcccgact ggaaagcggg cagtgagcgc aacgcaatta
4381 atgtgagtta gcgcgaattg atctg
//
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文献和实验相关实验
广义的细胞色素 c是含有血红素 c的细胞色素之总称。通常是指主要存在于高等动植物、酵母、霉菌等线粒体中的细胞色素 c，起着传递电子的作用。为分子量约 1.3万的碱性蛋白质，还原型的吸收带为 550、 520、 415毫微米，氧化型为 407毫微米。α 因此很早便被研究，并获得结晶。不仅一级结构而且立体结构也已清楚（图 1）。其结构基本上是生物界共有的。由细胞色素 c氧化酶〔细胞色素（ a＋ a3 ）复合体〕氧化。在呼吸链中从细胞色素 c开始接受电子，各种还原酶都已清楚
1.常用载体分类：2,常用载体元件介绍：(1) 启动子：启动子是位于结构基因5'端上游的DNA序列，能活化RNA聚合酶，使之与模板DNA准确的结合并具有转录起始的特异性。启动子主要分为广谱启动子和特异性启动子。广谱启动子为在各种组织、细胞上广泛表达的启动子；特异性启动子为在特异性组织或者细胞类型上表达的启动子。常用广谱启动子有CAG，CMV，EF1a，PGK等，还包含启动RNA的III型启动子U6和H1启动子；常用特异性启动子如成熟神经元特异性启动子hSyn，投射神经元特异性启动
恰当的选择固相载体和连接分子是固相反应的重要组成部分. 固相载体和连接分子都应该在多种遗传反应过程期间是保持稳定, 而且产品的切除应该是在温和的条件下实现. 本章将介绍各种不同的载体和连接分子. 如果反应后可以用过滤的方法去除, 从理论上讲, 任何材料都可用作固相载体. 聚乙烯, 聚苯乙烯-外涂聚乙烯薄层, 平滑的纸或棉花等都可以. 最常用的载体是: 聚苯乙烯(PS), 聚乙二醇(PEG), Tentagel (TG), 和PEGA (聚乙二醇-聚丙
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pTrcHis2 C载体
¥1000
