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质粒(载体)
pET-32b
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产物名称:
pET-32b
产物型号:
产物展商:
XYbscience
产物文档:
无相关文档
简单介绍
pET-32b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。pET-32b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。
pET-32b
的详细介绍
pET-32b
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
XY0034
pET-32b
20μ濒
?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:
T7/lac
复制子:
ColE1 ori,F1 ori
终止子:
T7 terminator
质粒分类:
大肠杆菌载体;笔贰罢系列表达质粒
质粒大小:
5899bp
质粒标签:
狈-罢谤虫,狈-6×贬颈蝉,狈-迟丑谤辞尘产颈苍,狈-厂,狈-别苍迟别谤辞办颈苍补蝉别,颁-6×贬颈蝉
原核抗性:
氨苄青霉素础尘辫
克隆菌株:
顿贬5α
培养条件:
37℃,有氧,尝叠
表达宿主:
BL21(DE3)
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:
T7:TAATACGACTCACTATAGGG
3'测序引物:
T7-ter:TGCTAGTTATTGCTCAGCGG
质粒介绍
The pET-32a-c series is designed for cloning and high-level expression of peptide sequences fused with the 109aa Trx?Tag? thioredoxin protein (1). Cloning sites are available for producing fusion proteins also containing cleavable His?Tag® and S?Tag? sequences for detection and purification. Unique sites are shown on the circle map. Note that the sequence is numbered by the pBR322 convention, so the T7 expression region is reversed on the circle map. The cloning/expression region of the coding strand transcribed by T7 RNA polymerase is shown below. The f1 origin is oriented so that infection with helper phage will produce virions containing single-stranded DNA that corresponds to the coding strand. Therefore, single-stranded sequencing should be performed using the T7 terminator primer.
质粒图谱
质粒序列
LOCUS Exported 5899 bp ds-DNA circular SYN 05-8-2015
DEFINITION synthetic circular DNA
ACCESSION .
VERSION .
KEYWORDS Untitled 9
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 5899)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported 2015-8-5 from MLCC
http://www.miaolingbio.com
FEATURES Location/Qualifiers
source 1..5899
/organism="synthetic DNA construct"
/mol_type="other DNA"
source 699..721
/organism="Enterobacteria phage T7"
/mol_type="genomic DNA"
/db_xref="taxon:10760"
terminator 26..73
/note="T7 terminator"
/note="transcription terminator for bacteriophage T7 RNA
polymerase"
CDS complement(140..157)
/codon_start=1
/product="6xHis affinity tag"
/note="6xHis"
/translation="HHHHHH"
CDS complement(218..232)
/codon_start=1
/product="enterokinase recognition and cleavage site"
/note="enterokinase site"
/translation="DDDDK"
CDS complement(248..292)
/codon_start=1
/product="affinity and epitope tag derived from pancreatic
ribonuclease A"
/note="S-Tag"
/translation="KETAAAKFERQHMDS"
CDS complement(299..316)
/codon_start=1
/product="thrombin recognition and cleavage site"
/note="thrombin site"
/translation="LVPRGS"
CDS complement(326..343)
/codon_start=1
/product="6xHis affinity tag"
/note="6xHis"
/translation="HHHHHH"
CDS complement(365..691)
/codon_start=1
/gene="trxA"
/product="E. coli thioredoxin"
/note="TrxA"
/translation="MSDKIIHLTDDSFDTDVLKADGAILVDFWAEWCGPCKMIAPILDE
IADEYQGKLTVAKLNIDQNPGTAPKYGIRGIPTLLLFKNGEVAATKVGALSKGQLKEFL
DANLA"
RBS 699..721
/note="efficient ribosome binding site from bacteriophage
T7 gene 10 (Olins and Rangwala, 1989)"
protein_bind 736..760
/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)."
promoter complement(761..779)
/note="T7 promoter"
/note="promoter for bacteriophage T7 RNA polymerase"
promoter 1092..1169
/gene="lacI"
/note="lacI promoter"
CDS 1170..2252
/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"
CDS 3061..3252
/codon_start=1
/gene="rop"
/product="Rop protein, which maintains plasmids at low copy
number"
/note="rop"
/translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
DELYRSCLARFGDDGENL"
misc_feature 3354..3496
/note="bom"
/note="basis of mobility region from pBR322"
rep_origin complement(3682..4270)
/direction=LEFT
/note="ori"
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
CDS complement(4441..5301)
/codon_start=1
/gene="bla"
/product="beta-lactamase"
/note="AmpR"
/note="confers resistance to ampicillin, carbenicillin, and
related antibiotics"
/translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
LIKHW"
promoter complement(5302..5406)
/gene="bla"
/note="AmpR promoter"
rep_origin complement(5433..5888)
/direction=LEFT
/note="f1 ori"
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
ORIGIN
1 atccggatat agttcctcct ttcagcaaaa aacccctcaa gacccgttta gaggccccaa
61 ggggttatgc tagttattgc tcagcggtgg cagcagccaa ctcagcttcc tttcgggctt
121 tgttagcagc cggatctcag tggtggtggt ggtggtgctc gagtgcggcc gcaagcttgt
181 cgacggagct cgaattcgga tccgatatcg ccatggcctt gtcgtcgtcg tcggtaccca
241 gatctgggct gtccatgtgc tggcgttcga atttagcagc agcggtttct ttcataccag
301 aaccgcgtgg caccagacca gaagaatgat gatgatgatg gtgcatatgg ccagaaccag
361 aaccggccag gttagcgtcg aggaactctt tcaactgacc tttagacagt gcacccactt
421 tggttgccgc cacttcaccg tttttgaaca gcagcagagt cgggatacca cggatgccat
481 atttcggcgc agtgccaggg ttttgatcga tgttcagttt tgcaacggtc agtttgccct
541 gatattcgtc agcgatttca tccagaatcg gggcgatcat tttgcacgga ccgcaccact
601 ctgcccagaa atcgacgagg atcgccccgt ccgctttgag tacatccgtg tcaaaactgt
661 cgtcagtcag gtgaataatt ttatcgctca tatgtatatc tccttcttaa agttaaacaa
721 aattatttct agaggggaat tgttatccgc tcacaattcc cctatagtga gtcgtattaa
781 tttcgcggga tcgagatcga tctcgatcct ctacgccgga cgcatcgtgg ccggcatcac
841 cggcgccaca ggtgcggttg ctggcgccta tatcgccgac atcaccgatg gggaagatcg
901 ggctcgccac ttcgggctca tgagcgcttg tttcggcgtg ggtatggtgg caggccccgt
961 ggccggggga ctgttgggcg ccatctcctt gcatgcacca ttccttgcgg cggcggtgct
1021 caacggcctc aacctactac tgggctgctt cctaatgcag gagtcgcata agggagagcg
1081 tcgagatccc ggacaccatc gaatggcgca aaacctttcg cggtatggca tgatagcgcc
1141 cggaagagag tcaattcagg gtggtgaatg tgaaaccagt aacgttatac gatgtcgcag
1201 agtatgccgg tgtctcttat cagaccgttt cccgcgtggt gaaccaggcc agccacgttt
1261 ctgcgaaaac gcgggaaaaa gtggaagcgg cgatggcgga gctgaattac attcccaacc
1321 gcgtggcaca acaactggcg ggcaaacagt cgttgctgat tggcgttgcc acctccagtc
1381 tggccctgca cgcgccgtcg caaattgtcg cggcgattaa atctcgcgcc gatcaactgg
1441 gtgccagcgt ggtggtgtcg atggtagaac gaagcggcgt cgaagcctgt aaagcggcgg
1501 tgcacaatct tctcgcgcaa cgcgtcagtg ggctgatcat taactatccg ctggatgacc
1561 aggatgccat tgctgtggaa gctgcctgca ctaatgttcc ggcgttattt cttgatgtct
1621 ctgaccagac acccatcaac agtattattt tctcccatga agacggtacg cgactgggcg
1681 tggagcatct ggtcgcattg ggtcaccagc aaatcgcgct gttagcgggc ccattaagtt
1741 ctgtctcggc gcgtctgcgt ctggctggct ggcataaata tctcactcgc aatcaaattc
1801 agccgatagc ggaacgggaa ggcgactgga gtgccatgtc cggttttcaa caaaccatgc
1861 aaatgctgaa tgagggcatc gttcccactg cgatgctggt tgccaacgat cagatggcgc
1921 tgggcgcaat gcgcgccatt accgagtccg ggctgcgcgt tggtgcggac atctcggtag
1981 tgggatacga cgataccgaa gacagctcat gttatatccc gccgttaacc accatcaaac
2041 aggattttcg cctgctgggg caaaccagcg tggaccgctt gctgcaactc tctcagggcc
2101 aggcggtgaa gggcaatcag ctgttgcccg tctcactggt gaaaagaaaa accaccctgg
2161 cgcccaatac gcaaaccgcc tctccccgcg cgttggccga ttcattaatg cagctggcac
2221 gacaggtttc ccgactggaa agcgggcagt gagcgcaacg caattaatgt aagttagctc
2281 actcattagg caccgggatc tcgaccgatg cccttgagag ccttcaaccc agtcagctcc
2341 ttccggtggg cgcggggcat gactatcgtc gccgcactta tgactgtctt ctttatcatg
2401 caactcgtag gacaggtgcc ggcagcgctc tgggtcattt tcggcgagga ccgctttcgc
2461 tggagcgcga cgatgatcgg cctgtcgctt gcggtattcg gaatcttgca cgccctcgct
2521 caagccttcg tcactggtcc cgccaccaaa cgtttcggcg agaagcaggc cattatcgcc
2581 ggcatggcgg ccccacgggt gcgcatgatc gtgctcctgt cgttgaggac ccggctaggc
2641 tggcggggtt gccttactgg ttagcagaat gaatcaccga tacgcgagcg aacgtgaagc
2701 gactgctgct gcaaaacgtc tgcgacctga gcaacaacat gaatggtctt cggtttccgt
2761 gtttcgtaaa gtctggaaac gcggaagtca gcgccctgca ccattatgtt ccggatctgc
2821 atcgcaggat gctgctggct accctgtgga acacctacat ctgtattaac gaagcgctgg
2881 cattgaccct gagtgatttt tctctggtcc cgccgcatcc ataccgccag ttgtttaccc
2941 tcacaacgtt ccagtaaccg ggcatgttca tcatcagtaa cccgtatcgt gagcatcctc
3001 tctcgtttca tcggtatcat tacccccatg aacagaaatc ccccttacac ggaggcatca
3061 gtgaccaaac aggaaaaaac cgcccttaac atggcccgct ttatcagaag ccagacatta
3121 acgcttctgg agaaactcaa cgagctggac gcggatgaac aggcagacat ctgtgaatcg
3181 cttcacgacc acgctgatga gctttaccgc agctgcctcg cgcgtttcgg tgatgacggt
3241 gaaaacctct gacacatgca gctcccggag acggtcacag cttgtctgta agcggatgcc
3301 gggagcagac aagcccgtca gggcgcgtca gcgggtgttg gcgggtgtcg gggcgcagcc
3361 atgacccagt cacgtagcga tagcggagtg tatactggct taactatgcg gcatcagagc
3421 agattgtact gagagtgcac catatatgcg gtgtgaaata ccgcacagat gcgtaaggag
3481 aaaataccgc atcaggcgct cttccgcttc ctcgctcact gactcgctgc gctcggtcgt
3541 tcggctgcgg cgagcggtat cagctcactc aaaggcggta atacggttat ccacagaatc
3601 aggggataac gcaggaaaga acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa
3661 aaaggccgcg ttgctggcgt ttttccatag gctccgcccc cctgacgagc atcacaaaaa
3721 tcgacgctca agtcagaggt ggcgaaaccc gacaggacta taaagatacc aggcgtttcc
3781 ccctggaagc tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg gatacctgtc
3841 cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta ggtatctcag
3901 ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga
3961 ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggtaagac acgacttatc
4021 gccactggca gcagccactg gtaacaggat tagcagagcg aggtatgtag gcggtgctac
4081 agagttcttg aagtggtggc ctaactacgg ctacactaga aggacagtat ttggtatctg
4141 cgctctgctg aagccagtta ccttcggaaa aagagttggt agctcttgat ccggcaaaca
4201 aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa
4261 aggatctcaa gaagatcctt tgatcttttc tacggggtct gacgctcagt ggaacgaaaa
4321 ctcacgttaa gggattttgg tcatgagatt atcaaaaagg atcttcacct agatcctttt
4381 aaattaaaaa tgaagtttta aatcaatcta aagtatatat gagtaaactt ggtctgacag
4441 ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc tgtctatttc gttcatccat
4501 agttgcctga ctccccgtcg tgtagataac tacgatacgg gagggcttac catctggccc
4561 cagtgctgca atgataccgc gagacccacg ctcaccggct ccagatttat cagcaataaa
4621 ccagccagcc ggaagggccg agcgcagaag tggtcctgca actttatccg cctccatcca
4681 gtctattaat tgttgccggg aagctagagt aagtagttcg ccagttaata gtttgcgcaa
4741 cgttgttgcc attgctgcag gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt
4801 cagctccggt tcccaacgat caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc
4861 ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag tgttatcact
4921 catggttatg gcagcactgc ataattctct tactgtcatg ccatccgtaa gatgcttttc
4981 tgtgactggt gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg
5041 ctcttgcccg gcgtcaatac gggataatac cgcgccacat agcagaactt taaaagtgct
5101 catcattgga aaacgttctt cggggcgaaa actctcaagg atcttaccgc tgttgagatc
5161 cagttcgatg taacccactc gtgcacccaa ctgatcttca gcatctttta ctttcaccag
5221 cgtttctggg tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa taagggcgac
5281 acggaaatgt tgaatactca tactcttcct ttttcaatat tattgaagca tttatcaggg
5341 ttattgtctc atgagcggat acatatttga atgtatttag aaaaataaac aaataggggt
5401 tccgcgcaca tttccccgaa aagtgccacc tgaaattgta aacgttaata ttttgttaaa
5461 attcgcgtta aatttttgtt aaatcagctc attttttaac caataggccg aaatcggcaa
5521 aatcccttat aaatcaaaag aatagaccga gatagggttg agtgttgttc cagtttggaa
5581 caagagtcca ctattaaaga acgtggactc caacgtcaaa gggcgaaaaa ccgtctatca
5641 gggcgatggc ccactacgtg aaccatcacc ctaatcaagt tttttggggt cgaggtgccg
5701 taaagcacta aatcggaacc ctaaagggag cccccgattt agagcttgac ggggaaagcc
5761 ggcgaacgtg gcgagaaagg aagggaagaa agcgaaagga gcgggcgcta gggcgctggc
5821 aagtgtagcg gtcacgctgc gcgtaaccac cacacccgcc gcgcttaatg cgccgctaca
5881 gggcgcgtcc cattcgcca
//
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1435 pSG5L Flag HA
1436 pcDNA3 Flag HA
401 pSG5L
476 p5E-mcs
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