中國微生物菌種查詢網(wǎng)pET-26b質(zhì)粒載體基本信息介紹！
中國微生物菌種查詢網(wǎng) / 2018-11-28 09:08:19
基本信息
啟動子 : T7/lac
復制子 : ColE1 ori，F(xiàn)1 ori
終止子 : T7 terminator
質(zhì)粒分類 : 大腸桿菌載體；PET系列表達質(zhì)粒
質(zhì)粒大小 : 5360bp
質(zhì)粒標簽 : C-6×His
原核抗性 : 卡那霉素Kan
克隆菌株 : DH5α
培養(yǎng)條件 : 37℃，有氧，LB
表達宿主 : BL21(DE3)
誘導方式:
IPTG或乳糖及其類似物
5'測序引物 : T7:TAATACGACTCACTATAGGG
3'測序引物 : T7-ter : TGCTAGTTATTGCTCAGCGG
備注 : 分泌表達
質(zhì)粒簡介
The pET-26b(+) vector carries an N-terminal pelB signal sequence for potential periplasmic localization, plus optional C-terminal His?Tag? sequence. 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 circular 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 .
質(zhì)粒圖譜
質(zhì)粒序列
LOCUS Exported 5360 bp ds-DNA circular SYN 31-7-2015
DEFINITION synthetic circular DNA
ACCESSION .
VERSION .
KEYWORDS Untitled 2
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 5360)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported 2015-7-31 MLCC
http://www.miaolingbio.com
FEATURES Location/Qualifiers
source 1..5360
/organism="synthetic DNA construct"
/mol_type="other DNA"
source 297..319
/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(224..289)
/codon_start=1
/gene="pelB (fragment)"
/product="leader peptide for secretion"
/note="pelB signal sequence"
/translation="MKYLLPTAAAGLLLLAAQPAMA"
RBS 297..319
/note="efficient ribosome binding site from bacteriophage
T7 gene 10 (Olins and Rangwala, 1989)"
protein_bind 334..358
/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(359..377)
/note="T7 promoter"
/note="promoter for bacteriophage T7 RNA polymerase"
promoter 686..763
/gene="lacI"
/note="lacI promoter"
CDS 764..1846
/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 2655..2846
/codon_start=1
/gene="rop"
/product="Rop protein, which maintains plasmids at low copy
number"
/note="rop"
/translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
DELYRSCLARFGDDGENL"
misc_feature 2948..3090
/note="bom"
/note="basis of mobility region from pBR322"
rep_origin complement(3276..3864)
/direction=LEFT
/note="ori"
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
CDS 3986..4801
/codon_start=1
/product="aminoglycoside phosphotransferase"
/note="KanR"
/note="confers resistance to kanamycin in bacteria or G418
(Geneticin(R)) in eukaryotes"
/translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
rep_origin complement(4894..5349)
/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 tccgaattaa ttccgatatc catggccatc gccggctggg
241 cagcgaggag cagcagacca gcagcagcgg tcggcagcag gtatttcata tgtatatctc
301 cttcttaaag ttaaacaaaa ttatttctag aggggaattg ttatccgctc acaattcccc
361 tatagtgagt cgtattaatt tcgcgggatc gagatctcga tcctctacgc cggacgcatc
421 gtggccggca tcaccggcgc cacaggtgcg gttgctggcg cctatatcgc cgacatcacc
481 gatggggaag atcgggctcg ccacttcggg ctcatgagcg cttgtttcgg cgtgggtatg
541 gtggcaggcc ccgtggccgg gggactgttg ggcgccatct ccttgcatgc accattcctt
601 gcggcggcgg tgctcaacgg cctcaaccta ctactgggct gcttcctaat gcaggagtcg
661 cataagggag agcgtcgaga tcccggacac catcgaatgg cgcaaaacct ttcgcggtat
721 ggcatgatag cgcccggaag agagtcaatt cagggtggtg aatgtgaaac cagtaacgtt
781 atacgatgtc gcagagtatg ccggtgtctc ttatcagacc gtttcccgcg tggtgaacca
841 ggccagccac gtttctgcga aaacgcggga aaaagtggaa gcggcgatgg cggagctgaa
901 ttacattccc aaccgcgtgg cacaacaact ggcgggcaaa cagtcgttgc tgattggcgt
961 tgccacctcc agtctggccc tgcacgcgcc gtcgcaaatt gtcgcggcga ttaaatctcg
1021 cgccgatcaa ctgggtgcca gcgtggtggt gtcgatggta gaacgaagcg gcgtcgaagc
1081 ctgtaaagcg gcggtgcaca atcttctcgc gcaacgcgtc agtgggctga tcattaacta
1141 tccgctggat gaccaggatg ccattgctgt ggaagctgcc tgcactaatg ttccggcgtt
1201 atttcttgat gtctctgacc agacacccat caacagtatt attttctccc atgaagacgg