pET-27b(+)

pET-27b(+)

pET27b载体基本信息

出品公司:	Ybscience
别名:	pET27b, pet 27b
质粒类型:	大肠杆菌蛋白表达
表达水平:	高
克隆方法:	多克隆位点，限制性内切酶
载体大小:	5414 bp
5' 测序引物:	T7
5' 测序引物序列:	5'-TAATACGACTCACTATAGGG-3'
载体标签:	N-pelB, C-His, C-HSV
载体抗性:	Kanamycin
表达宿主菌:	BL21(DE3) 、BLR(DE3)、BL21(DE3)pLysS
备注:	含有pelB信号肽，能够将目的蛋白定位于细胞周质腔，除抗性不同外，其它方面均与pET-25b相同。
稳定性:	瞬时表达 Transient
组成型:	组成型 Constitutive
病毒/非病毒:	非病毒

 

pET27b载体质粒图谱和多克隆位点信息

pET27b载体简介

 

The pET-27b(+) vector (Cat. No. 69863-3) carries an N-terminal pelB signal sequence for potential periplasmic localization, plus optional C-terminal HSV•Tag® and His•Tag® sequences. This vector differs from pET-25b(+) only by its selectable marker (kanamycin vs. ampicillin resistance).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 (Cat. No. 69337-3).

 

 

pET27b载体序列

LOCUS       pET-27b(+)    5414 bp     DNA Circular    SYN
DEFINITION  pET-27b(+)
ACCESSION   
KEYWORDS    
SOURCE      
  ORGANISM  other sequences; artificial sequences; vectors.
COMMENT     This file is created by Vector NTI
            http://www.biofeng.com/
COMMENT     ORIGDB|GenBank
COMMENT     VNTAUTHORNAME|biofeng.com|
COMMENT     VNTOAUTHORAD1|Novagen Inc.|
FEATURES             Location/Qualifiers
     source          1..5414
                     /organism="pET-27b(+)"
                     /mol_type="other DNA"
     terminator      complement(1..129)
                     /label="T7_terminator"
     misc_feature    69..87
                     /label="T7_Terminal_primer"
     misc_feature    complement(352..368)
                     /label="T7_transl_en_RBS"
     misc_feature    complement(386..413)
                     /label="lacO"
     promoter        complement(413..431)
                     /label="T7_promoter"
     gene            463..726
                     /label="tet (300 - 563)"
                     /gene="tet (300 - 563)"
     misc_feature    complement(515..534)
                     /label="pBRrevBam_primer"
     CDS             568..1068
                     /label="ORF frame 1"
                     /translation="MSACFGVGMVAGPVAGGLLGAISLHAPFLAAAVLNGLNLLLGCF
                     LMQESHKGERRDPGHHRMAQNLSRYGMIAPGRESIQGGECETSNVIRCRRVCRCLLSD
                     RFPRGEPGQPRFCENAGKSGSGDGGAELHSQPRGTTTGGQTVVADWRCHLQSGPARAV
                     ANCRGD*"
     misc_feature    809..1900
                     /label="lacI"
                     /translation="MSACFGVGMVAGPVAGGLLGAISLHAPFLAAAVLNGLNLLLGCF
                     LMQESHKGERRDPGHHRMAQNLSRYGMIAPGRESIQGGECETSNVIRCRRVCRCLLSD
                     RFPRGEPGQPRFCENAGKSGSGDGGAELHSQPRGTTTGGQTVVADWRCHLQSGPARAV
                     ANCRGD*"
     CDS             941..1900
                     /label="ORF frame 2"
                     /translation="MAELNYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKS
                     RADQLGASVVVSMVERSGVEACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNV
                     PALFLDVSDQTPINSIIFSHEDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGW
                     HKYLTRNQIQPIAEREGDWSAMSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITE
                     SGLRVGADISVVGYDDTEDSSCYIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQ
                     LLPVSLVKRKTTLAPNTQTASPRALADSLMQLARQVSRLESGQ*"
     gene            1959..2234
                     /label="tet (576 - 851)"
                     /gene="tet (576 - 851)"
                     /translation="MAELNYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKS
                     RADQLGASVVVSMVERSGVEACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNV
                     PALFLDVSDQTPINSIIFSHEDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGW
                     HKYLTRNQIQPIAEREGDWSAMSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITE
                     SGLRVGADISVVGYDDTEDSSCYIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQ
                     LLPVSLVKRKTTLAPNTQTASPRALADSLMQLARQVSRLESGQ*"
     misc_feature    2709..2900
                     /label="ROP"
                     /translation="MAELNYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKS
                     RADQLGASVVVSMVERSGVEACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNV
                     PALFLDVSDQTPINSIIFSHEDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGW
                     HKYLTRNQIQPIAEREGDWSAMSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITE
                     SGLRVGADISVVGYDDTEDSSCYIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQ
                     LLPVSLVKRKTTLAPNTQTASPRALADSLMQLARQVSRLESGQ*"
     misc_feature    complement(2894..2916)
                     /label="pGEX_3_primer"
                     /translation="MAELNYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKS
                     RADQLGASVVVSMVERSGVEACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNV
                     PALFLDVSDQTPINSIIFSHEDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGW
                     HKYLTRNQIQPIAEREGDWSAMSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITE
                     SGLRVGADISVVGYDDTEDSSCYIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQ
                     LLPVSLVKRKTTLAPNTQTASPRALADSLMQLARQVSRLESGQ*"
     rep_origin      complement(3315..3934)
                     /label="pBR322_origin"
                     /translation="MAELNYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKS
                     RADQLGASVVVSMVERSGVEACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNV
                     PALFLDVSDQTPINSIIFSHEDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGW
                     HKYLTRNQIQPIAEREGDWSAMSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITE
                     SGLRVGADISVVGYDDTEDSSCYIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQ
                     LLPVSLVKRKTTLAPNTQTASPRALADSLMQLARQVSRLESGQ*"
     gene            4040..4855
                     /label="KanR2"
                     /gene="KanR2"
                     /translation="MAELNYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKS
                     RADQLGASVVVSMVERSGVEACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNV
                     PALFLDVSDQTPINSIIFSHEDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGW
                     HKYLTRNQIQPIAEREGDWSAMSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITE
                     SGLRVGADISVVGYDDTEDSSCYIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQ
                     LLPVSLVKRKTTLAPNTQTASPRALADSLMQLARQVSRLESGQ*"
     CDS             4040..4855
                     /label="ORF frame 2"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLY
                     GKPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIP
                     GKTAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGL
                     VDASDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVG
                     RVGIADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF*"
ORIGIN
    1 atccggatat agttcctcct ttcagcaaaa aacccctcaa gacccgttta gaggccccaa
   61 ggggttatgc tagttattgc tcagcggtgg cagcagccaa ctcagcttcc tttcgggctt
  121 tgttagcagc cggatctcag tggtggtggt ggtggtgctc gacatcctcg gggtcttccg
  181 gggcgagttc tggctggcta gcccgtttga tctcgagtgc ggccgcaagc ttgtcgacgg
  241 agctcgaatt cggatccgaa ttaattccga tatccatggc catcgccggc tgggcagcga
  301 ggagcagcag accagcagca gcggtcggca gcaggtattt catatgtata tctccttctt
  361 aaagttaaac aaaattattt ctagagggga attgttatcc gctcacaatt cccctatagt
  421 gagtcgtatt aatttcgcgg gatcgagatc tcgatcctct acgccggacg catcgtggcc
  481 ggcatcaccg gcgccacagg tgcggttgct ggcgcctata tcgccgacat caccgatggg
  541 gaagatcggg ctcgccactt cgggctcatg agcgcttgtt tcggcgtggg tatggtggca
  601 ggccccgtgg ccgggggact gttgggcgcc atctccttgc atgcaccatt ccttgcggcg
  661 gcggtgctca acggcctcaa cctactactg ggctgcttcc taatgcagga gtcgcataag
  721 ggagagcgtc gagatcccgg acaccatcga atggcgcaaa acctttcgcg gtatggcatg
  781 atagcgcccg gaagagagtc aattcagggt ggtgaatgtg aaaccagtaa cgttatacga
  841 tgtcgcagag tatgccggtg tctcttatca gaccgtttcc cgcgtggtga accaggccag
  901 ccacgtttct gcgaaaacgc gggaaaaagt ggaagcggcg atggcggagc tgaattacat
  961 tcccaaccgc gtggcacaac aactggcggg caaacagtcg ttgctgattg gcgttgccac
 1021 ctccagtctg gccctgcacg cgccgtcgca aattgtcgcg gcgattaaat ctcgcgccga
 1081 tcaactgggt gccagcgtgg tggtgtcgat ggtagaacga agcggcgtcg aagcctgtaa
 1141 agcggcggtg cacaatcttc tcgcgcaacg cgtcagtggg ctgatcatta actatccgct
 1201 ggatgaccag gatgccattg ctgtggaagc tgcctgcact aatgttccgg cgttatttct
 1261 tgatgtctct gaccagacac ccatcaacag tattattttc tcccatgaag acggtacgcg
 1321 actgggcgtg gagcatctgg tcgcattggg tcaccagcaa atcgcgctgt tagcgggccc
 1381 attaagttct gtctcggcgc gtctgcgtct ggctggctgg cataaatatc tcactcgcaa
 1441 tcaaattcag ccgatagcgg aacgggaagg cgactggagt gccatgtccg gttttcaaca
 1501 aaccatgcaa atgctgaatg agggcatcgt tcccactgcg atgctggttg ccaacgatca
 1561 gatggcgctg ggcgcaatgc gcgccattac cgagtccggg ctgcgcgttg gtgcggatat
 1621 ctcggtagtg ggatacgacg ataccgaaga cagctcatgt tatatcccgc cgttaaccac
 1681 catcaaacag gattttcgcc tgctggggca aaccagcgtg gaccgcttgc tgcaactctc
 1741 tcagggccag gcggtgaagg gcaatcagct gttgcccgtc tcactggtga aaagaaaaac
 1801 caccctggcg cccaatacgc aaaccgcctc tccccgcgcg ttggccgatt cattaatgca
 1861 gctggcacga caggtttccc gactggaaag cgggcagtga gcgcaacgca attaatgtaa
 1921 gttagctcac tcattaggca ccgggatctc gaccgatgcc cttgagagcc ttcaacccag
 1981 tcagctcctt ccggtgggcg cggggcatga ctatcgtcgc cgcacttatg actgtcttct
 2041 ttatcatgca actcgtagga caggtgccgg cagcgctctg ggtcattttc ggcgaggacc
 2101 gctttcgctg gagcgcgacg atgatcggcc tgtcgcttgc ggtattcgga atcttgcacg
 2161 ccctcgctca agccttcgtc actggtcccg ccaccaaacg tttcggcgag aagcaggcca
 2221 ttatcgccgg catggcggcc ccacgggtgc gcatgatcgt gctcctgtcg ttgaggaccc
 2281 ggctaggctg gcggggttgc cttactggtt agcagaatga atcaccgata cgcgagcgaa
 2341 cgtgaagcga ctgctgctgc aaaacgtctg cgacctgagc aacaacatga atggtcttcg
 2401 gtttccgtgt ttcgtaaagt ctggaaacgc ggaagtcagc gccctgcacc attatgttcc
 2461 ggatctgcat cgcaggatgc tgctggctac cctgtggaac acctacatct gtattaacga
 2521 agcgctggca ttgaccctga gtgatttttc tctggtcccg ccgcatccat accgccagtt
 2581 gtttaccctc acaacgttcc agtaaccggg catgttcatc atcagtaacc cgtatcgtga
 2641 gcatcctctc tcgtttcatc ggtatcatta cccccatgaa cagaaatccc ccttacacgg
 2701 aggcatcagt gaccaaacag gaaaaaaccg cccttaacat ggcccgcttt atcagaagcc
 2761 agacattaac gcttctggag aaactcaacg agctggacgc ggatgaacag gcagacatct
 2821 gtgaatcgct tcacgaccac gctgatgagc tttaccgcag ctgcctcgcg cgtttcggtg
 2881 atgacggtga aaacctctga cacatgcagc tcccggagac ggtcacagct tgtctgtaag
 2941 cggatgccgg gagcagacaa gcccgtcagg gcgcgtcagc gggtgttggc gggtgtcggg
 3001 gcgcagccat gacccagtca cgtagcgata gcggagtgta tactggctta actatgcggc
 3061 atcagagcag attgtactga gagtgcacca tatatgcggt gtgaaatacc gcacagatgc
 3121 gtaaggagaa aataccgcat caggcgctct tccgcttcct cgctcactga ctcgctgcgc
 3181 tcggtcgttc ggctgcggcg agcggtatca gctcactcaa aggcggtaat acggttatcc
 3241 acagaatcag gggataacgc aggaaagaac atgtgagcaa aaggccagca aaaggccagg
 3301 aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc tccgcccccc tgacgagcat
 3361 cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga caggactata aagataccag
 3421 gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga
 3481 tacctgtccg cctttctccc ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg
 3541 tatctcagtt cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga accccccgtt
 3601 cagcccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc ggtaagacac
 3661 gacttatcgc cactggcagc agccactggt aacaggatta gcagagcgag gtatgtaggc
 3721 ggtgctacag agttcttgaa gtggtggcct aactacggct acactagaag gacagtattt
 3781 ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa gagttggtag ctcttgatcc
 3841 ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt gcaagcagca gattacgcgc
 3901 agaaaaaaag gatctcaaga agatcctttg atcttttcta cggggtctga cgctcagtgg
 3961 aacgaaaact cacgttaagg gattttggtc atgaacaata aaactgtctg cttacataaa
 4021 cagtaataca aggggtgtta tgagccatat tcaacgggaa acgtcttgct ctaggccgcg
 4081 attaaattcc aacatggatg ctgatttata tgggtataaa tgggctcgcg ataatgtcgg
 4141 gcaatcaggt gcgacaatct atcgattgta tgggaagccc gatgcgccag agttgtttct
 4201 gaaacatggc aaaggtagcg ttgccaatga tgttacagat gagatggtca gactaaactg
 4261 gctgacggaa tttatgcctc ttccgaccat caagcatttt atccgtactc ctgatgatgc
 4321 atggttactc accactgcga tccccgggaa aacagcattc caggtattag aagaatatcc
 4381 tgattcaggt gaaaatattg ttgatgcgct ggcagtgttc ctgcgccggt tgcattcgat
 4441 tcctgtttgt aattgtcctt ttaacagcga tcgcgtattt cgtctcgctc aggcgcaatc
 4501 acgaatgaat aacggtttgg ttgatgcgag tgattttgat gacgagcgta atggctggcc
 4561 tgttgaacaa gtctggaaag aaatgcataa acttttgcca ttctcaccgg attcagtcgt
 4621 cactcatggt gatttctcac ttgataacct tatttttgac gaggggaaat taataggttg
 4681 tattgatgtt ggacgagtcg gaatcgcaga ccgataccag gatcttgcca tcctatggaa
 4741 ctgcctcggt gagttttctc cttcattaca gaaacggctt tttcaaaaat atggtattga
 4801 taatcctgat atgaataaat tgcagtttca tttgatgctc gatgagtttt tctaagaatt
 4861 aattcatgag cggatacata tttgaatgta tttagaaaaa taaacaaata ggggttccgc
 4921 gcacatttcc ccgaaaagtg ccacctgaaa ttgtaaacgt taatattttg ttaaaattcg
 4981 cgttaaattt ttgttaaatc agctcatttt ttaaccaata ggccgaaatc ggcaaaatcc
 5041 cttataaatc aaaagaatag accgagatag ggttgagtgt tgttccagtt tggaacaaga
 5101 gtccactatt aaagaacgtg gactccaacg tcaaagggcg aaaaaccgtc tatcagggcg
 5161 atggcccact acgtgaacca tcaccctaat caagtttttt ggggtcgagg tgccgtaaag
 5221 cactaaatcg gaaccctaaa gggagccccc gatttagagc ttgacgggga aagccggcga
 5281 acgtggcgag aaaggaaggg aagaaagcga aaggagcggg cgctagggcg ctggcaagtg
 5341 tagcggtcac gctgcgcgta accaccacac ccgccgcgct taatgcgccg ctacagggcg
 5401 cgtcccattc gcca
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