pET-30 Xa/LIC

pET-30 Xa/LIC

pET30 Xa/LIC载体基本信息

出品公司:	Ybscience
别名:	pET30 Xa/LIC, pet 30a Xa/LIC
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:	5448 bp
5' 测序引物:	T7
5' 测序引物序列:	5'-TAATACGACTCACTATAGGG-3'
载体标签:	N-His, N-S, N-Xa Factor, C-His
载体抗性:	Kanamycin
表达宿主菌:	BL21(DE3) 、 Rosetta2(DE3)、C43(DE3)
备注:	For directional cloning of PCR-amplified DNA; ligation independent cloning; Factor Xa cleavage site
稳定性:	瞬时表达 Transient
组成型:	组成型 Constitutive
病毒/非病毒:	非病毒

pET30 Xa/LIC载体质粒图谱和多克隆位点信息

 

Feature Name	Start	End
T7_terminator	129	1
T7_Terminal_primer	69	87
lacI	328	1934
T7_transl_en_RBS	398	382
lacO	443	416
T7_promoter	461	443
tet (300 - 563)	497	760
pBRrevBam_primer	568	549
tet (576 - 851)	1993	2268
ROP	2743	2934
pGEX_3_primer	2950	2928
pBR322_origin	3968	3349
KanR2	4074	4889

ORF	Start	End
ORF frame 2	353	1102
ORF frame 3	975	1934
ORF frame 3	4074	4889

Enzyme Name	Cut
XhoI	158
NotI	166
EagI	166
HindIII	173
SalI	179
SacI	190
EcoRI	192
BamHI	198
EcoRV	206
NcoI	211
MscI	216
AgeI	257
KpnI	264
BglII	267
BstBI	294
NdeI	372
XbaI	410
BclI	1216
ApaI	1413
HpaI	1708
FspI	2284
NruI	4162
SmaI	4379
XmaI	4377

 

pET30 Xa/LIC载体简介

The pET-30 Xa/LIC vector is designed for rapid, directional cloning of PCR-amplified DNA for high-level expression of polypeptides containing N-terminal His•Tag® and S•Tag™ sequences for detection and purification. Using specifically designed primers for amplification and the pET-30Xa/LIC Vector Kit (Cat. No. 69073-3), inserts can be efficiently cloned without the need for restriction digestion or ligation, and the resulting fusion proteins can be cleaved with Factor Xa to precisely remove all vector-encoded amino acids. 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 polymeraseis 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).

pET30 Xa/LIC载体序列

LOCUS       pET-30 Xa/LIC    5448 bp     DNA Circular    SYN
DEFINITION  pET-30 Xa/LIC
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..5448
                     /organism="pET-30 Xa/LIC"
                     /mol_type="other DNA"
     terminator      complement(1..129)
                     /label="T7_terminator"
     misc_feature    69..87
                     /label="T7_Terminal_primer"
     misc_feature    328..1934
                     /label="lacI"
     CDS             353..1102
                     /label="ORF frame 2"
                     /translation="MMMMMVHMYISFLKLNKIISRGELLSAHNSPIVSRINFAGSRSI
                     SILYAGRIVAGITGATGAVAGAYIADITDGEDRARHFGLMSACFGVGMVAGPVAGGLL
                     GAISLHAPFLAAAVLNGLNLLLGCFLMQESHKGERRDPGHHRMAQNLSRYGMIAPGRE
                     SIQGGECETSNVIRCRRVCRCLLSDRFPRGEPGQPRFCENAGKSGSGDGGAELHSQPR
                     GTTTGGQTVVADWRCHLQSGPARAVANCRGD*"
     misc_feature    complement(382..398)
                     /label="T7_transl_en_RBS"
                     /translation="MMMMMVHMYISFLKLNKIISRGELLSAHNSPIVSRINFAGSRSI
                     SILYAGRIVAGITGATGAVAGAYIADITDGEDRARHFGLMSACFGVGMVAGPVAGGLL
                     GAISLHAPFLAAAVLNGLNLLLGCFLMQESHKGERRDPGHHRMAQNLSRYGMIAPGRE
                     SIQGGECETSNVIRCRRVCRCLLSDRFPRGEPGQPRFCENAGKSGSGDGGAELHSQPR
                     GTTTGGQTVVADWRCHLQSGPARAVANCRGD*"
     misc_feature    complement(416..443)
                     /label="lacO"
                     /translation="MMMMMVHMYISFLKLNKIISRGELLSAHNSPIVSRINFAGSRSI
                     SILYAGRIVAGITGATGAVAGAYIADITDGEDRARHFGLMSACFGVGMVAGPVAGGLL
                     GAISLHAPFLAAAVLNGLNLLLGCFLMQESHKGERRDPGHHRMAQNLSRYGMIAPGRE
                     SIQGGECETSNVIRCRRVCRCLLSDRFPRGEPGQPRFCENAGKSGSGDGGAELHSQPR
                     GTTTGGQTVVADWRCHLQSGPARAVANCRGD*"
     promoter        complement(443..461)
                     /label="T7_promoter"
                     /translation="MMMMMVHMYISFLKLNKIISRGELLSAHNSPIVSRINFAGSRSI
                     SILYAGRIVAGITGATGAVAGAYIADITDGEDRARHFGLMSACFGVGMVAGPVAGGLL
                     GAISLHAPFLAAAVLNGLNLLLGCFLMQESHKGERRDPGHHRMAQNLSRYGMIAPGRE
                     SIQGGECETSNVIRCRRVCRCLLSDRFPRGEPGQPRFCENAGKSGSGDGGAELHSQPR
                     GTTTGGQTVVADWRCHLQSGPARAVANCRGD*"
     gene            497..760
                     /label="tet (300 - 563)"
                     /gene="tet (300 - 563)"
                     /translation="MMMMMVHMYISFLKLNKIISRGELLSAHNSPIVSRINFAGSRSI
                     SILYAGRIVAGITGATGAVAGAYIADITDGEDRARHFGLMSACFGVGMVAGPVAGGLL
                     GAISLHAPFLAAAVLNGLNLLLGCFLMQESHKGERRDPGHHRMAQNLSRYGMIAPGRE
                     SIQGGECETSNVIRCRRVCRCLLSDRFPRGEPGQPRFCENAGKSGSGDGGAELHSQPR
                     GTTTGGQTVVADWRCHLQSGPARAVANCRGD*"
     misc_feature    complement(549..568)
                     /label="pBRrevBam_primer"
                     /translation="MMMMMVHMYISFLKLNKIISRGELLSAHNSPIVSRINFAGSRSI
                     SILYAGRIVAGITGATGAVAGAYIADITDGEDRARHFGLMSACFGVGMVAGPVAGGLL
                     GAISLHAPFLAAAVLNGLNLLLGCFLMQESHKGERRDPGHHRMAQNLSRYGMIAPGRE
                     SIQGGECETSNVIRCRRVCRCLLSDRFPRGEPGQPRFCENAGKSGSGDGGAELHSQPR
                     GTTTGGQTVVADWRCHLQSGPARAVANCRGD*"
     CDS             975..1934
                     /label="ORF frame 3"
                     /translation="MAELNYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKS
                     RADQLGASVVVSMVERSGVEACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNV
                     PALFLDVSDQTPINSIIFSHEDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGW
                     HKYLTRNQIQPIAEREGDWSAMSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITE
                     SGLRVGADISVVGYDDTEDSSCYIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQ
                     LLPVSLVKRKTTLAPNTQTASPRALADSLMQLARQVSRLESGQ*"
     gene            1993..2268
                     /label="tet (576 - 851)"
                     /gene="tet (576 - 851)"
                     /translation="MAELNYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKS
                     RADQLGASVVVSMVERSGVEACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNV
                     PALFLDVSDQTPINSIIFSHEDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGW
                     HKYLTRNQIQPIAEREGDWSAMSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITE
                     SGLRVGADISVVGYDDTEDSSCYIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQ
                     LLPVSLVKRKTTLAPNTQTASPRALADSLMQLARQVSRLESGQ*"
     misc_feature    2743..2934
                     /label="ROP"
                     /translation="MAELNYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKS
                     RADQLGASVVVSMVERSGVEACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNV
                     PALFLDVSDQTPINSIIFSHEDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGW
                     HKYLTRNQIQPIAEREGDWSAMSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITE
                     SGLRVGADISVVGYDDTEDSSCYIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQ
                     LLPVSLVKRKTTLAPNTQTASPRALADSLMQLARQVSRLESGQ*"
     misc_feature    complement(2928..2950)
                     /label="pGEX_3_primer"
                     /translation="MAELNYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKS
                     RADQLGASVVVSMVERSGVEACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNV
                     PALFLDVSDQTPINSIIFSHEDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGW
                     HKYLTRNQIQPIAEREGDWSAMSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITE
                     SGLRVGADISVVGYDDTEDSSCYIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQ
                     LLPVSLVKRKTTLAPNTQTASPRALADSLMQLARQVSRLESGQ*"
     rep_origin      complement(3349..3968)
                     /label="pBR322_origin"
                     /translation="MAELNYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKS
                     RADQLGASVVVSMVERSGVEACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNV
                     PALFLDVSDQTPINSIIFSHEDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGW
                     HKYLTRNQIQPIAEREGDWSAMSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITE
                     SGLRVGADISVVGYDDTEDSSCYIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQ
                     LLPVSLVKRKTTLAPNTQTASPRALADSLMQLARQVSRLESGQ*"
     gene            4074..4889
                     /label="KanR2"
                     /gene="KanR2"
                     /translation="MAELNYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKS
                     RADQLGASVVVSMVERSGVEACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNV
                     PALFLDVSDQTPINSIIFSHEDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGW
                     HKYLTRNQIQPIAEREGDWSAMSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITE
                     SGLRVGADISVVGYDDTEDSSCYIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQ
                     LLPVSLVKRKTTLAPNTQTASPRALADSLMQLARQVSRLESGQ*"
     CDS             4074..4889
                     /label="ORF frame 3"
                     /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 gagtgcggcc gcaagcttgt
  181 cgacggagct cgaattcgga tccgatatcg ccatggccag aggagagtta gagcgaccct
  241 caataccgga gccaccaccg gtacccagat ctgggctgtc catgtgctgg cgttcgaatt
  301 tagcagcagc ggtttctttc ataccagaac cgcgtggcac cagaccagaa gaatgatgat
  361 gatgatggtg catatgtata tctccttctt aaagttaaac aaaattattt ctagagggga
  421 attgttatcc gctcacaatt cccctatagt gagtcgtatt aatttcgcgg gatcgagatc
  481 gatctcgatc ctctacgccg gacgcatcgt ggccggcatc accggcgcca caggtgcggt
  541 tgctggcgcc tatatcgccg acatcaccga tggggaagat cgggctcgcc acttcgggct
  601 catgagcgct tgtttcggcg tgggtatggt ggcaggcccc gtggccgggg gactgttggg
  661 cgccatctcc ttgcatgcac cattccttgc ggcggcggtg ctcaacggcc tcaacctact
  721 actgggctgc ttcctaatgc aggagtcgca taagggagag cgtcgagatc ccggacacca
  781 tcgaatggcg caaaaccttt cgcggtatgg catgatagcg cccggaagag agtcaattca
  841 gggtggtgaa tgtgaaacca gtaacgttat acgatgtcgc agagtatgcc ggtgtctctt
  901 atcagaccgt ttcccgcgtg gtgaaccagg ccagccacgt ttctgcgaaa acgcgggaaa
  961 aagtggaagc ggcgatggcg gagctgaatt acattcccaa ccgcgtggca caacaactgg
 1021 cgggcaaaca gtcgttgctg attggcgttg ccacctccag tctggccctg cacgcgccgt
 1081 cgcaaattgt cgcggcgatt aaatctcgcg ccgatcaact gggtgccagc gtggtggtgt
 1141 cgatggtaga acgaagcggc gtcgaagcct gtaaagcggc ggtgcacaat cttctcgcgc
 1201 aacgcgtcag tgggctgatc attaactatc cgctggatga ccaggatgcc attgctgtgg
 1261 aagctgcctg cactaatgtt ccggcgttat ttcttgatgt ctctgaccag acacccatca
 1321 acagtattat tttctcccat gaagacggta cgcgactggg cgtggagcat ctggtcgcat
 1381 tgggtcacca gcaaatcgcg ctgttagcgg gcccattaag ttctgtctcg gcgcgtctgc
 1441 gtctggctgg ctggcataaa tatctcactc gcaatcaaat tcagccgata gcggaacggg
 1501 aaggcgactg gagtgccatg tccggttttc aacaaaccat gcaaatgctg aatgagggca
 1561 tcgttcccac tgcgatgctg gttgccaacg atcagatggc gctgggcgca atgcgcgcca
 1621 ttaccgagtc cgggctgcgc gttggtgcgg acatctcggt agtgggatac gacgataccg
 1681 aagacagctc atgttatatc ccgccgttaa ccaccatcaa acaggatttt cgcctgctgg
 1741 ggcaaaccag cgtggaccgc ttgctgcaac tctctcaggg ccaggcggtg aagggcaatc
 1801 agctgttgcc cgtctcactg gtgaaaagaa aaaccaccct ggcgcccaat acgcaaaccg
 1861 cctctccccg cgcgttggcc gattcattaa tgcagctggc acgacaggtt tcccgactgg
 1921 aaagcgggca gtgagcgcaa cgcaattaat gtaagttagc tcactcatta ggcaccggga
 1981 tctcgaccga tgcccttgag agccttcaac ccagtcagct ccttccggtg ggcgcggggc
 2041 atgactatcg tcgccgcact tatgactgtc ttctttatca tgcaactcgt aggacaggtg
 2101 ccggcagcgc tctgggtcat tttcggcgag gaccgctttc gctggagcgc gacgatgatc
 2161 ggcctgtcgc ttgcggtatt cggaatcttg cacgccctcg ctcaagcctt cgtcactggt
 2221 cccgccacca aacgtttcgg cgagaagcag gccattatcg ccggcatggc ggccccacgg
 2281 gtgcgcatga tcgtgctcct gtcgttgagg acccggctag gctggcgggg ttgccttact
 2341 ggttagcaga atgaatcacc gatacgcgag cgaacgtgaa gcgactgctg ctgcaaaacg
 2401 tctgcgacct gagcaacaac atgaatggtc ttcggtttcc gtgtttcgta aagtctggaa
 2461 acgcggaagt cagcgccctg caccattatg ttccggatct gcatcgcagg atgctgctgg
 2521 ctaccctgtg gaacacctac atctgtatta acgaagcgct ggcattgacc ctgagtgatt
 2581 tttctctggt cccgccgcat ccataccgcc agttgtttac cctcacaacg ttccagtaac
 2641 cgggcatgtt catcatcagt aacccgtatc gtgagcatcc tctctcgttt catcggtatc
 2701 attaccccca tgaacagaaa tcccccttac acggaggcat cagtgaccaa acaggaaaaa
 2761 accgccctta acatggcccg ctttatcaga agccagacat taacgcttct ggagaaactc
 2821 aacgagctgg acgcggatga acaggcagac atctgtgaat cgcttcacga ccacgctgat
 2881 gagctttacc gcagctgcct cgcgcgtttc ggtgatgacg gtgaaaacct ctgacacatg
 2941 cagctcccgg agacggtcac agcttgtctg taagcggatg ccgggagcag acaagcccgt
 3001 cagggcgcgt cagcgggtgt tggcgggtgt cggggcgcag ccatgaccca gtcacgtagc
 3061 gatagcggag tgtatactgg cttaactatg cggcatcaga gcagattgta ctgagagtgc
 3121 accatatatg cggtgtgaaa taccgcacag atgcgtaagg agaaaatacc gcatcaggcg
 3181 ctcttccgct tcctcgctca ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt
 3241 atcagctcac tcaaaggcgg taatacggtt atccacagaa tcaggggata acgcaggaaa
 3301 gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc
 3361 gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag
 3421 gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt
 3481 gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg
 3541 aagcgtggcg ctttctcata gctcacgctg taggtatctc agttcggtgt aggtcgttcg
 3601 ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg
 3661 taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac
 3721 tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg
 3781 gcctaactac ggctacacta gaaggacagt atttggtatc tgcgctctgc tgaagccagt
 3841 taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg
 3901 tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc
 3961 tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt
 4021 ggtcatgaac aataaaactg tctgcttaca taaacagtaa tacaaggggt gttatgagcc
 4081 atattcaacg ggaaacgtct tgctctaggc cgcgattaaa ttccaacatg gatgctgatt
 4141 tatatgggta taaatgggct cgcgataatg tcgggcaatc aggtgcgaca atctatcgat
 4201 tgtatgggaa gcccgatgcg ccagagttgt ttctgaaaca tggcaaaggt agcgttgcca
 4261 atgatgttac agatgagatg gtcagactaa actggctgac ggaatttatg cctcttccga
 4321 ccatcaagca ttttatccgt actcctgatg atgcatggtt actcaccact gcgatccccg
 4381 ggaaaacagc attccaggta ttagaagaat atcctgattc aggtgaaaat attgttgatg
 4441 cgctggcagt gttcctgcgc cggttgcatt cgattcctgt ttgtaattgt ccttttaaca
 4501 gcgatcgcgt atttcgtctc gctcaggcgc aatcacgaat gaataacggt ttggttgatg
 4561 cgagtgattt tgatgacgag cgtaatggct ggcctgttga acaagtctgg aaagaaatgc
 4621 ataaactttt gccattctca ccggattcag tcgtcactca tggtgatttc tcacttgata
 4681 accttatttt tgacgagggg aaattaatag gttgtattga tgttggacga gtcggaatcg
 4741 cagaccgata ccaggatctt gccatcctat ggaactgcct cggtgagttt tctccttcat
 4801 tacagaaacg gctttttcaa aaatatggta ttgataatcc tgatatgaat aaattgcagt
 4861 ttcatttgat gctcgatgag tttttctaag aattaattca tgagcggata catatttgaa
 4921 tgtatttaga aaaataaaca aataggggtt ccgcgcacat ttccccgaaa agtgccacct
 4981 gaaattgtaa acgttaatat tttgttaaaa ttcgcgttaa atttttgtta aatcagctca
 5041 ttttttaacc aataggccga aatcggcaaa atcccttata aatcaaaaga atagaccgag
 5101 atagggttga gtgttgttcc agtttggaac aagagtccac tattaaagaa cgtggactcc
 5161 aacgtcaaag ggcgaaaaac cgtctatcag ggcgatggcc cactacgtga accatcaccc
 5221 taatcaagtt ttttggggtc gaggtgccgt aaagcactaa atcggaaccc taaagggagc
 5281 ccccgattta gagcttgacg gggaaagccg gcgaacgtgg cgagaaagga agggaagaaa
 5341 gcgaaaggag cgggcgctag ggcgctggca agtgtagcgg tcacgctgcg cgtaaccacc
 5401 acacccgccg cgcttaatgc gccgctacag ggcgcgtccc attcgcca
//