The pET-41 series is designed for cloning and high-level expression of peptide sequences fused with the 220 aa GST•Tag™ protein. 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 (cat. no. 69337-3).Vector encoded sequence can be completely removed when cloning into the PshAI site (as shown below) and then cleaving the GST fusion protein with Enterokinase.
LOCUS pET-41 c(+) 5934 bp DNA Circular SYN
DEFINITION pET-41 c(+)
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..5934
/organism="pET-41 c(+)"
/mol_type="other DNA"
terminator complement(1..120)
/label="T7_terminator"
misc_feature 69..87
/label="T7_Terminal_primer"
CDS complement(143..1096)
/label="ORF frame 3"
/translation="MSPILGYWKIKGLVQPTRLLLEYLEEKYEEHLYERDEGDKWRNK
KFELGLEFPNLPYYIDGDVKLTQSMAIIRYIADKHNMLGGCPKERAEISMLEGAVLDI
RYGVSRIAYSKDFETLKVDFLSKLPEMLKMFEDRLCHKTYLNGDHVTHPDFMLYDALD
VVLYMDPMCLDAFPKLVCFKKRIEAIPQIDKYLKSSKYIAWPLQGWQATFGGGDHPPK
SDGSTSGSGHHHHHHSAGLVPRGSTAIGMKETAAAKFERQHMDSPDLGTGGGSGDDDD
KSPMAISGIRILYRPWRACRRAPSTSLRPHSSTTTTTTTTN*"
misc_feature complement(266..280)
/label="EK"
/translation="MSPILGYWKIKGLVQPTRLLLEYLEEKYEEHLYERDEGDKWRNK
KFELGLEFPNLPYYIDGDVKLTQSMAIIRYIADKHNMLGGCPKERAEISMLEGAVLDI
RYGVSRIAYSKDFETLKVDFLSKLPEMLKMFEDRLCHKTYLNGDHVTHPDFMLYDALD
VVLYMDPMCLDAFPKLVCFKKRIEAIPQIDKYLKSSKYIAWPLQGWQATFGGGDHPPK
SDGSTSGSGHHHHHHSAGLVPRGSTAIGMKETAAAKFERQHMDSPDLGTGGGSGDDDD
KSPMAISGIRILYRPWRACRRAPSTSLRPHSSTTTTTTTTN*"
misc_feature complement(311..355)
/label="S15"
/translation="MSPILGYWKIKGLVQPTRLLLEYLEEKYEEHLYERDEGDKWRNK
KFELGLEFPNLPYYIDGDVKLTQSMAIIRYIADKHNMLGGCPKERAEISMLEGAVLDI
RYGVSRIAYSKDFETLKVDFLSKLPEMLKMFEDRLCHKTYLNGDHVTHPDFMLYDALD
VVLYMDPMCLDAFPKLVCFKKRIEAIPQIDKYLKSSKYIAWPLQGWQATFGGGDHPPK
SDGSTSGSGHHHHHHSAGLVPRGSTAIGMKETAAAKFERQHMDSPDLGTGGGSGDDDD
KSPMAISGIRILYRPWRACRRAPSTSLRPHSSTTTTTTTTN*"
misc_feature complement(398..415)
/label="6xHIS"
/translation="MSPILGYWKIKGLVQPTRLLLEYLEEKYEEHLYERDEGDKWRNK
KFELGLEFPNLPYYIDGDVKLTQSMAIIRYIADKHNMLGGCPKERAEISMLEGAVLDI
RYGVSRIAYSKDFETLKVDFLSKLPEMLKMFEDRLCHKTYLNGDHVTHPDFMLYDALD
VVLYMDPMCLDAFPKLVCFKKRIEAIPQIDKYLKSSKYIAWPLQGWQATFGGGDHPPK
SDGSTSGSGHHHHHHSAGLVPRGSTAIGMKETAAAKFERQHMDSPDLGTGGGSGDDDD
KSPMAISGIRILYRPWRACRRAPSTSLRPHSSTTTTTTTTN*"
gene complement(446..1096)
/label="GST (variant)"
/gene="GST (variant)"
/translation="MSPILGYWKIKGLVQPTRLLLEYLEEKYEEHLYERDEGDKWRNK
KFELGLEFPNLPYYIDGDVKLTQSMAIIRYIADKHNMLGGCPKERAEISMLEGAVLDI
RYGVSRIAYSKDFETLKVDFLSKLPEMLKMFEDRLCHKTYLNGDHVTHPDFMLYDALD
VVLYMDPMCLDAFPKLVCFKKRIEAIPQIDKYLKSSKYIAWPLQGWQATFGGGDHPPK
SDGSTSGSGHHHHHHSAGLVPRGSTAIGMKETAAAKFERQHMDSPDLGTGGGSGDDDD
KSPMAISGIRILYRPWRACRRAPSTSLRPHSSTTTTTTTTN*"
CDS 1097..1825
/label="ORF frame 2"
/translation="MYISFLKLNKIISRGELLSAHNSPIVSRINFAGSRSISILYAGR
IVAGITGATGAVAGAYIADITDGEDRARHFGLMSACFGVGMVAGPVAGGLLGAISLHA
PFLAAAVLNGLNLLLGCFLMQESHKGERRDPGHHRMAQNLSRYGMIAPGRESIQGGEC
ETSNVIRCRRVCRCLLSDRFPRGEPGQPRFCENAGKSGSGDGGAELHSQPRGTTTGGQ
TVVADWRCHLQSGPARAVANCRGD*"
misc_feature complement(1105..1121)
/label="T7_transl_en_RBS"
/translation="MYISFLKLNKIISRGELLSAHNSPIVSRINFAGSRSISILYAGR
IVAGITGATGAVAGAYIADITDGEDRARHFGLMSACFGVGMVAGPVAGGLLGAISLHA
PFLAAAVLNGLNLLLGCFLMQESHKGERRDPGHHRMAQNLSRYGMIAPGRESIQGGEC
ETSNVIRCRRVCRCLLSDRFPRGEPGQPRFCENAGKSGSGDGGAELHSQPRGTTTGGQ
TVVADWRCHLQSGPARAVANCRGD*"
misc_feature complement(1139..1166)
/label="lacO"
/translation="MYISFLKLNKIISRGELLSAHNSPIVSRINFAGSRSISILYAGR
IVAGITGATGAVAGAYIADITDGEDRARHFGLMSACFGVGMVAGPVAGGLLGAISLHA
PFLAAAVLNGLNLLLGCFLMQESHKGERRDPGHHRMAQNLSRYGMIAPGRESIQGGEC
ETSNVIRCRRVCRCLLSDRFPRGEPGQPRFCENAGKSGSGDGGAELHSQPRGTTTGGQ
TVVADWRCHLQSGPARAVANCRGD*"
promoter complement(1166..1184)
/label="T7_promoter"
/translation="MYISFLKLNKIISRGELLSAHNSPIVSRINFAGSRSISILYAGR
IVAGITGATGAVAGAYIADITDGEDRARHFGLMSACFGVGMVAGPVAGGLLGAISLHA
PFLAAAVLNGLNLLLGCFLMQESHKGERRDPGHHRMAQNLSRYGMIAPGRESIQGGEC
ETSNVIRCRRVCRCLLSDRFPRGEPGQPRFCENAGKSGSGDGGAELHSQPRGTTTGGQ
TVVADWRCHLQSGPARAVANCRGD*"
gene 1220..1483
/label="tet (300 - 563)"
/gene="tet (300 - 563)"
/translation="MYISFLKLNKIISRGELLSAHNSPIVSRINFAGSRSISILYAGR
IVAGITGATGAVAGAYIADITDGEDRARHFGLMSACFGVGMVAGPVAGGLLGAISLHA
PFLAAAVLNGLNLLLGCFLMQESHKGERRDPGHHRMAQNLSRYGMIAPGRESIQGGEC
ETSNVIRCRRVCRCLLSDRFPRGEPGQPRFCENAGKSGSGDGGAELHSQPRGTTTGGQ
TVVADWRCHLQSGPARAVANCRGD*"
misc_feature complement(1272..1291)
/label="pBRrevBam_primer"
/translation="MYISFLKLNKIISRGELLSAHNSPIVSRINFAGSRSISILYAGR
IVAGITGATGAVAGAYIADITDGEDRARHFGLMSACFGVGMVAGPVAGGLLGAISLHA
PFLAAAVLNGLNLLLGCFLMQESHKGERRDPGHHRMAQNLSRYGMIAPGRESIQGGEC
ETSNVIRCRRVCRCLLSDRFPRGEPGQPRFCENAGKSGSGDGGAELHSQPRGTTTGGQ
TVVADWRCHLQSGPARAVANCRGD*"
misc_feature 1566..2657
/label="lacI"
/translation="MYISFLKLNKIISRGELLSAHNSPIVSRINFAGSRSISILYAGR
IVAGITGATGAVAGAYIADITDGEDRARHFGLMSACFGVGMVAGPVAGGLLGAISLHA
PFLAAAVLNGLNLLLGCFLMQESHKGERRDPGHHRMAQNLSRYGMIAPGRESIQGGEC
ETSNVIRCRRVCRCLLSDRFPRGEPGQPRFCENAGKSGSGDGGAELHSQPRGTTTGGQ
TVVADWRCHLQSGPARAVANCRGD*"
CDS 1698..2657
/label="ORF frame 3"
/translation="MAELNYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKS
RADQLGASVVVSMVERSGVEACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNV
PALFLDVSDQTPINSIIFSHEDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGW
HKYLTRNQIQPIAEREGDWSAMSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITE
SGLRVGADISVVGYDDTEDSSCYIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQ
LLPVSLVKRKTTLAPNTQTASPRALADSLMQLARQVSRLESGQ*"
misc_feature 3229..3420
/label="ROP"
/translation="MAELNYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKS
RADQLGASVVVSMVERSGVEACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNV
PALFLDVSDQTPINSIIFSHEDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGW
HKYLTRNQIQPIAEREGDWSAMSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITE
SGLRVGADISVVGYDDTEDSSCYIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQ
LLPVSLVKRKTTLAPNTQTASPRALADSLMQLARQVSRLESGQ*"
misc_feature complement(3414..3436)
/label="pGEX_3_primer"
/translation="MAELNYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKS
RADQLGASVVVSMVERSGVEACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNV
PALFLDVSDQTPINSIIFSHEDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGW
HKYLTRNQIQPIAEREGDWSAMSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITE
SGLRVGADISVVGYDDTEDSSCYIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQ
LLPVSLVKRKTTLAPNTQTASPRALADSLMQLARQVSRLESGQ*"
rep_origin complement(3835..4454)
/label="pBR322_origin"
/translation="MAELNYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKS
RADQLGASVVVSMVERSGVEACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNV
PALFLDVSDQTPINSIIFSHEDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGW
HKYLTRNQIQPIAEREGDWSAMSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITE
SGLRVGADISVVGYDDTEDSSCYIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQ
LLPVSLVKRKTTLAPNTQTASPRALADSLMQLARQVSRLESGQ*"
gene 4560..5375
/label="KanR2"
/gene="KanR2"
/translation="MAELNYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKS
RADQLGASVVVSMVERSGVEACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNV
PALFLDVSDQTPINSIIFSHEDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGW
HKYLTRNQIQPIAEREGDWSAMSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITE
SGLRVGADISVVGYDDTEDSSCYIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQ
LLPVSLVKRKTTLAPNTQTASPRALADSLMQLARQVSRLESGQ*"
CDS 4560..5375
/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 tgtttagcag cctaggtatt aatcaattag tggtggtggt ggtggtggtg gtgctcgagt
181 gcggccgcaa gcttgtcgac ggagctcgcc tgcaggcgcg ccaaggcctg tacagaattc
241 ggatccccga tatcgccatg ggactcttgt cgtcgtcatc accggagcca ccaccggtac
301 ccagatctgg gctgtccatg tgctggcgtt cgaatttagc agcagcggtt tctttcatac
361 caattgcagt actaccgcgt ggcaccagac ccgcggagtg atggtgatgg tgatgaccag
421 aaccactagt tgaaccatcc gattttggag gatggtcgcc accaccaaac gtggcttgcc
481 agccctgcaa aggccatgct atatacttgc tggatttcaa gtacttatca atttgtggga
541 tagcttcaat acgtttttta aaacaaacta attttgggaa cgcatccagg cacattgggt
601 ccatgtataa aacaacatca agagcgtcat acaacatgaa gtcaggatgg gttacatgat
661 caccatttaa atatgtttta tgacataaac gatcttcgaa cattttcagc atttcaggta
721 gcttgctaag aaaatcaact ttgagagttt caaagtcttt actatatgca attctcgaaa
781 caccgtatct aatatccaaa accgctcctt caagcattga aatctctgca cgctcttttg
841 gacaaccacc caacatgttg tgcttgtcag ctatataacg tatgatggcc atagactgtg
901 ttaatttaac atcaccatca atataataag gaagattggg aaactccaaa cccaattcaa
961 actttttgtt tcgccattta tcaccttcat cgcgctcata caaatgctct tcatattttt
1021 cttcaagata ttccaaaaga agtcgagtgg gttgcacaag gcccttaatt ttccaataac
1081 ctagtatagg ggacatatgt atatctcctt cttaaagtta aacaaaatta tttctagagg
1141 ggaattgtta tccgctcaca attcccctat agtgagtcgt attaatttcg cgggatcgag
1201 atcgatctcg atcctctacg ccggacgcat cgtggccggc atcaccggcg ccacaggtgc
1261 ggttgctggc gcctatatcg ccgacatcac cgatggggaa gatcgggctc gccacttcgg
1321 gctcatgagc gcttgtttcg gcgtgggtat ggtggcaggc cccgtggccg ggggactgtt
1381 gggcgccatc tccttgcatg caccattcct tgcggcggcg gtgctcaacg gcctcaacct
1441 actactgggc tgcttcctaa tgcaggagtc gcataaggga gagcgtcgag atcccggaca
1501 ccatcgaatg gcgcaaaacc tttcgcggta tggcatgata gcgcccggaa gagagtcaat
1561 tcagggtggt gaatgtgaaa ccagtaacgt tatacgatgt cgcagagtat gccggtgtct
1621 cttatcagac cgtttcccgc gtggtgaacc aggccagcca cgtttctgcg aaaacgcggg
1681 aaaaagtgga agcggcgatg gcggagctga attacattcc caaccgcgtg gcacaacaac
1741 tggcgggcaa acagtcgttg ctgattggcg ttgccacctc cagtctggcc ctgcacgcgc
1801 cgtcgcaaat tgtcgcggcg attaaatctc gcgccgatca actgggtgcc agcgtggtgg
1861 tgtcgatggt agaacgaagc ggcgtcgaag cctgtaaagc ggcggtgcac aatcttctcg
1921 cgcaacgcgt cagtgggctg atcattaact atccgctgga tgaccaggat gccattgctg
1981 tggaagctgc ctgcactaat gttccggcgt tatttcttga tgtctctgac cagacaccca
2041 tcaacagtat tattttctcc catgaagacg gtacgcgact gggcgtggag catctggtcg
2101 cattgggtca ccagcaaatc gcgctgttag cgggcccatt aagttctgtc tcggcgcgtc
2161 tgcgtctggc tggctggcat aaatatctca ctcgcaatca aattcagccg atagcggaac