pET-41c
Catalog No. PVT10581
Packing 2ug
pET-41c Information
Promoter: T7/lac promoter
Copier: ColE1 ori, F1 ori
Terminator: T7 Terminator
Plasmid size: 5934bp
Plasmid Label: N-GST, N-6*His, N-Thrombin
Prokaryotic resistance: kanamycin Kan
Clonal strain: DH5 alpha
Culture conditions: 37 C, aerobic, LB
Expression host: BL21 (DE3)
Culture conditions: 37 C, aerobic, LB
Induction: IPTG or lactose and its analogues
5'Sequencing Primers: T7:TAATACGACTCACTATAGG
3'Sequencing Primer: T7-ter: TGCTAGTTATTGCTCAGCGG
pET-41c Description
PET41 series vectors are designed to clone and express proteins at high level. The vectors fuse and express a purified fragment of GST tagged protein containing 220 amino acids. The single polyclonal site of pET41a vector is shown in the circular plasmid map above. Note: The vector sequence is encoded by the coding rules of the pBR322 plasmid, so the expression region of T7 protein is reversed on the plasmid map.
The cloning and expression regions initiated by T7 RNA polymerase were also labeled in the plasmid map. The F1 replicon of the plasmid is directed, so viral particles containing protein coding sequence can be produced by T7 phage polymerase, and the protein expression will be terminated by T7 terminator sequence.
The pET-41a vector contains an EK protease cleavage site. When the target gene is inserted into the vector using the PSAI restriction endonuclease site on the vector, all the fusion amino acid sequences on the vector, including GST tag sequences, can be removed by EK protease.
pET-41c Multiple cloning site
pET-41c Sequence
LOCUS Exported 5934 bp ds-DNA circular SYN 31-AUG-2016
DEFINITION synthetic circular DNA
ACCESSION .
VERSION .
KEYWORDS pET-41c
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 5934)
AUTHORS .
TITLE Direct Submission
FEATURES Location/Qualifiers
source 1..5934
/organism="synthetic DNA construct"
/mol_type="other DNA"
terminator 26..73
/note="T7 terminator"
/note="transcription terminator for bacteriophage T7 RNA
polymerase"
CDS complement(150..173)
/codon_start=1
/product="8xHis affinity tag"
/note="8xHis"
/translation="HHHHHHHH"
CDS complement(266..280)
/codon_start=1
/product="enterokinase recognition and cleavage site"
/note="enterokinase site"
/translation="DDDDK"
CDS complement(311..355)
/codon_start=1
/product="affinity and epitope tag derived from pancreatic
ribonuclease A"
/note="S-Tag"
/translation="KETAAAKFERQHMDS"
CDS complement(371..388)
/codon_start=1
/product="thrombin recognition and cleavage site"
/note="thrombin site"
/translation="LVPRGS"
CDS complement(398..415)
/codon_start=1
/product="6xHis affinity tag"
/note="6xHis"
/translation="HHHHHH"
CDS complement(443..1096)
/codon_start=1
/product="glutathione S-transferase from Schistosoma
japonicum"
/note="GST"
/translation="MSPILGYWKIKGLVQPTRLLLEYLEEKYEEHLYERDEGDKWRNKK
FELGLEFPNLPYYIDGDVKLTQSMAIIRYIADKHNMLGGCPKERAEISMLEGAVLDIRY
GVSRIAYSKDFETLKVDFLSKLPEMLKMFEDRLCHKTYLNGDHVTHPDFMLYDALDVVL
YMDPMCLDAFPKLVCFKKRIEAIPQIDKYLKSSKYIAWPLQGWQATFGGGDHPPK"
protein_bind 1141..1165
/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(1166..1184)
/note="T7 promoter"
/note="promoter for bacteriophage T7 RNA polymerase"
promoter 1497..1574
/gene="lacI"
/note="lacI promoter"
CDS 1575..2657
/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 3229..3420
/codon_start=1
/gene="rop"
/product="Rop protein"
/note="rop"
/translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
DELYRSCLARFGDDGENL"
rep_origin complement(3850..4438)
/direction=LEFT
/note="ori"
/note="high-copy-number colE1/pMB1/pBR322/pUC origin of
replication"
CDS 4560..5375
/codon_start=1
/gene="aph(3')-Ia"
/product="aminoglycoside phosphotransferase"
/note="KanR"
/note="confers resistance to kanamycin"
/translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
rep_origin complement(5468..5923)
/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 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
2221 gggaaggcga ctggagtgcc atgtccggtt ttcaacaaac catgcaaatg ctgaatgagg
2281 gcatcgttcc cactgcgatg ctggttgcca acgatcagat ggcgctgggc gcaatgcgcg
2341 ccattaccga gtccgggctg cgcgttggtg cggacatctc ggtagtggga tacgacgata
2401 ccgaagacag ctcatgttat atcccgccgt taaccaccat caaacaggat tttcgcctgc
2461 tggggcaaac cagcgtggac cgcttgctgc aactctctca gggccaggcg gtgaagggca
2521 atcagctgtt gcccgtctca ctggtgaaaa gaaaaaccac cctggcgccc aatacgcaaa
2581 ccgcctctcc ccgcgcgttg gccgattcat taatgcagct ggcacgacag gtttcccgac
2641 tggaaagcgg gcagtgagcg caacgcaatt aatgtaagtt agctcactca ttaggcaccg
2701 ggatctcgac cgatgccctt gagagccttc aacccagtca gctccttccg gtgggcgcgg
2761 ggcatgacta gcatgatcgt gctcctgtcg ttgaggaccc ggctaggctg gcggggttgc
2821 cttactggtt agcagaatga atcaccgata cgcgagcgaa cgtgaagcga ctgctgctgc
2881 aaaacgtctg cgacctgagc aacaacatga atggtcttcg gtttccgtgt ttcgtaaagt
2941 ctggaaacgc ggaagtcagc gccctgcacc attatgttcc ggatctgcat cgcaggatgc
3001 tgctggctac cctgtggaac acctacatct gtattaacga agcgctggca ttgaccctga
3061 gtgatttttc tctggtcccg ccgcatccat accgccagtt gtttaccctc acaacgttcc
3121 agtaaccggg catgttcatc atcagtaacc cgtatcgtga gcatcctctc tcgtttcatc
3181 ggtatcatta cccccatgaa cagaaatccc ccttacacgg aggcatcagt gaccaaacag
3241 gaaaaaaccg cccttaacat ggcccgcttt atcagaagcc agacattaac gcttctggag
3301 aaactcaacg agctggacgc ggatgaacag gcagacatct gtgaatcgct tcacgaccac
3361 gctgatgagc tttaccgcag ctgcctcgcg cgtttcggtg atgacggtga aaacctctga
3421 cacatgcagc tcccggagac ggtcacagct tgtctgtaag cggatgccgg gagcagacaa
3481 gcccgtcagg gcgcgtcagc gggtgttggc gggtgtcggg gcgcagccat gacccagtca
3541 cgtagcgata gcggagtgta tactggctta actatgcggc atcagagcag attgtactga
3601 gagtgcacca tatatgcggt gtgaaatacc gcacagatgc gtaaggagaa aataccgcat
3661 caggcgctct tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg
3721 agcggtatca gctcactcaa aggcggtaat acggttatcc acagaatcag gggataacgc
3781 aggaaagaac atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt
3841 gctggcgttt ttccataggc tccgcccccc tgacgagcat cacaaaaatc gacgctcaag
3901 tcagaggtgg cgaaacccga caggactata aagataccag gcgtttcccc ctggaagctc
3961 cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga tacctgtccg cctttctccc
4021 ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg tatctcagtt cggtgtaggt
4081 cgttcgctcc aagctgggct gtgtgcacga accccccgtt cagcccgacc gctgcgcctt
4141 atccggtaac tatcgtcttg agtccaaccc ggtaagacac gacttatcgc cactggcagc
4201 agccactggt aacaggatta gcagagcgag gtatgtaggc ggtgctacag agttcttgaa
4261 gtggtggcct aactacggct acactagaag gacagtattt ggtatctgcg ctctgctgaa
4321 gccagttacc ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg
4381 tagcggtggt ttttttgttt gcaagcagca gattacgcgc agaaaaaaag gatctcaaga
4441 agatcctttg atcttttcta cggggtctga cgctcagtgg aacgaaaact cacgttaagg
4501 gattttggtc atgaacaata aaactgtctg cttacataaa cagtaataca aggggtgtta
4561 tgagccatat tcaacgggaa acgtcttgct ctaggccgcg attaaattcc aacatggatg
4621 ctgatttata tgggtataaa tgggctcgcg ataatgtcgg gcaatcaggt gcgacaatct
4681 atcgattgta tgggaagccc gatgcgccag agttgtttct gaaacatggc aaaggtagcg
4741 ttgccaatga tgttacagat gagatggtca gactaaactg gctgacggaa tttatgcctc
4801 ttccgaccat caagcatttt atccgtactc ctgatgatgc atggttactc accactgcga
4861 tccccgggaa aacagcattc caggtattag aagaatatcc tgattcaggt gaaaatattg
4921 ttgatgcgct ggcagtgttc ctgcgccggt tgcattcgat tcctgtttgt aattgtcctt
4981 ttaacagcga tcgcgtattt cgtctcgctc aggcgcaatc acgaatgaat aacggtttgg
5041 ttgatgcgag tgattttgat gacgagcgta atggctggcc tgttgaacaa gtctggaaag
5101 aaatgcataa acttttgcca ttctcaccgg attcagtcgt cactcatggt gatttctcac
5161 ttgataacct tatttttgac gaggggaaat taataggttg tattgatgtt ggacgagtcg
5221 gaatcgcaga ccgataccag gatcttgcca tcctatggaa ctgcctcggt gagttttctc
5281 cttcattaca gaaacggctt tttcaaaaat atggtattga taatcctgat atgaataaat
5341 tgcagtttca tttgatgctc gatgagtttt tctaagaatt aattcatgag cggatacata
5401 tttgaatgta tttagaaaaa taaacaaata ggggttccgc gcacatttcc ccgaaaagtg
5461 ccacctgaaa ttgtaaacgt taatattttg ttaaaattcg cgttaaattt ttgttaaatc
5521 agctcatttt ttaaccaata ggccgaaatc ggcaaaatcc cttataaatc aaaagaatag
5581 accgagatag ggttgagtgt tgttccagtt tggaacaaga gtccactatt aaagaacgtg
5641 gactccaacg tcaaagggcg aaaaaccgtc tatcagggcg atggcccact acgtgaacca
5701 tcaccctaat caagtttttt ggggtcgagg tgccgtaaag cactaaatcg gaaccctaaa
5761 gggagccccc gatttagagc ttgacgggga aagccggcga acgtggcgag aaaggaaggg
5821 aagaaagcga aaggagcggg cgctagggcg ctggcaagtg tagcggtcac gctgcgcgta
5881 accaccacac ccgccgcgct taatgcgccg ctacagggcg cgtcccattc gcca
//
