pET-37b
PVT0094 2ug
pET-37b Information
Promoter: T7/lac
Replicator: ColE1 ori, F1 ori
Terminator: T7 Terminator
Plasmid classification: Escherichia coli vector; PET series expression plasmid
Plasmid size: 5932bp
Plasmid Tags: N-CBD, N-thrombin, N-S, N-Factor Xa, C-8*His
Prokaryotic resistance: kanamycin Kan
Cloned strain: DH5 alpha
Culture conditions: 37 centigrade, aerobic, LB
Expression host: BL21 (DE3)
Culture conditions: 37 centigrade, aerobic, LB
Inducement: IPTG or lactose and its analogues
5'sequencing primers: T7:TAATACGACTCACTATAGGG
3'sequencing primers: T7-ter:TGCTAGTTATTGCTCAGCGG
Remarks: secretory expression, cellulose binding domain
pET-37b Description
pET-37b Sequence
LOCUS Exported 5932 bp ds-DNA circular SYN 10-8-2015
DEFINITION synthetic circular DNA
ACCESSION .
VERSION .
KEYWORDS Untitled
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 5932)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported 2015-8-1
FEATURES Location/Qualifiers
source 1..5932
/organism="synthetic DNA construct"
/mol_type="other DNA"
source 865..887
/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(150..173)
/codon_start=1
/product="8xHis affinity tag"
/note="8xHis"
/translation="HHHHHHHH"
CDS complement(249..260)
/codon_start=1
/product="Factor Xa recognition and cleavage site"
/note="Factor Xa site"
/translation="IEGR"
CDS complement(291..335)
/codon_start=1
/product="affinity and epitope tag derived from pancreatic
ribonuclease A"
/note="S-Tag"
/translation="KETAAAKFERQHMDS"
CDS complement(351..368)
/codon_start=1
/product="thrombin recognition and cleavage site"
/note="thrombin site"
/translation="LVPRGS"
RBS 865..887
/note="efficient ribosome binding site from bacteriophage
T7 gene 10 (Olins and Rangwala, 1989)"
protein_bind 902..926
/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(927..945)
/note="T7 promoter"
/note="promoter for bacteriophage T7 RNA polymerase"
promoter 1258..1335
/gene="lacI"
/note="lacI promoter"
CDS 1336..2418
/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 3227..3418
/codon_start=1
/gene="rop"
/product="Rop protein, which maintains plasmids at low copy
number"
/note="rop"
/translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
DELYRSCLARFGDDGENL"
misc_feature 3520..3662
/note="bom"
/note="basis of mobility region from pBR322"
rep_origin complement(3848..4436)
/direction=LEFT
/note="ori"
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
CDS 4558..5373
/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(5466..5921)
/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 ggagctcgaa ttcggatccg atatcgccat ggccagagga
241 gagttagagc gaccctcaat accggagcca ccaccggtac ccagatctgg gctgtccatg
301 tgctggcgtt cgaatttagc agcagcggtt tctttcatac caattgcagt actaccgcgt
361 ggcaccagac ccgcggaacc tggtgccgtg gggctggtcg tcggcacggt gcccgtgcag
421 gtggtgccgt tgagcgagaa cgacgccggc gtcgggttgg acccggccca cgagccgttg
481 aacccgaacg acgccgtgcc gccggtcggg atgctgccgt tccagggcag gctggtgacg
541 gagacctggc cgccgttggt cgaggcggtg ccgttccaca gctgctggat acgctggcct
601 gcggtgtagg tccagtcgag cttccacgac gagacggggt cgccgaggtt ggtgatcgtg
661 acgttggcgc cgaagccgcc gggccactgg ttggtgacgg cgtagtcgac gcggcagccg
721 ggactagcgg cctgggcggg cagcacgacc gtggcgccga cgacgagcgt cgccgccgcg
781 gcggcgagcg tcgtgcgcag agcggtgcgg gcgccgcggg ccgggtggcc gggtgcgggc
841 gtggtacgag ggtccatatg tatatctcct tcttaaagtt aaacaaaatt atttctagag
901 gggaattgtt atccgctcac aattccccta tagtgagtcg tattaatttc gcgggatcga
961 gatcgatctc gatcctctac gccggacgca tcgtggccgg catcaccggc gccacaggtg
1021 cggttgctgg cgcctatatc gccgacatca ccgatgggga agatcgggct cgccacttcg
1081 ggctcatgag cgcttgtttc ggcgtgggta tggtggcagg ccccgtggcc gggggactgt
1141 tgggcgccat ctccttgcat gcaccattcc ttgcggcggc ggtgctcaac ggcctcaacc
1201 tactactggg ctgcttccta atgcaggagt cgcataaggg agagcgtcga gatcccggac
1261 accatcgaat ggcgcaaaac ctttcgcggt atggcatgat agcgcccgga agagagtcaa
1321 ttcagggtgg tgaatgtgaa accagtaacg ttatacgatg tcgcagagta tgccggtgtc
1381 tcttatcaga ccgtttcccg cgtggtgaac caggccagcc acgtttctgc gaaaacgcgg
1441 gaaaaagtgg aagcggcgat ggcggagctg aattacattc ccaaccgcgt ggcacaacaa
1501 ctggcgggca aacagtcgtt gctgattggc gttgccacct ccagtctggc cctgcacgcg
1561 ccgtcgcaaa ttgtcgcggc gattaaatct cgcgccgatc aactgggtgc cagcgtggtg
1621 gtgtcgatgg tagaacgaag cggcgtcgaa gcctgtaaag cggcggtgca caatcttctc
1681 gcgcaacgcg tcagtgggct gatcattaac tatccgctgg atgaccagga tgccattgct
1741 gtggaagctg cctgcactaa tgttccggcg ttatttcttg atgtctctga ccagacaccc
1801 atcaacagta ttattttctc ccatgaagac ggtacgcgac tgggcgtgga gcatctggtc
1861 gcattgggtc accagcaaat cgcgctgtta gcgggcccat taagttctgt ctcggcgcgt
1921 ctgcgtctgg ctggctggca taaatatctc actcgcaatc aaattcagcc gatagcggaa
1981 cgggaaggcg actggagtgc catgtccggt tttcaacaaa ccatgcaaat gctgaatgag
2041 ggcatcgttc ccactgcgat gctggttgcc aacgatcaga tggcgctggg cgcaatgcgc
2101 gccattaccg agtccgggct gcgcgttggt gcggacatct cggtagtggg atacgacgat
2161 accgaagaca gctcatgtta tatcccgccg ttaaccacca tcaaacagga ttttcgcctg
2221 ctggggcaaa ccagcgtgga ccgcttgctg caactctctc agggccaggc ggtgaagggc
2281 aatcagctgt tgcccgtctc actggtgaaa agaaaaacca ccctggcgcc caatacgcaa
2341 accgcctctc cccgcgcgtt ggccgattca ttaatgcagc tggcacgaca ggtttcccga
2401 ctggaaagcg ggcagtgagc gcaacgcaat taatgtaagt tagctcactc attaggcacc
2461 gggatctcga ccgatgccct tgagagcctt caacccagtc agctccttcc ggtgggcgcg
2521 gggcatgact atcgtcgccg cacttatgac tgtcttcttt atcatgcaac tcgtaggaca
2581 ggtgccggca gcgctctggg tcattttcgg cgaggaccgc tttcgctgga gcgcgacgat
2641 gatcggcctg tcgcttgcgg tattcggaat cttgcacgcc ctcgctcaag ccttcgtcac
2701 tggtcccgcc accaaacgtt tcggcgagaa gcaggccatt atcgccggca tggcggcccc
2761 acgggtgcgc atgatcgtgc tcctgtcgtt gaggacccgg ctaggctggc ggggttgcct
2821 tactggttag cagaatgaat caccgatacg cgagcgaacg tgaagcgact gctgctgcaa
2881 aacgtctgcg acctgagcaa caacatgaat ggtcttcggt ttccgtgttt cgtaaagtct
2941 ggaaacgcgg aagtcagcgc cctgcaccat tatgttccgg atctgcatcg caggatgctg
3001 ctggctaccc tgtggaacac ctacatctgt attaacgaag cgctggcatt gaccctgagt
3061 gatttttctc tggtcccgcc gcatccatac cgccagttgt ttaccctcac aacgttccag
3121 taaccgggca tgttcatcat cagtaacccg tatcgtgagc atcctctctc gtttcatcgg
3181 tatcattacc cccatgaaca gaaatccccc ttacacggag gcatcagtga ccaaacagga
3241 aaaaaccgcc cttaacatgg cccgctttat cagaagccag acattaacgc ttctggagaa
3301 actcaacgag ctggacgcgg atgaacaggc agacatctgt gaatcgcttc acgaccacgc
3361 tgatgagctt taccgcagct gcctcgcgcg tttcggtgat gacggtgaaa acctctgaca
3421 catgcagctc ccggagacgg tcacagcttg tctgtaagcg gatgccggga gcagacaagc
3481 ccgtcagggc gcgtcagcgg gtgttggcgg gtgtcggggc gcagccatga cccagtcacg
3541 tagcgatagc ggagtgtata ctggcttaac tatgcggcat cagagcagat tgtactgaga
3601 gtgcaccata tatgcggtgt gaaataccgc acagatgcgt aaggagaaaa taccgcatca
3661 ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag
3721 cggtatcagc tcactcaaag gcggtaatac ggttatccac agaatcaggg gataacgcag
3781 gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc
3841 tggcgttttt ccataggctc cgcccccctg acgagcatca caaaaatcga cgctcaagtc
3901 agaggtggcg aaacccgaca ggactataaa gataccaggc gtttccccct ggaagctccc
3961 tcgtgcgctc tcctgttccg accctgccgc ttaccggata cctgtccgcc tttctccctt
4021 cgggaagcgt ggcgctttct catagctcac gctgtaggta tctcagttcg gtgtaggtcg
4081 ttcgctccaa gctgggctgt gtgcacgaac cccccgttca gcccgaccgc tgcgccttat
4141 ccggtaacta tcgtcttgag tccaacccgg taagacacga cttatcgcca ctggcagcag
4201 ccactggtaa caggattagc agagcgaggt atgtaggcgg tgctacagag ttcttgaagt
4261 ggtggcctaa ctacggctac actagaagga cagtatttgg tatctgcgct ctgctgaagc
4321 cagttacctt cggaaaaaga gttggtagct cttgatccgg caaacaaacc accgctggta
4381 gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag
4441 atcctttgat cttttctacg gggtctgacg ctcagtggaa cgaaaactca cgttaaggga
4501 ttttggtcat gaacaataaa actgtctgct tacataaaca gtaatacaag gggtgttatg
4561 agccatattc aacgggaaac gtcttgctct aggccgcgat taaattccaa catggatgct
4621 gatttatatg ggtataaatg ggctcgcgat aatgtcgggc aatcaggtgc gacaatctat
4681 cgattgtatg ggaagcccga tgcgccagag ttgtttctga aacatggcaa aggtagcgtt
4741 gccaatgatg ttacagatga gatggtcaga ctaaactggc tgacggaatt tatgcctctt
4801 ccgaccatca agcattttat ccgtactcct gatgatgcat ggttactcac cactgcgatc
4861 cccgggaaaa cagcattcca ggtattagaa gaatatcctg attcaggtga aaatattgtt
4921 gatgcgctgg cagtgttcct gcgccggttg cattcgattc ctgtttgtaa ttgtcctttt
4981 aacagcgatc gcgtatttcg tctcgctcag gcgcaatcac gaatgaataa cggtttggtt
5041 gatgcgagtg attttgatga cgagcgtaat ggctggcctg ttgaacaagt ctggaaagaa
5101 atgcataaac ttttgccatt ctcaccggat tcagtcgtca ctcatggtga tttctcactt
5161 gataacctta tttttgacga ggggaaatta ataggttgta ttgatgttgg acgagtcgga
5221 atcgcagacc gataccagga tcttgccatc ctatggaact gcctcggtga gttttctcct
5281 tcattacaga aacggctttt tcaaaaatat ggtattgata atcctgatat gaataaattg
5341 cagtttcatt tgatgctcga tgagtttttc taagaattaa ttcatgagcg gatacatatt
5401 tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc
5461 acctgaaatt gtaaacgtta atattttgtt aaaattcgcg ttaaattttt gttaaatcag
5521 ctcatttttt aaccaatagg ccgaaatcgg caaaatccct tataaatcaa aagaatagac
5581 cgagataggg ttgagtgttg ttccagtttg gaacaagagt ccactattaa agaacgtgga
5641 ctccaacgtc aaagggcgaa aaaccgtcta tcagggcgat ggcccactac gtgaaccatc
5701 accctaatca agttttttgg ggtcgaggtg ccgtaaagca ctaaatcgga accctaaagg
5761 gagcccccga tttagagctt gacggggaaa gccggcgaac gtggcgagaa aggaagggaa
5821 gaaagcgaaa ggagcgggcg ctagggcgct ggcaagtgta gcggtcacgc tgcgcgtaac
5881 caccacaccc gccgcgctta atgcgccgct acagggcgcg tcccattcgc ca
//
Caution:
Product is for research use only!
