LOCUS CVU78874 4983 bp DNA circular SYN 03-DEC-1996
DEFINITION pGEX-6P-3 cloning vector, complete sequence.
ACCESSION U78874
VERSION U78874.1 GI:1699068
KEYWORDS .
SOURCE Cloning vector pGEX-6P-3
ORGANISM Cloning vector pGEX-6P-3
artificial sequences; vectors.
REFERENCE 1 (bases 1 to 4983)
AUTHORS Smith,D.B. and Johnson,K.S.
TITLE Single-step purification of polypeptides expressed in Escherichia
coli as fusions with glutathione S-transferase
JOURNAL Gene 67 (1), 31-40 (1988)
MEDLINE 88329742
PUBMED 3047011
REFERENCE 2 (bases 918 to 938)
AUTHORS Cordingley,M.G., Callahan,P.L., Sardana,V.V., Garsky,V.M. and
Colonno,R.J.
TITLE Substrate requirements of human rhinovirus 3C protease for peptide
cleavage in vitro
JOURNAL J. Biol. Chem. 265 (16), 9062-9065 (1990)
MEDLINE 90264387
PUBMED 2160953
REFERENCE 3 (bases 1 to 4983)
AUTHORS Dunst,R.W., Bell,P.A. and English,J.D.
TITLE Pharmacia Biotech PreScission GST Gene Fusion Expression Vector
JOURNAL Unpublished
REFERENCE 4 (bases 1 to 4983)
AUTHORS Dunst,R.W., Bell,P.A. and English,J.D.
TITLE Direct Submission
JOURNAL Submitted (20-NOV-1996) Product Center Molecular Biology Reagents,
Pharmacia Biotech Inc., 2202 North Bartlett Avenue, Milwaukee, WI
53202-1009, USA
FEATURES Location/Qualifiers
source 1..4983
/organism="Cloning vector pGEX-6P-3"
/db_xref="taxon:54766"
/lab_host="Escherichia coli"
/plasmid="pGEX-6P-3"
-35_signal 183..188
/note="tac promoter"
-10_signal 205..211
/note="tac promoter; for inducible GST gene fusion
expression"
gene 244..995
/gene="GST"
RBS 244
/gene="GST"
CDS 258..995
/gene="GST"
/codon_start=1
/transl_table=11
/product="glutathione S-transferase"
/protein_id="AAB37352.1"
/db_xref="GI:1699069"
/translation="MSPILGYWKIKGLVQPTRLLLEYLEEKYEEHLYERDEGDKWRNK
KFELGLEFPNLPYYIDGDVKLTQSMAIIRYIADKHNMLGGCPKERAEISMLEGAVLDI
RYGVSRIAYSKDFETLKVDFLSKLPEMLKMFEDRLCHKTYLNGDHVTHPDFMLYDALD
VVLYMDPMCLDAFPKLVCFKKRIEAIPQIDKYLKSSKYIAWPLQGWQATFGGGDHPPK
SDLEVLFQGPLGSPNSRVDSSGRIVTD"
misc_feature 918..938
/gene="GST"
/note="prescission protease recognition/cleavage region"
misc_feature 945..980
/gene="GST"
/note="multiple cloning site (MCS); contains unique
restriction sites for BamH I, EcoR I, Sma I, Sal I, Xho I,
and Not I"
-35_signal 1321..1326
-10_signal 1344..1349
/note="beta-lactamase promoter"
CDS 1391..2251
/note="ampicillin resistance gene"
/codon_start=1
/transl_table=11
/product="beta-lactamase"
/protein_id="AAB37353.1"
/db_xref="GI:1699070"
/translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGY
IELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVE
YSPVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRL
DRWEPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPL
LRSALPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIA
EIGASLIKHW"
misc_feature 2316..3012
/note="region necessary for plasmid replication"
rep_origin 3009
/note="site of plasmid replication initiation"
CDS 3332..4414
/note="overproducing allele"
/codon_start=1
/transl_table=11
/product="lac repressor"
/protein_id="AAB37354.1"
/db_xref="GI:1699071"
/translation="MKPVTLYDVAEYAGVSYQTVSRVVNQASHVSAKTREKVEAAMAE
LNYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSRADQLGASVVVSMVERS
GVEACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSII
FSHEDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREG
DWSAMSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDT
EDSSCYIPPSTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNT
QTASPRALADSLMQLARQVSRLESGQ"
BASE COUNT 1228 a 1207 c 1296 g 1252 t
ORIGIN
1 acgttatcga ctgcacggtg caccaatgct tctggcgtca ggcagccatc ggaagctgtg
61 gtatggctgt gcaggtcgta aatcactgca taattcgtgt cgctcaaggc gcactcccgt
121 tctggataat gttttttgcg ccgacatcat aacggttctg gcaaatattc tgaaatgagc
181 tgttgacaat taatcatcgg ctcgtataat gtgtggaatt gtgagcggat aacaatttca
241 cacaggaaac agtattcatg tcccctatac taggttattg gaaaattaag ggccttgtgc
301 aacccactcg acttcttttg gaatatcttg aagaaaaata tgaagagcat ttgtatgagc
361 gcgatgaagg tgataaatgg cgaaacaaaa agtttgaatt gggtttggag tttcccaatc
421 ttccttatta tattgatggt gatgttaaat taacacagtc tatggccatc atacgttata
481 tagctgacaa gcacaacatg ttgggtggtt gtccaaaaga gcgtgcagag atttcaatgc
541 ttgaaggagc ggttttggat attagatacg gtgtttcgag aattgcatat agtaaagact
601 ttgaaactct caaagttgat tttcttagca agctacctga aatgctgaaa atgttcgaag
661 atcgtttatg tcataaaaca tatttaaatg gtgatcatgt aacccatcct gacttcatgt
721 tgtatgacgc tcttgatgtt gttttataca tggacccaat gtgcctggat gcgttcccaa
781 aattagtttg ttttaaaaaa cgtattgaag ctatcccaca aattgataag tacttgaaat
841 ccagcaagta tatagcatgg cctttgcagg gctggcaagc cacgtttggt ggtggcgacc
901 atcctccaaa atcggatctg gaagttctgt tccaggggcc cctgggatcc ccgaattccc
961 gggtcgactc gagcggccgc atcgtgactg actgacgatc tgcctcgcgc gtttcggtga
1021 tgacggtgaa aacctctgac acatgcagct cccggagacg gtcacagctt gtctgtaagc
1081 ggatgccggg agcagacaag cccgtcaggg cgcgtcagcg ggtgttggcg ggtgtcgggg
1141 cgcagccatg acccagtcac gtagcgatag cggagtgtat aattcttgaa gacgaaaggg
1201 cctcgtgata cgcctatttt tataggttaa tgtcatgata ataatggttt cttagacgtc
1261 aggtggcact tttcggggaa atgtgcgcgg aacccctatt tgtttatttt tctaaataca
1321 ttcaaatatg tatccgctca tgagacaata accctgataa atgcttcaat aatattgaaa
1381 aaggaagagt atgagtattc aacatttccg tgtcgccctt attccctttt ttgcggcatt
1441 ttgccttcct gtttttgctc acccagaaac gctggtgaaa gtaaaagatg ctgaagatca
1501 gttgggtgca cgagtgggtt acatcgaact ggatctcaac agcggtaaga tccttgagag
1561 ttttcgcccc gaagaacgtt ttccaatgat gagcactttt aaagttctgc tatgtggcgc
1621 ggtattatcc cgtgttgacg ccgggcaaga gcaactcggt cgccgcatac actattctca
1681 gaatgacttg gttgagtact caccagtcac agaaaagcat cttacggatg gcatgacagt
1741 aagagaatta tgcagtgctg ccataaccat gagtgataac actgcggcca acttacttct
1801 gacaacgatc ggaggaccga aggagctaac cgcttttttg cacaacatgg gggatcatgt
1861 aactcgcctt gatcgttggg aaccggagct gaatgaagcc ataccaaacg acgagcgtga
1921 caccacgatg cctgcagcaa tggcaacaac gttgcgcaaa ctattaactg gcgaactact
1981 tactctagct tcccggcaac aattaataga ctggatggag gcggataaag ttgcaggacc
2041 acttctgcgc tcggcccttc cggctggctg gtttattgct gataaatctg gagccggtga
2101 gcgtgggtct cgcggtatca ttgcagcact ggggccagat ggtaagccct cccgtatcgt
2161 agttatctac acgacgggga gtcaggcaac tatggatgaa cgaaatagac agatcgctga
2221 gataggtgcc tcactgatta agcattggta actgtcagac caagtttact catatatact
2281 ttagattgat ttaaaacttc atttttaatt taaaaggatc taggtgaaga tcctttttga
2341 taatctcatg accaaaatcc cttaacgtga gttttcgttc cactgagcgt cagaccccgt
2401 agaaaagatc aaaggatctt cttgagatcc tttttttctg cgcgtaatct gctgcttgca
2461 aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg gatcaagagc taccaactct
2521 ttttccgaag gtaactggct tcagcagagc gcagatacca aatactgtcc ttctagtgta
2581 gccgtagtta ggccaccact tcaagaactc tgtagcaccg cctacatacc tcgctctgct
2641 aatcctgtta ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg ggttggactc
2701 aagacgatag ttaccggata aggcgcagcg gtcgggctga acggggggtt cgtgcacaca
2761 gcccagcttg gagcgaacga cctacaccga actgagatac ctacagcgtg agctatgaga
2821 aagcgccacg cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg
2881 aacaggagag cgcacgaggg agcttccagg gggaaacgcc tggtatcttt atagtcctgt
2941 cgggtttcgc cacctctgac ttgagcgtcg atttttgtga tgctcgtcag gggggcggag
3001 cctatggaaa aacgccagca acgcggcctt tttacggttc ctggcctttt gctggccttt
3061 tgctcacatg ttctttcctg cgttatcccc tgattctgtg gataaccgta ttaccgcctt
3121 tgagtgagct gataccgctc gccgcagccg aacgaccgag cgcagcgagt cagtgagcga
3181 ggaagcggaa gagcgcctga tgcggtattt tctccttacg catctgtgcg gtatttcaca
3241 ccgcataaat tccgacacca tcgaatggtg caaaaccttt cgcggtatgg catgatagcg
3301 cccggaagag agtcaattca gggtggtgaa tgtgaaacca gtaacgttat acgatgtcgc
3361 agagtatgcc ggtgtctctt atcagaccgt ttcccgcgtg gtgaaccagg ccagccacgt
3421 ttctgcgaaa acgcgggaaa aagtggaagc ggcgatggcg gagctgaatt acattcccaa
3481 ccgcgtggca caacaactgg cgggcaaaca gtcgttgctg attggcgttg ccacctccag
3541 tctggccctg cacgcgccgt cgcaaattgt cgcggcgatt aaatctcgcg ccgatcaact
3601 gggtgccagc gtggtggtgt cgatggtaga acgaagcggc gtcgaagcct gtaaagcggc
3661 ggtgcacaat cttctcgcgc aacgcgtcag tgggctgatc attaactatc cgctggatga
3721 ccaggatgcc attgctgtgg aagctgcctg cactaatgtt ccggcgttat ttcttgatgt
3781 ctctgaccag acacccatca acagtattat tttctcccat gaagacggta cgcgactggg
3841 cgtggagcat ctggtcgcat tgggtcacca gcaaatcgcg ctgttagcgg gcccattaag
3901 ttctgtctcg gcgcgtctgc gtctggctgg ctggcataaa tatctcactc gcaatcaaat
3961 tcagccgata gcggaacggg aaggcgactg gagtgccatg tccggttttc aacaaaccat
4021 gcaaatgctg aatgagggca tcgttcccac tgcgatgctg gttgccaacg atcagatggc
4081 gctgggcgca atgcgcgcca ttaccgagtc cgggctgcgc gttggtgcgg atatctcggt
4141 agtgggatac gacgataccg aagacagctc atgttatatc ccgccgtcaa ccaccatcaa
4201 acaggatttt cgcctgctgg ggcaaaccag cgtggaccgc ttgctgcaac tctctcaggg
4261 ccaggcggtg aagggcaatc agctgttgcc cgtctcactg gtgaaaagaa aaaccaccct
4321 ggcgcccaat acgcaaaccg cctctccccg cgcgttggcc gattcattaa tgcagctggc
4381 acgacaggtt tcccgactgg aaagcgggca gtgagcgcaa cgcaattaat gtgagttagc
4441 tcactcatta ggcaccccag gctttacact ttatgcttcc ggctcgtatg ttgtgtggaa
4501 ttgtgagcgg ataacaattt cacacaggaa acagctatga ccatgattac ggattcactg
4561 gccgtcgttt tacaacgtcg tgactgggaa aaccctggcg ttacccaact taatcgcctt
4621 gcagcacatc cccctttcgc cagctggcgt aatagcgaag aggcccgcac cgatcgccct
4681 tcccaacagt tgcgcagcct gaatggcgaa tggcgctttg cctggtttcc ggcaccagaa
4741 gcggtgccgg aaagctggct ggagtgcgat cttcctgagg ccgatactgt cgtcgtcccc
4801 tcaaactggc agatgcacgg ttacgatgcg cccatctaca ccaacgtaac ctatcccatt
4861 acggtcaatc cgccgtttgt tcccacggag aatccgacgg gttgttactc gctcacattt
4921 aatgttgatg aaagctggct acaggaaggc cagacgcgaa ttatttttga tggcgttgga
4981 att