LOCUS XXU13855 4968 bp DNA circular SYN 13-DEC-1994
DEFINITION pGEX-4T3 cloning vector, complete sequence.
ACCESSION U13855
VERSION U13855.1
KEYWORDS glutathione S-transferase; beta-lactamase; lac repressor.
SOURCE unidentified cloning vector
ORGANISM unidentified cloning vector
other sequences; artificial sequences; vectors.
REFERENCE 1 (bases 1 to 4968)
AUTHORS Malone,J.A.
TITLE pGEX-4T3: A cloning vector for the inducible expression of genes as
glutathione S-transferase fusion proteins containing a thrombin
cleavage site
JOURNAL Unpublished
REFERENCE 2 (bases 1 to 4968)
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)
PUBMED 3047011
REFERENCE 3 (bases 264 to 911)
AUTHORS Smith,D.B., Davern,K.M., Board,P.G., Tiu,W.U., Garcia,E.G. and
Mitchell,G.F.
TITLE Mr 26,000 antigen of Schistosoma japonicum recognized by resistant
WEHI 129/J mice is a parasite glutathione S-transferase
JOURNAL Proc. Natl. Acad. Sci. U.S.A. 83 (22), 8703-8707 (1986)
PUBMED 3095841
REMARK Erratum:[Proc Natl Acad Sci U S A 1987 Sep;84(18):6541]
REFERENCE 4 (bases 881 to 911)
AUTHORS Smith,D.B., Davern,K.M., Board,P.G., Tiu,W.U., Garcia,E.G. and
Mitchell,G.F.
TITLE Correction: Mr 26,000 antigen of Schistosoma japonicum recognized
by resistant WEHI 129/J mice is a parasite glutathione
S-transferase
JOURNAL Proc. Natl. Acad. Sci. U.S.A. 84, 6541-6541 (1987)
REFERENCE 5 (bases 1 to 4968)
AUTHORS Malone,J.A.
TITLE Direct Submission
JOURNAL Submitted (19-AUG-1994) James A. Malone, International Technical
Services, Molecular Biology Reagents Division, Pharmacia Biotech
Inc., 2202 N. Bartlett Ave., Milwaukee, WI 53202-1009, USA
FEATURES Location/Qualifiers
source 1..4968
/organism="unidentified cloning vector"
/mol_type="genomic DNA"
/db_xref="taxon:45196"
/lab_host="Escherichia coli"
regulatory 183..211
/regulatory_class="promoter"
/standard_name="tac"
/note="tac promoter for inducible expression of
glutathione S-transferase"
protein_bind 217..237
/bound_moiety="lac repressor protein"
CDS 258..980
/citation=[3]
/citation=[4]
/codon_start=1
/transl_table=11
/product="glutathione S-transferase"
/protein_id="AAA57104.1"
/translation="MSPILGYWKIKGLVQPTRLLLEYLEEKYEEHLYERDEGDKWRNK
KFELGLEFPNLPYYIDGDVKLTQSMAIIRYIADKHNMLGGCPKERAEISMLEGAVLDI
RYGVSRIAYSKDFETLKVDFLSKLPEMLKMFEDRLCHKTYLNGDHVTHPDFMLYDALD
VVLYMDPMCLDAFPKLVCFKKRIEAIPQIDKYLKSSKYIAWPLQGWQATFGGGDHPPK
SDLVPRGSPNSRVDSSGRIVTD"
misc_feature 918..935
/note="encodes thrombin recognition site"
misc_feature 930..965
/note="Multiple Cloning Site (MCS); contains the unique
restriction sites BamHI, EcoRI, SmaI, SalI, XhoI, and
NotI"
gene 1306..2236
/gene="bla"
regulatory 1306..1334
/regulatory_class="promoter"
/gene="bla"
CDS 1376..2236
/gene="bla"
/codon_start=1
/transl_table=11
/product="beta-lactamase"
/protein_id="AAA57105.1"
/translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGY
IELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVE
YSPVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRL
DRWEPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPL
LRSALPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIA
EIGASLIKHW"
rep_origin 2994
/note="base 2994 represents the first base of the newly
synthesized single strand"
/direction=right
gene 3317..4399
/gene="lacIq"
CDS 3317..4399
/gene="lacIq"
/codon_start=1
/transl_table=11
/product="lac repressor"
/protein_id="AAA57106.1"
/translation="MKPVTLYDVAEYAGVSYQTVSRVVNQASHVSAKTREKVEAAMAE
LNYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSRADQLGASVVVSMVERS
GVEACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSII
FSHEDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREG
DWSAMSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDT
EDSSCYIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNT
QTASPRALADSLMQLARQVSRLESGQ"
BASE COUNT 1225 a 1202 c 1291 g 1250 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 gttccgcgtg gatccccgaa ttcccgggtc gactcgagcg
961 gccgcatcgt gactgactga cgatctgcct cgcgcgtttc ggtgatgacg gtgaaaacct
1021 ctgacacatg cagctcccgg agacggtcac agcttgtctg taagcggatg ccgggagcag
1081 acaagcccgt cagggcgcgt cagcgggtgt tggcgggtgt cggggcgcag ccatgaccca
1141 gtcacgtagc gatagcggag tgtataattc ttgaagacga aagggcctcg tgatacgcct
1201 atttttatag gttaatgtca tgataataat ggtttcttag acgtcaggtg gcacttttcg
1261 gggaaatgtg cgcggaaccc ctatttgttt atttttctaa atacattcaa atatgtatcc
1321 gctcatgaga caataaccct gataaatgct tcaataatat tgaaaaagga agagtatgag
1381 tattcaacat ttccgtgtcg cccttattcc cttttttgcg gcattttgcc ttcctgtttt
1441 tgctcaccca gaaacgctgg tgaaagtaaa agatgctgaa gatcagttgg gtgcacgagt
1501 gggttacatc gaactggatc tcaacagcgg taagatcctt gagagttttc gccccgaaga
1561 acgttttcca atgatgagca cttttaaagt tctgctatgt ggcgcggtat tatcccgtgt
1621 tgacgccggg caagagcaac tcggtcgccg catacactat tctcagaatg acttggttga
1681 gtactcacca gtcacagaaa agcatcttac ggatggcatg acagtaagag aattatgcag
1741 tgctgccata accatgagtg ataacactgc ggccaactta cttctgacaa cgatcggagg
1801 accgaaggag ctaaccgctt ttttgcacaa catgggggat catgtaactc gccttgatcg
1861 ttgggaaccg gagctgaatg aagccatacc aaacgacgag cgtgacacca cgatgcctgc
1921 agcaatggca acaacgttgc gcaaactatt aactggcgaa ctacttactc tagcttcccg
1981 gcaacaatta atagactgga tggaggcgga taaagttgca ggaccacttc tgcgctcggc
2041 ccttccggct ggctggttta ttgctgataa atctggagcc ggtgagcgtg ggtctcgcgg
2101 tatcattgca gcactggggc cagatggtaa gccctcccgt atcgtagtta tctacacgac
2161 ggggagtcag gcaactatgg atgaacgaaa tagacagatc gctgagatag gtgcctcact
2221 gattaagcat tggtaactgt cagaccaagt ttactcatat atactttaga ttgatttaaa
2281 acttcatttt taatttaaaa ggatctaggt gaagatcctt tttgataatc tcatgaccaa
2341 aatcccttaa cgtgagtttt cgttccactg agcgtcagac cccgtagaaa agatcaaagg
2401 atcttcttga gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc
2461 gctaccagcg gtggtttgtt tgccggatca agagctacca actctttttc cgaaggtaac
2521 tggcttcagc agagcgcaga taccaaatac tgtccttcta gtgtagccgt agttaggcca
2581 ccacttcaag aactctgtag caccgcctac atacctcgct ctgctaatcc tgttaccagt
2641 ggctgctgcc agtggcgata agtcgtgtct taccgggttg gactcaagac gatagttacc
2701 ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc acacagccca gcttggagcg
2761 aacgacctac accgaactga gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc
2821 cgaagggaga aaggcggaca ggtatccggt aagcggcagg gtcggaacag gagagcgcac
2881 gagggagctt ccagggggaa acgcctggta tctttatagt cctgtcgggt ttcgccacct
2941 ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc
3001 cagcaacgcg gcctttttac ggttcctggc cttttgctgg ccttttgctc acatgttctt
3061 tcctgcgtta tcccctgatt ctgtggataa ccgtattacc gcctttgagt gagctgatac
3121 cgctcgccgc agccgaacga ccgagcgcag cgagtcagtg agcgaggaag cggaagagcg
3181 cctgatgcgg tattttctcc ttacgcatct gtgcggtatt tcacaccgca taaattccga
3241 caccatcgaa tggtgcaaaa cctttcgcgg tatggcatga tagcgcccgg aagagagtca
3301 attcagggtg gtgaatgtga aaccagtaac gttatacgat gtcgcagagt atgccggtgt
3361 ctcttatcag accgtttccc gcgtggtgaa ccaggccagc cacgtttctg cgaaaacgcg
3421 ggaaaaagtg gaagcggcga tggcggagct gaattacatt cccaaccgcg tggcacaaca
3481 actggcgggc aaacagtcgt tgctgattgg cgttgccacc tccagtctgg ccctgcacgc
3541 gccgtcgcaa attgtcgcgg cgattaaatc tcgcgccgat caactgggtg ccagcgtggt
3601 ggtgtcgatg gtagaacgaa gcggcgtcga agcctgtaaa gcggcggtgc acaatcttct
3661 cgcgcaacgc gtcagtgggc tgatcattaa ctatccgctg gatgaccagg atgccattgc
3721 tgtggaagct gcctgcacta atgttccggc gttatttctt gatgtctctg accagacacc
3781 catcaacagt attattttct cccatgaaga cggtacgcga ctgggcgtgg agcatctggt
3841 cgcattgggt caccagcaaa tcgcgctgtt agcgggccca ttaagttctg tctcggcgcg
3901 tctgcgtctg gctggctggc ataaatatct cactcgcaat caaattcagc cgatagcgga
3961 acgggaaggc gactggagtg ccatgtccgg ttttcaacaa accatgcaaa tgctgaatga
4021 gggcatcgtt cccactgcga tgctggttgc caacgatcag atggcgctgg gcgcaatgcg
4081 cgccattacc gagtccgggc tgcgcgttgg tgcggatatc tcggtagtgg gatacgacga
4141 taccgaagac agctcatgtt atatcccgcc gttaaccacc atcaaacagg attttcgcct
4201 gctggggcaa accagcgtgg accgcttgct gcaactctct cagggccagg cggtgaaggg
4261 caatcagctg ttgcccgtct cactggtgaa aagaaaaacc accctggcgc ccaatacgca
4321 aaccgcctct ccccgcgcgt tggccgattc attaatgcag ctggcacgac aggtttcccg
4381 actggaaagc gggcagtgag cgcaacgcaa ttaatgtgag ttagctcact cattaggcac
4441 cccaggcttt acactttatg cttccggctc gtatgttgtg tggaattgtg agcggataac
4501 aatttcacac aggaaacagc tatgaccatg attacggatt cactggccgt cgttttacaa
4561 cgtcgtgact gggaaaaccc tggcgttacc caacttaatc gccttgcagc acatccccct
4621 ttcgccagct ggcgtaatag cgaagaggcc cgcaccgatc gcccttccca acagttgcgc
4681 agcctgaatg gcgaatggcg ctttgcctgg tttccggcac cagaagcggt gccggaaagc
4741 tggctggagt gcgatcttcc tgaggccgat actgtcgtcg tcccctcaaa ctggcagatg
4801 cacggttacg atgcgcccat ctacaccaac gtaacctatc ccattacggt caatccgccg
4861 tttgttccca cggagaatcc gacgggttgt tactcgctca catttaatgt tgatgaaagc
4921 tggctacagg aaggccagac gcgaattatt tttgatggcg ttggaatt
//