pGEX-2TK Bacterial expression vector
LOCUS XXU13851 4969 bp DNA circular SYN 13-DEC-1994
DEFINITION pGEX-2TK cloning vector, complete sequence.
ACCESSION U13851
NID g595713
KEYWORDS glutathione S-transferase; beta-lactamase; lac repressor.
SOURCE unidentified cloning vector.
ORGANISM unidentified cloning vector
artificial sequence; cloning vectors.
REFERENCE 1 (bases 1 to 4969)
AUTHORS Malone,J.A.
TITLE pGEX-2TK: A cloning vector for the inducible expression of genes as
glutathione S-transferase fusion proteins containing a thrombin
cleavage site and a cAMP kinase recognition site
JOURNAL Unpublished (1994)
REFERENCE 2 (bases 1 to 4969)
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
REFERENCE 3 (bases 936 to 950)
AUTHORS Kaelin,W.G., Krek,W., Sellers,W.R., DeCaprio,J.A., Ajchenbaum,F.,
Fuchs,C.S., Chittenden,T., Li,Y., Farnham,P.J., Blanar,M.A.,
Livingston,D.M. and Flemington,E.K.
TITLE Expression cloning of a cDNA encoding a retinoblastoma-binding
protein with E2F-like properties
JOURNAL Cell 70 (2), 351-364 (1992)
MEDLINE 92346721
REFERENCE 4 (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 [published
erratum appears in Proc Natl Acad Sci U S A 1987 Sep;84(18):6541]
JOURNAL Proc. Natl. Acad. Sci. U.S.A. 83 (22), 8703-8707 (1986)
MEDLINE 87041520
REFERENCE 5 (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 6 (bases 1 to 4969)
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..4969
/organism="unidentified cloning vector"
/lab_host="Escherichia coli"
promoter 183..211
/standard_name="tac"
/note="tac promoter for inducible expression of
glutathione S-transferase"
protein_bind 217..237
/gene="lac"
/bound_moiety="lac repressor protein"
CDS 258..977
/citation=[4]
/citation=[5]
/codon_start=1
/product="glutathione S-transferase"
/db_xref="PID:g595714"
/transl_table=11
/translation="MSPILGYWKIKGLVQPTRLLLEYLEEKYEEHLYERDEGDKWRNK
KFELGLEFPNLPYYIDGDVKLTQSMAIIRYIADKHNMLGGCPKERAEISMLEGAVLDI
RYGVSRIAYSKDFETLKVDFLSKLPEMLKMFEDRLCHKTYLNGDHVTHPDFMLYDALD
VVLYMDPMCLDAFPKLVCFKKRIEAIPQIDKYLKSSKYIAWPLQGWQATFGGGDHPPK
SDLVPRGSRRASVGSPGIHRD"
misc_feature 918..935
/note="encodes thrombin recognition site"
misc_feature 936..950
/note="encodes cAMP-dependent protein kinase recognition
site"
/citation=[2]
misc_feature 951..966
/note="Multiple Cloning Site (MCS); contains the unique
restriction sites BamHI, SmaI and EcoRI"
promoter 1307..1335
/gene="bla"
CDS 1377..2237
/gene="bla"
/codon_start=1
/product="beta-lactamase"
/db_xref="PID:g595715"
/transl_table=11
/translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGY
IELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVE
YSPVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRL
DRWEPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPL
LRSALPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIA
EIGASLIKHW"
rep_origin 2995
/note="base 2995 represents the first base of the newly
synthesized single strand"
/direction=right
CDS 3318..4400
/gene="lacIq"
/codon_start=1
/product="lac repressor"
/db_xref="PID:g595716"
/transl_table=11
/translation="MKPVTLYDVAEYAGVSYQTVSRVVNQASHVSAKTREKVEAAMAE
LNYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSRADQLGASVVVSMVERS
GVEACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSII
FSHEDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREG
DWSAMSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDT
EDSSCYIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNT
QTASPRALADSLMQLARQVSRLESGQ"
BASE COUNT 1225 a 1198 c 1290 g 1256 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 gatctcgtcg tgcatctgtt ggatccccgg
961 gaattcatcg tgactgactg acgatctgcc tcgcgcgttt cggtgatgac ggtgaaaacc
1021 tctgacacat gcagctcccg gagacggtca cagcttgtct gtaagcggat gccgggagca
1081 gacaagcccg tcagggcgcg tcagcgggtg ttggcgggtg tcggggcgca gccatgaccc
1141 agtcacgtag cgatagcgga gtgtataatt cttgaagacg aaagggcctc gtgatacgcc
1201 tatttttata ggttaatgtc atgataataa tggtttctta gacgtcaggt ggcacttttc
1261 ggggaaatgt gcgcggaacc cctatttgtt tatttttcta aatacattca aatatgtatc
1321 cgctcatgag acaataaccc tgataaatgc ttcaataata ttgaaaaagg aagagtatga
1381 gtattcaaca tttccgtgtc gcccttattc ccttttttgc ggcattttgc cttcctgttt
1441 ttgctcaccc agaaacgctg gtgaaagtaa aagatgctga agatcagttg ggtgcacgag
1501 tgggttacat cgaactggat ctcaacagcg gtaagatcct tgagagtttt cgccccgaag
1561 aacgttttcc aatgatgagc acttttaaag ttctgctatg tggcgcggta ttatcccgtg
1621 ttgacgccgg gcaagagcaa ctcggtcgcc gcatacacta ttctcagaat gacttggttg
1681 agtactcacc agtcacagaa aagcatctta cggatggcat gacagtaaga gaattatgca
1741 gtgctgccat aaccatgagt gataacactg cggccaactt acttctgaca acgatcggag
1801 gaccgaagga gctaaccgct tttttgcaca acatggggga tcatgtaact cgccttgatc
1861 gttgggaacc ggagctgaat gaagccatac caaacgacga gcgtgacacc acgatgcctg
1921 cagcaatggc aacaacgttg cgcaaactat taactggcga actacttact ctagcttccc
1981 ggcaacaatt aatagactgg atggaggcgg ataaagttgc aggaccactt ctgcgctcgg
2041 cccttccggc tggctggttt attgctgata aatctggagc cggtgagcgt gggtctcgcg
2101 gtatcattgc agcactgggg ccagatggta agccctcccg tatcgtagtt atctacacga
2161 cggggagtca ggcaactatg gatgaacgaa atagacagat cgctgagata ggtgcctcac
2221 tgattaagca ttggtaactg tcagaccaag tttactcata tatactttag attgatttaa
2281 aacttcattt ttaatttaaa aggatctagg tgaagatcct ttttgataat ctcatgacca
2341 aaatccctta acgtgagttt tcgttccact gagcgtcaga ccccgtagaa aagatcaaag
2401 gatcttcttg agatcctttt tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac
2461 cgctaccagc ggtggtttgt ttgccggatc aagagctacc aactcttttt ccgaaggtaa
2521 ctggcttcag cagagcgcag ataccaaata ctgtccttct agtgtagccg tagttaggcc
2581 accacttcaa gaactctgta gcaccgccta catacctcgc tctgctaatc ctgttaccag
2641 tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac
2701 cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc agcttggagc
2761 gaacgaccta caccgaactg agatacctac agcgtgagct atgagaaagc gccacgcttc
2821 ccgaagggag aaaggcggac aggtatccgg taagcggcag ggtcggaaca ggagagcgca
2881 cgagggagct tccaggggga aacgcctggt atctttatag tcctgtcggg tttcgccacc
2941 tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg
3001 ccagcaacgc ggccttttta cggttcctgg ccttttgctg gccttttgct cacatgttct
3061 ttcctgcgtt atcccctgat tctgtggata accgtattac cgcctttgag tgagctgata
3121 ccgctcgccg cagccgaacg accgagcgca gcgagtcagt gagcgaggaa gcggaagagc
3181 gcctgatgcg gtattttctc cttacgcatc tgtgcggtat ttcacaccgc ataaattccg
3241 acaccatcga atggtgcaaa acctttcgcg gtatggcatg atagcgcccg gaagagagtc
3301 aattcagggt ggtgaatgtg aaaccagtaa cgttatacga tgtcgcagag tatgccggtg
3361 tctcttatca gaccgtttcc cgcgtggtga accaggccag ccacgtttct gcgaaaacgc
3421 gggaaaaagt ggaagcggcg atggcggagc tgaattacat tcccaaccgc gtggcacaac
3481 aactggcggg caaacagtcg ttgctgattg gcgttgccac ctccagtctg gccctgcacg
3541 cgccgtcgca aattgtcgcg gcgattaaat ctcgcgccga tcaactgggt gccagcgtgg
3601 tggtgtcgat ggtagaacga agcggcgtcg aagcctgtaa agcggcggtg cacaatcttc
3661 tcgcgcaacg cgtcagtggg ctgatcatta actatccgct ggatgaccag gatgccattg
3721 ctgtggaagc tgcctgcact aatgttccgg cgttatttct tgatgtctct gaccagacac
3781 ccatcaacag tattattttc tcccatgaag acggtacgcg actgggcgtg gagcatctgg
3841 tcgcattggg tcaccagcaa atcgcgctgt tagcgggccc attaagttct gtctcggcgc
3901 gtctgcgtct ggctggctgg cataaatatc tcactcgcaa tcaaattcag ccgatagcgg
3961 aacgggaagg cgactggagt gccatgtccg gttttcaaca aaccatgcaa atgctgaatg
4021 agggcatcgt tcccactgcg atgctggttg ccaacgatca gatggcgctg ggcgcaatgc
4081 gcgccattac cgagtccggg ctgcgcgttg gtgcggatat ctcggtagtg ggatacgacg
4141 ataccgaaga cagctcatgt tatatcccgc cgttaaccac catcaaacag gattttcgcc
4201 tgctggggca aaccagcgtg gaccgcttgc tgcaactctc tcagggccag gcggtgaagg
4261 gcaatcagct gttgcccgtc tcactggtga aaagaaaaac caccctggcg cccaatacgc
4321 aaaccgcctc tccccgcgcg ttggccgatt cattaatgca gctggcacga caggtttccc
4381 gactggaaag cgggcagtga gcgcaacgca attaatgtga gttagctcac tcattaggca
4441 ccccaggctt tacactttat gcttccggct cgtatgttgt gtggaattgt gagcggataa
4501 caatttcaca caggaaacag ctatgaccat gattacggat tcactggccg tcgttttaca
4561 acgtcgtgac tgggaaaacc ctggcgttac ccaacttaat cgccttgcag cacatccccc
4621 tttcgccagc tggcgtaata gcgaagaggc ccgcaccgat cgcccttccc aacagttgcg
4681 cagcctgaat ggcgaatggc gctttgcctg gtttccggca ccagaagcgg tgccggaaag
4741 ctggctggag tgcgatcttc ctgaggccga tactgtcgtc gtcccctcaa actggcagat
4801 gcacggttac gatgcgccca tctacaccaa cgtaacctat cccattacgg tcaatccgcc
4861 gtttgttccc acggagaatc cgacgggttg ttactcgctc acatttaatg ttgatgaaag
4921 ctggctacag gaaggccaga cgcgaattat ttttgatggc gttggaatt
//