LOCUS       Exported                5329 bp ds-DNA     circular SYN 18-DEC-2018
DEFINITION  Cloning vector pGATA, complete sequence.
ACCESSION   DQ997052
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5329)
  AUTHORS   Trudel P, Provost S, Massie B, Chartrand P, Wall L.
  TITLE     pGATA: a positive selection vector based on the toxicity of the 
            transcription factor GATA-1 to bacteria
  JOURNAL   BioTechniques 20 (4), 684-693 (1996)
  PUBMED    8800690
REFERENCE   2  (bases 1 to 5329)
  AUTHORS   Trudel P, Chartrand P, Wall L, Ching RW.
  TITLE     Direct Submission
  JOURNAL   Submitted (15-SEP-2006) Biochemistry, University of Toronto, 555 
            University Avenue, Toronto, ON M5G 1X8, Canada
REFERENCE   3  (bases 1 to 5329)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..5329
                     /organism="Cloning vector pGATA"
                     /mol_type="other DNA"
                     /label=vector based on pGEX-2T
                     /note="vector based on pGEX-2T"
                     /db_xref="taxon:407393"
     promoter        183..211
                     /label=tac promoter
                     /note="strong E. coli promoter; hybrid between the trp and 
                     lac UV5 promoters"
     protein_bind    219..235
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by 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)."
     CDS             258..1337
                     /codon_start=1
                     /product="glutathione S-transferase/GATA-1 fusion protein"
                     /label=glutathione S-transferase/GATA-1 fusion protein
                     /protein_id="ABJ91394.1"
                     /translation="MSPILGYWKIKGLVQPTRLLLEYLEEKYEEHLYERDEGDKWRNKK
                     FELGLEFPNLPYYIDGDVKLTQSMAIIRYIADKHNMLGGCPKERAEISMLEGAVLDIRY
                     GVSRIAYSKDFETLKVDFLSKLPEMLKMFEDRLCHKTYFAQLTLTKGNKSWSSTAVAAA
                     LELVDPPGCRNSISSLSIPSTSRGGPVPNSPYSESYYARNGDHVTHPDFMLYDALDVVL
                     YMDPMCLDAFPKLVCFKKRIEAIPQIDKYLKSSKYIAWPLQGWQATFGGGDHPPKSDLV
                     PRGSKRAGTQCTNCQTTTTTLWRRNASGDPVCNACGLYFKLHQVNRPLTMRKDGIQTRN
                     RKASGKGKKKRGSNGIHRD"
     misc_feature    join(258..721,830..1095)
                     /note="coding sequence of GST is split by an in-frame 
                     multiple cloning site from pBlueScript"
     promoter        691..709
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     misc_structure  722..829
                     /label=multiple cloning site (MCS)
                     /note="multiple cloning site (MCS)"
     misc_feature    722..829
                     /label=MCS
                     /note="pBluescript multiple cloning site"
     primer_bind     746..762
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(796..812)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        complement(838..856)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     misc_feature    1095..1112
                     /label=encodes thrombin protease cut site
                     /note="encodes thrombin protease cut site"
     CDS             1095..1112
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
     misc_feature    1110..1319
                     /note="fragment of murine GATA-1; contains the second zinc 
                     finger and basic regions of the protein"
     promoter        1632..1736
                     /gene="bla"
                     /label=AmpR promoter
     CDS             1737..2597
                     /codon_start=1
                     /product="beta lactamase"
                     /label=beta lactamase
                     /note="ampicillin resistance; AmpR"
                     /protein_id="ABJ91392.1"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     rep_origin      2768..3356
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     promoter        3600..3677
                     /gene="lacI (mutant)"
                     /label=lacIq promoter
                     /note="In the lacIq allele, a single base change in the 
                     promoter boosts expression of the lacI gene about 10-fold."
     CDS             3678..4760
                     /codon_start=1
                     /gene="LacI"
                     /product="LacI"
                     /label=LacI
                     /note="Lac repressor"
                     /protein_id="ABJ91393.1"
                     /translation="MKPVTLYDVAEYAGVSYQTVSRVVNQASHVSAKTREKVEAAMAEL
                     NYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSRADQLGASVVVSMVERSGV
                     EACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSIIFSH
                     EDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREGDWSA
                     MSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTEDSSC
                     YIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR
                     ALADSLMQLARQVSRLESGQ"
     gene            3678..4760
                     /gene="LacI"
                     /label=LacI
     protein_bind    4773..4794
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        4809..4839
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4847..4863
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by 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)."
     primer_bind     4871..4887
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     CDS             4883..81
                     /codon_start=1
                     /gene="lacZ fragment"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /label=lacZ-alpha
                     /translation="MTMITDSLAVVLQRRDWENPGVTQLNRLAAHPPFASWRNSEEART
                     DRPSQQLRSLNGEWRFAWFPAPEAVPESWLECDLPEADTVVVPSNWQMHGYDAPIYTNV
                     TYPITVNPPFVPTENPTGCYSLTFNVDESWLQEGQTRIIFDGVGITLSTARCTNASGVR
                     QPSEAVVWLCRS"
     primer_bind     complement(4903..4919)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"