LOCUS       Exported                5664 bp ds-DNA     circular SYN 18-DEC-2018
DEFINITION  Expression vector pT7MT, complete sequence.
ACCESSION   DQ504436
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5664)
  AUTHORS   Marikar FM, Fang L, Jiang SH, Hua ZC.
  TITLE     pT7MT, a metallothionein 2A-tagged novel prokaryotic fusion 
            expression vector
  JOURNAL   J. Microbiol. Biotechnol. 17 (5), 728-732 (2007)
  PUBMED    18051292
REFERENCE   2  (bases 1 to 5664)
  AUTHORS   Marikar FMM.T., Hua Z-C.
  TITLE     Direct Submission
  JOURNAL   Submitted (20-APR-2006) State Key Lab of Pharmaceutical 
            Biotechnology, Nanjing University, 20 HanKou Road, Nanjing, Jiangsu 
            210093, P.R.China
REFERENCE   3  (bases 1 to 5664)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..5664
                     /organism="Expression vector pT7MT"
                     /lab_host="Escherichia coli"
                     /mol_type="other DNA"
                     /db_xref="taxon:387955"
     rep_origin      12..467
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     CDS             599..1459
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIDYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     rep_origin      1630..2218
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     misc_feature    2404..2546
                     /label=bom
                     /note="basis of mobility region from pBR322"
     CDS             complement(2648..2839)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(3648..4730)
                     /codon_start=1
                     /gene="lacI"
                     /product="lac repressor"
                     /label=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
                     ALADSLMRLARQVSRLESGQ"
     promoter        5121..5139
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      5121..5137
                     /regulatory_class="promoter"
                     /note="T7 promoter"
     misc_feature    5140..5166
                     /label=Lac operator
                     /note="Lac operator"
     protein_bind    5140..5164
                     /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)."
     misc_feature    5173..5238
                     /note="Omega sequence; leader sequence from tobacco mosaic 
                     virus; MTV U2 RAA; tobacco mosaic virus U2"
     RBS             5254..5276
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     regulatory      5269..5281
                     /regulatory_class="ribosome_binding_site"
     CDS             5283..5462
                     /codon_start=1
                     /product="metallothionein 2A"
                     /label=metallothionein 2A
                     /note="used as fusion partner; derived from Homo sapiens 
                     MT2A; similar to NM_005953"
                     /protein_id="ABG24572.1"
                     /translation="MENCSCAAGDSCTCAGSCKCKECKCTSCKKSCCSCCPVGCAKCAQ
                     GCICKGASDKCSCCA"
     CDS             5463..5474
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     misc_feature    5463..5465
                     /product="polylinker"
                     /note="factor Xa sequence; polylinker region; included for 
                     subsequent fusion protein cleavage"
     misc_feature    5476..5526
                     /note="multiple cloning site; MCS; designed for inserting 
                     fusion partners"
     regulatory      5592..5664
                     /regulatory_class="terminator"
                     /note="T7 terminator"
     terminator      5592..5639
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"