LOCUS       Exported                4012 bp ds-DNA     circular SYN 18-DEC-2018
DEFINITION  Expression vector pETIA, complete sequence.
ACCESSION   GQ421429
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4012)
  AUTHORS   Tanaka S, Onda M.
  TITLE     Bacterial expression with pETIA
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 4012)
  AUTHORS   Tanaka S, Onda M.
  TITLE     Direct Submission
  JOURNAL   Submitted (29-JUL-2009) BioDynamics Laboratory Inc., 9-7 Hongo 
            2-Chome, Bunkyo-Ku, Tokyo 113-0033, Japan
REFERENCE   3  (bases 1 to 4012)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..4012
                     /organism="Expression vector pETIA"
                     /mol_type="other DNA"
                     /db_xref="taxon:670878"
     promoter        213..231
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      213..229
                     /regulatory_class="promoter"
                     /note="T7 promoter"
     RBS             263..285
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     misc_feature    305..322
                     /label=histidine tag
                     /note="histidine tag"
     CDS             305..322
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             326..358
                     /codon_start=1
                     /product="leader peptide from bacteriophage T7 gene 10"
                     /label=T7 tag (gene 10 leader)
                     /note="promotes efficient translation in E. coli"
                     /translation="MASMTGGQQMG"
     CDS             362..376
                     /codon_start=1
                     /product="enterokinase recognition and cleavage site"
                     /label=enterokinase site
                     /translation="DDDDK"
     misc_feature    376..422
                     /label=multiple cloning site
                     /note="multiple cloning site"
     regulatory      498..545
                     /regulatory_class="terminator"
                     /note="T7 terminator"
     terminator      498..545
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     promoter        588..679
                     /gene="bla"
                     /label=AmpR promoter
     CDS             680..1540
                     /codon_start=1
                     /gene="bla"
                     /product="beta lactamase"
                     /label=bla
                     /note="confers ampicillin resistance"
                     /protein_id="ACV53162.1"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPTAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     gene            680..1540
                     /gene="bla"
                     /label=bla
     rep_origin      1695..2314
                     /label=pMB1 ori
                     /note="pMB1 ori"
     rep_origin      1711..2299
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     misc_feature    2485..2627
                     /label=bom
                     /note="basis of mobility region from pBR322"
     promoter        2813..2890
                     /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             2891..3973
                     /codon_start=1
                     /gene="lacI"
                     /product="lac repressor"
                     /label=lacI
                     /protein_id="ACV53163.1"
                     /translation="MKPVTLYDVAEYAGVSYQTVSRVVNQASHVSAKTREKVEAAMAEL
                     NYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSRADQLGASVVVSMVERSGV
                     EACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSIIFSH
                     EDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREGDWSA
                     MSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTEDSSC
                     YIPPSTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR
                     ALADSLMQLARQVSRLESGQ"
     gene            2891..3973
                     /gene="lacI"
                     /label=lacI