LOCUS       Exported                6510 bp ds-DNA     circular SYN 18-DEC-2018
DEFINITION  Expression vector pET151-OmyCasp3 DNA, complete sequence.
ACCESSION   AB477343
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6510)
  AUTHORS   Ojima N.
  TITLE     Characterization of rainbow trout caspase-3
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 6510)
  AUTHORS   Ojima N.
  TITLE     Direct Submission
  JOURNAL   Submitted (15-JAN-2009) Contact:Nobuhiko Ojima National Research 
            Institute of Fisheries Science, Fisheries Research Agency, Aquatic 
            Genomics Research Center; 2-12-4 Fukuura, Kanazawa, Yokohama 
            236-8648, Japan
REFERENCE   3  (bases 1 to 6510)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..6510
                     /organism="Expression vector pET151-OmyCasp3"
                     /mol_type="other DNA"
                     /db_xref="taxon:590940"
     promoter        209..227
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    228..252
                     /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)."
     RBS             267..289
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             297..1142
                     /codon_start=1
                     /gene="casp3"
                     /product="histidine-tagged truncated caspase-3"
                     /label=casp3
                     /protein_id="BAU69681.1"
                     /translation="MHHHHHHGKPIPNPLLGLDSTENLYFQGIDPFTAKPQSHCFRYSL
                     SYPSIGQCIIINNKNFDRRTGMNVRNGTDVDAGNVMKVFGKLGYKVKVYNDQKVDQIKH
                     VLTTASKEDHSCSASFVCVLLSHGDDGVFFGTDASIELKSLTSLFRGDRCNSLVGKPKL
                     FFIQACRGTDLDGGIEADSSSDGSSTKIPVEADFLYAYSTAPGYYSWRNTQTGSWFIQS
                     LCEMIGKYSKELEVQHILTRVNHKVATEFESASNSPGFDAKKQIPCIVSMLTKEMYFTP
                     "
     gene            297..1142
                     /gene="casp3"
                     /label=casp3
     CDS             300..317
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             318..359
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     CDS             360..380
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     CDS             1142..1153
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     terminator      1222..1269
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     promoter        1593..1697
                     /gene="bla"
                     /label=AmpR promoter
     CDS             1698..2558
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=bla
                     /protein_id="BAU69682.1"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     gene            1698..2558
                     /gene="bla"
                     /label=bla
     rep_origin      2729..3317
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     misc_feature    3503..3645
                     /label=bom
                     /note="basis of mobility region from pBR322"
     CDS             complement(3747..3938)
                     /codon_start=1
                     /gene="rop"
                     /product="regulatory protein rop"
                     /label=rop
                     /protein_id="BAU69683.1"
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     gene            complement(3747..3938)
                     /gene="rop"
                     /label=rop
     CDS             complement(5250..6332)
                     /codon_start=1
                     /gene="lacI"
                     /product="lactose operon repressor"
                     /label=lacI
                     /protein_id="BAU69684.1"
                     /translation="MKPVTLYDVAEYAGVSYQTVSRVVNQASHVSAKTREKVEAAMAEL
                     NYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSRADQLGASVVVSMVERSGV
                     EACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSIIFSH
                     EDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREGDWSA
                     MSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTEDSSC
                     YIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR
                     ALADSLMQLARQVSRLESGQ"
     gene            complement(5250..6332)
                     /gene="lacI"
                     /label=lacI
     promoter        complement(6333..6410)
                     /gene="lacI"
                     /label=lacI promoter