LOCUS       Exported                5680 bp ds-DNA     circular SYN 18-DEC-2018
DEFINITION  Expression vector pETS2SUL, complete sequence.
ACCESSION   EF205334
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5680)
  AUTHORS   Weeks SD, Drinker M, Loll PJ.
  TITLE     Ligation independent cloning vectors for expression of SUMO fusions
  JOURNAL   Protein Expr. Purif. 53 (1), 40-50 (2007)
  PUBMED    17251035
REFERENCE   2  (bases 1 to 5680)
  AUTHORS   Weeks SD, Drinker M, Loll PJ.
  TITLE     Direct Submission
  JOURNAL   Submitted (03-JAN-2007) Biochemistry and Molecular Biology, Drexel 
            University College of Medicine, 245 N 15th Street, Mailstop 497, 
            Philadelphia, PA 19102, USA
REFERENCE   3  (bases 1 to 5680)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..5680
                     /organism="Expression vector pETS2SUL"
                     /mol_type="other DNA"
                     /db_xref="taxon:421738"
     terminator      26..73
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     regulatory      complement(26..72)
                     /regulatory_class="terminator"
                     /note="T7 terminator"
     CDS             complement(154..477)
                     /codon_start=1
                     /product="Strep-II SUMO"
                     /label=Strep-II SUMO
                     /protein_id="ABN58903.1"
                     /translation="MASWSHPQFEKSDSEVNQEAKPEVKPEVKPETHINLKVSDGSSEI
                     FFKIKKTTPLRRLMEAFAKRQGKEMDSLRFLYDGIRIQADQTPEDLDMEDNDIIEAHRE
                     QIGG"
     CDS             complement(157..441)
                     /codon_start=1
                     /gene="S. cerevisiae SMT3 (truncated)"
                     /product="cleavable ubiquitin-like protein tag"
                     /label=SUMO
                     /translation="DSEVNQEAKPEVKPEVKPETHINLKVSDGSSEIFFKIKKTTPLRR
                     LMEAFAKRQGKEMDSLRFLYDGIRIQADQTPEDLDMEDNDIIEAHREQIG"
     CDS             complement(445..468)
                     /codon_start=1
                     /product="peptide that binds Strep-Tactin(R), an engineered
                     form of streptavidin"
                     /label=Strep-Tag II
                     /translation="WSHPQFEK"
     RBS             484..506
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     protein_bind    521..545
                     /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    complement(523..543)
                     /label=Lac operator
                     /note="Lac operator"
     promoter        complement(546..564)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      complement(548..570)
                     /regulatory_class="promoter"
                     /note="T7 promoter"
     promoter        873..950
                     /gene="lacI"
                     /label=lacI promoter
     CDS             951..2033
                     /codon_start=1
                     /product="LacI"
                     /label=LacI
                     /protein_id="ABN58906.1"
                     /translation="MKPVTLYDVAEYAGVSYQTVSRVVNQASHVSAKTREKVEAAMAEL
                     NYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSRADQLGASVVVSMVERSGV
                     EACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSIIFSH
                     EDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREGDWSA
                     MSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTEDSSC
                     YIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR
                     ALADSLMQLARQVSRLESGQ"
     CDS             2842..3033
                     /codon_start=1
                     /product="rop"
                     /label=rop
                     /protein_id="ABN58905.1"
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     misc_feature    3135..3277
                     /label=bom
                     /note="basis of mobility region from pBR322"
     rep_origin      complement(3463..4051)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     rep_origin      complement(3643..4051)
                     /direction=LEFT
     CDS             complement(4222..5082)
                     /codon_start=1
                     /product="beta-lactamase"
                     /label=beta-lactamase
                     /protein_id="ABN58904.1"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     promoter        complement(5083..5187)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(5214..5669)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     rep_origin      5289..5669
                     /label=f1 origin
                     /note="f1 origin"