LOCUS       Exported                3476 bp ds-DNA     circular SYN 18-DEC-2018
DEFINITION  Expression vector pASS2SUL, complete sequence.
ACCESSION   EF205336
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3476)
  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 3476)
  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 3476)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..3476
                     /organism="Expression vector pASS2SUL"
                     /mol_type="other DNA"
                     /db_xref="taxon:421740"
     regulatory      19..71
                     /regulatory_class="promoter"
                     /note="Tet promoter/operator region"
     promoter        36..91
                     /gene="tetR"
                     /label=tetR/tetA promoters
                     /note="overlapping promoters for bacterial tetR and tetA"
     protein_bind    42..60
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O1 for the tetR and tetA genes"
     protein_bind    72..90
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     CDS             99..122
                     /codon_start=1
                     /product="TetA fragment"
                     /label=TetA fragment
                     /protein_id="ABN58911.1"
                     /translation="MNSSTKI"
     RBS             130..152
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             159..482
                     /codon_start=1
                     /product="Strep-II SUMO"
                     /label=Strep-II SUMO
                     /protein_id="ABN58912.1"
                     /translation="MASWSHPQFEKSDSEVNQEAKPEVKPEVKPETHINLKVSDGSSEI
                     FFKIKKTTPLRRLMEAFAKRQGKEMDSLRFLYDGIRIQADQTPEDLDMEDNDIIEAHRE
                     QIGG"
     CDS             168..191
                     /codon_start=1
                     /product="peptide that binds Strep-Tactin(R), an engineered
                     form of streptavidin"
                     /label=Strep-Tag II
                     /translation="WSHPQFEK"
     CDS             195..479
                     /codon_start=1
                     /gene="S. cerevisiae SMT3 (truncated)"
                     /product="cleavable ubiquitin-like protein tag"
                     /label=SUMO
                     /translation="DSEVNQEAKPEVKPEVKPETHINLKVSDGSSEIFFKIKKTTPLRR
                     LMEAFAKRQGKEMDSLRFLYDGIRIQADQTPEDLDMEDNDIIEAHREQIG"
     regulatory      508..579
                     /regulatory_class="terminator"
                     /note="lpp terminator region"
     rep_origin      587..1042
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     rep_origin      587..967
                     /label=f1 origin
                     /note="f1 origin"
     promoter        1069..1173
                     /gene="bla"
                     /label=AmpR promoter
     CDS             1174..2034
                     /codon_start=1
                     /product="beta-lactamase"
                     /label=beta-lactamase
                     /protein_id="ABN58913.1"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     CDS             2041..2667
                     /codon_start=1
                     /product="Tet repressor"
                     /label=Tet repressor
                     /protein_id="ABN58914.1"
                     /translation="MMSRLDKSKVINSALELLNEVGIEGLTTRKLAQKLGVEQPTLYWH
                     VKNKRALLDALAIEMLDRHHTHFCPLEGESWQDFLRNNAKSFRCALLSHRDGAKVHLGT
                     RPTEKQYETLENQLAFLCQQGFSLENALYALSAVGHFTLGCVLEDQEHQVAKEERETPT
                     TDSMPPLLRQAIELFDHQGAEPAFLFGLELIICGLEKQLKCESGS"
     rep_origin      2820..3408
                     /direction=RIGHT