LOCUS       Exported                3473 bp ds-DNA     circular SYN 18-DEC-2018
DEFINITION  Expression vector pASHSUL, complete sequence.
ACCESSION   EF205335
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3473)
  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 3473)
  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 3473)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..3473
                     /organism="Expression vector pASHSUL"
                     /mol_type="other DNA"
                     /db_xref="taxon:421739"
     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="ABN58907.1"
                     /translation="MNSSTKI"
     RBS             130..152
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             159..479
                     /codon_start=1
                     /product="His6-SUMO"
                     /label=His6-SUMO
                     /protein_id="ABN58908.1"
                     /translation="MGHHHHHHGGSDSEVNQEAKPEVKPEVKPETHINLKVSDGSSEIF
                     FKIKKTTPLRRLMEAFAKRQGKEMDSLRFLYDGIRIQADQTPEDLDMEDNDIIEAHREQ
                     IGG"
     CDS             165..182
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             192..476
                     /codon_start=1
                     /gene="S. cerevisiae SMT3 (truncated)"
                     /product="cleavable ubiquitin-like protein tag"
                     /label=SUMO
                     /translation="DSEVNQEAKPEVKPEVKPETHINLKVSDGSSEIFFKIKKTTPLRR
                     LMEAFAKRQGKEMDSLRFLYDGIRIQADQTPEDLDMEDNDIIEAHREQIG"
     regulatory      505..576
                     /regulatory_class="terminator"
                     /note="lpp terminator region"
     rep_origin      584..1039
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     rep_origin      584..964
                     /label=f1 origin
                     /note="f1 origin"
     promoter        1066..1170
                     /gene="bla"
                     /label=AmpR promoter
     CDS             1171..2031
                     /codon_start=1
                     /product="beta-lactamase"
                     /label=beta-lactamase
                     /protein_id="ABN58909.1"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     CDS             2038..2664
                     /codon_start=1
                     /product="Tet repressor"
                     /label=Tet repressor
                     /protein_id="ABN58910.1"
                     /translation="MMSRLDKSKVINSALELLNEVGIEGLTTRKLAQKLGVEQPTLYWH
                     VKNKRALLDALAIEMLDRHHTHFCPLEGESWQDFLRNNAKSFRCALLSHRDGAKVHLGT
                     RPTEKQYETLENQLAFLCQQGFSLENALYALSAVGHFTLGCVLEDQEHQVAKEERETPT
                     TDSMPPLLRQAIELFDHQGAEPAFLFGLELIICGLEKQLKCESGS"
     rep_origin      2817..3405
                     /direction=RIGHT