LOCUS       Exported                5540 bp ds-DNA     circular SYN 18-DEC-2018
DEFINITION  Expression vector pML1381, complete sequence.
ACCESSION   KR232336
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5540)
  AUTHORS   DeLoache WC, Russ ZN, Narcross L, Gonzales AM, Martin VJ, Dueber JE.
  TITLE     An enzyme-coupled biosensor enables (S)-reticuline production in 
            yeast from glucose
  JOURNAL   Nat. Chem. Biol. 11 (7), 465-471 (2015)
  PUBMED    25984720
REFERENCE   2  (bases 1 to 5540)
  AUTHORS   DeLoache WC, Russ ZN, Narcross L, Gonzales AM, Martin VJJ., Dueber 
            JE.
  TITLE     Direct Submission
  JOURNAL   Submitted (22-APR-2015) Department of Bioengineering, University of 
            California, Berkeley, 2151 Berkeley Way Room 542, Berkeley, CA 
            94704, USA
REFERENCE   3  (bases 1 to 5540)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..5540
                     /organism="Expression vector pML1381"
                     /mol_type="other DNA"
                     /db_xref="taxon:1655360"
     misc_feature    5..524
                     /label=similar to LEU2 5
                     /note="similar to LEU2 5"
     misc_feature    687..692
                     /label=BsmBI recognition site
                     /note="BsmBI recognition site"
     misc_feature    694..697
                     /label=BsmBI recognition site
                     /note="BsmBI recognition site"
     misc_feature    complement(764..767)
                     /label=BsmBI recognition site
                     /note="BsmBI recognition site"
     misc_feature    complement(769..774)
                     /label=BsmBI recognition site
                     /note="BsmBI recognition site"
     regulatory      937..1584
                     /regulatory_class="promoter"
     promoter        1177..1584
                     /gene="S. cerevisiae LEU2"
                     /label=LEU2 promoter
     CDS             1585..2679
                     /codon_start=1
                     /product="LEU2p auxotrophy marker"
                     /label=LEU2p auxotrophy marker
                     /protein_id="AKH61619.1"
                     /translation="MSAPKKIVVLPGDHVGQEITAEAIKVLKAISDVRSNVKFDFENHL
                     IGGAAIDATGVPLPDEALEASKKADAVLLGAVGGPKWGTGSVRPEQGLLKIRKELQLYA
                     NLRPCNFASDSLLDLSPIKPQFAKGTDFVVVRELVGGIYFGKRKEDDGDGVAWDSEQYT
                     VPEVQRITRMAAFMALQHEPPLPIWSLDKANVLASSRLWRKTVEETIKNEFPTLKVQHQ
                     LIDSAAMILVKNPTHLNGIIITSNMFGDIISDEASVIPGSLGLLPSASLASLPDKNTAF
                     GLYEPCHGSAPDLPKNKVNPIATILSAAMMLKLSLNLPEEGKAIEDAVKKVLDAGIRTG
                     DLGGSNSTTEVGDAVAEEVKKILA"
     regulatory      2680..3156
                     /regulatory_class="terminator"
     misc_feature    3161..3670
                     /label=similar to LEU2 3
                     /note="similar to LEU2 3"
     regulatory      complement(3683..3779)
                     /regulatory_class="terminator"
     CDS             complement(3777..4637)
                     /codon_start=1
                     /product="beta-lactamase"
                     /label=beta-lactamase
                     /protein_id="AKH61618.1"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     regulatory      complement(4638..4768)
                     /regulatory_class="promoter"
     promoter        complement(4638..4742)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(4769..5532)
                     /direction=LEFT
     rep_origin      4845..5433
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"