LOCUS       Exported               10263 bp ds-DNA     circular SYN 18-DEC-2018
DEFINITION  Cloning vector pLUMCL, complete sequence.
ACCESSION   KC999859
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 10263)
  AUTHORS   Shahana S, Childers DS, Ballou ER, Bohovych I, Odds FC, Gow NA, 
            Brown AJ.
  TITLE     New Clox Systems for Rapid and Efficient Gene Disruption in Candida 
            albicans
  JOURNAL   PLoS ONE 9 (6), E100390 (2014)
  PUBMED    24940603
REFERENCE   2  (bases 1 to 10263)
  AUTHORS   Shahana S, Childers DS, Brown AJP.
  TITLE     Direct Submission
  JOURNAL   Submitted (01-MAY-2013) Microbiology, School of Medical Sciences, 
            Institute of Medical Sciences - Foresterhill, Aberdeen, 
            Aberdeenshire AB25 2ZD, United Kingdom
REFERENCE   3  (bases 1 to 10263)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     ##Assembly-Data-START##
            Sequencing Technology :: Sanger dideoxy sequencing 
            ##Assembly-Data-END##
FEATURES             Location/Qualifiers
     source          1..10263
                     /organism="Cloning vector pLUMCL"
                     /mol_type="other DNA"
                     /db_xref="taxon:1415791"
     CDS             complement(663..1757)
                     /codon_start=1
                     /gene="S. cerevisiae LEU2"
                     /product="3-isopropylmalate dehydrogenase, required for 
                     leucine biosynthesis"
                     /label=LEU2
                     /note="yeast auxotrophic marker"
                     /translation="MSAPKKIVVLPGDHVGQEITAEAIKVLKAISDVRSNVKFDFENHL
                     IGGAAIDATGVPLPDEALEASKKVDAVLLGAVGGPKWGTGSVRPEQGLLKIRKELQLYA
                     NLRPCNFASDSLLDLSPIKPQFAKGTDFVVVRELVGGIYFGKRKEDDGDGVAWDSEQYT
                     VPEVQRITRMAAFMALQHEPPLPIWSLDKANVLASSRLWRKTVEETIKNEFPTLKVQHQ
                     LIDSAAMILVKNPTHLNGIIITSNMFGDIISDEASVIPGSLGLLPSASLASLPDKNTAF
                     GLYEPCHGSAPDLPKNKVDPIATILSAAMMLKLSLNLPEEGKAIEDAVKKVLDAGIRTG
                     DLGGSNSTTEVGDAVAEEVKKILA"
     promoter        complement(1770..2174)
                     /gene="S. cerevisiae LEU2"
                     /label=LEU2 promoter
     rep_origin      complement(2474..2929)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     3074..3090
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        3097..3115
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     3148..3164
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     misc_feature    join(3178..3211,7365..7398)
                     /label=loxP
                     /note="loxP"
     protein_bind    3178..3211
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (GCATACAT)."
     CDS             3641..4453
                     /codon_start=1
                     /product="URA3"
                     /label=URA3
                     /protein_id="AGZ63436.1"
                     /translation="MTVNTKTYSERAETHASPVAQRLFRLMELKKTNLCASIDVDTTKE
                     FLELIDKLGPYVCLIKTHIDIINDFSYESTIEPLLELSRKHQFMIFEDRKFADIGNTVK
                     KQYIGGVYKISSWADITNAHGVTGNGVVEGLKQGAKETTTNQEPRGLLMLAELSSVGSL
                     AYGEYSQKTVEIAKSDKEFVIGFIAQRDMGGQEEGFDWLIMTPGVGLDDKGDGLGQQYR
                     TVDEVVSTGTDIIIVGRGLFGKGRDPDIEGKRYRDAGWNAYLKKTGQL"
     regulatory      4595..5926
                     /regulatory_class="promoter"
                     /gene="pMET3"
                     /note="MET3 promoter"
     gene            4595..5926
                     /gene="pMET3"
                     /label=pMET3
     CDS             5954..6361
                     /codon_start=1
                     /product="Cre"
                     /label=Cre
                     /note="Cre recombinase"
                     /protein_id="AGZ63437.1"
                     /translation="MSNLLTVHQNLPALPVDATSDEVRKNLMDMFRDRQAFSEHTWKML
                     LSVCRSWAAWCKLNNRKWFPAEPEDVRDYLLYLQARGLAVKTIQQHLGQLNMLHRRSGL
                     PRPSDSNAVSLVMRRIRKENVDAGERAKQAL"
     protein_bind    7365..7398
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (GCATACAT)."
     primer_bind     complement(7443..7459)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        complement(7489..7507)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(7528..7544)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    7552..7568
                     /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)."
     promoter        complement(7576..7606)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     rep_origin      complement(7930..8518)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(8689..9549)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFFHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     promoter        complement(9550..9654)
                     /gene="bla"
                     /label=AmpR promoter
     misc_feature    9691..10194
                     /label=CEN/ARS
                     /note="S. cerevisiae CEN6 centromere fused to an 
                     autonomously replicating sequence"