LOCUS       Exported                6482 bp ds-DNA     circular SYN 18-DEC-2018
DEFINITION  Cloning vector pYC50, complete sequence.
ACCESSION   KP238578
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6482)
  AUTHORS   Yanez Carrillo P, Castano Navarro I.
  TITLE     Expression vectors for C-terminal fusions with fluorescent proteins 
            and epitope tags in C. glabrata
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 6482)
  AUTHORS   Yanez Carrillo P, Castano Navarro I.
  TITLE     Direct Submission
  JOURNAL   Submitted (05-DEC-2014) Division de Biologia Molecular, Instituto 
            Potosino de Investigacion Cientifica y Tecnologica, Camino a la 
            Presa San Jose 2055 Col. Lomas 4 seccion, San Luis Potosi 78216, 
            Mexico
REFERENCE   3  (bases 1 to 6482)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..6482
                     /organism="Cloning vector pYC50"
                     /mol_type="other DNA"
                     /label=derived from Candida glabrata
                     /note="derived from Candida glabrata"
                     /db_xref="taxon:1629574"
     promoter        195..410
                     /gene="S. cerevisiae URA3"
                     /label=URA3 promoter
     CDS             411..1214
                     /codon_start=1
                     /gene="S. cerevisiae URA3"
                     /product="orotidine-5-phosphate decarboxylase, required 
                     for uracil biosynthesis"
                     /label=URA3
                     /note="yeast auxotrophic marker, counterselectable with 
                     5-fluoroorotic acid (5-FOA)"
                     /translation="MSKATYKERAATHPSPVAAKLFNIMHEKQTNLCASLDVRTTKELL
                     ELVEALGPKICLLKTHVDILTDFSMEGTVKPLKALSAKYNFLLFEDRKFADIGNTVKLQ
                     YSAGVYRIAEWADITNAHGVVGPGIVSGLKQAAEEVTKEPRGLLMLAELSCKGSLSTGE
                     YTKGTVDIAKSDKDFVIGFIAQRDMGGRDEGYDWLIMTPGVGLDDKGDALGQQYRTVDD
                     VVSTGSDIIIVGRGLFAKGRDAKVEGERYRKAGWEAYLRRCGQQN"
     rep_origin      complement(1343..1798)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     1943..1959
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        1966..1984
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     2017..2033
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     misc_feature    2040..2579
                     /label=MYC sequence
                     /note="MYC sequence"
     CDS             2052..2549
                     /codon_start=1
                     /product="13 tandem Myc epitope tags"
                     /label=13xMyc
                     /translation="EQKLISEEDLNGEQKLISEEDLNGLDGEQKLISEEDLNGEQKLIS
                     EEDLNGEQKLISEEDLNGLDGEQKLISEEDLNGEQKLISEEDLNGEQKLISEEDLNGLD
                     GEQKLISEEDLNGEQKLISEEDLNGEQKLISEEDLNGLDGEQKLISEEDLNGEQKLISE
                     EDL"
     misc_feature    2602..2635
                     /label=FRT sequence
                     /note="FRT sequence"
     protein_bind    complement(2602..2635)
                     /label=FRT (minimal)
                     /bound_moiety="FLP recombinase from the Saccharomyces 
                     cerevisiae 2u plasmid"
                     /note="supports FLP-mediated excision but not integration 
                     (Turan and Bode, 2011)"
     3UTR           2636..2924
                     /label=3UTR of CTA1 orf in Candida glabrata BG2
                     /note="3UTR of CTA1 orf in Candida glabrata BG2"
     regulatory      2932..3306
                     /regulatory_class="promoter"
                     /note="region upstream of TEF1 orf in Ashbya gossypii"
     promoter        2965..3308
                     /label=TEF promoter
                     /note="Ashbya gossypii TEF promoter"
     CDS             3326..3898
                     /codon_start=1
                     /gene="NAT1"
                     /product="nourseothricin-resistance protein"
                     /label=NAT1
                     /protein_id="AJW76806.1"
                     /translation="MSTTLDDTAYRYRTSVPGDAEAIEALDGSFTTDTVFRVTATGDGF
                     TLREVPVDPPLTKVFPDDESDDESDAGEDGDPDSRTFVAYGDDGDLAGFVVVSYSGWNR
                     RLTVEDIEVAPEHRGHGVGRALMGLATEFARERGAGHLWLEVTNVNAPAIHAYRRMGFT
                     LCGLDTALYDGTASDGEQALYMSMPCP"
     gene            3326..3898
                     /gene="NAT1"
                     /label=NAT1
     regulatory      3909..4144
                     /regulatory_class="terminator"
                     /note="region downstream of TEF1 orf of Ashbya gossypii"
     terminator      3912..4109
                     /label=TEF terminator
                     /note="Ashbya gossypii TEF terminator"
     misc_feature    complement(4152..4185)
                     /label=FRT sequence
                     /note="FRT sequence"
     protein_bind    complement(4152..4185)
                     /label=FRT (minimal)
                     /bound_moiety="FLP recombinase from the Saccharomyces 
                     cerevisiae 2u plasmid"
                     /note="supports FLP-mediated excision but not integration 
                     (Turan and Bode, 2011)"
     promoter        complement(4222..4240)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(4261..4277)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    4285..4301
                     /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(4309..4339)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     rep_origin      complement(4663..5251)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(5422..6282)
                     /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(6283..6387)
                     /gene="bla"
                     /label=AmpR promoter