LOCUS       40924_47928        6482 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Cloning vector pYC50, complete sequence.
ACCESSION   .
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  JOURNAL   Unpublished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)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 6482)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: 
            "Unpublished"
COMMENT     SGRef: number: 2; type: "Journal Article"; journalName: "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"
COMMENT     SGRef: number: 3; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..6482
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        190..410
                     /label=URA3 promoter
     CDS             411..1211
                     /codon_start=1
                     /label=URA3
                     /note="orotidine-5'-phosphate decarboxylase, required for
                     uracil biosynthesis"
                     /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"
     CDS             2052..2081
                     /codon_start=1
                     /label=Myc
                     /note="Myc (human c-Myc proto-oncogene) epitope tag"
                     /translation="EQKLISEEDL"
     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)"
     3'UTR           2636..2924
                     /label=3'UTR of CTA1 orf in Candida glabrata BG2
                     /note="3'UTR of CTA1 orf in Candida glabrata BG2"
     promoter        2965..3308
                     /label=TEF promoter
                     /note="Ashbya gossypii TEF promoter"
     CDS             3332..3895
                     /codon_start=1
                     /label=NrsR
                     /note="nourseothricin acetyltransferase"
                     /translation="TTLDDTAYRYRTSVPGDAEAIEALDGSFTTDTVFRVTATGDGFTL
                     REVPVDPPLTKVFPDDESDDESDAGEDGDPDSRTFVAYGDDGDLAGFVVVSYSGWNRRL
                     TVEDIEVAPEHRGHGVGRALMGLATEFARERGAGHLWLEVTNVNAPAIHAYRRMGFTLC
                     GLDTALYDGTASDGEQALYMSMPCP"
     terminator      3912..4109
                     /label=TEF terminator
                     /note="Ashbya gossypii TEF terminator"
     protein_bind    complement(4152..4185)
                     /label=FRT (minimal)
                     /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    complement(4285..4301)
                     /label=lac operator
                     /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"
     protein_bind    complement(4354..4375)
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     rep_origin      complement(4663..5251)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(5425..6282)
                     /codon_start=1
                     /label=AmpR
                     /note="beta-lactamase"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFFHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     promoter        complement(6283..6387)
                     /label=AmpR promoter
ORIGIN
        1 tcgcgcgttt cggtgatgac ggtgaaaacc tctgacacat gcagctcccg gagacggtca
       61 cagcttgtct gtaagcggat gccgggagca gacaagcccg tcagggcgcg tcagcgggtg
      121 ttggcgggtg tcggggctgg cttaactatg cggcatcaga gcagattgta ctgagagtgc
      181 accacgcttt tcaattcaat tcatcatttt ttttttattc ttttttttga tttcggtttc
      241 tttgaaattt ttttgattcg gtaatctccg aacagaagga agaacgaagg aaggagcaca
      301 gacttagatt ggtatatata cgcatatgta gtgttgaaga aacatgaaat tgcccagtat
      361 tcttaaccca actgcacaga acaaaaacct gcaggaaacg aagataaatc atgtcgaaag
      421 ctacatataa ggaacgtgct gctactcatc ctagtcctgt tgctgccaag ctatttaata
      481 tcatgcacga aaagcaaaca aacttgtgtg cttcattgga tgttcgtacc accaaggaat
      541 tactggagtt agttgaagca ttaggtccca aaatttgttt actaaaaaca catgtggata
      601 tcttgactga tttttccatg gagggcacag ttaagccgct aaaggcatta tccgccaagt
      661 acaatttttt actcttcgaa gacagaaaat ttgctgacat tggtaataca gtcaaattgc
      721 agtactctgc gggtgtatac agaatagcag aatgggcaga cattacgaat gcacacggtg
      781 tggtgggccc aggtattgtt agcggtttga agcaggcggc agaagaagta acaaaggaac
      841 ctagaggcct tttgatgtta gcagaattgt catgcaaggg ctccctatct actggagaat
      901 atactaaggg tactgttgac attgcgaaga gcgacaaaga ttttgttatc ggctttattg
      961 ctcaaagaga catgggtgga agagatgaag gttacgattg gttgattatg acacccggtg
     1021 tgggtttaga tgacaaggga gacgcattgg gtcaacagta tagaaccgtg gatgatgtgg
     1081 tctctacagg atctgacatt attattgttg gaagaggact atttgcaaag ggaagggatg
     1141 ctaaggtaga gggtgaacgt tacagaaaag caggctggga agcatatttg agaagatgcg
     1201 gccagcaaaa ctaaaaaact gtattataag taaatgcatg tatactaaac tcacaaatta
     1261 gagcttcaat ttaattatat cagttattac cctgcggtgt gaaataccgc acagatgcgt
     1321 aaggagaaaa taccgcatca ggaaattgta aacgttaata ttttgttaaa attcgcgtta
     1381 aatttttgtt aaatcagctc attttttaac caataggccg aaatcggcaa aatcccttat
     1441 aaatcaaaag aatagaccga gatagggttg agtgttgttc cagtttggaa caagagtcca
     1501 ctattaaaga acgtggactc caacgtcaaa gggcgaaaaa ccgtctatca gggcgatggc
     1561 ccactacgtg aaccatcacc ctaatcaagt tttttggggt cgaggtgccg taaagcacta
     1621 aatcggaacc ctaaagggag cccccgattt agagcttgac ggggaaagcc ggcgaacgtg
     1681 gcgagaaagg aagggaagaa agcgaaagga gcgggcgcta gggcgctggc aagtgtagcg
     1741 gtcacgctgc gcgtaaccac cacacccgcc gcgcttaatg cgccgctaca gggcgcgtcg
     1801 cgccattcgc cattcaggct gcgcaactgt tgggaagggc gatcggtgcg ggcctcttcg
     1861 ctattacgcc agctggcgaa ggggggatgt gctgcaaggc gattaagttg ggtaacgcca
     1921 gggttttccc agtcacgacg ttgtaaaacg acggccagtg aattgtaata cgactcacta
     1981 tagggcgaat tggagctcca ccgcggtggc ggccgctcta gaactagtgg atcctagggt
     2041 taattaacgg tgaacaaaag ctaatctccg aggaagactt gaacggtgaa caaaaattaa
     2101 tctcagaaga agacttgaac ggactcgacg gtgaacaaaa gttgatttct gaagaagatt
     2161 tgaacggtga acaaaagcta atctccgagg aagacttgaa cggtgaacaa aaattaatct
     2221 cagaagaaga cttgaacgga ctcgacggtg aacaaaagtt gatttctgaa gaagatttga
     2281 acggtgaaca aaagctaatc tccgaggaag acttgaacgg tgaacaaaaa ttaatctcag
     2341 aagaagactt gaacggactc gacggtgaac aaaagttgat ttctgaagaa gatttgaacg
     2401 gtgaacaaaa gctaatctcc gaggaagact tgaacggtga acaaaaatta atctcagaag
     2461 aagacttgaa cggactcgac ggtgaacaaa agttgatttc tgaagaagat ttgaacggtg
     2521 aacaaaagct aatctccgag gaagacttga acggtgaaca aaaattaatc aatcactagc
     2581 tagtgaattc gcgccagatc cgaagttcct atactttcta gagaatagga acttcgtgcg
     2641 cttttgaacc acgtaaagtg ctgtaaaaca gctctattca atgaggtaaa tgataaagtt
     2701 tataagagac caaaaaattc aaaaaaaaaa ttgaaaaatt atattggtat ttatatgaaa
     2761 caatagttca gttcaaagcg ctagcttttc tcatctcaac taaccggtat taggtgggta
     2821 tttatttaat atgggaacaa gaaatgtgac actaatgttc atgtttttat tttcattatt
     2881 tattctatat ggtttgggac ttgatgttat gtcttaattc atagagatcc gcttgcctcg
     2941 tccccgccgg gtcacccggc cagcgacatg gaggcccaga ataccctcct tgacagtctt
     3001 gacgtgcgca gctcaggggc atgatgtgac tgtcgcccgt acatttagcc catacatccc
     3061 catgtataat catttgcatc catacatttt gatggccgca cggcgcgaag caaaaattac
     3121 ggctcctcgc tgcagacctg cgagcaggga aacgctcccc tcacagacgc gttgaattgt
     3181 ccccacgccg cgcccctgta gagaaatata aaaggttagg atttgccact gaggttcttc
     3241 tttcatatac ttccttttaa aatcttgcta ggatacagtt ctcacatcac atccgaacat
     3301 aaacaaccgg atcgatccta caattatgtc tactactttg gatgatactg cttatagata
     3361 cagaacttct gttccaggtg atgctgaagc tattgaagct ttggatggtt ctttcactac
     3421 cgatactgtt tttagagtta ctgctactgg tgatggtttc actttgagag aagttccagt
     3481 tgatccacca ttgactaagg ttttcccaga tgatgaatcc gatgatgaat ccgatgctgg
     3541 tgaagatggt gatccagatt ctagaacttt cgttgcttat ggtgatgatg gtgatttggc
     3601 tggtttcgtt gttgtttctt attctggttg gaacagaaga ttgactgttg aagatattga
     3661 agttgctcca gaacatagag gtcatggtgt tggtagagct ttgatgggtt tggctactga
     3721 attcgccaga gaaagaggtg ctggtcattt gtggttggaa gttaccaatg ttaatgctcc
     3781 agctattcat gcttatagaa gaatgggttt cactttgtgt ggtttggata ctgctttata
     3841 cgatggtact gcttccgatg gtgaacaagc tttgtatatg tctatgccat gtccataata
     3901 actcgaccag tactgacaat aaaaagattc ttgttttcaa gaacttgtca tttgtatagt
     3961 ttttttatat tgtagttgtt ctattttaat caaatgttag cgtgatttat attttttttc
     4021 gcctcgacat catctgccca gatgcgaagt taagtgcgca gaaagtaata tcatgcgtca
     4081 atcgtatgtg aatgctggtc gctatactgc tgtcgattcg atactaacgc cgccatccag
     4141 tgtcgatcga cgaagttcct atactttcta gagaatagga acttcgtcct cgaggggggg
     4201 cccggtaccc agcttttgtt ccctttagtg agggttaatt ccgagcttgg cgtaatcatg
     4261 gtcatagctg tttcctgtgt gaaattgtta tccgctcaca attccacaca acataggagc
     4321 cggaagcata aagtgtaaag cctggggtgc ctaatgagtg aggtaactca cattaattgc
     4381 gttgcgctca ctgcccgctt tccagtcggg aaacctgtcg tgccagctgc attaatgaat
     4441 cggccaacgc gcggggagag gcggtttgcg tattgggcgc tcttccgctt cctcgctcac
     4501 tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta tcagctcact caaaggcggt
     4561 aatacggtta tccacagaat caggggataa cgcaggaaag aacatgtgag caaaaggcca
     4621 gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctcggccc
     4681 ccctgacgag catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact
     4741 ataaagatac caggcgttcc cccctggaag ctccctcgtg cgctctcctg ttccgaccct
     4801 gccgcttacc ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcaatg
     4861 ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca
     4921 cgaacccccc gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa
     4981 cccggtaaga cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc
     5041 gaggtatgta ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag
     5101 aaggacagta tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg
     5161 tagctcttga tccggcaaac aaaccaccgc tggtagcggt ggtttttttg tttgcaagca
     5221 gcagattacg cgcagaaaaa aaggatctca agaagatcct ttgatctttt ctacggggtc
     5281 tgacgctcag tggaacgaaa actcacgtta agggattttg gtcatgagat tatcaaaaag
     5341 gatcttcacc tagatccttt taaattaaaa atgaagtttt aaatcaatct aaagtatata
     5401 tgagtaaact tggtctgaca gttaccaatg cttaatcagt gaggcaccta tctcagcgat
     5461 ctgtctattt cgttcatcca tagttgcctg actgcccgtc gtgtagataa ctacgatacg
     5521 ggagggctta ccatctggcc ccagtgctgc aatgataccg cgagacccac gctcaccggc
     5581 tccagattta tcagcaataa accagccagc cggaagggcc gagcgcagaa gtggtcctgc
     5641 aactttatcc gcctccatcc agtctattaa ttgttgccgg gaagctagag taagtagttc
     5701 gccagttaat agtttgcgca acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc
     5761 gtcgtttggt atggcttcat tcagctccgg ttcccaacga tcaaggcgag ttacatgatc
     5821 ccccatgttg tgaaaaaaag cggttagctc cttcggtcct ccgatcgttg tcagaagtaa
     5881 gttggccgca gtgttatcac tcatggttat ggcagcactg cataattctc ttactgtcat
     5941 gccatccgta agatgctttt ctgtgactgg tgagtactca accaagtcat tctgagaata
     6001 gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata cgggataata ccgcgccaca
     6061 tagcagaact ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa aactctcaag
     6121 gatcttaccg ctgttgagat ccagttcgat gtaacccact cgtgcaccca actgatcttc
     6181 agcatctttt actttcacca gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc
     6241 aaaaaaggga ataagggcga cacggaaatg ttgaatactc atactcttcc tttttcaata
     6301 ttattgaagc atttatcagg gttattgtct catgagcgga tacatatttg aatgtattta
     6361 gaaaaataaa caaatagggg ttccgcgcac atttccccga aaagtgccac ctgacgtcta
     6421 agaaaccatt attatcatga cattaaccta taaaaatagg cgtatcacga ggccctttcg
     6481 tc
//
