LOCUS       40924_47938        6640 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Cloning vector pYC54, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6640)
  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  JOURNAL   Unpublished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 6640)
  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 6640)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 6640)
  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            "Unpublished"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..6640
                     /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             2034..2750
                     /codon_start=1
                     /label=Venus
                     /note="yellow fluorescent protein (YFP) with fast and
                     efficient maturation (Nagai et al., 2002)"
                     /translation="LMSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KLICTTGKLPVPWPTLVTTLGYGLMCFARYPDHMKRHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYITADKQKNGIK
                     ANFKIRHNIEDGGVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALSKDPNEKRDHMVLL
                     EFVTAAGITHGMDELYK"
     misc_feature    2760..2793
                     /label=FRT sequence
                     /note="FRT sequence"
     protein_bind    complement(2760..2793)
                     /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           2794..3082
                     /label=3'UTR of CTA1 orf in Candida glabrata BG2
                     /note="3'UTR of CTA1 orf in Candida glabrata BG2"
     promoter        3123..3466
                     /label=TEF promoter
                     /note="Ashbya gossypii TEF promoter"
     CDS             3490..4053
                     /codon_start=1
                     /label=NrsR
                     /note="nourseothricin acetyltransferase"
                     /translation="TTLDDTAYRYRTSVPGDAEAIEALDGSFTTDTVFRVTATGDGFTL
                     REVPVDPPLTKVFPDDESDDESDAGEDGDPDSRTFVAYGDDGDLAGFVVVSYSGWNRRL
                     TVEDIEVAPEHRGHGVGRALMGLATEFARERGAGHLWLEVTNVNAPAIHAYRRMGFTLC
                     GLDTALYDGTASDGEQALYMSMPCP"
     terminator      4070..4267
                     /label=TEF terminator
                     /note="Ashbya gossypii TEF terminator"
     protein_bind    complement(4310..4343)
                     /label=FRT (minimal)
                     /note="supports FLP-mediated excision but not integration
                     (Turan and Bode, 2011)"
     promoter        complement(4380..4398)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(4419..4435)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    complement(4443..4459)
                     /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(4467..4497)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    complement(4512..4533)
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     rep_origin      complement(4821..5409)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(5583..6440)
                     /codon_start=1
                     /label=AmpR
                     /note="beta-lactamase"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFFHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     promoter        complement(6441..6545)
                     /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 atcctaatga
     2041 gtaaaggaga agaacttttc actggagttg tcccaattct tgttgaatta gatggtgatg
     2101 ttaatgggca caaattttct gtcagtggag agggtgaagg tgatgcaaca tacggaaaac
     2161 ttacccttaa attgatttgc actactggaa aactacctgt tccatggcca acacttgtca
     2221 ctactttggg ttatggtcta atgtgctttg ctagataccc agatcatatg aaacggcatg
     2281 actttttcaa gagtgccatg cccgaaggtt atgtacagga aagaactata tttttcaaag
     2341 atgacgggaa ctacaagaca cgtgctgaag tcaagtttga aggtgatacc cttgttaata
     2401 gaatcgagtt aaaaggtatt gattttaaag aagatggaaa cattcttgga cacaaattgg
     2461 aatacaacta taactcacac aatgtataca tcactgcaga caaacaaaag aatggaatca
     2521 aagctaactt caaaattaga cacaacattg aagatggagg tgttcaacta gcagaccatt
     2581 atcaacaaaa tactccaatt ggcgatggcc ctgtcctttt accagacaac cattacctgt
     2641 cctatcaatc tgccctttcg aaagatccca acgaaaagag agaccacatg gtccttcttg
     2701 agtttgtaac agctgctggg attacacatg gcatggatga actatacaaa tagagatccg
     2761 aagttcctat actttctaga gaataggaac ttcgtgcgct tttgaaccac gtaaagtgct
     2821 gtaaaacagc tctattcaat gaggtaaatg ataaagttta taagagacca aaaaattcaa
     2881 aaaaaaaatt gaaaaattat attggtattt atatgaaaca atagttcagt tcaaagcgct
     2941 agcttttctc atctcaacta accggtatta ggtgggtatt tatttaatat gggaacaaga
     3001 aatgtgacac taatgttcat gtttttattt tcattattta ttctatatgg tttgggactt
     3061 gatgttatgt cttaattcat agagatccgc ttgcctcgtc cccgccgggt cacccggcca
     3121 gcgacatgga ggcccagaat accctccttg acagtcttga cgtgcgcagc tcaggggcat
     3181 gatgtgactg tcgcccgtac atttagccca tacatcccca tgtataatca tttgcatcca
     3241 tacattttga tggccgcacg gcgcgaagca aaaattacgg ctcctcgctg cagacctgcg
     3301 agcagggaaa cgctcccctc acagacgcgt tgaattgtcc ccacgccgcg cccctgtaga
     3361 gaaatataaa aggttaggat ttgccactga ggttcttctt tcatatactt ccttttaaaa
     3421 tcttgctagg atacagttct cacatcacat ccgaacataa acaaccggat cgatcctaca
     3481 attatgtcta ctactttgga tgatactgct tatagataca gaacttctgt tccaggtgat
     3541 gctgaagcta ttgaagcttt ggatggttct ttcactaccg atactgtttt tagagttact
     3601 gctactggtg atggtttcac tttgagagaa gttccagttg atccaccatt gactaaggtt
     3661 ttcccagatg atgaatccga tgatgaatcc gatgctggtg aagatggtga tccagattct
     3721 agaactttcg ttgcttatgg tgatgatggt gatttggctg gtttcgttgt tgtttcttat
     3781 tctggttgga acagaagatt gactgttgaa gatattgaag ttgctccaga acatagaggt
     3841 catggtgttg gtagagcttt gatgggtttg gctactgaat tcgccagaga aagaggtgct
     3901 ggtcatttgt ggttggaagt taccaatgtt aatgctccag ctattcatgc ttatagaaga
     3961 atgggtttca ctttgtgtgg tttggatact gctttatacg atggtactgc ttccgatggt
     4021 gaacaagctt tgtatatgtc tatgccatgt ccataataac tcgaccagta ctgacaataa
     4081 aaagattctt gttttcaaga acttgtcatt tgtatagttt ttttatattg tagttgttct
     4141 attttaatca aatgttagcg tgatttatat tttttttcgc ctcgacatca tctgcccaga
     4201 tgcgaagtta agtgcgcaga aagtaatatc atgcgtcaat cgtatgtgaa tgctggtcgc
     4261 tatactgctg tcgattcgat actaacgccg ccatccagtg tcgatcgacg aagttcctat
     4321 actttctaga gaataggaac ttcgtcctcg agggggggcc cggtacccag cttttgttcc
     4381 ctttagtgag ggttaattcc gagcttggcg taatcatggt catagctgtt tcctgtgtga
     4441 aattgttatc cgctcacaat tccacacaac ataggagccg gaagcataaa gtgtaaagcc
     4501 tggggtgcct aatgagtgag gtaactcaca ttaattgcgt tgcgctcact gcccgctttc
     4561 cagtcgggaa acctgtcgtg ccagctgcat taatgaatcg gccaacgcgc ggggagaggc
     4621 ggtttgcgta ttgggcgctc ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt
     4681 cggctgcggc gagcggtatc agctcactca aaggcggtaa tacggttatc cacagaatca
     4741 ggggataacg caggaaagaa catgtgagca aaaggccagc aaaaggccag gaaccgtaaa
     4801 aaggccgcgt tgctggcgtt tttccatagg ctcggccccc ctgacgagca tcacaaaaat
     4861 cgacgctcaa gtcagaggtg gcgaaacccg acaggactat aaagatacca ggcgttcccc
     4921 cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc
     4981 gcctttctcc cttcgggaag cgtggcgctt tctcaatgct cacgctgtag gtatctcagt
     5041 tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt tcagcccgac
     5101 cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggtaagaca cgacttatcg
     5161 ccactggcag cagccactgg taacaggatt agcagagcga ggtatgtagg cggtgctaca
     5221 gagttcttga agtggtggcc taactacggc tacactagaa ggacagtatt tggtatctgc
     5281 gctctgctga agccagttac cttcggaaaa agagttggta gctcttgatc cggcaaacaa
     5341 accaccgctg gtagcggtgg tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa
     5401 ggatctcaag aagatccttt gatcttttct acggggtctg acgctcagtg gaacgaaaac
     5461 tcacgttaag ggattttggt catgagatta tcaaaaagga tcttcaccta gatcctttta
     5521 aattaaaaat gaagttttaa atcaatctaa agtatatatg agtaaacttg gtctgacagt
     5581 taccaatgct taatcagtga ggcacctatc tcagcgatct gtctatttcg ttcatccata
     5641 gttgcctgac tgcccgtcgt gtagataact acgatacggg agggcttacc atctggcccc
     5701 agtgctgcaa tgataccgcg agacccacgc tcaccggctc cagatttatc agcaataaac
     5761 cagccagccg gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag
     5821 tctattaatt gttgccggga agctagagta agtagttcgc cagttaatag tttgcgcaac
     5881 gttgttgcca ttgctacagg catcgtggtg tcacgctcgt cgtttggtat ggcttcattc
     5941 agctccggtt cccaacgatc aaggcgagtt acatgatccc ccatgttgtg aaaaaaagcg
     6001 gttagctcct tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt gttatcactc
     6061 atggttatgg cagcactgca taattctctt actgtcatgc catccgtaag atgcttttct
     6121 gtgactggtg agtactcaac caagtcattc tgagaatagt gtatgcggcg accgagttgc
     6181 tcttgcccgg cgtcaatacg ggataatacc gcgccacata gcagaacttt aaaagtgctc
     6241 atcattggaa aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc
     6301 agttcgatgt aacccactcg tgcacccaac tgatcttcag catcttttac tttcaccagc
     6361 gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca
     6421 cggaaatgtt gaatactcat actcttcctt tttcaatatt attgaagcat ttatcagggt
     6481 tattgtctca tgagcggata catatttgaa tgtatttaga aaaataaaca aataggggtt
     6541 ccgcgcacat ttccccgaaa agtgccacct gacgtctaag aaaccattat tatcatgaca
     6601 ttaacctata aaaataggcg tatcacgagg ccctttcgtc
//
