LOCUS       sequence-110444-        6639 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Recipient integrative vector to construct CFP tagged fusions.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6639)
  AUTHORS   Yanez-Carrillo P, Orta-Zavalza E, Gutierrez-Escobedo G, 
            Patron-Soberano A, De Las Penas A, Castano I
  TITLE     Expression vectors for C-terminal fusions with fluorescent proteins 
            and epitope tags in Candida glabrata.
  JOURNAL   Fungal Genet Biol. 2015 May 15. pii: S1087-1845(15)00087-0. doi: 
            10.1016/j.fgb.2015.04.020.
  PUBMED    25986172
REFERENCE   2  (bases 1 to 6639)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Fungal 
            Genet Biol. 2015 May 15. pii: S1087-1845(15)00087-0. doi: 
            10.1016/j.fgb.2015.04.020."
COMMENT     Sequence Label: pYC52
FEATURES             Location/Qualifiers
     source          1..6639
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(29..51)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     151..170
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        190..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     complement(1480..1501)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     1692..1711
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1928..1950
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     1942..1959
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     1943..1959
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     1966..1985
                     /label=T7
                     /note="T7 promoter, forward primer"
     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"
     primer_bind     2017..2033
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     CDS             2037..2750
                     /codon_start=1
                     /product="S65T variant of Aequorea victoria green 
                     fluorescent protein (Heim et al., 1995)"
                     /label=GFP (S65T)
                     /note="excitable with blue light"
                     /translation="MSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLTWGVQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNAISDNVYITADKQKNGIKA
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELYK"
     CDS             2037..2750
                     /codon_start=1
                     /product="green fluorescent protein"
                     /label=GFP
                     /translation="MSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLTWGVQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNAISDNVYITADKQKNGIKA
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELYK"
     primer_bind     complement(2067..2095)
                     /label=GFP-R
                     /note="GFP, reverse primer. Does NOT anneal to EGFP"
     primer_bind     2690..2710
                     /label=GFP-F
                     /note="GFP, forward primer. Does NOT anneal to EGFP"
     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)"
     promoter        3123..3466
                     /label=TEF promoter
                     /note="Ashbya gossypii TEF promoter"
     terminator      4070..4267
                     /label=TEF terminator
                     /note="Ashbya gossypii TEF terminator"
     protein_bind    complement(4310..4343)
                     /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(4379..4397)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(4418..4434)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4418..4434)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4431..4453)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4442..4458
                     /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(join(4466..4472,4473..4490,4491..4496))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     primer_bind     complement(4649..4666)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4820..5408)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(join(5579..6370,6371..6439))
                     /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"
     primer_bind     6202..6221
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6440..6544)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     6612..6630
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
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 atttatttgc actactggaa aactacctgt tccatggcca acacttgtca
     2221 ctactttgac ttggggtgtt caatgctttg ctagataccc agatcatatg aaacagcatg
     2281 actttttcaa gagtgccatg cccgaaggtt atgtacagga aagaactata tttttcaaag
     2341 atgacgggaa ctacaagaca cgtgctgaag tcaagtttga aggtgatacc cttgttaata
     2401 gaatcgagtt aaaaggtatt gattttaaag aagatggaaa cattcttgga cacaaattgg
     2461 aatacaacgc tatttcagat aatgtataca tcactgcaga caaacaaaag aatggaatca
     2521 aagctaattt caaaattaga cacaacattg aagatggaag cgttcaacta gcagaccatt
     2581 atcaacaaaa tactccaatt ggcgatggcc ctgtcctttt accagacaac cattacctgt
     2641 ccacacaatc tgccctttcg aaagatccca acgaaaagag agaccacatg gtccttcttg
     2701 agtttgtaac agctgctggg attacactag 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 cggtaccagc ttttgttccc
     4381 tttagtgagg gttaattccg agcttggcgt aatcatggtc atagctgttt cctgtgtgaa
     4441 attgttatcc gctcacaatt ccacacaaca taggagccgg aagcataaag tgtaaagcct
     4501 ggggtgccta atgagtgagg taactcacat taattgcgtt gcgctcactg cccgctttcc
     4561 agtcgggaaa cctgtcgtgc cagctgcatt aatgaatcgg ccaacgcgcg gggagaggcg
     4621 gtttgcgtat tgggcgctct tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc
     4681 ggctgcggcg agcggtatca gctcactcaa aggcggtaat acggttatcc acagaatcag
     4741 gggataacgc aggaaagaac atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa
     4801 aggccgcgtt gctggcgttt ttccataggc tcggcccccc tgacgagcat cacaaaaatc
     4861 gacgctcaag tcagaggtgg cgaaacccga caggactata aagataccag gcgttccccc
     4921 ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga tacctgtccg
     4981 cctttctccc ttcgggaagc gtggcgcttt ctcaatgctc acgctgtagg tatctcagtt
     5041 cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga accccccgtt cagcccgacc
     5101 gctgcgcctt atccggtaac tatcgtcttg agtccaaccc ggtaagacac gacttatcgc
     5161 cactggcagc agccactggt aacaggatta gcagagcgag gtatgtaggc ggtgctacag
     5221 agttcttgaa gtggtggcct aactacggct acactagaag gacagtattt ggtatctgcg
     5281 ctctgctgaa gccagttacc ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa
     5341 ccaccgctgg tagcggtggt ttttttgttt gcaagcagca gattacgcgc agaaaaaaag
     5401 gatctcaaga agatcctttg atcttttcta cggggtctga cgctcagtgg aacgaaaact
     5461 cacgttaagg gattttggtc atgagattat caaaaaggat cttcacctag atccttttaa
     5521 attaaaaatg aagttttaaa tcaatctaaa gtatatatga gtaaacttgg tctgacagtt
     5581 accaatgctt aatcagtgag gcacctatct cagcgatctg tctatttcgt tcatccatag
     5641 ttgcctgact gcccgtcgtg tagataacta cgatacggga gggcttacca tctggcccca
     5701 gtgctgcaat gataccgcga gacccacgct caccggctcc agatttatca gcaataaacc
     5761 agccagccgg aagggccgag cgcagaagtg gtcctgcaac tttatccgcc tccatccagt
     5821 ctattaattg ttgccgggaa gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg
     5881 ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg gcttcattca
     5941 gctccggttc ccaacgatca aggcgagtta catgatcccc catgttgtga aaaaaagcgg
     6001 ttagctcctt cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg ttatcactca
     6061 tggttatggc agcactgcat aattctctta ctgtcatgcc atccgtaaga tgcttttctg
     6121 tgactggtga gtactcaacc aagtcattct gagaatagtg tatgcggcga ccgagttgct
     6181 cttgcccggc gtcaatacgg gataataccg cgccacatag cagaacttta aaagtgctca
     6241 tcattggaaa acgttcttcg gggcgaaaac tctcaaggat cttaccgctg ttgagatcca
     6301 gttcgatgta acccactcgt gcacccaact gatcttcagc atcttttact ttcaccagcg
     6361 tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata agggcgacac
     6421 ggaaatgttg aatactcata ctcttccttt ttcaatatta ttgaagcatt tatcagggtt
     6481 attgtctcat gagcggatac atatttgaat gtatttagaa aaataaacaa ataggggttc
     6541 cgcgcacatt tccccgaaaa gtgccacctg acgtctaaga aaccattatt atcatgacat
     6601 taacctataa aaataggcgt atcacgaggc cctttcgtc
//
