LOCUS       sequence-110406-        6017 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Recipient integrative vector to construct FLAG tagged fusions.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6017)
  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 6017)
  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: pYC46
FEATURES             Location/Qualifiers
     source          1..6017
                     /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             2043..2066
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             2073..2096
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             2103..2126
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     protein_bind    complement(2138..2171)
                     /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        2501..2844
                     /label=TEF promoter
                     /note="Ashbya gossypii TEF promoter"
     terminator      3448..3645
                     /label=TEF terminator
                     /note="Ashbya gossypii TEF terminator"
     protein_bind    complement(3688..3721)
                     /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(3757..3775)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(3796..3812)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3796..3812)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3809..3831)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3820..3836
                     /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(3844..3850,3851..3868,3869..3874))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     primer_bind     complement(4027..4044)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4198..4786)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(join(4957..5748,5749..5817))
                     /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     5580..5599
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5818..5922)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     5990..6008
                     /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 atcctaggag
     2041 cagactacaa agatgacgat gacaaaggag ccgattataa ggacgatgac gataaagggg
     2101 ctgactacaa agatgacgat gacaagtgat aagatccgaa gttcctatac tttctagaga
     2161 ataggaactt cgtgcgcttt tgaaccacgt aaagtgctgt aaaccagctc tattcaatga
     2221 ggtaaatgat aaagtttata agagaccaaa aaattcaaaa aaaaaattga aaaattatat
     2281 tggtatttat atgaaacaat agttcagttc aaagcgctag cttttctcat ctcaactaac
     2341 cggtattagg tgggtattta tttaatatgg gaacaagaaa tgtgacatta atgttcatgt
     2401 ttttattttc attatttatt ctatatggtt tgggacttga tgttatgtct taattcatag
     2461 agatccgctt gcctcgtccc cgccgggtca cccggccagc gacatggagg cccagaatac
     2521 cctccttgac agtcttgacg tgcgcagctc aggggcatga tgtgactgtc gcccgtacat
     2581 ttagcccata catccccatg tataatcatt tgcatccata cattttgatg gccgcacggc
     2641 gcgaagcaaa aattacggct cctcgctgca gacctgcgag cagggaaacg ctcccctcac
     2701 agacgcgttg aattgtcccc acgccgcgcc cctgtagaga aatataaaag gttaggattt
     2761 gccactgagg ttcttctttc atatacttcc ttttaaaatc ttgctaggat acagttctca
     2821 catcacatcc gaacataaac aaccggatcg atcctacaat tatgtctact actttggatg
     2881 atactgctta tagatacaga acttctgttc caggtgatgc tgaagctatt gaagctttgg
     2941 atggttcttt cactaccgat actgttttta gagttactgc tactggtgat ggtttcactt
     3001 tgagagaagt tccagttgat ccaccattga ctaaggtttt cccagatgat gaatccgatg
     3061 atgaatccga tgctggtgaa gatggtgatc cagattctag aactttcgtt gcttatggtg
     3121 atgatggtga tttggctggt ttcgttgttg tttcttattc tggttggaac agaagattga
     3181 ctgttgaaga tattgaagtt gctccagaac atagaggtca tggtgttggt agagctttga
     3241 tgggtttggc tactgaattc gccagagaaa gaggtgctgg tcatttgtgg ttggaagtta
     3301 ccaatgttaa tgctccagct attcatgctt atagaagaat gggtttcact ttgtgtggtt
     3361 tggatactgc tttatacgat ggtactgctt ccgatggtga acaagctttg tatatgtcta
     3421 tgccatgtcc ataataactc gaccagtact gacaataaaa agattcttgt tttcaagaac
     3481 ttgtcatttg tatagttttt ttatattgta gttgttctat tttaatcaaa tgttagcgtg
     3541 atttatattt tttttcgcct cgacatcatc tgcccagatg cgaagttaag tgcgcagaaa
     3601 gtaatatcat gcgtcaatcg tatgtgaatg ctggtcgcta tactgctgtc gattcgatac
     3661 taacgccgcc atccagtgtc gatcgacgaa gttcctatac tttctagaga ataggaactt
     3721 cgtcctcgag ggggggcccg gtaccagctt ttgttccctt tagtgagggt taattccgag
     3781 cttggcgtaa tcatggtcat agctgtttcc tgtgtgaaat tgttatccgc tcacaattcc
     3841 acacaacata ggagccggaa gcataaagtg taaagcctgg ggtgcctaat gagtgaggta
     3901 actcacatta attgcgttgc gctcactgcc cgctttccag tcgggaaacc tgtcgtgcca
     3961 gctgcattaa tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg ggcgctcttc
     4021 cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc
     4081 tcactcaaag gcggtaatac ggttatccac agaatcaggg gataacgcag gaaagaacat
     4141 gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt
     4201 ccataggctc ggcccccctg acgagcatca caaaaatcga cgctcaagtc agaggtggcg
     4261 aaacccgaca ggactataaa gataccaggc gttcccccct ggaagctccc tcgtgcgctc
     4321 tcctgttccg accctgccgc ttaccggata cctgtccgcc tttctccctt cgggaagcgt
     4381 ggcgctttct caatgctcac gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa
     4441 gctgggctgt gtgcacgaac cccccgttca gcccgaccgc tgcgccttat ccggtaacta
     4501 tcgtcttgag tccaacccgg taagacacga cttatcgcca ctggcagcag ccactggtaa
     4561 caggattagc agagcgaggt atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa
     4621 ctacggctac actagaagga cagtatttgg tatctgcgct ctgctgaagc cagttacctt
     4681 cggaaaaaga gttggtagct cttgatccgg caaacaaacc accgctggta gcggtggttt
     4741 ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag atcctttgat
     4801 cttttctacg gggtctgacg ctcagtggaa cgaaaactca cgttaaggga ttttggtcat
     4861 gagattatca aaaaggatct tcacctagat ccttttaaat taaaaatgaa gttttaaatc
     4921 aatctaaagt atatatgagt aaacttggtc tgacagttac caatgcttaa tcagtgaggc
     4981 acctatctca gcgatctgtc tatttcgttc atccatagtt gcctgactgc ccgtcgtgta
     5041 gataactacg atacgggagg gcttaccatc tggccccagt gctgcaatga taccgcgaga
     5101 cccacgctca ccggctccag atttatcagc aataaaccag ccagccggaa gggccgagcg
     5161 cagaagtggt cctgcaactt tatccgcctc catccagtct attaattgtt gccgggaagc
     5221 tagagtaagt agttcgccag ttaatagttt gcgcaacgtt gttgccattg ctacaggcat
     5281 cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc tccggttccc aacgatcaag
     5341 gcgagttaca tgatccccca tgttgtgaaa aaaagcggtt agctccttcg gtcctccgat
     5401 cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg gttatggcag cactgcataa
     5461 ttctcttact gtcatgccat ccgtaagatg cttttctgtg actggtgagt actcaaccaa
     5521 gtcattctga gaatagtgta tgcggcgacc gagttgctct tgcccggcgt caatacggga
     5581 taataccgcg ccacatagca gaactttaaa agtgctcatc attggaaaac gttcttcggg
     5641 gcgaaaactc tcaaggatct taccgctgtt gagatccagt tcgatgtaac ccactcgtgc
     5701 acccaactga tcttcagcat cttttacttt caccagcgtt tctgggtgag caaaaacagg
     5761 aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa tactcatact
     5821 cttccttttt caatattatt gaagcattta tcagggttat tgtctcatga gcggatacat
     5881 atttgaatgt atttagaaaa ataaacaaat aggggttccg cgcacatttc cccgaaaagt
     5941 gccacctgac gtctaagaaa ccattattat catgacatta acctataaaa ataggcgtat
     6001 cacgaggccc tttcgtc
//
