LOCUS       sequence-111239-        6024 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Recipient integrative vector to construct HA tagged fusions.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6024)
  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 6024)
  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: pYC48
FEATURES             Location/Qualifiers
     source          1..6024
                     /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..2126
                     /codon_start=1
                     /product="three tandem HA epitope tags"
                     /label=3xHA
                     /translation="YPYDVPDYAGYPYDVPDYAGSYPYDVPDYA"
     protein_bind    complement(2145..2178)
                     /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        2508..2851
                     /label=TEF promoter
                     /note="Ashbya gossypii TEF promoter"
     terminator      3455..3652
                     /label=TEF terminator
                     /note="Ashbya gossypii TEF terminator"
     protein_bind    complement(3695..3728)
                     /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(3764..3782)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(3803..3819)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3803..3819)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3816..3838)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3827..3843
                     /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(3851..3857,3858..3875,3876..3881))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     primer_bind     complement(4034..4051)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4205..4793)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(join(4964..5755,5756..5824))
                     /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     5587..5606
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5825..5929)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     5997..6015
                     /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 atcctatacc
     2041 catacgatgt tcctgactat gcgggctatc cgtatgacgt cccggactat gcaggctcct
     2101 atccatatga cgttccagat tacgctgctc agtgctgaag atccgaagtt cctatacttt
     2161 ctagagaata ggaacttcgt gcgcttttga accacgtaaa gtgctgtaaa ccagctctat
     2221 tcaatgaggt aaatgataaa gtttataaga gaccaaaaaa ttcaaaaaaa aaattgaaaa
     2281 attatattgg tatttatatg aaacaatagt tcagttcaaa gcgctagctt ttctcatctc
     2341 aactaaccgg tattaggtgg gtatttattt aatatgggaa caagaaatgt gacattaatg
     2401 ttcatgtttt tattttcatt atttattcta tatggtttgg gacttgatgt tatgtcttaa
     2461 ttcatagaga tccgcttgcc tcgtccccgc cgggtcaccc ggccagcgac atggaggccc
     2521 agaataccct ccttgacagt cttgacgtgc gcagctcagg ggcatgatgt gactgtcgcc
     2581 cgtacattta gcccatacat ccccatgtat aatcatttgc atccatacat tttgatggcc
     2641 gcacggcgcg aagcaaaaat tacggctcct cgctgcagac ctgcgagcag ggaaacgctc
     2701 ccctcacaga cgcgttgaat tgtccccacg ccgcgcccct gtagagaaat ataaaaggtt
     2761 aggatttgcc actgaggttc ttctttcata tacttccttt taaaatcttg ctaggataca
     2821 gttctcacat cacatccgaa cataaacaac cggatcgatc ctacaattat gtctactact
     2881 ttggatgata ctgcttatag atacagaact tctgttccag gtgatgctga agctattgaa
     2941 gctttggatg gttctttcac taccgatact gtttttagag ttactgctac tggtgatggt
     3001 ttcactttga gagaagttcc agttgatcca ccattgacta aggttttccc agatgatgaa
     3061 tccgatgatg aatccgatgc tggtgaagat ggtgatccag attctagaac tttcgttgct
     3121 tatggtgatg atggtgattt ggctggtttc gttgttgttt cttattctgg ttggaacaga
     3181 agattgactg ttgaagatat tgaagttgct ccagaacata gaggtcatgg tgttggtaga
     3241 gctttgatgg gtttggctac tgaattcgcc agagaaagag gtgctggtca tttgtggttg
     3301 gaagttacca atgttaatgc tccagctatt catgcttata gaagaatggg tttcactttg
     3361 tgtggtttgg atactgcttt atacgatggt actgcttccg atggtgaaca agctttgtat
     3421 atgtctatgc catgtccata ataactcgac cagtactgac aataaaaaga ttcttgtttt
     3481 caagaacttg tcatttgtat agttttttta tattgtagtt gttctatttt aatcaaatgt
     3541 tagcgtgatt tatatttttt ttcgcctcga catcatctgc ccagatgcga agttaagtgc
     3601 gcagaaagta atatcatgcg tcaatcgtat gtgaatgctg gtcgctatac tgctgtcgat
     3661 tcgatactaa cgccgccatc cagtgtcgat cgacgaagtt cctatacttt ctagagaata
     3721 ggaacttcgt cctcgagggg gggcccggta ccagcttttg ttccctttag tgagggttaa
     3781 ttccgagctt ggcgtaatca tggtcatagc tgtttcctgt gtgaaattgt tatccgctca
     3841 caattccaca caacatagga gccggaagca taaagtgtaa agcctggggt gcctaatgag
     3901 tgaggtaact cacattaatt gcgttgcgct cactgcccgc tttccagtcg ggaaacctgt
     3961 cgtgccagct gcattaatga atcggccaac gcgcggggag aggcggtttg cgtattgggc
     4021 gctcttccgc ttcctcgctc actgactcgc tgcgctcggt cgttcggctg cggcgagcgg
     4081 tatcagctca ctcaaaggcg gtaatacggt tatccacaga atcaggggat aacgcaggaa
     4141 agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg
     4201 cgtttttcca taggctcggc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga
     4261 ggtggcgaaa cccgacagga ctataaagat accaggcgtt cccccctgga agctccctcg
     4321 tgcgctctcc tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg
     4381 gaagcgtggc gctttctcaa tgctcacgct gtaggtatct cagttcggtg taggtcgttc
     4441 gctccaagct gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg
     4501 gtaactatcg tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca
     4561 ctggtaacag gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt
     4621 ggcctaacta cggctacact agaaggacag tatttggtat ctgcgctctg ctgaagccag
     4681 ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg
     4741 gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct caagaagatc
     4801 ctttgatctt ttctacgggg tctgacgctc agtggaacga aaactcacgt taagggattt
     4861 tggtcatgag attatcaaaa aggatcttca cctagatcct tttaaattaa aaatgaagtt
     4921 ttaaatcaat ctaaagtata tatgagtaaa cttggtctga cagttaccaa tgcttaatca
     4981 gtgaggcacc tatctcagcg atctgtctat ttcgttcatc catagttgcc tgactgcccg
     5041 tcgtgtagat aactacgata cgggagggct taccatctgg ccccagtgct gcaatgatac
     5101 cgcgagaccc acgctcaccg gctccagatt tatcagcaat aaaccagcca gccggaaggg
     5161 ccgagcgcag aagtggtcct gcaactttat ccgcctccat ccagtctatt aattgttgcc
     5221 gggaagctag agtaagtagt tcgccagtta atagtttgcg caacgttgtt gccattgcta
     5281 caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc ggttcccaac
     5341 gatcaaggcg agttacatga tcccccatgt tgtgaaaaaa agcggttagc tccttcggtc
     5401 ctccgatcgt tgtcagaagt aagttggccg cagtgttatc actcatggtt atggcagcac
     5461 tgcataattc tcttactgtc atgccatccg taagatgctt ttctgtgact ggtgagtact
     5521 caaccaagtc attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa
     5581 tacgggataa taccgcgcca catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt
     5641 cttcggggcg aaaactctca aggatcttac cgctgttgag atccagttcg atgtaaccca
     5701 ctcgtgcacc caactgatct tcagcatctt ttactttcac cagcgtttct gggtgagcaa
     5761 aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa tgttgaatac
     5821 tcatactctt cctttttcaa tattattgaa gcatttatca gggttattgt ctcatgagcg
     5881 gatacatatt tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc acatttcccc
     5941 gaaaagtgcc acctgacgtc taagaaacca ttattatcat gacattaacc tataaaaata
     6001 ggcgtatcac gaggcccttt cgtc
//
