LOCUS       sequence-106683-        5234 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Recipient replicative vector to construct CFP tagged fusions.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5234)
  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 5234)
  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: pYC61
FEATURES             Location/Qualifiers
     source          1..5234
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     151..170
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        341..747
                     /gene="S. cerevisiae TEF1"
                     /label=TEF1 promoter
                     /note="promoter for EF-1-alpha"
     CDS             761..1333
                     /codon_start=1
                     /gene="Streptomyces noursei nat1"
                     /product="nourseothricin acetyltransferase"
                     /label=NrsR
                     /note="confers resistance to nourseothricin"
                     /translation="MGTTLDDTAYRYRTSVPGDAEAIEALDGSFTTDTVFRVTATGDGF
                     TLREVPVDPPLTKVFPDDESDDESDDGEDGDPDSRTFVAYGDDGDLAGFVVVSYSGWNR
                     RLTVEDIEVAPEHRGHGVGRALMGLATEFARELGAGHLWLEVTNVNAPAIHAYRRMGFT
                     LCGLDTALYDGTASDGEQALYMSMPCP"
     terminator      1340..1587
                     /gene="S. cerevisiae CYC1"
                     /label=CYC1 terminator
                     /note="transcription terminator for CYC1"
     primer_bind     complement(1356..1374)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     primer_bind     1608..1624
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     primer_bind     1609..1625
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     CDS             1666..2379
                     /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             1666..2379
                     /codon_start=1
                     /product="green fluorescent protein"
                     /label=GFP
                     /translation="MSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLTWGVQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNAISDNVYITADKQKNGIKA
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELYK"
     primer_bind     complement(1696..1724)
                     /label=GFP-R
                     /note="GFP, reverse primer. Does NOT anneal to EGFP"
     primer_bind     2319..2339
                     /label=GFP-F
                     /note="GFP, forward primer. Does NOT anneal to EGFP"
     primer_bind     complement(2384..2400)
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2384..2400)
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     primer_bind     complement(2437..2457)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(2437..2455)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(2476..2492)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2476..2492)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2489..2511)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2500..2516
                     /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(2524..2530,2531..2548,2549..2554))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2569..2590
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2707..2724)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2878..3466)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2958..2977)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3637..4428,4429..4497))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     4260..4279
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4498..4569)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     5207..5225
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
ORIGIN
        1 tcgcgcgttt cggtgatgac ggtgaaaacc tctgacacat gcacctcccg gagacggtca
       61 cagcttgtct gtaagcggat gccgggagca gacaagcccg tcagggcgcg tcagcgggtg
      121 ttggcgggtg tcggggctgg cttaactatg cggcatcaga gcagattgta ctgagagtgc
      181 accataccac agcttttcaa ttcaattcat catttttttt ttattctttt ttttgatttc
      241 ggtttctttg aaattttttt gattcggtaa tctccgaaca gaaggaagaa cgaaggaagg
      301 agcacagact tagattggta tatatacgca taccataaag cacagctcca tagcttcaaa
      361 atgtttctac tcctttttta ctcttccaga ttttctcgga ctccgcgcat cgccgtacca
      421 cttcaaaaca cccaagcaca gcatactaaa tttcccctct ttcttcctct agggtgtcgt
      481 taattacccg tactaaaggt ttggaaaaga aaaaagagac cgcctcgttt ctttttcttc
      541 gtcgaaaaag gcaataaaaa tttttatcac gtttcttttt cttgaaaatt ttttttttga
      601 tttttttctc tttcgatgac ctcccattga tatttaagtt aataaacggt cttcaatttc
      661 tcaagtttca gtttcatttt tcttgttcta ttacaacttt ttttacttct tgctcattag
      721 aaagaaagca tagcaatcta atctaagttt tggatctaaa atgggcacca ctcttgacga
      781 cacggcttac cgataccgca ccagtgtccc gggggacgcc gaggccatcg aggcactgga
      841 tgggtccttc accaccgaca ccgtcttccg cgtcaccgcc accggggacg gcttcaccct
      901 gcgggaggtg ccggtggacc cgcccctgac caaggtgttc cccgacgacg aatcggacga
      961 cgaatcggac gacggggagg acggcgaccc tgactcccgg acgttcgtcg cgtacgggga
     1021 cgacggcgac ctggcgggct tcgtggtcgt ctcgtactcc ggctggaacc gccggctgac
     1081 cgtcgaggac atcgaggtcg ccccggagca ccgggggcac ggggtcgggc gcgcgttgat
     1141 ggggctcgcg acggagttcg cccgcgagct gggcgccggg cacctctggc tggaggtcac
     1201 caacgtcaac gcaccggcga tccacgcgta ccggcggatg gggttcaccc tctgcggcct
     1261 ggacaccgcc ctgtacgacg gcaccgcctc ggacggcgag caggcgctct acatgagcat
     1321 gccctgcccc taagtcgagt catgtaatta gttatgtcac gcttacattc acgccctccc
     1381 cccacatccg ctctaaccga aaaggaagga gttagacaac ctgaagtcta ggtccctatt
     1441 tattttttta tagttatgtt agtattaaga acgttattta tatttcaaat ttttcttttt
     1501 tttctgtaca gacgcgtgta cgcatgtaac attatactga aaaccttgct tgagaaggtt
     1561 ttgggacgct cgaaggcttt aatttgcggc cggtaccggg ccccccctcg aggtcgacgg
     1621 tatcgataag cttgatatcg aattcctgca gcccggggga tcctaatgag taaaggagaa
     1681 gaacttttca ctggagttgt cccaattctt gttgaattag atggtgatgt taatgggcac
     1741 aaattttctg tcagtggaga gggtgaaggt gatgcaacat acggaaaact tacccttaaa
     1801 tttatttgca ctactggaaa actacctgtt ccatggccaa cacttgtcac tactttgact
     1861 tggggtgttc aatgctttgc tagataccca gatcatatga aacagcatga ctttttcaag
     1921 agtgccatgc ccgaaggtta tgtacaggaa agaactatat ttttcaaaga tgacgggaac
     1981 tacaagacac gtgctgaagt caagtttgaa ggtgataccc ttgttaatag aatcgagtta
     2041 aaaggtattg attttaaaga agatggaaac attcttggac acaaattgga atacaacgct
     2101 atttcagata atgtatacat cactgcagac aaacaaaaga atggaatcaa agctaatttc
     2161 aaaattagac acaacattga agatggaagc gttcaactag cagaccatta tcaacaaaat
     2221 actccaattg gcgatggccc tgtcctttta ccagacaacc attacctgtc cacacaatct
     2281 gccctttcga aagatcccaa cgaaaagaga gaccacatgg tccttcttga gtttgtaaca
     2341 gctgctggga ttacactagg catggatgaa ctatacaaat agagatccac tagttctaga
     2401 gcggccgcca ccgcggtgga gctccagctt ttgttccctt tagtgagggt taattgcgcg
     2461 cttggcgtaa tcatggtcat agctgtttcc tgtgtgaaat tgttatccgc tcacaattcc
     2521 acacaacata cgagccggaa gcataaagtg taaagcctgg ggtgcctaat gagtgagcta
     2581 actcacatta attgcgttgc gctcactgcc cgctttccag tcgggaaacc tgtcgtgcca
     2641 gctgcattaa tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg ggcgctcttc
     2701 cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc
     2761 tcactcaaag gcggtaatac ggttatccac agaatcaggg gataacgcag gaaagaacat
     2821 gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt
     2881 ccataggctc cgcccccctg acgagcatca caaaaatcga cgctcaagtc agaggtggcg
     2941 aaacccgaca ggactataaa gataccaggc gtttccccct ggaagctccc tcgtgcgctc
     3001 tcctgttccg accctgccgc ttaccggata cctgtccgcc tttctccctt cgggaagcgt
     3061 ggcgctttct catagctcac gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa
     3121 gctgggctgt gtgcacgaac cccccgttca gcccgaccgc tgcgccttat ccggtaacta
     3181 tcgtcttgag tccaacccgg taagacacga cttatcgcca ctggcagcag ccactggtaa
     3241 caggattagc agagcgaggt atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa
     3301 ctacggctac actagaagga cagtatttgg tatctgcgct ctgctgaagc cagttacctt
     3361 cggaaaaaga gttggtagct cttgatccgg caaacaaacc accgctggta gcggtggttt
     3421 ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag atcctttgat
     3481 cttttctacg gggtctgacg ctcagtggaa cgaaaactca cgttaaggga ttttggtcat
     3541 gagattatca aaaaggatct tcacctagat ccttttaaat taaaaatgaa gttttaaatc
     3601 aatctaaagt atatatgagt aaacttggtc tgacagttac caatgcttaa tcagtgaggc
     3661 acctatctca gcgatctgtc tatttcgttc atccatagtt gcctgactcc ccgtcgtgta
     3721 gataactacg atacgggagg gcttaccatc tggccccagt gctgcaatga taccgcgaga
     3781 cccacgctca ccggctccag atttatcagc aataaaccag ccagccggaa gggccgagcg
     3841 cagaagtggt cctgcaactt tatccgcctc catccagtct attaattgtt gccgggaagc
     3901 tagagtaagt agttcgccag ttaatagttt gcgcaacgtt gttgccattg ctacaggcat
     3961 cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc tccggttccc aacgatcaag
     4021 gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat
     4081 cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg gttatggcag cactgcataa
     4141 ttctcttact gtcatgccat ccgtaagatg cttttctgtg actggtgagt actcaaccaa
     4201 gtcattctga gaatagtgta tgcggcgacc gagttgctct tgcccggcgt caatacggga
     4261 taataccgcg ccacatagca gaactttaaa agtgctcatc attggaaaac gttcttcggg
     4321 gcgaaaactc tcaaggatct taccgctgtt gagatccagt tcgatgtaac ccactcgtgc
     4381 acccaactga tcttcagcat cttttacttt caccagcgtt tctgggtgag caaaaacagg
     4441 aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa tactcatact
     4501 cttccttttt caatattatt gaagcattta tcagggttat tgtctcatga gcggatacat
     4561 atttgaatgt ggggatctct aatgatgcaa tttttgtttt gtacactaag attctatttc
     4621 ggaactctta cccatgtcat tatttacttt attttcatta ttgtaatttt ttaagctttt
     4681 attatgaatt atttaaatac aattcttttt gatcatgtga cagacatcac tgaaagatgt
     4741 gctctgcgtt attagtcaga ttgaaatata cattttcatg tagatatgat gttcttactt
     4801 taaatgttgt tccatttgct agcttaaata gacgatatta taaatttaag tattcactaa
     4861 aaatctgacc cattaatgtt agaattagta ttattttttg tgatattaca tttttatatt
     4921 ttggatttca accaatgctc tgcgatatga ttcaacagaa taatatgtac aagctacttt
     4981 aattcatata tactaataga ttcacagatc cagtacgact gcaagaattt acgttttgtt
     5041 tgaaatggta gtagctacac taaattaaca atgatcgatt aatccacatc cacagtgtgt
     5101 aatcatccta atgaaaacaa gaataactac aaaaatgaat ataaactttt tgcatgttag
     5161 tctaacctgc taagaaacca ttattatcat gacattaacc tataaaaata ggcgtatcac
     5221 gaggcccttt cgtc
//
