LOCUS       sequence-127170-        4719 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  N-terminal tagging with mCitrine using URA3 as a selection marker in
            S. cerevisiae..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4719)
  AUTHORS   Higuchi-Sanabria R, Garcia EJ, Tomoiaga D, Munteanu EL, Feinstein P,
            Pon LA
  TITLE     Characterization of Fluorescent Proteins for Three- and Four-Color 
            Live-Cell Imaging in S. cerevisiae.
  JOURNAL   PLoS One. 2016 Jan 4;11(1):e0146120. doi: 
            10.1371/journal.pone.0146120. eCollection 2016.
  PUBMED    26727004
REFERENCE   2  (bases 1 to 4719)
  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: "PLoS One.";
            date: "2016-01-4"; volume: "11(1):e0146120. doi"; pages: " 
            10.1371/journal.pone.0146120. eCollection 2016"
COMMENT     Sequence Label: POM42-mCitrine
FEATURES             Location/Qualifiers
     source          1..4719
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    53..86
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     protein_bind    1538..1571
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     CDS             1610..2320
                     /codon_start=1
                     /product="yeast-enhanced green fluorescent protein"
                     /label=yeGFP
                     /note="codons optimized for translation in S. cerevisiae 
                     and C. albicans"
                     /translation="SKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLKF
                     ICTTGKLPVPWPTLVTTLGYGLMCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDDGN
                     YKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKVN
                     FKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSYQSRLSKDPNEKRDHMVLLEF
                     VTAAGITHGIDELYK"
     primer_bind     complement(2356..2375)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2357..2375)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2462..2479)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2633..3221)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2713..2732)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3392..4183,4184..4252))
                     /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     4015..4034
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4253..4357)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     4425..4443
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(4481..4503)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     4603..4622
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        join(4703..4719,1..2)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     join(4703..4719,1)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
ORIGIN
        1 gaacgcggcc gccagctgaa gcttcgtacg ctgcaggtcg acaaccctta atataacttc
       61 gtataatgta tgctatacga agttattagg tctagagatc ccaatacaac agatcacgtg
      121 atcttttgta agatgaagtt gaagtgagtg ttgcaccgtg ccaatgcagg tggctattag
      181 attaaatatg tgatttgttc tattaagttt cctgtataat taatggggag cgctgattct
      241 cttttggtac gcttcccatc cagcatttct gtatctttca ccttcaacct taggatctct
      301 acccttggcg aaaagtcctc tgccaacaat gatgatatct gatccaccac ttacaacttc
      361 gtcgacggtt ctgtactgct gacccaatgc atcgcctttg tcgtctaaac ctacacctgg
      421 ggtcatgatt agccaatcaa acccttcttc tcttcctccc atatcgttct gagcaatgaa
      481 cccaataacg aaatctttat cactctttgc aatatcaacg gtacccttag tatattcacc
      541 gtgtgctaga gaacccttgg aagacaattc agcaagcatc aataatcccc ttggttcttt
      601 ggtgacctct tgcgcacctt gtttcaagcc agcaacaata ccagcaccag taaccccgtg
      661 ggcgttggtg atatcagacc attctgcgat acggtaaacg cccgatgtat attgtaattt
      721 gactgtgtta ccgatatcgg cgaattttct gtcctcaaat atcaagaact tgtatttctc
      781 tgccaatgct ttcaatggaa cgacagtacc ctcataactg aaatcatcca agatatcaac
      841 gtgtgttttc aaaaggcaaa tgtatggacc caacgtttca acaagtttca atagctcatc
      901 agtcgaacga acgtcaagag aagcacacaa attggtcttc ttttcatcca ttaaacgtaa
      961 aagtttcgat gcaaccggac ttgcatgagt ctcagctcta ctggtatatg attttgtgga
     1021 catggtgcaa ctaattgacg ggagtgtatt gacgctggcg tactggcttt cacaaaatgg
     1081 cccaatcaca accacatctt agatagttga aatgacttta gataacatca attgagatga
     1141 gcttaatcat gtcaaagcta aaagtgtcac catgaacgac aattcttaag caaatcacgt
     1201 gatatagatc cacgaataac caccatttga tgctcgaggc aagtaatgtg tgtaaaaaaa
     1261 tgcgttacca ccatccaatg cagaccgatc ttctacccag aatcacatat atttatgtac
     1321 cgagtacctt ttttctatct tccaattgct tctcccatat gattgtctcc gtaagctcga
     1381 aatttctaag ttggatttta atcttcacgc aggatgacag ttcgatgagc ttctgaggag
     1441 tgtttagaac ataatcattt atccatggtc tatctcttct tgtcgctttt tctcctcgat
     1501 agaacctaaa taaaacgagc tctcgagaac ccttaatata acttcgtata atgtatgcta
     1561 tacgaagtta ttaggtgata tcatgcgtac gctgcaggtc gacggatcct ctaaaggtga
     1621 agaattattc actggtgttg tcccaatttt ggttgaatta gatggtgatg ttaatggtca
     1681 caaattttct gtctccggtg aaggtgaagg tgatgctact tacggtaaat tgaccttaaa
     1741 atttatttgt actactggta aattgccagt tccatggcca accttagtca ctactttagg
     1801 ttatggtttg atgtgttttg ctagataccc agatcatatg aaacaacatg actttttcaa
     1861 gtctgccatg ccagaaggtt atgttcaaga aagaactatt tttttcaaag atgacggtaa
     1921 ctacaagacc agagctgaag tcaagtttga aggtgatacc ttagttaata gaatcgaatt
     1981 aaaaggtatt gattttaaag aagatggtaa cattttaggt cacaaattgg aatacaacta
     2041 taactctcac aatgtttaca tcatggctga caaacaaaag aatggtatca aagttaactt
     2101 caaaattaga cacaacattg aagatggttc tgttcaatta gctgaccatt atcaacaaaa
     2161 tactccaatt ggtgatggtc cagtcttgtt accagacaac cattacttat cctatcaatc
     2221 tagattatcc aaagatccaa acgaaaagag agaccacatg gtcttgttag aatttgttac
     2281 tgctgctggt attacccatg gtattgatga attgtacaaa actagtggcc tatgcggccg
     2341 cggatctgcc ggtctcccta tagtgagtcg tattaatttc gataagccag gttaacctgc
     2401 attaatgaat cggccaacgc gcggggagag gcggtttgcg tattgggcgc tcttccgctt
     2461 cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta tcagctcact
     2521 caaaggcggt aatacggtta tccacagaat caggggataa cgcaggaaag aacatgtgag
     2581 caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg tttttccata
     2641 ggctccgccc ccctgacgag catcacaaaa atcgacgctc aagtcagagg tggcgaaacc
     2701 cgacaggact ataaagatac caggcgtttc cccctggaag ctccctcgtg cgctctcctg
     2761 ttccgaccct gccgcttacc ggatacctgt ccgcctttct cccttcggga agcgtggcgc
     2821 tttctcaatg ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg
     2881 gctgtgtgca cgaacccccc gttcagcccg accgctgcgc cttatccggt aactatcgtc
     2941 ttgagtccaa cccggtaaga cacgacttat cgccactggc agcagccact ggtaacagga
     3001 ttagcagagc gaggtatgta ggcggtgcta cagagttctt gaagtggtgg cctaactacg
     3061 gctacactag aaggacagta tttggtatct gcgctctgct gaagccagtt accttcggaa
     3121 aaagagttgg tagctcttga tccggcaaac aaaccaccgc tggtagcggt ggtttttttg
     3181 tttgcaagca gcagattacg cgcagaaaaa aaggatctca agaagatcct ttgatctttt
     3241 ctacggggtc tgacgctcag tggaacgaaa actcacgtta agggattttg gtcatgagat
     3301 tatcaaaaag gatcttcacc tagatccttt taaattaaaa atgaagtttt aaatcaatct
     3361 aaagtatata tgagtaaact tggtctgaca gttaccaatg cttaatcagt gaggcaccta
     3421 tctcagcgat ctgtctattt cgttcatcca tagttgcctg actccccgtc gtgtagataa
     3481 ctacgatacg ggagggctta ccatctggcc ccagtgctgc aatgataccg cgagacccac
     3541 gctcaccggc tccagattta tcagcaataa accagccagc cggaagggcc gagcgcagaa
     3601 gtggtcctgc aactttatcc gcctccatcc agtctattaa ttgttgccgg gaagctagag
     3661 taagtagttc gccagttaat agtttgcgca acgttgttgc cattgctaca ggcatcgtgg
     3721 tgtcacgctc gtcgtttggt atggcttcat tcagctccgg ttcccaacga tcaaggcgag
     3781 ttacatgatc ccccatgttg tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg
     3841 tcagaagtaa gttggccgca gtgttatcac tcatggttat ggcagcactg cataattctc
     3901 ttactgtcat gccatccgta agatgctttt ctgtgactgg tgagtactca accaagtcat
     3961 tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata cgggataata
     4021 ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa
     4081 aactctcaag gatcttaccg ctgttgagat ccagttcgat gtaacccact cgtgcaccca
     4141 actgatcttc agcatctttt actttcacca gcgtttctgg gtgagcaaaa acaggaaggc
     4201 aaaatgccgc aaaaaaggga ataagggcga cacggaaatg ttgaatactc atactcttcc
     4261 tttttcaata ttattgaagc atttatcagg gttattgtct catgagcgga tacatatttg
     4321 aatgtattta gaaaaataaa caaatagggg ttccgcgcac atttccccga aaagtgccac
     4381 ctgacgtcta agaaaccatt attatcatga cattaaccta taaaaatagg cgtatcacga
     4441 ggccctttcg tctcgcgcgt ttcggtgatg acggtgaaaa cctctgacac atgcagctcc
     4501 cggagacggt cacagcttgt ctgtaagcgg atgccgggag cagacaagcc cgtcagggcg
     4561 cgtcagcggg tgttggcggg tgtcggggct ggcttaacta tgcggcatca gagcagattg
     4621 tactgagagt gcaccatatg gacatattgt cgttagaacg cggctacaat taatacataa
     4681 ccttatgtat catacacata cgatttaggt gacactata
//
