LOCUS       sequence-127185-        5556 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  N-terminal tagging with mCitrine using LEU2 as a selection marker in
            S. cerevisiae..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5556)
  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 5556)
  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: POM43-mCitrine
FEATURES             Location/Qualifiers
     source          1..5556
                     /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    2375..2408
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     CDS             2447..3157
                     /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(3193..3212)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(3194..3212)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(3299..3316)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3470..4058)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3550..3569)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4229..5020,5021..5089))
                     /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     4852..4871
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5090..5194)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     5262..5280
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(5318..5340)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     5440..5459
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        join(5540..5556,1..2)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     join(5540..5556,1)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
ORIGIN
        1 gaacgcggcc gccagctgaa gcttcgtacg ctgcaggtcg acaaccctta atataacttc
       61 gtataatgta tgctatacga agttattagg tctagagatc cgcaggctaa ccggaacctg
      121 tattatttag tttatgctac gttaaataaa gacctttcgt tcacataact gaatgtgtaa
      181 tggccttgag atttcaagca taccaagttg gtggagacgg ggtcgttaca aaagactctt
      241 taagccaaga tttccttgac agccttggcg atagcatcgc caacctcagt ggtagagtta
      301 gaaccaccaa ggtcaccggt tctgacacct gcatccaaga catttctaac agcttcttca
      361 agagccctac cttcttcaac caaatccaag gataacttca acatcatagc tgcagataag
      421 atggtagcaa ttgggttaac cttgtttgct ggtaaatctg gggcagaacc atgacatggt
      481 tcgtacaaac cgaatgcctt gttagtgtca ggtagggaag ctagagatgc agaaggtaat
      541 aaacccaaag aacctggaat aacagaggct tcatcggaga taatatcacc aaacatgttg
      601 ttggtaataa caacaccgtt tagcttagtt ggtgatttaa ccaaaatcat agcagcagag
      661 tcgatcaatt ggtgctgaac agttaattgt gggaactcag tcttgatggt ttcttcaaca
      721 gtctttctcc acaatctgga agaggcaagc acgttagcct tgtcaagaga ccagattggt
      781 aatggtgggt tttgttgcaa tgccaagaaa gcagccattc ttgtaattct ttgaacttca
      841 ggaacactgt atttctcaga gtcccaagca actccgtcac cttcatcttc ttttctttca
      901 ccaaagtaga taccaccaac caattctcta acgacgacga aatcggtacc ctttgcatat
      961 tcaggcttca aaggagaaag atctagtaaa gaatcagaag caaagttaca tggtctcaag
     1021 ttggcgtata gacccaattc ctttctgatc ttcaatagac cttgttctgg tctaactgcg
     1081 cccgtacccc attttggacc accaacagca cctagtaaga cagcatcggc tttcttagag
     1141 gcttctagag cttcatctgg taaaggagtg ccagtggcat cgatggcagc acccccgatc
     1201 aagtgatgtt ggaaattgaa cttaatttct ggacggactt cagcaatggc attcaagacc
     1261 ttaatagctt cgtcagtaac ttctttaccg acgtgatcac ccggtaggac aacgatattc
     1321 ttagacatgg tataatctgt gtagtgtggg atacttttta cttcttcaaa taggtatcaa
     1381 caataaaaat aaatcgaagc aaatgtagga atgcgttaaa gcagatgtac ttacttagag
     1441 tacatagata tatttatata attcaatata taaaagtata tgaagcatct gatgttgaac
     1501 ctgtcatgac tctaaactgg aatgggtagt tatgcttctg aaggttttcc gttagaaaat
     1561 gagtgcaggg atcaggcccg gaaccggttt tagcctgaaa ggaaaagaaa aaaatgcggt
     1621 gattactagc acgtgactgc gctgaattgg aatccaccac aagtacagcg aatggcaaga
     1681 gggaagggga atatattgtg cctggcggta gtctgctata tcaaatgtta aacataacta
     1741 aatgcagtag tgaatagagt aatgtatttc tacatatgtg ggtagggcca atgggagccc
     1801 gatgtctatt gtacagatat tttcctttta tagttaatca attcaacttg gaaatcattg
     1861 agtcgatatc tacttggata actggagccg tggcagcgtt tgaaatttga cgagtcttct
     1921 tgttttgttc gcgcattttc agtttattca aacgggagat ggcaccgaat ccactgactt
     1981 ctctgttcaa tttgaatggt aataggaaga aagtacattt cgagaaattt tcgcccaacg
     2041 tcgaaatctc atggatgata tcctcgatac aaatgacacc ttcatcacct aatttctctt
     2101 caacgatatt gttgtcattc aagattatct cttttggttc tttatcttct ggcctttgcc
     2161 acattattct agatctcttt tggattaaag aacggataga tgcaagtgaa ggttgtccga
     2221 taacaatata tggtgataca agtttcaaca atgggtacac gtccttggta agtttgatga
     2281 aaacaccagt gttcaactca acaagtctca aaagagataa cactttgtaa gcctttgctg
     2341 ggatcttcac agcgagctct cgagaaccct taatataact tcgtataatg tatgctatac
     2401 gaagttatta ggtgatatca tgcgtacgct gcaggtcgac ggatcctcta aaggtgaaga
     2461 attattcact ggtgttgtcc caattttggt tgaattagat ggtgatgtta atggtcacaa
     2521 attttctgtc tccggtgaag gtgaaggtga tgctacttac ggtaaattga ccttaaaatt
     2581 tatttgtact actggtaaat tgccagttcc atggccaacc ttagtcacta ctttaggtta
     2641 tggtttgatg tgttttgcta gatacccaga tcatatgaaa caacatgact ttttcaagtc
     2701 tgccatgcca gaaggttatg ttcaagaaag aactattttt ttcaaagatg acggtaacta
     2761 caagaccaga gctgaagtca agtttgaagg tgatacctta gttaatagaa tcgaattaaa
     2821 aggtattgat tttaaagaag atggtaacat tttaggtcac aaattggaat acaactataa
     2881 ctctcacaat gtttacatca tggctgacaa acaaaagaat ggtatcaaag ttaacttcaa
     2941 aattagacac aacattgaag atggttctgt tcaattagct gaccattatc aacaaaatac
     3001 tccaattggt gatggtccag tcttgttacc agacaaccat tacttatcct atcaatctag
     3061 attatccaaa gatccaaacg aaaagagaga ccacatggtc ttgttagaat ttgttactgc
     3121 tgctggtatt acccatggta ttgatgaatt gtacaaaact agtggcctat gcggccgcgg
     3181 atctgccggt ctccctatag tgagtcgtat taatttcgat aagccaggtt aacctgcatt
     3241 aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat tgggcgctct tccgcttcct
     3301 cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca gctcactcaa
     3361 aggcggtaat acggttatcc acagaatcag gggataacgc aggaaagaac atgtgagcaa
     3421 aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc
     3481 tccgcccccc tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga
     3541 caggactata aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc
     3601 cgaccctgcc gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt
     3661 ctcaatgctc acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct
     3721 gtgtgcacga accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg
     3781 agtccaaccc ggtaagacac gacttatcgc cactggcagc agccactggt aacaggatta
     3841 gcagagcgag gtatgtaggc ggtgctacag agttcttgaa gtggtggcct aactacggct
     3901 acactagaag gacagtattt ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa
     3961 gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt
     4021 gcaagcagca gattacgcgc agaaaaaaag gatctcaaga agatcctttg atcttttcta
     4081 cggggtctga cgctcagtgg aacgaaaact cacgttaagg gattttggtc atgagattat
     4141 caaaaaggat cttcacctag atccttttaa attaaaaatg aagttttaaa tcaatctaaa
     4201 gtatatatga gtaaacttgg tctgacagtt accaatgctt aatcagtgag gcacctatct
     4261 cagcgatctg tctatttcgt tcatccatag ttgcctgact ccccgtcgtg tagataacta
     4321 cgatacggga gggcttacca tctggcccca gtgctgcaat gataccgcga gacccacgct
     4381 caccggctcc agatttatca gcaataaacc agccagccgg aagggccgag cgcagaagtg
     4441 gtcctgcaac tttatccgcc tccatccagt ctattaattg ttgccgggaa gctagagtaa
     4501 gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt
     4561 cacgctcgtc gtttggtatg gcttcattca gctccggttc ccaacgatca aggcgagtta
     4621 catgatcccc catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca
     4681 gaagtaagtt ggccgcagtg ttatcactca tggttatggc agcactgcat aattctctta
     4741 ctgtcatgcc atccgtaaga tgcttttctg tgactggtga gtactcaacc aagtcattct
     4801 gagaatagtg tatgcggcga ccgagttgct cttgcccggc gtcaatacgg gataataccg
     4861 cgccacatag cagaacttta aaagtgctca tcattggaaa acgttcttcg gggcgaaaac
     4921 tctcaaggat cttaccgctg ttgagatcca gttcgatgta acccactcgt gcacccaact
     4981 gatcttcagc atcttttact ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa
     5041 atgccgcaaa aaagggaata agggcgacac ggaaatgttg aatactcata ctcttccttt
     5101 ttcaatatta ttgaagcatt tatcagggtt attgtctcat gagcggatac atatttgaat
     5161 gtatttagaa aaataaacaa ataggggttc cgcgcacatt tccccgaaaa gtgccacctg
     5221 acgtctaaga aaccattatt atcatgacat taacctataa aaataggcgt atcacgaggc
     5281 cctttcgtct cgcgcgtttc ggtgatgacg gtgaaaacct ctgacacatg cagctcccgg
     5341 agacggtcac agcttgtctg taagcggatg ccgggagcag acaagcccgt cagggcgcgt
     5401 cagcgggtgt tggcgggtgt cggggctggc ttaactatgc ggcatcagag cagattgtac
     5461 tgagagtgca ccatatggac atattgtcgt tagaacgcgg ctacaattaa tacataacct
     5521 tatgtatcat acacatacga tttaggtgac actata
//
