LOCUS       sequence-399725-        1923 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  Expression of yoTagRFP657-T2A-mtq2 in yeast.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 1923)
  AUTHORS   Botman D, de Groot DH, Wehrens M, van Rossum K, Yska S, van der 
            Kieft B, Albert V, Minghetti N, Goedhart J, Teusink B
  TITLE     Side-by-side systematic characterization of novel FPs in budding 
            yeast
  JOURNAL   bioRxiv 2025.06.27.661975; doi: 
            https://doi.org/10.1101/2025.06.27.661975
REFERENCE   2  (bases 1 to 1923)
  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: "bioRxiv 
            2025.06.27.661975; doi: https://doi.org/10.1101/2025.06.27.661975"
COMMENT     Sequence Label: pDRF1-GW yoTagRFP657-T2A-mtq2
FEATURES             Location/Qualifiers
     source          1..1923
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     protein_bind    81..105
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             817..870
                     /codon_start=1
                     /product="2A peptide from Thosea asigna virus capsid 
                     protein"
                     /label=T2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="EGRGSLLTCGDVEENPGP"
     regulatory      886..895
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(892..894,895..897,898..1611)
                     /codon_start=1
                     /product="enhanced monomeric variant of CFP (Goedhart et 
                     al., 2012)"
                     /label=mTurquoise2
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLSWGVQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYFSDNVYITADKQKNGIK
                     ANFKIRHNIEDGGVQLADHYQQNTPIGDGPVLLPDNHYLSTQSKLSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(937..958)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1198..1217)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1545..1566
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     protein_bind    complement(1621..1645)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     primer_bind     complement(1707..1726)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1708..1726)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
ORIGIN
        1 ttttttctct cttttataca cacattcaaa agaaagaaaa aaaatatacc ccagcctcga
       61 ctagtgaatt cgcggccatc acaagtttgt acaaaaaagc aggctgctag catgagcgaa
      121 ttgatcacag aaaatatgca catgaagttg tatatggagg gaaccgtcaa taaccatcat
      181 gtcaaatgca cttctgaagg tgagggcaaa ccttacgaag ggacccaaac acagagaatt
      241 aaggtagtag aagggggtcc actgccattt gcctttgaca tcctagcaac atccttcatg
      301 tacggttccc acacattcat aaaccacact caaggaatac cagacttttg gaagcagagt
      361 tttccagaag gttttacgtg ggaaagagtt actacttacg aggacggtgg tgtattaaca
      421 gccacacaag atacatcatt acaagatggt tgtctgatct acaacgttaa aatcagaggt
      481 gttaactttc cttcaaatgg gccagttatg caaaagaaaa ctttaggctg ggaggcccat
      541 actgaaatgc tttacccagc agatggcggc ttggaaggca ggacggcttt ggctttaaag
      601 ctagttggtg gaggacattt gatttgtaat ttcaaaacaa cttacagaag caaaaagcct
      661 gctaaaaatc ttaagatgcc aggagtgtac tatgtcgatt acagattgga acgtattaaa
      721 gaggctgata aggaaaccta tgtggaacaa catgaagtgg cagtcgcgag atattgtgat
      781 cttccttcta aactaggtca taaactcaat tccggagagg gcagaggaag tcttctaaca
      841 tgcggtgacg tggaggagaa tcccggccct ggggatccac cggtcgccac catggtgagc
      901 aagggcgagg agctgttcac cggggtggtg cccatcctgg tcgagctgga cggcgacgta
      961 aacggccaca agttcagcgt gtccggcgag ggcgagggcg atgccaccta cggcaagctg
     1021 accctgaagt tcatctgcac caccggcaag ctgcccgtgc cctggcccac cctcgtgacc
     1081 accctgtcct ggggcgtgca gtgcttcgcc cgctaccccg accacatgaa gcagcacgac
     1141 ttcttcaagt ccgccatgcc cgaaggctac gtccaggagc gcaccatctt cttcaaggac
     1201 gacggcaact acaagacccg cgccgaggtg aagttcgagg gcgacaccct ggtgaaccgc
     1261 atcgagctga agggcatcga cttcaaggag gacggcaaca tcctggggca caagctggag
     1321 tacaactact ttagcgacaa cgtctatatc accgccgaca agcagaagaa cggcatcaag
     1381 gccaacttca agatccgcca caacatcgag gacggcggcg tgcagctcgc cgaccactac
     1441 cagcagaaca cccccatcgg cgacggcccc gtgctgctgc ccgacaacca ctacctgagc
     1501 acccagtcca agctgagcaa agaccccaac gagaagcgcg atcacatggt cctgctggag
     1561 ttcgtgaccg ccgccgggat cactctcggc atggacgagc tgtacaagta aagcggccgc
     1621 acccagcttt cttgtacaaa gtggtgtagt taattcatga tggccgctgc aggtcgacct
     1681 cgaggggggg cccggtaccc aattcgccct atagtgagtc gtattacgcg cggatccagc
     1741 tttggacttc ttcgccagag gtttggtcaa gtctccaatc aaggttgtcg gcttgtctac
     1801 cttgccagaa atttacgaaa agatggaaaa gggtcaaatc gttggtagat acgttgttga
     1861 cacttctaaa taagcgaatt tcttatgatt tatgattttt attattaaat aagtttaaaa
     1921 aac
//
