LOCUS       sequence-399726-        1868 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  Expression of mKelly-T2A-mtq2 in yeast.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 1868)
  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 1868)
  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 mKelly-T2A-mtq2
FEATURES             Location/Qualifiers
     source          1..1868
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     protein_bind    89..113
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             762..815
                     /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      831..840
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(837..839,840..842,843..1556)
                     /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(882..903)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1143..1162)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1490..1511
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     protein_bind    complement(1566..1590)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     primer_bind     complement(1652..1671)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1653..1671)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
ORIGIN
        1 aaaccatttt ttttctctct tttatacaca cattcaaaag aaagaaaaaa aatatacccc
       61 agcctcgact agtgaattcg cggccatcac aagtttgtac aaaaaagcag gctgctagca
      121 tgcgtatgaa actgtacatg gaaggtaccg ttggcaacca tcacttcaaa tgcaccgcgg
      181 aaggtgaggg taaaccgtac gaaggcaccc agacccagcg tattaaagtt gttgaaggtg
      241 gtccgctgcc gttcgcgttc gacatcctgg cgacctgctt catgtatggt tctaaaactt
      301 ttatcaacta ccctcagggt atcccggact tcttcaaaca gtctttccca gaaggtttca
      361 cctgggaacg tgttaccacc tacgaagacg gcggtgttct gaccgttacc caggacacct
      421 ctctgcagga cggttacctg atctacaacg ttaaactgcg tggtgttaac ttcccgtcta
      481 acggccctgt tatgcagaaa aagactctgg gttgggaacc gaccaccgag tacctggtcc
      541 cggttgacgg tggcctggag ggtacatgtg atatggcgct gcgtctggtt ggtggcggtc
      601 acctgaagtg caatctgaaa accacgtatc gttccaaaaa accagcgaaa aacctgaaga
      661 tgccgggtga gcaccaggtt gaccgtaagc tggagcgtat caaagaggcg gacaacgaaa
      721 cctacgttga acagcacgaa gttgcggttg cgagatccgg agagggcaga ggaagtcttc
      781 taacatgcgg tgacgtggag gagaatcccg gccctgggga tccaccggtc gccaccatgg
      841 tgagcaaggg cgaggagctg ttcaccgggg tggtgcccat cctggtcgag ctggacggcg
      901 acgtaaacgg ccacaagttc agcgtgtccg gcgagggcga gggcgatgcc acctacggca
      961 agctgaccct gaagttcatc tgcaccaccg gcaagctgcc cgtgccctgg cccaccctcg
     1021 tgaccaccct gtcctggggc gtgcagtgct tcgcccgcta ccccgaccac atgaagcagc
     1081 acgacttctt caagtccgcc atgcccgaag gctacgtcca ggagcgcacc atcttcttca
     1141 aggacgacgg caactacaag acccgcgccg aggtgaagtt cgagggcgac accctggtga
     1201 accgcatcga gctgaagggc atcgacttca aggaggacgg caacatcctg gggcacaagc
     1261 tggagtacaa ctactttagc gacaacgtct atatcaccgc cgacaagcag aagaacggca
     1321 tcaaggccaa cttcaagatc cgccacaaca tcgaggacgg cggcgtgcag ctcgccgacc
     1381 actaccagca gaacaccccc atcggcgacg gccccgtgct gctgcccgac aaccactacc
     1441 tgagcaccca gtccaagctg agcaaagacc ccaacgagaa gcgcgatcac atggtcctgc
     1501 tggagttcgt gaccgccgcc gggatcactc tcggcatgga cgagctgtac aagtaaagcg
     1561 gccgcaccca gctttcttgt acaaagtggt gtagttaatt catgatggcc gctgcaggtc
     1621 gacctcgagg gggggcccgg tacccaattc gccctatagt gagtcgtatt acgcgcggat
     1681 ccagctttgg acttcttcgc cagaggtttg gtcaagtctc caatcaaggt tgtcggcttg
     1741 tctaccttgc cagaaattta cgaaaagatg gaaaagggtc aaatcgttgg tagatacgtt
     1801 gttgacactt ctaaataagc gaatttctta tgatttatga tttttattat aaataagtta
     1861 aaaaaaat
//
