LOCUS       sequence-407763-        2172 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  Expression of miFP663-T2A-mtq2 in yeast.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 2172)
  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 2172)
  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 miFP663-T2A-mtq2
FEATURES             Location/Qualifiers
     source          1..2172
                     /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             1081..1134
                     /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      1150..1159
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(1156..1158,1159..1161,1162..1875)
                     /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(1201..1222)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1462..1481)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1809..1830
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     protein_bind    complement(1885..1909)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     primer_bind     complement(1971..1990)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1972..1990)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
ORIGIN
        1 ttttttctct cttttataca cacattcaaa agaaagaaaa aaaatatacc ccagcctcga
       61 ctagtgaatt cgcggccatc acaagtttgt acaaaaaagc aggctgctag catggtgagc
      121 accgctaccg atcccctcga cctggatgtg tgtgccagag agcccatcca catcccagga
      181 ctgatccagc cttacggcgc gctgctcgtg gtcgaccctg ccgacggaag aatcgtgcag
      241 gcctctacaa cagcagccga tcttctggga gtgcctatgg ctgcgctgct gggcatgcct
      301 tacacccagg tgctgacact gcctgaggca cagcccttcg ccgtggacga tcagccacag
      361 cacctgatgc atgccgaggt gcggttccct cggagagcca cacctcctgc ctctgcttat
      421 gtggctgcct ggcatcttta tccccagcag tggctggtgg aaattgaacc cggggacgcc
      481 agactgctgg acaccgtgct gagagaagcc ctgcctctcc tgcggagcgt ggagagagat
      541 cccggaattg ccgaagccgc tgtgcgggtg gccaagggcc tgagatctct gatcggcttc
      601 gaccgcgtga tgatctacag attcgacgag gaatggaacg gcgacatcat tgccgaggct
      661 cggaagcctg agctggaagc ctatctggga ctccactacc ctgccagcct gacccctgct
      721 ctggccagag ccctgtacct gcggaataga gtgcggcaga tcgccgacgt gggctaccag
      781 cctagcccta tccagcctac catgcaccct cagctgggca cccctgtgga tctgtccgat
      841 gtgtccctga gaagcatgtc tccatgccac ctggaattcc tggccaacat cggtgtgagc
      901 gctaccctgg tcgccagcat cgtcgtgaac gatgccctgt ggggactgat cgtctgccac
      961 cactacagcc caagattcac caaccacgcc ctgcgcgcct tgacagacgc cgtggctaaa
     1021 acactggctg ccagaattgg cgccctgcag gctgaggcca gagccaggct ggagtccgga
     1081 gagggcagag gaagtcttct aacatgcggt gacgtggagg agaatcccgg ccctggggat
     1141 ccaccggtcg ccaccatggt gagcaagggc gaggagctgt tcaccggggt ggtgcccatc
     1201 ctggtcgagc tggacggcga cgtaaacggc cacaagttca gcgtgtccgg cgagggcgag
     1261 ggcgatgcca cctacggcaa gctgaccctg aagttcatct gcaccaccgg caagctgccc
     1321 gtgccctggc ccaccctcgt gaccaccctg tcctggggcg tgcagtgctt cgcccgctac
     1381 cccgaccaca tgaagcagca cgacttcttc aagtccgcca tgcccgaagg ctacgtccag
     1441 gagcgcacca tcttcttcaa ggacgacggc aactacaaga cccgcgccga ggtgaagttc
     1501 gagggcgaca ccctggtgaa ccgcatcgag ctgaagggca tcgacttcaa ggaggacggc
     1561 aacatcctgg ggcacaagct ggagtacaac tactttagcg acaacgtcta tatcaccgcc
     1621 gacaagcaga agaacggcat caaggccaac ttcaagatcc gccacaacat cgaggacggc
     1681 ggcgtgcagc tcgccgacca ctaccagcag aacaccccca tcggcgacgg ccccgtgctg
     1741 ctgcccgaca accactacct gagcacccag tccaagctga gcaaagaccc caacgagaag
     1801 cgcgatcaca tggtcctgct ggagttcgtg accgccgccg ggatcactct cggcatggac
     1861 gagctgtaca agtaaagcgg ccgcacccag ctttcttgta caaagtggtg tagttaattc
     1921 atgatggccg ctgcaggtcg acctcgaggg ggggcccggt acccaattcg ccctatagtg
     1981 agtcgtatta cgcgcggatc cagctttgga cttcttcgcc agaggtttgg tcaagtctcc
     2041 aatcaaggtt gtcggcttgt ctaccttgcc agaaatttac gaaaagatgg aaaagggtca
     2101 aatcgttggt agatacgttg ttgacacttc taaataagcg aatttcttat gatttatgat
     2161 ttttattatt aa
//
