LOCUS       sequence-399723-        2169 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  Expression of miRFP2-T2A-mtq2 in yeast.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 2169)
  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 2169)
  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 miRFP2-T2A-mtq2
FEATURES             Location/Qualifiers
     source          1..2169
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     protein_bind    86..110
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             1062..1115
                     /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      1131..1140
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(1137..1139,1140..1142,1143..1856)
                     /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(1182..1203)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1443..1462)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1790..1811
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     protein_bind    complement(1866..1890)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     primer_bind     complement(1952..1971)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1953..1971)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
ORIGIN
        1 aaccattttt tctctctttt atacacacat tcaaaagaaa gaaaaaaaat ataccccagc
       61 ctcgactagt gaattcgcgg ccatcacaag tttgtacaaa aaagcaggct gctagcatgg
      121 tcgcaggaca cgcaagtgga agccccgact ttggaaccgc cagtccgagc gactgtgagc
      181 acgaagaaat ccatctggcc ggatccatcc agccacacgg aacactgctg gtggtctccg
      241 agcctgacca tagaataatt caggcatctg ccaacgccgc tgaattcctg aatctgggaa
      301 gtgtgctggg aatccccctg gctgagatcg acggggatct actgatcaag attctgcccc
      361 acctggacct taccgcagaa ggaatgccag tggccgtcag gtgccgcatt ggaaacccct
      421 ccacagagta cgacggactg atgcatcgac cacctgaagg aggactgatc attgagctgg
      481 aacgggctgg gccatccatt gatctgagcg gaacactggc tcccgcactg gagcgaattc
      541 gaactgcagg atccctgagg gctctgtgcg acgatactgt actgctgttc cagcagtgta
      601 ccggctacga ccgagtgatg gtctatcggt tcgatgaacg ggggcacgga gaggtgtatt
      661 cagaaattca tgtgactgga ctggagagct actttggcaa caggtatcct agctccctcg
      721 tgccacagat ggcccggcgg ctgtacgtga gacagagggt gcgcgtcctg gtagatgtca
      781 cctatcagcc tgtgccactg gagccaagac tgagtcccct gactggcagg gacctggata
      841 tgtcagggtg ttttctgcgc tctatgagtc ctactcacct gcagttcctg aagaacatgg
      901 gcgtgcgggc aacactggtg gtcagcctgg tggtcggagg caaactgtgg gggctggtga
      961 tctgccacca ttatctgcca cgcttcatcc attttgagct gcgagcaatt tgtgtgctgc
     1021 tggccgaagc tatcgcaacc cgcattacag ccctgtccgg agagggcaga ggaagtcttc
     1081 taacatgcgg tgacgtggag gagaatcccg gccctgggga tccaccggtc gccaccatgg
     1141 tgagcaaggg cgaggagctg ttcaccgggg tggtgcccat cctggtcgag ctggacggcg
     1201 acgtaaacgg ccacaagttc agcgtgtccg gcgagggcga gggcgatgcc acctacggca
     1261 agctgaccct gaagttcatc tgcaccaccg gcaagctgcc cgtgccctgg cccaccctcg
     1321 tgaccaccct gtcctggggc gtgcagtgct tcgcccgcta ccccgaccac atgaagcagc
     1381 acgacttctt caagtccgcc atgcccgaag gctacgtcca ggagcgcacc atcttcttca
     1441 aggacgacgg caactacaag acccgcgccg aggtgaagtt cgagggcgac accctggtga
     1501 accgcatcga gctgaagggc atcgacttca aggaggacgg caacatcctg gggcacaagc
     1561 tggagtacaa ctactttagc gacaacgtct atatcaccgc cgacaagcag aagaacggca
     1621 tcaaggccaa cttcaagatc cgccacaaca tcgaggacgg cggcgtgcag ctcgccgacc
     1681 actaccagca gaacaccccc atcggcgacg gccccgtgct gctgcccgac aaccactacc
     1741 tgagcaccca gtccaagctg agcaaagacc ccaacgagaa gcgcgatcac atggtcctgc
     1801 tggagttcgt gaccgccgcc gggatcactc tcggcatgga cgagctgtac aagtaaagcg
     1861 gccgcaccca gctttcttgt acaaagtggt gtagttaatt catgatggcc gctgcaggtc
     1921 gacctcgagg gggggcccgg tacccaattc gccctatagt gagtcgtatt acgcgcggat
     1981 ccagctttgg acttcttcgc cagaggtttg gtcaagtctc caatcaaggt tgtcggcttg
     2041 tctaccttgc cagaaattta cgaaaagatg gaaaagggtc aaatcgttgg tagatacgtt
     2101 gttgacactt ctaaataagc gaatttctta tgatttatga tttttatatt aaataaagta
     2161 taaaaaaac
//
