LOCUS       sequence-399722-        2179 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  Expression of miRFP680-T2A-mtq2 in yeast.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 2179)
  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 2179)
  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 miRFP680-T2A-mtq2
FEATURES             Location/Qualifiers
     source          1..2179
                     /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             120..1067
                     /codon_start=1
                     /product="phytochrome-based near-infrared fluorescent 
                     protein, originally termed iRFP (Filonov et al., 2011)"
                     /label=iRFP713
                     /note="name was changed to iRFP713 (Shcherbakova and 
                     Verkhusha, 2013)"
                     /translation="MAEGSVARQPDLLTCDDEPIHIPGAIQPHGLLLALAADMTIVAGS
                     DNLPELTGLAIGALIGRSAADVFDSETHNRLTIALAEPGAAVGAPITVGFTMRKDAGFI
                     GSWHRHDQLIFLELEPPQRDVAEPQAFFRRTNSAIRRLQAAETLESACAAAAQEVRKIT
                     GFDRVMIYRFASDFSGEVIAEDRCAEVESKLGLHYPASTVPAQARRLYTINPVRIIPDI
                     NYRPVPVTPDLNPVTGRPIDLSFAILRSVSPCHLEFMRNIGMHGTMSISILRGERLWGL
                     IVCHHRTPYYVDLDGRQACKRVAERLATQIGVMEE"
     CDS             1074..1127
                     /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      1143..1152
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(1149..1151,1152..1154,1155..1868)
                     /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(1194..1215)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1455..1474)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1802..1823
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     protein_bind    complement(1878..1902)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     primer_bind     complement(1964..1983)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1965..1983)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
ORIGIN
        1 aacccatttt ttttctctct tttatacaca cattcaaaag aaagaaaaaa aatatacccc
       61 agcctcgact agtgaattcg cggccatcac aagtttgtac aaaaaagcag gctgctagca
      121 tggcggaagg ctccgtcgcc aggcagcctg acctcttgac ctgcgacgat gagccgatcc
      181 atatccccgg tgccatccaa ccgcatggac tgctgctcgc cctcgccgcc gacatgacga
      241 tcgttgccgg cagcgacaac cttcccgaac tcaccggact ggcgatcggc gccctgatcg
      301 gccgctctgc ggccgatgtc ttcgactcgg agacgcacaa ccgtctgacg atcgccttgg
      361 ccgagcccgg ggcggccgtc ggagcaccga tcactgtcgg cttcacgatg cgaaaggacg
      421 caggcttcat cggctcctgg catcgccatg atcagctcat cttcctcgag ctcgagcctc
      481 cccagcggga cgtcgccgag ccgcaggcgt tcttccgccg caccaacagc gccatccgcc
      541 gcctgcaggc cgccgaaacc ttggaaagcg cctgcgccgc cgcggcgcaa gaggtgcgga
      601 agattaccgg cttcgatcgg gtgatgatct atcgcttcgc ctccgacttc agcggcgaag
      661 tgatcgcaga ggatcggtgc gccgaggtcg agtcaaaact aggcctgcac tatcctgcct
      721 caaccgtgcc ggcgcaggcc cgtcggctct ataccatcaa cccggtacgg atcattcccg
      781 atatcaatta tcggccggtg ccggtcaccc cagacctcaa tccggtcacc gggcggccga
      841 ttgatcttag cttcgccatc ctgcgcagcg tctcgccctg ccatctggag ttcatgcgca
      901 acataggcat gcacggcacg atgtcgatct cgattttgcg cggcgagcga ctgtggggat
      961 tgatcgtttg ccatcaccga acgccgtact acgtcgatct cgatggccgc caagcctgca
     1021 agagggtcgc cgagaggctg gccactcaga tcggcgtgat ggaagagtcc ggagagggca
     1081 gaggaagtct tctaacatgc ggtgacgtgg aggagaatcc cggccctggg gatccaccgg
     1141 tcgccaccat ggtgagcaag ggcgaggagc tgttcaccgg ggtggtgccc atcctggtcg
     1201 agctggacgg cgacgtaaac ggccacaagt tcagcgtgtc cggcgagggc gagggcgatg
     1261 ccacctacgg caagctgacc ctgaagttca tctgcaccac cggcaagctg cccgtgccct
     1321 ggcccaccct cgtgaccacc ctgtcctggg gcgtgcagtg cttcgcccgc taccccgacc
     1381 acatgaagca gcacgacttc ttcaagtccg ccatgcccga aggctacgtc caggagcgca
     1441 ccatcttctt caaggacgac ggcaactaca agacccgcgc cgaggtgaag ttcgagggcg
     1501 acaccctggt gaaccgcatc gagctgaagg gcatcgactt caaggaggac ggcaacatcc
     1561 tggggcacaa gctggagtac aactacttta gcgacaacgt ctatatcacc gccgacaagc
     1621 agaagaacgg catcaaggcc aacttcaaga tccgccacaa catcgaggac ggcggcgtgc
     1681 agctcgccga ccactaccag cagaacaccc ccatcggcga cggccccgtg ctgctgcccg
     1741 acaaccacta cctgagcacc cagtccaagc tgagcaaaga ccccaacgag aagcgcgatc
     1801 acatggtcct gctggagttc gtgaccgccg ccgggatcac tctcggcatg gacgagctgt
     1861 acaagtaaag cggccgcacc cagctttctt gtacaaagtg gtgtagttaa ttcatgatgg
     1921 ccgctgcagg tcgacctcga gggggggccc ggtacccaat tcgccctata gtgagtcgta
     1981 ttacgcgcgg atccagcttt ggacttcttc gccagaggtt tggtcaagtc tccaatcaag
     2041 gttgtcggct tgtctacctt gccagaaatt tacgaaaaga tggaaaaggg tcaaatcgtt
     2101 ggtagatacg ttgttgacac ttctaaataa gcgaatttct tatgatttat gatttttatt
     2161 attaaattaa gtaataaaa
//
