LOCUS       sequence-399721-        1674 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  Expression of miRFp670nano3-T2A-mtq2 in yeast.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 1674)
  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 1674)
  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 miRFp670nano3-T2A-mtq2
FEATURES             Location/Qualifiers
     source          1..1674
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     protein_bind    88..112
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             566..619
                     /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      635..644
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(641..643,644..646,647..1360)
                     /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(686..707)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(947..966)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1294..1315
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     protein_bind    complement(1370..1394)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     primer_bind     complement(1456..1475)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1457..1475)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
ORIGIN
        1 aaaccatttt tttctctctt ttatacacac attcaaaaga aagaaaaaaa atatacccca
       61 gcctcgacta gtgaattcgc ggccatcaca agtttgtaca aaaaagcagg ctgctagcat
      121 ggcaaacctg gacaagatgc tgaacaccac cgtgaccgag gtgcgcaagt tcctgcaagc
      181 ggacagagtg tgcgtgttca agttcgagga agattactcc ggcaccgtct cgcacgaagc
      241 cgtggacgac agatggatta gcatcctgaa gacccaggtg caggacagat acttcatgga
      301 aaccagaggc gaggaatacg tccacggcag ataccaggcc atcgccgaca tctacacagc
      361 caatctggtc gagtgctaca gagacctgct gatcgagttt caggtgcggg ccattctggc
      421 tgtccccatc ctgcaaggca agaagctgtg gggcctgctg gtggcccacc aactggccgg
      481 ccctcgggag tggcagacct gggaaatcga cttcctgaaa cagcaagccg tggtgatggg
      541 catcgccatc cagcagagct ccggagaggg cagaggaagt cttctaacat gcggtgacgt
      601 ggaggagaat cccggccctg gggatccacc ggtcgccacc atggtgagca agggcgagga
      661 gctgttcacc ggggtggtgc ccatcctggt cgagctggac ggcgacgtaa acggccacaa
      721 gttcagcgtg tccggcgagg gcgagggcga tgccacctac ggcaagctga ccctgaagtt
      781 catctgcacc accggcaagc tgcccgtgcc ctggcccacc ctcgtgacca ccctgtcctg
      841 gggcgtgcag tgcttcgccc gctaccccga ccacatgaag cagcacgact tcttcaagtc
      901 cgccatgccc gaaggctacg tccaggagcg caccatcttc ttcaaggacg acggcaacta
      961 caagacccgc gccgaggtga agttcgaggg cgacaccctg gtgaaccgca tcgagctgaa
     1021 gggcatcgac ttcaaggagg acggcaacat cctggggcac aagctggagt acaactactt
     1081 tagcgacaac gtctatatca ccgccgacaa gcagaagaac ggcatcaagg ccaacttcaa
     1141 gatccgccac aacatcgagg acggcggcgt gcagctcgcc gaccactacc agcagaacac
     1201 ccccatcggc gacggccccg tgctgctgcc cgacaaccac tacctgagca cccagtccaa
     1261 gctgagcaaa gaccccaacg agaagcgcga tcacatggtc ctgctggagt tcgtgaccgc
     1321 cgccgggatc actctcggca tggacgagct gtacaagtaa agcggccgca cccagctttc
     1381 ttgtacaaag tggtgtagtt aattcatgat ggccgctgca ggtcgacctc gagggggggc
     1441 ccggtaccca attcgcccta tagtgagtcg tattacgcgc ggatccagct ttggacttct
     1501 tcgccagagg tttggtcaag tctccaatca aggttgtcgg cttgtctacc ttgccagaaa
     1561 tttacgaaaa gatggaaaag ggtcaaatcg ttggtagata cgttgttgac acttctaaat
     1621 aagcgaattt cttatgattt atgattttta ttataaataa gttataaaaa aaat
//
