LOCUS       sequence-478994-        1687 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  Produces an mTurquoise-antiFRET-mScarlet3 fusion protein in bacteria
            and in mamallian cells it produces 1:1 unfused mTurquoise2 and 
            mScarlet3-H FPs for ratiometric analysis.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 1687)
  TITLE     mScarlet3-H plasmids
REFERENCE   2  (bases 1 to 1687)
  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"
COMMENT     Sequence Label: pDRESS_mTurquoise2_spatial-linker-P2A_mScarlet3-H
FEATURES             Location/Qualifiers
     source          1..1687
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     CDS             13..30
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             40..57
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
     regulatory      76..85
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(82..84,85..87,88..798)
                     /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(127..148)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(388..407)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     735..756
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     CDS             919..975
                     /codon_start=1
                     /product="2A peptide from porcine teschovirus-1 
                     polyprotein"
                     /label=P2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="ATNFSLLKQAGDVEENPGP"
     regulatory      994..1003
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1000..1686
                     /codon_start=1
                     /product="bright cysteine-free monomeric red fluorescent 
                     protein with fast and complete maturation (Gadella et al., 
                     2023)"
                     /label=mScarlet3
                     /translation="MDSTEAVIKEFMRFKVHMEGSMNGHEFEIEGEGEGRPYEGTQTAK
                     LRVTKGGPLPFSWDILSPQFMYGSRAFTKHPADIPDYWKQSFPEGFKWERVMNFEDGGA
                     VSVAQDTSLEDGTLIYKVKLRGTNFPPDGPVMQKKTMGWEASTERLYPEDVVLKGDIKH
                     ALRLKDGGRYLADFKTTYRAKKPVQMPGAFNIDRKLDITSHNEDYTVVEQYERSVARHS
                     TGGSGGS"
     primer_bind     1398..1417
                     /label=mCherry-F
                     /note="mCherry, forward primer"
ORIGIN
        1 atgcggggtt ctcatcatca ccaccatcac agcagcggcc tggtgccgcg cggcagctcg
       61 ctagcgctgc cggtcgccac catggtgagc aagggcgagg agctgttcac cggggtggtg
      121 cccatcctgg tcgagctgga cggcgacgta aacggccaca agttcagcgt gtccggcgag
      181 ggcgagggcg atgccaccta cggcaagctg accctgaagt tcatctgcac caccggcaag
      241 ctgcccgtgc cctggcccac cctcgtgacc accctgtcct ggggcgtgca gtgcttcgcc
      301 cgctaccccg accacatgaa gcagcacgac ttcttcaagt ccgccatgcc cgaaggctac
      361 gtccaggagc gcaccatctt cttcaaggac gacggcaact acaagacccg cgccgaggtg
      421 aagttcgagg gcgacaccct ggtgaaccgc atcgagctga agggcatcga cttcaaggag
      481 gacggcaaca tcctggggca caagctggag tacaactact ttagcgacaa cgtctatatc
      541 accgccgaca agcagaagaa cggcatcaag gccaacttca agatccgcca caacatcgag
      601 gacggcggcg tgcagctcgc cgaccactac cagcagaaca cccccatcgg cgacggcccc
      661 gtgctgctgc ccgacaacca ctacctgagc acccagtcca agctgagcaa agaccccaac
      721 gagaagcgcg atcacatggt cctgctggag ttcgtgaccg ccgccgggat cactctcggc
      781 atggacgagc tctacaagtc gaccagatct ctggccgagg cggcagctaa ggaggccgcg
      841 gcgaaggagg cagccgcgaa agaagcagcc gcaaaggaag cggccgccaa ggctgctgca
      901 agatctctcg gatcgggtgc tactaacttc agcctgctga agcaggctgg agacgtggag
      961 gagaaccctg gaccttcgct agcgctaccg gtcgccacca tggatagcac cgaggcagtg
     1021 atcaaggagt tcatgcggtt caaggtgcac atggagggct ccatgaacgg ccacgagttc
     1081 gagatcgagg gcgagggcga gggccgcccc tacgagggca cccagaccgc caagctgagg
     1141 gtgaccaagg gtggccccct gcccttctcc tgggacatcc tgtcccctca gttcatgtac
     1201 ggctccaggg ccttcacgaa gcaccccgcc gacatccccg actactggaa gcagtccttc
     1261 cccgagggct tcaagtggga gcgcgtgatg aacttcgagg acggcggcgc cgtgtccgtg
     1321 gcccaggaca cctccctgga ggacggcacc ctgatctaca aggtgaagct ccgcggcacc
     1381 aacttccctc ctgacggccc cgtaatgcag aagaagacaa tgggctggga agcatccacc
     1441 gagcggttgt accccgagga cgtcgtgctg aagggcgaca ttaagcacgc cctgcgcctg
     1501 aaggacggcg gccgctacct ggcggacttc aagaccacct acagggccaa gaagcccgtg
     1561 cagatgcccg gcgccttcaa catcgaccgc aagttggaca tcacatccca caacgaggac
     1621 tacaccgtgg tggaacagta cgaacgctcc gtggcccgcc actccaccgg cggctccggt
     1681 ggctcct
//
