LOCUS       sequence-484397-        1521 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  Expression plasmid for rtTA-2A-mRuby2 driven from the EFS promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 1521)
  AUTHORS   Kabaria SR, Bae Y, Ehmann ME, Lende-Dorn BA, Beitz AM, Peterman EL, 
            Love KS, Ploessl DS, Galloway KE
  TITLE     Programmable promoter editing for precise control of transgene 
            expression.
  JOURNAL   Nat Biotechnol. 2025 Oct 13. doi: 10.1038/s41587-025-02854-y.
  PUBMED    41083901
REFERENCE   2  (bases 1 to 1521)
  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: "Nat 
            Biotechnol. 2025 Oct 13. doi: 10.1038/s41587-025-02854-y."
COMMENT     Sequence Label: EFS-rtTA-2A-mRuby2-bGH_pKG2064
FEATURES             Location/Qualifiers
     source          1..1521
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     CDS             1..744
                     /codon_start=1
                     /product="modified rtTA protein that binds tightly to 
                     promoters containing the tet operator in the presence of 
                     doxycycline"
                     /label=Tet-On(R) 3G
                     /translation="MSRLDKSKVINSALELLNGVGIEGLTTRKLAQKLGVEQPTLYWHV
                     KNKRALLDALPIEMLDRHHTHSCPLEGESWQDFLRNNAKSYRCALLSHRDGAKVHLGTR
                     PTEKQYETLENQLAFLCQQGFSLENALYALSAVGHFTLGCVLEEQEHQVAKEERETPTT
                     DSMPPLLKQAIELFDRQGAEPAFLFGLELIICGLEKQLKCESGGPTDALDDFDLDMLPA
                     DALDDFDLDMLPADALDDFDLDMLPG"
     CDS             754..810
                     /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"
     CDS             811..1521
                     /codon_start=1
                     /product="monomeric red fluorescent protein derived from 
                     mRuby, with improved photophysical properties (Lam et al., 
                     2012)"
                     /label=mRuby2
                     /translation="MVSKGEELIKENMRMKVVMEGSVNGHQFKCTGEGEGNPYMGTQTM
                     RIKVIEGGPLPFAFDILATSFMYGSRTFIKYPKGIPDFFKQSFPEGFTWERVTRYEDGG
                     VVTVMQDTSLEDGCLVYHVQVRGVNFPSNGPVMQKKTKGWEPNTEMMYPADGGLRGYTH
                     MALKVDGGGHLSCSFVTTYRSKKTVGNIKMPGIHAVDHRLERLEESDNEMFVVQREHAV
                     AKFAGLGGGMDELYK"
ORIGIN
        1 atgagccggc tggacaagag caaagtgatc aacagcgccc tggaactgct gaacggcgtg
       61 ggcatcgagg gcctgaccac ccggaagctg gcccagaaac tgggcgtgga acagcccacc
      121 ctgtactggc acgtgaagaa caagcgggcc ctgctggacg ccctgcccat cgagatgctg
      181 gaccggcacc acacccacag ctgccctctg gaaggcgaga gctggcagga cttcctgcgg
      241 aacaacgcca agagctacag atgcgccctg ctgagccacc gggacggcgc caaagtgcac
      301 ctgggcacca gacccaccga gaagcagtac gagacactgg aaaaccagct ggccttcctg
      361 tgccagcagg gcttcagcct ggaaaacgcc ctgtacgccc tgagcgccgt gggccacttc
      421 accctgggct gcgtgctgga agaacaggaa caccaggtcg ccaaagagga aagagagaca
      481 cccaccaccg acagcatgcc ccccctgctg aagcaggcca tcgagctgtt cgacagacag
      541 ggcgccgagc ccgccttcct gttcggcctg gaactgatca tctgcggcct cgagaagcag
      601 ctgaagtgcg agagcggcgg acccaccgac gccctggacg acttcgacct ggacatgctg
      661 cccgccgatg ccctggatga ttttgatctg gatatgctcc ctgctgacgc tctcgatgac
      721 ttcgatctcg atatgctgcc aggcggaagc ggagctacta acttcagcct gctgaagcag
      781 gctggagacg tggaggagaa ccctggacct atggtgtcta agggcgaaga gctgatcaag
      841 gaaaatatgc gtatgaaggt ggtcatggaa ggttcggtca acggccacca attcaaatgc
      901 acaggtgaag gagaaggcaa tccgtacatg ggaactcaaa ccatgaggat caaagtcatc
      961 gagggaggac ccctgccatt tgcctttgac attcttgcca cgtcgttcat gtatggcagc
     1021 cgtactttta tcaagtaccc gaaaggcatt cctgatttct ttaaacagtc ctttcctgag
     1081 ggttttactt gggaaagagt tacgagatac gaagatggtg gagtcgtcac cgtcatgcag
     1141 gacaccagcc ttgaggatgg ctgtctcgtt taccacgtcc aagtcagagg ggtaaacttt
     1201 ccctccaatg gtcccgtgat gcagaagaag accaagggtt gggagcctaa tacagagatg
     1261 atgtatccag cagatggtgg tctgagggga tacactcata tggcactgaa agttgatggt
     1321 ggtggccatc tgtcttgctc tttcgtaaca acttacaggt caaaaaagac cgtcgggaac
     1381 atcaagatgc ccggtatcca tgccgttgat caccgcctgg aaaggttaga ggaaagtgac
     1441 aatgaaatgt tcgtagtaca acgcgaacac gcagttgcca agttcgccgg gcttggtggt
     1501 gggatggacg agctgtacaa g
//
