LOCUS       sequence-491500-        4070 bp    DNA     circular SYN 04-NOV-2025
DEFINITION  203-bp DIAL Reporter Plasmid with YB_TATA expressing 
            mCherry-GS-HRasG12V in the presence of ZFa and editable by Cre 
            recombinase.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4070)
  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 4070)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Nov 4, 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: (203-bp-DIAL-YB_TATA)-mCherry-HRasG12V-BGH_pKG...
FEATURES             Location/Qualifiers
     source          1..4070
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      1..589
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     490..509
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     terminator      754..840
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      932..959
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     primer_bind     967..990
                     /label=pENTR-F
                     /note="pENTR vectors, forward primer"
     protein_bind    complement(1158..1191)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     primer_bind     1328..1347
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     protein_bind    complement(1395..1428)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     CDS             1507..2214
                     /codon_start=1
                     /product="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
                     /label=mCherry
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEG
                     TQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNF
                     EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK
                     GEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERA
                     EGRHSTGGMDELYK"
     primer_bind     complement(1652..1670)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     1920..1939
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     CDS             join(2245..2277,2278..2280,2281..2814)
                     /codon_start=1
                     /product="human oncoprotein generated by the G12V mutation 
                     in the small GTPase H-Ras"
                     /label=H-Ras (G12V)
                     /translation="MTEYKLVVVGAVGVGKSALTIQLIQNHFVDEYDPTIEDSYRKQVV
                     IDGETCLLDILDTAGQEEYSAMRDQYMRTGEGFLCVFAINNTKSFEDIHQYREQIKRVK
                     DSDDVPMVLVGNKCDLAARTVESRQAQDLARSYGIPYIETSAKTRQGVEDAFYTLVREI
                     RQHKLRKLNPPDESGPGCMSCKCVLS"
     polyA_signal    2826..3050
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(3161..3180)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             3172..3981
                     /codon_start=1
                     /gene="aph(3')-Ia"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYGKP
                     DAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPGDAWLLTTAIPGKTA
                     FQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDASD
                     FDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGIAD
                     RYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     complement(3239..3258)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
ORIGIN
        1 ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc
       61 agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt
      121 cagcagagcg cagataccaa atactgttct tctagtgtag ccgtagttag gccaccactt
      181 caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc
      241 tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa
      301 ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac
      361 ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg
      421 gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga
      481 gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact
      541 tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa
      601 cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgt tctttcctgc
      661 gttatcccct gattctgtgg ataaccgtat taccgctagc atggatctcg gggacgtcta
      721 actactaagc gagagtaggg aactgccagg catcaaataa aacgaaaggc tcagtcggaa
      781 gactgggcct ttcgttttat ctgttgtttg tcggtgaacg ctctcctgag taggacaaat
      841 ccgccgggag cggatttgaa cgttgtgaag caacggcccg gagggtggcg ggcaggacgc
      901 ccgccataaa ctgccaggca tcaaactaag cagaaggcca tcctgacgga tggccttttt
      961 gcgtttctac aaactcttcc tgttagttag ttacttaagc tcgggcccac ctgcagtcta
     1021 cttactcgga tcgggagatc tgaattggag tgaggacgtg tagagtgagg acgtgcggag
     1081 tgaggacgtg tagagtgagg acgtgcggag tgaggacgtg tagagtgagg acgtgccctc
     1141 ctctggatca aatccgaata acttcgtata gcatacatta tacgaagtta tctcgagtcg
     1201 ctcggtacga tttaaattga attctgcagt cgacggtacc gcgggcccgg gactttgtac
     1261 aaaaaagcag gctccaccat gggaaccaat tcagtcgact ggatccggta cctcctacag
     1321 ctcctgggca acgtgctggt tattgtgctg tctcatcatt ttggcaaaga attgattaat
     1381 tcgagcgaac gcgtataact tcgtatagca tacattatac gaagttatcg gatttgatct
     1441 gatcaagcaa ctctagaggg tatataatgg gggccactag tctactacca gagctcatcg
     1501 ctagcaatgg tgagcaaggg cgaggaggat aacatggcca tcatcaagga gttcatgcgc
     1561 ttcaaggtgc acatggaggg ctccgtgaac ggccacgagt tcgagatcga gggcgagggc
     1621 gagggccgcc cctacgaggg cacccagacc gccaagctga aggtgaccaa gggtggcccc
     1681 ctgcccttcg cctgggacat cctgtcccct cagttcatgt acggctccaa ggcctacgtg
     1741 aagcaccccg ccgacatccc cgactacttg aagctgtcct tccccgaggg cttcaagtgg
     1801 gagcgcgtga tgaacttcga ggacggcggc gtggtgaccg tgacccagga ctcctccctg
     1861 caggacggcg agttcatcta caaggtgaag ctgcgcggca ccaacttccc ctccgacggc
     1921 cccgtaatgc agaagaagac catgggctgg gaggcctcct ccgagcggat gtaccccgag
     1981 gacggcgccc tgaagggcga gatcaagcag aggctgaagc tgaaggacgg cggccactac
     2041 gacgctgagg tcaagaccac ctacaaggcc aagaagcccg tgcagctgcc cggcgcctac
     2101 aacgtcaaca tcaagttgga catcacctcc cacaacgagg actacaccat cgtggaacag
     2161 tacgaacgcg ccgagggccg ccactccacc ggcggcatgg acgagctgta caagggtggt
     2221 gggggaagcg gaggaggagg ttcgatgacc gaatacaagc tggtggtggt gggcgccgtc
     2281 ggtgtgggca agagtgcgct gaccatccag ctgatccaga accattttgt ggacgaatac
     2341 gaccccacta tagaggattc ctaccggaag caggtggtca ttgatgggga gacgtgcctg
     2401 ttggacatcc tggataccgc cggccaggag gagtacagcg ccatgcggga ccagtacatg
     2461 cgcaccgggg agggcttcct gtgtgtgttt gccatcaaca acaccaagtc ttttgaggac
     2521 atccaccagt acagggagca gatcaaacgg gtgaaggact cggatgacgt gcccatggtg
     2581 ctggtgggga acaagtgtga cctggctgca cgcactgtgg aatctcggca ggctcaggac
     2641 ctcgcccgaa gctacggcat cccctacatc gagacctcgg ccaagacccg gcagggagtg
     2701 gaggatgcct tctacacgtt ggtgcgtgag atccggcagc acaagctgcg gaagctgaac
     2761 cctcctgatg agagtggccc cggctgcatg agctgcaagt gtgtgctctc ctaacaacgt
     2821 tgcacctgtg ccttctagtt gccagccatc tgttgtttgc ccctcccccg tgccttcctt
     2881 gaccctggaa ggtgccactc ccactgtcct ttcctaataa aatgaggaaa ttgcatcgca
     2941 ttgtctgagt aggtgtcatt ctattctggg gggtggggtg gggcaggaca gcaaggggga
     3001 ggattgggaa gacaatagca ggcatgctgg ggatgcggtg ggctctatgg cgctcgctga
     3061 ctgcaggtgc ccgtgtctca aaatctctga tgttacattg cacaagataa aaatatatca
     3121 tcatgaacaa taaaactgtc tgcttacata aacagtaata caaggggtgt tatgagccat
     3181 attcaacggg aaacgtcgag gccgcgatta aattccaaca tggatgctga tttatatggg
     3241 tataaatggg ctcgcgataa tgtcgggcaa tcaggtgcga caatctatcg cttgtatggg
     3301 aagcccgatg cgccagagtt gtttctgaaa catggcaaag gtagcgttgc caatgatgtt
     3361 acagatgaga tggtcagact aaactggctg acggaattta tgcctcttcc gaccatcaag
     3421 cattttatcc gtactcctgg tgatgcatgg ttactcacca ctgcgatccc cggaaaaaca
     3481 gcattccagg tattagaaga atatcctgat tcaggtgaaa atattgttga tgcgctggca
     3541 gtgttcctgc gccggttgca ttcgattcct gtttgtaatt gtccttttaa cagcgatcgc
     3601 gtatttcgtc tagctcaggc gcaatcacga atgaataacg gtttggttga tgcgagtgat
     3661 tttgatgacg agcgtaatgg ctggcctgtt gaacaagtct ggaaagaaat gcataaactt
     3721 ttgccattct caccggattc agtcgtcact catggtgatt tctcacttga taaccttatt
     3781 tttgacgagg ggaaattaat aggttgtatt gatgttggac gagtcggaat cgcagaccga
     3841 taccaggatc ttgccatcct atggaactgc ctcggtgagt tttctccttc attacagaaa
     3901 cggctttttc aaaaatatgg tattgataat cctgatatga ataaattgca gtttcatttg
     3961 atgctcgatg agtttttcta atcagaattg gttaattggt tgtaacatta ttcagattgg
     4021 gccccgttcc actgagcgtc agaccccgta gaaaagatca aaggatcttc
//
