LOCUS       sequence-492408-        4304 bp    DNA     circular SYN 10-NOV-2025
DEFINITION  203-bp DIAL Reporter Plasmid with YB_TATA expressing mGreenLantern 
            and mCherry 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 4304)
  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 4304)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Nov 10, 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)-mGL-2A-mCherry-BGH_pKG2848
FEATURES             Location/Qualifiers
     source          1..4304
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             67..774
                     /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(212..230)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     480..499
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     polyA_signal    815..1039
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(1150..1169)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             1161..1970
                     /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(1228..1247)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      2060..2648
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2549..2568
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     terminator      2813..2899
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      2991..3018
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     primer_bind     3026..3049
                     /label=pENTR-F
                     /note="pENTR vectors, forward primer"
     protein_bind    complement(3217..3250)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     primer_bind     3387..3406
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     protein_bind    complement(3454..3487)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     regulatory      3570..3579
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(3576..3578,3579..3581,3582..4292)
                     /codon_start=1
                     /product="monomeric GFP derivative that displays 
                     exceptionally high brightness in cells (Campbell et al., 
                     2020)"
                     /label=mGreenLantern
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVRGEGEGDATNGKLTL
                     KFICTTGKLPVPWPTLVTTLGYGVACFARYPDHMKQHDFFKSAMPEGYVQERTISFKDD
                     GTYKTRAEVKFEGDTLVNRIVLKGIDFKEDGNILGHKLEYNFNSHKVYITADKQKNGIK
                     ANFKTRHNVEDGGVQLADHYQQNTPIGDGPVLLPDNHYLSHQSKLSKDPNEKRDHMVLK
                     ERVTAAGITHDMDELYK"
     primer_bind     complement(3621..3642)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     CDS             join(4302..4304,1..54)
                     /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"
ORIGIN
        1 actaacttca gcctgctgaa gcaggctgga gacgtggagg agaaccctgg acctgcacca
       61 ggaagtatgg tgagcaaggg cgaggaggat aacatggcca tcatcaagga gttcatgcgc
      121 ttcaaggtgc acatggaggg ctccgtgaac ggccacgagt tcgagatcga gggcgagggc
      181 gagggccgcc cctacgaggg cacccagacc gccaagctga aggtgaccaa gggtggcccc
      241 ctgcccttcg cctgggacat cctgtcccct cagttcatgt acggctccaa ggcctacgtg
      301 aagcaccccg ccgacatccc cgactacttg aagctgtcct tccccgaggg cttcaagtgg
      361 gagcgcgtga tgaacttcga ggacggcggc gtggtgaccg tgacccagga ctcctccctg
      421 caggacggcg agttcatcta caaggtgaag ctgcgcggca ccaacttccc ctccgacggc
      481 cccgtaatgc agaagaagac catgggctgg gaggcctcct ccgagcggat gtaccccgag
      541 gacggcgccc tgaagggcga gatcaagcag aggctgaagc tgaaggacgg cggccactac
      601 gacgctgagg tcaagaccac ctacaaggcc aagaagcccg tgcagctgcc cggcgcctac
      661 aacgtcaaca tcaagttgga catcacctcc cacaacgagg actacaccat cgtggaacag
      721 tacgaacgcg ccgagggccg ccactccacc ggcggcatgg acgagctgta caagcaattg
      781 ccaaaaaaga agagaaggta taacaacgtt gcacctgtgc cttctagttg ccagccatct
      841 gttgtttgcc cctcccccgt gccttccttg accctggaag gtgccactcc cactgtcctt
      901 tcctaataaa atgaggaaat tgcatcgcat tgtctgagta ggtgtcattc tattctgggg
      961 ggtggggtgg ggcaggacag caagggggag gattgggaag acaatagcag gcatgctggg
     1021 gatgcggtgg gctctatggc gctcgctgac tgcaggtgcc cgtgtctcaa aatctctgat
     1081 gttacattgc acaagataaa aatatatcat catgaacaat aaaactgtct gcttacataa
     1141 acagtaatac aaggggtgtt atgagccata ttcaacggga aacgtcgagg ccgcgattaa
     1201 attccaacat ggatgctgat ttatatgggt ataaatgggc tcgcgataat gtcgggcaat
     1261 caggtgcgac aatctatcgc ttgtatggga agcccgatgc gccagagttg tttctgaaac
     1321 atggcaaagg tagcgttgcc aatgatgtta cagatgagat ggtcagacta aactggctga
     1381 cggaatttat gcctcttccg accatcaagc attttatccg tactcctggt gatgcatggt
     1441 tactcaccac tgcgatcccc ggaaaaacag cattccaggt attagaagaa tatcctgatt
     1501 caggtgaaaa tattgttgat gcgctggcag tgttcctgcg ccggttgcat tcgattcctg
     1561 tttgtaattg tccttttaac agcgatcgcg tatttcgtct agctcaggcg caatcacgaa
     1621 tgaataacgg tttggttgat gcgagtgatt ttgatgacga gcgtaatggc tggcctgttg
     1681 aacaagtctg gaaagaaatg cataaacttt tgccattctc accggattca gtcgtcactc
     1741 atggtgattt ctcacttgat aaccttattt ttgacgaggg gaaattaata ggttgtattg
     1801 atgttggacg agtcggaatc gcagaccgat accaggatct tgccatccta tggaactgcc
     1861 tcggtgagtt ttctccttca ttacagaaac ggctttttca aaaatatggt attgataatc
     1921 ctgatatgaa taaattgcag tttcatttga tgctcgatga gtttttctaa tcagaattgg
     1981 ttaattggtt gtaacattat tcagattggg ccccgttcca ctgagcgtca gaccccgtag
     2041 aaaagatcaa aggatcttct tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa
     2101 caaaaaaacc accgctacca gcggtggttt gtttgccgga tcaagagcta ccaactcttt
     2161 ttccgaaggt aactggcttc agcagagcgc agataccaaa tactgttctt ctagtgtagc
     2221 cgtagttagg ccaccacttc aagaactctg tagcaccgcc tacatacctc gctctgctaa
     2281 tcctgttacc agtggctgct gccagtggcg ataagtcgtg tcttaccggg ttggactcaa
     2341 gacgatagtt accggataag gcgcagcggt cgggctgaac ggggggttcg tgcacacagc
     2401 ccagcttgga gcgaacgacc tacaccgaac tgagatacct acagcgtgag ctatgagaaa
     2461 gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa
     2521 caggagagcg cacgagggag cttccagggg gaaacgcctg gtatctttat agtcctgtcg
     2581 ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc
     2641 tatggaaaaa cgccagcaac gcggcctttt tacggttcct ggccttttgc tggccttttg
     2701 ctcacatgtt ctttcctgcg ttatcccctg attctgtgga taaccgtatt accgctagca
     2761 tggatctcgg ggacgtctaa ctactaagcg agagtaggga actgccaggc atcaaataaa
     2821 acgaaaggct cagtcggaag actgggcctt tcgttttatc tgttgtttgt cggtgaacgc
     2881 tctcctgagt aggacaaatc cgccgggagc ggatttgaac gttgtgaagc aacggcccgg
     2941 agggtggcgg gcaggacgcc cgccataaac tgccaggcat caaactaagc agaaggccat
     3001 cctgacggat ggcctttttg cgtttctaca aactcttcct gttagttagt tacttaagct
     3061 cgggcccacc tgcagtctac ttactcggat cgggagatct gaattggagt gaggacgtgt
     3121 agagtgagga cgtgcggagt gaggacgtgt agagtgagga cgtgcggagt gaggacgtgt
     3181 agagtgagga cgtgccctcc tctggatcaa atccgaataa cttcgtatag catacattat
     3241 acgaagttat ctcgagtcgc tcggtacgat ttaaattgaa ttctgcagtc gacggtaccg
     3301 cgggcccggg actttgtaca aaaaagcagg ctccaccatg ggaaccaatt cagtcgactg
     3361 gatccggtac ctcctacagc tcctgggcaa cgtgctggtt attgtgctgt ctcatcattt
     3421 tggcaaagaa ttgattaatt cgagcgaacg cgtataactt cgtatagcat acattatacg
     3481 aagttatcgg atttgatctg atcaagcaac tctagagggt atataatggg ggccactagt
     3541 ctactaccag agctcatcgc tagcaatgag ccaccatggt gagcaagggc gaggagctgt
     3601 tcaccggggt ggtgcccatc ctggtcgagc tggacggcga cgtaaacggc cacaagttca
     3661 gcgtccgcgg cgagggcgag ggcgatgcca ccaacggcaa gctgaccctg aagttcatct
     3721 gcaccaccgg caagctgccc gtgccctggc ccaccctcgt gaccacctta ggctacggcg
     3781 tggcctgctt cgcccgctac cccgaccaca tgaagcagca cgacttcttc aagtccgcca
     3841 tgcccgaagg ctacgtccag gagcgcacca tctctttcaa ggacgacggt acctacaaga
     3901 cccgcgccga ggtgaagttc gagggcgaca ccctggtgaa ccgcatcgtg ctgaagggca
     3961 tcgacttcaa ggaggacggc aacatcctgg ggcacaagct ggagtacaac ttcaacagcc
     4021 acaaggtcta tatcacggcc gacaagcaga agaacggcat caaggctaac ttcaagaccc
     4081 gccacaacgt tgaggacggc ggcgtgcagc tcgccgacca ctaccagcag aacaccccca
     4141 tcggcgacgg ccccgtgctg ctgcccgaca accactacct gagccatcag tccaaactga
     4201 gcaaagaccc caacgagaag cgcgatcaca tggtcctgaa ggagagggtg accgccgccg
     4261 ggattacaca tgacatggac gagctgtaca agggaagcgg agct
//
