LOCUS       sequence-491506-        4896 bp    DNA     circular SYN 04-NOV-2025
DEFINITION  203-bp DIAL Reporter Plasmid with YB_TATA expressing Halo-p53 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 4896)
  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 4896)
  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)-Halo-p53-BGH_pKG2745
FEATURES             Location/Qualifiers
     source          1..4896
                     /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)."
     regulatory      1511..1520
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1517..2407
                     /codon_start=1
                     /product="modified bacterial dehalogenase that forms 
                     covalent bonds with chloroalkane derivatives"
                     /label=HaloTag(R)
                     /translation="MAEIGTGFPFDPHYVEVLGERMHYVDVGPRDGTPVLFLHGNPTSS
                     YVWRNIIPHVAPTHRCIAPDLIGMGKSDKPDLGYFFDDHVRFMDAFIEALGLEEVVLVI
                     HDWGSALGFHWAKRNPERVKGIAFMEFIRPIPTWDEWPEFARETFQAFRTTDVGRKLII
                     DQNVFIEGTLPMGVVRPLTEVEMDHYREPFLNPVDREPLWRFPNELPIAGEPANIVALV
                     EEYMDWLHQSPVPKLLFWGTPGVLIPPAEAARLAKSLPNCKAVDIGPGLNLLQEDNPDL
                     IGSEIARWLSTLEISG"
     CDS             2420..2440
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="EDLYFQS"
     CDS             complement(2840..2851)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    3652..3876
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(3987..4006)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             3998..4807
                     /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(4065..4084)
                     /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 ctagcaatga gccaccatgg cagaaatcgg tactggcttt ccattcgacc cccattatgt
     1561 ggaagtcctg ggcgagcgca tgcactacgt cgatgttggt ccgcgcgatg gcacccctgt
     1621 gctgttcctg cacggtaacc cgacctcctc ctacgtgtgg cgcaacatca tcccgcatgt
     1681 tgcaccgacc catcgctgca ttgctccaga cctgatcggt atgggcaaat ccgacaaacc
     1741 agacctgggt tatttcttcg acgaccacgt ccgcttcatg gatgccttca tcgaagccct
     1801 gggtctggaa gaggtcgtcc tggtcattca cgactggggc tccgctctgg gtttccactg
     1861 ggccaagcgc aatccagagc gcgtcaaagg tattgcattt atggagttca tccgccctat
     1921 cccgacctgg gacgaatggc cagaatttgc ccgcgagacc ttccaggcct tccgcaccac
     1981 cgacgtcggc cgcaagctga tcatcgatca gaacgttttt atcgagggta cgctgccgat
     2041 gggtgtcgtc cgcccgctga ctgaagtcga gatggaccat taccgcgagc cgttcctgaa
     2101 tcctgttgac cgcgagccac tgtggcgctt cccaaacgag ctgccaatcg ccggtgagcc
     2161 agcgaacatc gtcgcgctgg tcgaagaata catggactgg ctgcaccagt cccctgtccc
     2221 gaagctgctg ttctggggca ccccaggcgt tctgatccca ccggccgaag ccgctcgcct
     2281 ggccaaaagc ctgcctaact gcaaggctgt ggacatcggc ccgggtctga atctgctgca
     2341 agaagacaac ccggacctga tcggcagcga gatcgcgcgc tggctgtcga cgctcgagat
     2401 ttccggcgag ccaaccactg aggatctgta ctttcagagc gataacgcga attcagtcga
     2461 cggatccatg actgccatgg aggagtcaca gtcggatatc agcctcgagc tccctctgag
     2521 ccaggagaca ttttcaggct tatggaaact acttcctcca gaagatatcc tgccatcacc
     2581 tcactgcatg gacgatctgt tgctgcccca ggatgttgag gagttttttg aaggcccaag
     2641 tgaagccctc cgagtgtcag gagctcctgc agcacaggac cctgtcaccg agacccctgg
     2701 gccagtggcc cctgccccag ccactccatg gcccctgtca tcttttgtcc cttctcaaaa
     2761 aacttaccag ggcaactatg gcttccacct gggcttcctg cagtctggga cagccaagtc
     2821 tgttatgtgc acgtactctc ctcccctcaa taagctattc tgccagctgg cgaagacgtg
     2881 ccctgtgcag ttgtgggtca gcgccacacc tccagctggg agccgtgtcc gcgccatggc
     2941 catctacaag aagtcacagc acatgacgga ggtcgtgaga cgctgccccc accatgagcg
     3001 ctgctccgat ggtgatggcc tggctcctcc ccagcatctt atccgggtgg aaggaaattt
     3061 gtatcccgag tatctggaag acaggcagac ttttcgccac agcgtggtgg taccttatga
     3121 gccacccgag gccggctctg agtataccac catccactac aagtacatgt gtaatagctc
     3181 ctgcatgggg ggcatgaacc gccgacctat ccttaccatc atcacactgg aagactccag
     3241 tgggaacctt ctgggacggg acagctttga ggttcgtgtt tgtgcctgcc ctgggagaga
     3301 ccgccgtaca gaagaagaaa atttccgcaa aaaggaagtc ctttgccctg aactgccccc
     3361 agggagcgca aagagagcgc tgcccacatg cacaagcgcc tctcccccgc aaaagaaaaa
     3421 accacttgat ggagagtatt tcaccctcaa gatccgcggg cgtaaacgct tcgagatgtt
     3481 ccgggagctg aatgaggcct tagagttaaa ggatgcccat gctacagagg agtctggaga
     3541 cagcagggct cactccagct acctgaagac caagaagggc cagtctactt cccgccataa
     3601 aaaaacaatg gtcaagaaag tggggcctga ctcagactga caacgttgca cctgtgcctt
     3661 ctagttgcca gccatctgtt gtttgcccct cccccgtgcc ttccttgacc ctggaaggtg
     3721 ccactcccac tgtcctttcc taataaaatg aggaaattgc atcgcattgt ctgagtaggt
     3781 gtcattctat tctggggggt ggggtggggc aggacagcaa gggggaggat tgggaagaca
     3841 atagcaggca tgctggggat gcggtgggct ctatggcgct cgctgactgc aggtgcccgt
     3901 gtctcaaaat ctctgatgtt acattgcaca agataaaaat atatcatcat gaacaataaa
     3961 actgtctgct tacataaaca gtaatacaag gggtgttatg agccatattc aacgggaaac
     4021 gtcgaggccg cgattaaatt ccaacatgga tgctgattta tatgggtata aatgggctcg
     4081 cgataatgtc gggcaatcag gtgcgacaat ctatcgcttg tatgggaagc ccgatgcgcc
     4141 agagttgttt ctgaaacatg gcaaaggtag cgttgccaat gatgttacag atgagatggt
     4201 cagactaaac tggctgacgg aatttatgcc tcttccgacc atcaagcatt ttatccgtac
     4261 tcctggtgat gcatggttac tcaccactgc gatccccgga aaaacagcat tccaggtatt
     4321 agaagaatat cctgattcag gtgaaaatat tgttgatgcg ctggcagtgt tcctgcgccg
     4381 gttgcattcg attcctgttt gtaattgtcc ttttaacagc gatcgcgtat ttcgtctagc
     4441 tcaggcgcaa tcacgaatga ataacggttt ggttgatgcg agtgattttg atgacgagcg
     4501 taatggctgg cctgttgaac aagtctggaa agaaatgcat aaacttttgc cattctcacc
     4561 ggattcagtc gtcactcatg gtgatttctc acttgataac cttatttttg acgaggggaa
     4621 attaataggt tgtattgatg ttggacgagt cggaatcgca gaccgatacc aggatcttgc
     4681 catcctatgg aactgcctcg gtgagttttc tccttcatta cagaaacggc tttttcaaaa
     4741 atatggtatt gataatcctg atatgaataa attgcagttt catttgatgc tcgatgagtt
     4801 tttctaatca gaattggtta attggttgta acattattca gattgggccc cgttccactg
     4861 agcgtcagac cccgtagaaa agatcaaagg atcttc
//
