LOCUS       sequence-492436-        4258 bp    DNA     circular SYN 10-NOV-2025
DEFINITION  610-bp DIAL Reporter Plasmid with YB_TATA expressing mGreenLantern 
            in the presence of DOX and rtTA and editable by Cre recombinase.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4258)
  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 4258)
  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: (610-bp-TET-DIAL-YB-TATA)-mGL-bGH_pKG3263
FEATURES             Location/Qualifiers
     source          1..4258
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     regulatory      65..74
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(71..73,74..76,77..790)
                     /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(116..137)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     polyA_signal    802..1026
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(1137..1156)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             1148..1957
                     /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(1215..1234)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      2047..2635
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2536..2555
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     terminator      2800..2886
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      2978..3005
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     primer_bind     3013..3036
                     /label=pENTR-F
                     /note="pENTR vectors, forward primer"
     promoter        3168..3546
                     /label=TRE3G promoter
                     /note="3rd-generation Tet-responsive promoter that can be 
                     activated by binding of Tet-On(R) 3G"
     protein_bind    3176..3194
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    3212..3230
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    3248..3266
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    3284..3302
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    3320..3338
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    3356..3374
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    3392..3410
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     primer_bind     3455..3479
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     protein_bind    complement(3564..3597)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     primer_bind     complement(3698..3717)
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     complement(3931..3950)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4050..4072
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(4110..4128)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     protein_bind    complement(4203..4236)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
ORIGIN
        1 gccactctag agggtatata atgggggcca ctagtctact accagagctc atcgctagca
       61 atgagccacc atggtgagca agggcgagga gctgttcacc ggggtggtgc ccatcctggt
      121 cgagctggac ggcgacgtaa acggccacaa gttcagcgtc cgcggcgagg gcgagggcga
      181 tgccaccaac ggcaagctga ccctgaagtt catctgcacc accggcaagc tgcccgtgcc
      241 ctggcccacc ctcgtgacca ccttaggcta cggcgtggcc tgcttcgccc gctaccccga
      301 ccacatgaag cagcacgact tcttcaagtc cgccatgccc gaaggctacg tccaggagcg
      361 caccatctct ttcaaggacg acggtaccta caagacccgc gccgaggtga agttcgaggg
      421 cgacaccctg gtgaaccgca tcgtgctgaa gggcatcgac ttcaaggagg acggcaacat
      481 cctggggcac aagctggagt acaacttcaa cagccacaag gtctatatca cggccgacaa
      541 gcagaagaac ggcatcaagg ctaacttcaa gacccgccac aacgttgagg acggcggcgt
      601 gcagctcgcc gaccactacc agcagaacac ccccatcggc gacggccccg tgctgctgcc
      661 cgacaaccac tacctgagcc atcagtccaa actgagcaaa gaccccaacg agaagcgcga
      721 tcacatggtc ctgaaggaga gggtgaccgc cgccgggatt acacatgaca tggacgagct
      781 gtacaagtaa caacgttgca cctgtgcctt ctagttgcca gccatctgtt gtttgcccct
      841 cccccgtgcc ttccttgacc ctggaaggtg ccactcccac tgtcctttcc taataaaatg
      901 aggaaattgc atcgcattgt ctgagtaggt gtcattctat tctggggggt ggggtggggc
      961 aggacagcaa gggggaggat tgggaagaca atagcaggca tgctggggat gcggtgggct
     1021 ctatggcgct cgctgactgc aggtgcccgt gtctcaaaat ctctgatgtt acattgcaca
     1081 agataaaaat atatcatcat gaacaataaa actgtctgct tacataaaca gtaatacaag
     1141 gggtgttatg agccatattc aacgggaaac gtcgaggccg cgattaaatt ccaacatgga
     1201 tgctgattta tatgggtata aatgggctcg cgataatgtc gggcaatcag gtgcgacaat
     1261 ctatcgcttg tatgggaagc ccgatgcgcc agagttgttt ctgaaacatg gcaaaggtag
     1321 cgttgccaat gatgttacag atgagatggt cagactaaac tggctgacgg aatttatgcc
     1381 tcttccgacc atcaagcatt ttatccgtac tcctggtgat gcatggttac tcaccactgc
     1441 gatccccgga aaaacagcat tccaggtatt agaagaatat cctgattcag gtgaaaatat
     1501 tgttgatgcg ctggcagtgt tcctgcgccg gttgcattcg attcctgttt gtaattgtcc
     1561 ttttaacagc gatcgcgtat ttcgtctagc tcaggcgcaa tcacgaatga ataacggttt
     1621 ggttgatgcg agtgattttg atgacgagcg taatggctgg cctgttgaac aagtctggaa
     1681 agaaatgcat aaacttttgc cattctcacc ggattcagtc gtcactcatg gtgatttctc
     1741 acttgataac cttatttttg acgaggggaa attaataggt tgtattgatg ttggacgagt
     1801 cggaatcgca gaccgatacc aggatcttgc catcctatgg aactgcctcg gtgagttttc
     1861 tccttcatta cagaaacggc tttttcaaaa atatggtatt gataatcctg atatgaataa
     1921 attgcagttt catttgatgc tcgatgagtt tttctaatca gaattggtta attggttgta
     1981 acattattca gattgggccc cgttccactg agcgtcagac cccgtagaaa agatcaaagg
     2041 atcttcttga gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc
     2101 gctaccagcg gtggtttgtt tgccggatca agagctacca actctttttc cgaaggtaac
     2161 tggcttcagc agagcgcaga taccaaatac tgttcttcta gtgtagccgt agttaggcca
     2221 ccacttcaag aactctgtag caccgcctac atacctcgct ctgctaatcc tgttaccagt
     2281 ggctgctgcc agtggcgata agtcgtgtct taccgggttg gactcaagac gatagttacc
     2341 ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc acacagccca gcttggagcg
     2401 aacgacctac accgaactga gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc
     2461 cgaagggaga aaggcggaca ggtatccggt aagcggcagg gtcggaacag gagagcgcac
     2521 gagggagctt ccagggggaa acgcctggta tctttatagt cctgtcgggt ttcgccacct
     2581 ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc
     2641 cagcaacgcg gcctttttac ggttcctggc cttttgctgg ccttttgctc acatgttctt
     2701 tcctgcgtta tcccctgatt ctgtggataa ccgtattacc gctagcatgg atctcgggga
     2761 cgtctaacta ctaagcgaga gtagggaact gccaggcatc aaataaaacg aaaggctcag
     2821 tcggaagact gggcctttcg ttttatctgt tgtttgtcgg tgaacgctct cctgagtagg
     2881 acaaatccgc cgggagcgga tttgaacgtt gtgaagcaac ggcccggagg gtggcgggca
     2941 ggacgcccgc cataaactgc caggcatcaa actaagcaga aggccatcct gacggatggc
     3001 ctttttgcgt ttctacaaac tcttcctgtt agttagttac ttaagctcgg gcccacctgc
     3061 agtctactta ctcggatcgg gcctgttagt tagttactta agctcgggcc cacctgcagt
     3121 ctactgggaa ccaattcagt cgactggatc cggtactact attcctcgag tttactccct
     3181 atcagtgata gagaacgtat gaagagttta ctccctatca gtgatagaga acgtatgcag
     3241 actttactcc ctatcagtga tagagaacgt ataaggagtt tactccctat cagtgataga
     3301 gaacgtatga ccagtttact ccctatcagt gatagagaac gtatctacag tttactccct
     3361 atcagtgata gagaacgtat atccagttta ctccctatca gtgatagaga acgtataagc
     3421 tttaggcgtg tacggtgggc gcctataaaa gcagagctcg tttagtgaac cgtcagatcg
     3481 cctggagcaa ttccacaaca cttttgtctt ataccaactt tccgtaccac ttcctaccct
     3541 cgtaaactct ggatcaaatc cgaataactt cgtatagcat acattatacg aagttatctc
     3601 gagtcgctcg gtacgattta aattgaattc tgcagtcgac ggtaccgcgg gcccgggact
     3661 gccggcatca ccggcgccac aggtgcggtt gctggcgcct atatcgccga catcaccgat
     3721 ggggaagatc gggctcgcca cttcgggctc atgagcgctt gtttcggcgt gggtatggtg
     3781 gcaggccccg tggccggggg actgttgggc gccatctcct tgcatgcacc attccttgcg
     3841 gcggcggtgc tcaacggcct caacctacta ctgggctgct tcctaatgca ggagtcgcat
     3901 aagggagagc gtcgaatggt gcactctcag tacaatctgc tctgatgccg catagttaag
     3961 ccagccccga cacccgccaa cacccgctga cgcgccctga cgggcttgtc tgctcccggc
     4021 atccgcttac agacaagctg tgaccgtctc cgggagctgc atgtgtcaga ggttttcacc
     4081 gtcatcaccg aaacgcgcga gacgaaaggg cctcgtgata cgcctatttt tataggttaa
     4141 tgtcatgata ataatggttt cttagacgtc aggtggcact tttcggggaa atgtgcgcat
     4201 aaataacttc gtatagcata cattatacga agttatcgga tttgatctga tcaagcaa
//
