LOCUS       sequence-491499-        3791 bp    DNA     circular SYN 04-NOV-2025
DEFINITION  203-bp TET-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 3791)
  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 3791)
  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-TET-DIAL-YB-TATA)-mGL-bGH_pKG3015
FEATURES             Location/Qualifiers
     source          1..3791
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(107..126)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             118..927
                     /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(185..204)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      1017..1605
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1506..1525
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     terminator      1770..1856
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      1948..1975
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     primer_bind     1983..2006
                     /label=pENTR-F
                     /note="pENTR vectors, forward primer"
     promoter        2081..2459
                     /label=TRE3G promoter
                     /note="3rd-generation Tet-responsive promoter that can be 
                     activated by binding of Tet-On(R) 3G"
     protein_bind    2089..2107
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    2125..2143
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    2161..2179
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    2197..2215
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    2233..2251
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    2269..2287
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    2305..2323
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     primer_bind     2368..2392
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     protein_bind    complement(2477..2510)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     primer_bind     2647..2666
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     protein_bind    complement(2714..2747)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     regulatory      2830..2839
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(2836..2838,2839..2841,2842..3555)
                     /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(2881..2902)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     polyA_signal    3567..3791
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
ORIGIN
        1 cgctgactgc aggtgcccgt gtctcaaaat ctctgatgtt acattgcaca agataaaaat
       61 atatcatcat gaacaataaa actgtctgct tacataaaca gtaatacaag gggtgttatg
      121 agccatattc aacgggaaac gtcgaggccg cgattaaatt ccaacatgga tgctgattta
      181 tatgggtata aatgggctcg cgataatgtc gggcaatcag gtgcgacaat ctatcgcttg
      241 tatgggaagc ccgatgcgcc agagttgttt ctgaaacatg gcaaaggtag cgttgccaat
      301 gatgttacag atgagatggt cagactaaac tggctgacgg aatttatgcc tcttccgacc
      361 atcaagcatt ttatccgtac tcctggtgat gcatggttac tcaccactgc gatccccgga
      421 aaaacagcat tccaggtatt agaagaatat cctgattcag gtgaaaatat tgttgatgcg
      481 ctggcagtgt tcctgcgccg gttgcattcg attcctgttt gtaattgtcc ttttaacagc
      541 gatcgcgtat ttcgtctagc tcaggcgcaa tcacgaatga ataacggttt ggttgatgcg
      601 agtgattttg atgacgagcg taatggctgg cctgttgaac aagtctggaa agaaatgcat
      661 aaacttttgc cattctcacc ggattcagtc gtcactcatg gtgatttctc acttgataac
      721 cttatttttg acgaggggaa attaataggt tgtattgatg ttggacgagt cggaatcgca
      781 gaccgatacc aggatcttgc catcctatgg aactgcctcg gtgagttttc tccttcatta
      841 cagaaacggc tttttcaaaa atatggtatt gataatcctg atatgaataa attgcagttt
      901 catttgatgc tcgatgagtt tttctaatca gaattggtta attggttgta acattattca
      961 gattgggccc cgttccactg agcgtcagac cccgtagaaa agatcaaagg atcttcttga
     1021 gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg
     1081 gtggtttgtt tgccggatca agagctacca actctttttc cgaaggtaac tggcttcagc
     1141 agagcgcaga taccaaatac tgttcttcta gtgtagccgt agttaggcca ccacttcaag
     1201 aactctgtag caccgcctac atacctcgct ctgctaatcc tgttaccagt ggctgctgcc
     1261 agtggcgata agtcgtgtct taccgggttg gactcaagac gatagttacc ggataaggcg
     1321 cagcggtcgg gctgaacggg gggttcgtgc acacagccca gcttggagcg aacgacctac
     1381 accgaactga gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga
     1441 aaggcggaca ggtatccggt aagcggcagg gtcggaacag gagagcgcac gagggagctt
     1501 ccagggggaa acgcctggta tctttatagt cctgtcgggt ttcgccacct ctgacttgag
     1561 cgtcgatttt tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg
     1621 gcctttttac ggttcctggc cttttgctgg ccttttgctc acatgttctt tcctgcgtta
     1681 tcccctgatt ctgtggataa ccgtattacc gctagcatgg atctcgggga cgtctaacta
     1741 ctaagcgaga gtagggaact gccaggcatc aaataaaacg aaaggctcag tcggaagact
     1801 gggcctttcg ttttatctgt tgtttgtcgg tgaacgctct cctgagtagg acaaatccgc
     1861 cgggagcgga tttgaacgtt gtgaagcaac ggcccggagg gtggcgggca ggacgcccgc
     1921 cataaactgc caggcatcaa actaagcaga aggccatcct gacggatggc ctttttgcgt
     1981 ttctacaaac tcttcctgtt agttagttac ttaagctcgg gcccacctgc agtctactgg
     2041 gaaccaattc agtcgactgg atccggtact actattcctc gagtttactc cctatcagtg
     2101 atagagaacg tatgaagagt ttactcccta tcagtgatag agaacgtatg cagactttac
     2161 tccctatcag tgatagagaa cgtataagga gtttactccc tatcagtgat agagaacgta
     2221 tgaccagttt actccctatc agtgatagag aacgtatcta cagtttactc cctatcagtg
     2281 atagagaacg tatatccagt ttactcccta tcagtgatag agaacgtata agctttaggc
     2341 gtgtacggtg ggcgcctata aaagcagagc tcgtttagtg aaccgtcaga tcgcctggag
     2401 caattccaca acacttttgt cttataccaa ctttccgtac cacttcctac cctcgtaaac
     2461 tctggatcaa atccgaataa cttcgtatag catacattat acgaagttat ctcgagtcgc
     2521 tcggtacgat ttaaattgaa ttctgcagtc gacggtaccg cgggcccggg actttgtaca
     2581 aaaaagcagg ctccaccatg ggaaccaatt cagtcgactg gatccggtac ctcctacagc
     2641 tcctgggcaa cgtgctggtt attgtgctgt ctcatcattt tggcaaagaa ttgattaatt
     2701 cgagcgaacg cgtataactt cgtatagcat acattatacg aagttatcgg atttgatctg
     2761 atcaagcaac tctagagggt atataatggg ggccactagt ctactaccag agctcatcgc
     2821 tagcaatgag ccaccatggt gagcaagggc gaggagctgt tcaccggggt ggtgcccatc
     2881 ctggtcgagc tggacggcga cgtaaacggc cacaagttca gcgtccgcgg cgagggcgag
     2941 ggcgatgcca ccaacggcaa gctgaccctg aagttcatct gcaccaccgg caagctgccc
     3001 gtgccctggc ccaccctcgt gaccacctta ggctacggcg tggcctgctt cgcccgctac
     3061 cccgaccaca tgaagcagca cgacttcttc aagtccgcca tgcccgaagg ctacgtccag
     3121 gagcgcacca tctctttcaa ggacgacggt acctacaaga cccgcgccga ggtgaagttc
     3181 gagggcgaca ccctggtgaa ccgcatcgtg ctgaagggca tcgacttcaa ggaggacggc
     3241 aacatcctgg ggcacaagct ggagtacaac ttcaacagcc acaaggtcta tatcacggcc
     3301 gacaagcaga agaacggcat caaggctaac ttcaagaccc gccacaacgt tgaggacggc
     3361 ggcgtgcagc tcgccgacca ctaccagcag aacaccccca tcggcgacgg ccccgtgctg
     3421 ctgcccgaca accactacct gagccatcag tccaaactga gcaaagaccc caacgagaag
     3481 cgcgatcaca tggtcctgaa ggagagggtg accgccgccg ggattacaca tgacatggac
     3541 gagctgtaca agtaacaacg ttgcacctgt gccttctagt tgccagccat ctgttgtttg
     3601 cccctccccc gtgccttcct tgaccctgga aggtgccact cccactgtcc tttcctaata
     3661 aaatgaggaa attgcatcgc attgtctgag taggtgtcat tctattctgg ggggtggggt
     3721 ggggcaggac agcaaggggg aggattggga agacaatagc aggcatgctg gggatgcggt
     3781 gggctctatg g
//
