LOCUS       sequence-491496-        3553 bp    DNA     circular SYN 04-NOV-2025
DEFINITION  loxPcon 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 3553)
  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 3553)
  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: (loxPcon-TET-DIAL-YB-TATA)-mGL-bGH_pKG3016
FEATURES             Location/Qualifiers
     source          1..3553
                     /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(2476..2509)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     regulatory      2592..2601
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(2598..2600,2601..2603,2604..3317)
                     /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(2643..2664)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     polyA_signal    3329..3553
                     /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 cctcgtaaaa
     2461 ttcgagcgaa cgcgtataac ttcgtatagc atacattata cgaagttatc ggatttgatc
     2521 tgatcaagca actctagagg gtatataatg ggggccacta gtctactacc agagctcatc
     2581 gctagcaatg agccaccatg gtgagcaagg gcgaggagct gttcaccggg gtggtgccca
     2641 tcctggtcga gctggacggc gacgtaaacg gccacaagtt cagcgtccgc ggcgagggcg
     2701 agggcgatgc caccaacggc aagctgaccc tgaagttcat ctgcaccacc ggcaagctgc
     2761 ccgtgccctg gcccaccctc gtgaccacct taggctacgg cgtggcctgc ttcgcccgct
     2821 accccgacca catgaagcag cacgacttct tcaagtccgc catgcccgaa ggctacgtcc
     2881 aggagcgcac catctctttc aaggacgacg gtacctacaa gacccgcgcc gaggtgaagt
     2941 tcgagggcga caccctggtg aaccgcatcg tgctgaaggg catcgacttc aaggaggacg
     3001 gcaacatcct ggggcacaag ctggagtaca acttcaacag ccacaaggtc tatatcacgg
     3061 ccgacaagca gaagaacggc atcaaggcta acttcaagac ccgccacaac gttgaggacg
     3121 gcggcgtgca gctcgccgac cactaccagc agaacacccc catcggcgac ggccccgtgc
     3181 tgctgcccga caaccactac ctgagccatc agtccaaact gagcaaagac cccaacgaga
     3241 agcgcgatca catggtcctg aaggagaggg tgaccgccgc cgggattaca catgacatgg
     3301 acgagctgta caagtaacaa cgttgcacct gtgccttcta gttgccagcc atctgttgtt
     3361 tgcccctccc ccgtgccttc cttgaccctg gaaggtgcca ctcccactgt cctttcctaa
     3421 taaaatgagg aaattgcatc gcattgtctg agtaggtgtc attctattct ggggggtggg
     3481 gtggggcagg acagcaaggg ggaggattgg gaagacaata gcaggcatgc tggggatgcg
     3541 gtgggctcta tgg
//
