LOCUS       sequence-169187-        4688 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Hybrid reporter construct with 6 transcription factor binding sites 
            (2 TetO, 2 UAS, 2 TetO) driving EYFP.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4688)
  AUTHORS   Hartfield RM, Schwarz KA, Muldoon JJ, Bagheri N, Leonard JN
  TITLE     Multiplexing Engineered Receptors for Multiparametric Evaluation of 
            Environmental Ligands.
  JOURNAL   ACS Synth Biol. 2017 Nov 17;6(11):2042-2055. doi: 
            10.1021/acssynbio.6b00279. Epub 2017 Aug 23.
  PUBMED    28771312
REFERENCE   2  (bases 1 to 4688)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; doi: 
            "10.1021/acssynbio.6b00279"; journalName: "ACS Synth Biol"; date: 
            "2017-11-17- 17"; volume: "6"; issue: "11"; pages: "2042-2055"
COMMENT     Sequence Label: pT2U2T2-YFP
FEATURES             Location/Qualifiers
     source          1..4688
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     intron          1..573
                     /label=beta-globin intron
                     /note="intron from rabbit beta-globin gene"
     primer_bind     complement(709..728)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    774..829
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(828..847)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(1220..1237)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1391..1979)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1471..1490)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2150..2941,2942..3010))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     2773..2792
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3011..3115)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     3183..3201
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     polyA_signal    3546..3680
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3583..3602)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3637..3656
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     CDS             complement(join(3705..4418,4419..4421,4422..4424))
                     /codon_start=1
                     /product="enhanced YFP"
                     /label=EYFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTFGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(3750..3771)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     4099..4118
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     complement(4532..4556)
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     protein_bind    4572..4590
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    4591..4609
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    4645..4663
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    4664..4682
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
ORIGIN
        1 gtgagtttgg ggacccttga ttgttctttc tttttcgcta ttgaaaaatt catgttatat
       61 ggagggggca aagttttcag ggtgttgttt agaatgggaa gatgtccctt gtatcaccat
      121 ggaccctcat gataattttg tttctttcac tttctactct gttgacaacc attgtctcct
      181 cttattttct tttcattttc tgtaactttt ttcgttaaac tttagcttgc atttgtaacg
      241 aatttttaaa ttcactttcg tttatttgtc agattgtaag tactttctct aatcactttt
      301 ttttcaaggc aatcagggta attatattgt acttcagcac agttttagag aacaattgtt
      361 ataattaaat gataaggtag aatatttctg catataaatt ctggctggcg tggaaatatt
      421 cttattggta gaaacaacta catcctggta atcatcctgc ctttctcttt atggttacaa
      481 tgatatacac tgtttgagat gaggataaaa tactctgagt ccaaaccggg cccctctgct
      541 aaccatgttc atgccttctt ctttttccta cagctcctgg gcaacgtgct ggttgttgtg
      601 ctgtctcatc attttggcaa agaattcact cctcaggtgc aggctgccta tcagaaggtg
      661 gtggctggtg tggccaatgc cctggctcac aaataccact gagatcgatc tttttccctc
      721 tgccaaaaat tatggggaca tcatgaagcc ccttgagcat ctgacttctg ggtaataaag
      781 gaaatttatt ttcattgcaa tagtgtgtgg gaattttttg tgtctctcac tcggaaggac
      841 atatgggagg gcaaatcatt taaaacatca gaatgagtat ttggtttaga gtttggcaac
      901 atatgccata tgctggctgc catgaacaaa ggtggctata aagaggtcat cagtatatga
      961 aacagccccc tgctgtccat tccttattcc atagaaaagc cttgacttga ggttagattt
     1021 tttttatatt ttgttttgtg ttattttttt ctttaacatc cctaaaattt tccttacatg
     1081 ttttactagc cagatttttc ctcctctcct gactactccc agtcatagct gtccctcttc
     1141 tcttatgaac tcgactgcat taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta
     1201 ttgggcgctc ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc
     1261 gagcggtatc agctcactca aaggcggtaa tacggttatc cacagaatca ggggataacg
     1321 caggaaagaa catgtgagca aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt
     1381 tgctggcgtt tttccatagg ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa
     1441 gtcagaggtg gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggaagct
     1501 ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc
     1561 cttcgggaag cgtggcgctt tctcaatgct cacgctgtag gtatctcagt tcggtgtagg
     1621 tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct
     1681 tatccggtaa ctatcgtctt gagtccaacc cggtaagaca cgacttatcg ccactggcag
     1741 cagccactgg taacaggatt agcagagcga ggtatgtagg cggtgctaca gagttcttga
     1801 agtggtggcc taactacggc tacactagaa ggacagtatt tggtatctgc gctctgctga
     1861 agccagttac cttcggaaaa agagttggta gctcttgatc cggcaaacaa accaccgctg
     1921 gtagcggtgg tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag
     1981 aagatccttt gatcttttct acggggtctg acgctcagtg gaacgaaaac tcacgttaag
     2041 ggattttggt catgagatta tcaaaaagga tcttcaccta gatcctttta aattaaaaat
     2101 gaagttttaa atcaatctaa agtatatatg agtaaacttg gtctgacagt taccaatgct
     2161 taatcagtga ggcacctatc tcagcgatct gtctatttcg ttcatccata gttgcctgac
     2221 tccccgtcgt gtagataact acgatacggg agggcttacc atctggcccc agtgctgcaa
     2281 tgataccgcg agacccacgc tcaccggctc cagatttatc agcaataaac cagccagccg
     2341 gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag tctattaatt
     2401 gttgccggga agctagagta agtagttcgc cagttaatag tttgcgcaac gttgttgcca
     2461 ttgctacagg catcgtggtg tcacgctcgt cgtttggtat ggcttcattc agctccggtt
     2521 cccaacgatc aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg gttagctcct
     2581 tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt gttatcactc atggttatgg
     2641 cagcactgca taattctctt actgtcatgc catccgtaag atgcttttct gtgactggtg
     2701 agtactcaac caagtcattc tgagaatagt gtatgcggcg accgagttgc tcttgcccgg
     2761 cgtcaatacg ggataatacc gcgccacata gcagaacttt aaaagtgctc atcattggaa
     2821 aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc agttcgatgt
     2881 aacccactcg tgcacccaac tgatcttcag catcttttac tttcaccagc gtttctgggt
     2941 gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt
     3001 gaatactcat actcttcctt tttcaatatt attgaagcat ttatcagggt tattgtctca
     3061 tgagcggata catatttgaa tgtatttaga aaaataaaca aataggggtt ccgcgcacat
     3121 ttccccgaaa agtgccacct gacgtctaag aaaccattat tatcatgaca ttaacctata
     3181 aaaataggcg tatcacgagg ccctttcgtc ttcactcgac gctggtcgag ctgatacttc
     3241 ccgtccgcca ggggacatgc cggcgatgct gaaggtcgcg cgcattcccg atgaagaggc
     3301 cggttaccgc ctgttgacct ggtgggacgg gcagggcgcc gcccgagtct tgcctcggcg
     3361 gcgggcgctc tgctcatgga gcgcgcgtcc ggggaccttg cacagatagc gtggtccggc
     3421 cagacgacga ggcttgcagg atcataatca gccataccac atttgtagag gttttacttg
     3481 ctttaaaaaa cctcccacac ctccccctga acctgaaaca taaaatgaat gcaattgttg
     3541 ttgttaactt gtttattgca gcttataatg gttacaaata aagcaatagc atcacaaatt
     3601 tcacaaataa agcatttttt tcactgcatt ctagttgtgg tttgtccaaa ctcatcaatg
     3661 tatcttatca tgtctggatc ctctagaact aggtcgacaa gcttttactt gtacagctcg
     3721 tccatgccga gagtgatccc ggcggcggtc acgaactcca gcaggaccat gtgatcgcgc
     3781 ttctcgttgg ggtctttgct cagggcggac tggtagctca ggtagtggtt gtcgggcagc
     3841 agcacggggc cgtcgccgat gggggtgttc tgctggtagt ggtcggcgag ctgcacgctg
     3901 ccgtcctcga tgttgtggcg gatcttgaag ttcaccttga tgccgttctt ctgcttgtcg
     3961 gccatgatat agacgttgtg gctgttgtag ttgtactcca gcttgtgccc caggatgttg
     4021 ccgtcctcct tgaagtcgat gcccttcagc tcgatgcggt tcaccagggt gtcgccctcg
     4081 aacttcacct cggcgcgggt cttgtagttg ccgtcgtcct tgaagaagat ggtgcgctcc
     4141 tggacgtagc cttcgggcat ggcggacttg aagaagtcgt gctgcttcat gtggtcgggg
     4201 tagcgggcga agcactgcag gccgtagccg aaggtggtca cgagggtggg ccagggcacg
     4261 ggcagcttgc cggtggtgca gatgaacttc agggtcagct tgccgtaggt ggcatcgccc
     4321 tcgccctcgc cggacacgct gaacttgtgg ccgtttacgt cgccgtccag ctcgaccagg
     4381 atgggcacca ccccggtgaa cagctcctcg cccttgctca ccatactagc gcggcctcga
     4441 cgatatctct agagctgaga acttcaggca ggctggatcg gtcccggtgt cttctatgga
     4501 ggtcaaaaca gcgtggatgg cgtctccagg cgatctgacg gttcactaaa cgagctctgc
     4561 ttataggtac ctccctatca gtgatagaga tccctatcag tgatagagac cggagtactg
     4621 tcctccgcgg agtactgtcc tccgtcccta tcagtgatag agatccctat cagtgataga
     4681 gaaccggt
//
