LOCUS       sequence-176278-        4713 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Hybrid reporter construct with 6 transcription factor binding sites 
            (2 UAS, 4 TetO) driving EYFP.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4713)
  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 4713)
  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: pU2T4-YFP
FEATURES             Location/Qualifiers
     source          1..4713
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     296..315
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     polyA_signal    314..369
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     415..434
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     intron          570..1142
                     /label=beta-globin intron
                     /note="intron from rabbit beta-globin gene"
     protein_bind    1183..1201
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    1202..1220
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    1222..1240
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    1241..1259
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     primer_bind     1275..1299
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     CDS             join(1407..1409,1410..1412,1413..2126)
                     /codon_start=1
                     /product="enhanced YFP"
                     /label=EYFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTFGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(1452..1473)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1713..1732)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2060..2081
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    2151..2285
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2175..2194)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2229..2248
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(2648..2666)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        2734..2838
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2839..2907,2908..3699)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(3057..3076)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3870..4458
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4359..4378
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4612..4629
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 gaagagggac agctatgact gggagtagtc aggagaggag gaaaaatctg gctagtaaaa
       61 catgtaagga aaattttagg gatgttaaag aaaaaaataa cacaaaacaa aatataaaaa
      121 aaatctaacc tcaagtcaag gcttttctat ggaataagga atggacagca gggggctgtt
      181 tcatatactg atgacctctt tatagccaac ctttgttcat ggcagccagc atatgggcat
      241 atgttgccaa actctaaacc aaatactcat tctgatgttt taaatgattt gccctcccat
      301 atgtccttcc gagtgagaga cacaaaaaat tccaacacac tattgcaatg aaaataaatt
      361 tcctttatta gccagaagtc agatgctcaa ggggcttcat gatgtcccca taatttttgg
      421 cagagggaaa aagatcgatc tcagtggtat ttgtgagcca gggcattggc cacaccagcc
      481 accaccttct gataggcagc ctgcacctga ggagtgaatt ctttgccaaa atgatgagac
      541 agcacaacaa ccagcacgtt gcccaggagc tgtaggaaag agaagaaggc atgaacatgg
      601 ttagcagagg ggcccggttt ggactcagag tattttatcc tcatctcaaa cagtgtatat
      661 cattgtaacc ataaagagaa aggcaggatg atgaccaggg tgtagttgtt tctaccaata
      721 agaatatttc cacgccagcc agaatttata tgcagaaata ttctacctta tcatttaatt
      781 ataacaattg ttctctaaaa ctgtgctgaa gtacaatata atataccctg attgccttga
      841 aaaaaaagtg attagagaaa gtacttacaa tctgacaaat aaacaaaagt gaatttaaaa
      901 attcgttaca aatgcaagct aaagtttaac gaaaaagtta cagaaaatga aaagaaaata
      961 agaggagaca atggttgtca acagagtaga aagtgaaaga aacaaaatta tcatgagggt
     1021 ccatggtgat acaagggaca tcttcccatt ctaaacaaca ccctgaaaac tttgccccct
     1081 ccatataaca tgaattttac aatagcgaaa aagaaagaac aatcaagggt ccccaaactc
     1141 acaccggtcg gaggacagta ctccgcggag gacagtactc cgtctctatc actgataggg
     1201 atctctatca ctgataggga gtctctatca ctgataggga tctctatcac tgatagggag
     1261 gtacctataa gcagagctcg tttagtgaac cgtcagatcg cctggagacg ccatccacgc
     1321 tgttttgacc tccatagaag acaccgggac cgatccagcc tgcctgaagt tctcagctct
     1381 agagatatcg tcgaggccgc gctagtatgg tgagcaaggg cgaggagctg ttcaccgggg
     1441 tggtgcccat cctggtcgag ctggacggcg acgtaaacgg ccacaagttc agcgtgtccg
     1501 gcgagggcga gggcgatgcc acctacggca agctgaccct gaagttcatc tgcaccaccg
     1561 gcaagctgcc cgtgccctgg cccaccctcg tgaccacctt cggctacggc ctgcagtgct
     1621 tcgcccgcta ccccgaccac atgaagcagc acgacttctt caagtccgcc atgcccgaag
     1681 gctacgtcca ggagcgcacc atcttcttca aggacgacgg caactacaag acccgcgccg
     1741 aggtgaagtt cgagggcgac accctggtga accgcatcga gctgaagggc atcgacttca
     1801 aggaggacgg caacatcctg gggcacaagc tggagtacaa ctacaacagc cacaacgtct
     1861 atatcatggc cgacaagcag aagaacggca tcaaggtgaa cttcaagatc cgccacaaca
     1921 tcgaggacgg cagcgtgcag ctcgccgacc actaccagca gaacaccccc atcggcgacg
     1981 gccccgtgct gctgcccgac aaccactacc tgagctacca gtccgccctg agcaaagacc
     2041 ccaacgagaa gcgcgatcac atggtcctgc tggagttcgt gaccgccgcc gggatcactc
     2101 tcggcatgga cgagctgtac aagtaaaagc ttgtcgacct agttctagag gatccagaca
     2161 tgataagata cattgatgag tttggacaaa ccacaactag aatgcagtga aaaaaatgct
     2221 ttatttgtga aatttgtgat gctattgctt tatttgtaac cattataagc tgcaataaac
     2281 aagttaacaa caacaattgc attcatttta tgtttcaggt tcagggggag gtgtgggagg
     2341 ttttttaaag caagtaaaac ctctacaaat gtggtatggc tgattatgat cctgcaagcc
     2401 tcgtcgtcct ggccggacca cgctatctgt gcaaggtccc cggccccgga cgcgcgctcc
     2461 atgagcagag cgcccgccgc cgaggcgaag actcgggcgg cgccctgccc gtcccaccag
     2521 gtcaacaggc ggtaaccggc ctcttcatcg ggaatgcgcg cgaccttcag catcgccggc
     2581 atgtccccct ggcggacggg aagtatccag ctcgaccaag ctggccgcgt cgagtgaaga
     2641 cgaaagggcc tcgtgatacg cctattttta taggttaatg tcatgataat aatggtttct
     2701 tagacgtcag gtggcacttt tcggggaaat gtgcgcggaa cccctatttg tttatttttc
     2761 taaatacatt caaatatgta tccgctcatg agacaataac cctgataaat gcttcaataa
     2821 tattgaaaaa ggaagagtat gagtattcaa catttccgtg tcgcccttat tccctttttt
     2881 gcggcatttt gccttcctgt ttttgctcac ccagaaacgc tggtgaaagt aaaagatgct
     2941 gaagatcagt tgggtgcacg agtgggttac atcgaactgg atctcaacag cggtaagatc
     3001 cttgagagtt ttcgccccga agaacgtttt ccaatgatga gcacttttaa agttctgcta
     3061 tgtggcgcgg tattatcccg tgttgacgcc gggcaagagc aactcggtcg ccgcatacac
     3121 tattctcaga atgacttggt tgagtactca ccagtcacag aaaagcatct tacggatggc
     3181 atgacagtaa gagaattatg cagtgctgcc ataaccatga gtgataacac tgcggccaac
     3241 ttacttctga caacgatcgg aggaccgaag gagctaaccg cttttttgca caacatgggg
     3301 gatcatgtaa ctcgccttga tcgttgggaa ccggagctga atgaagccat accaaacgac
     3361 gagcgtgaca ccacgatgcc tgtagcaatg gcaacaacgt tgcgcaaact attaactggc
     3421 gaactactta ctctagcttc ccggcaacaa ttaatagact ggatggaggc ggataaagtt
     3481 gcaggaccac ttctgcgctc ggcccttccg gctggctggt ttattgctga taaatctgga
     3541 gccggtgagc gtgggtctcg cggtatcatt gcagcactgg ggccagatgg taagccctcc
     3601 cgtatcgtag ttatctacac gacggggagt caggcaacta tggatgaacg aaatagacag
     3661 atcgctgaga taggtgcctc actgattaag cattggtaac tgtcagacca agtttactca
     3721 tatatacttt agattgattt aaaacttcat ttttaattta aaaggatcta ggtgaagatc
     3781 ctttttgata atctcatgac caaaatccct taacgtgagt tttcgttcca ctgagcgtca
     3841 gaccccgtag aaaagatcaa aggatcttct tgagatcctt tttttctgcg cgtaatctgc
     3901 tgcttgcaaa caaaaaaacc accgctacca gcggtggttt gtttgccgga tcaagagcta
     3961 ccaactcttt ttccgaaggt aactggcttc agcagagcgc agataccaaa tactgttctt
     4021 ctagtgtagc cgtagttagg ccaccacttc aagaactctg tagcaccgcc tacatacctc
     4081 gctctgctaa tcctgttacc agtggctgct gccagtggcg ataagtcgtg tcttaccggg
     4141 ttggactcaa gacgatagtt accggataag gcgcagcggt cgggctgaac ggggggttcg
     4201 tgcacacagc ccagcttgga gcgaacgacc tacaccgaac tgagatacct acagcgtgag
     4261 ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc
     4321 agggtcggaa caggagagcg cacgagggag cttccagggg gaaacgcctg gtatctttat
     4381 agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg
     4441 gggcggagcc tatggaaaaa cgccagcaac gcggcctttt tacggttcct ggccttttgc
     4501 tggccttttg ctcacatgtt ctttcctgcg ttatcccctg attctgtgga taaccgtatt
     4561 accgcctttg agtgagctga taccgctcgc cgcagccgaa cgaccgagcg cagcgagtca
     4621 gtgagcgagg aagcggaaga gcgcccaata cgcaaaccgc ctctccccgc gcgttggccg
     4681 attcattaat gcagtcgagg gatctccata aga
//
