LOCUS       sequence-169173-        4662 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Hybrid reporter construct with 4 transcription factor binding sites 
            (2 TetO, 2 UAS) driving EYFP.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4662)
  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 4662)
  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: pT2U2-YFP
FEATURES             Location/Qualifiers
     source          1..4662
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(64..88)
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     protein_bind    139..157
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    158..176
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     intron          183..755
                     /label=beta-globin intron
                     /note="intron from rabbit beta-globin gene"
     primer_bind     complement(891..910)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    956..1011
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(1010..1029)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(1402..1419)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1573..2161)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1653..1672)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2332..3123,3124..3192))
                     /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     2955..2974
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3193..3297)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     3365..3383
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     polyA_signal    3728..3862
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3765..3784)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3819..3838
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     CDS             complement(join(3899..4612,4613..4615,4616..4618))
                     /codon_start=1
                     /product="enhanced YFP"
                     /label=EYFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTFGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(3944..3965)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     4293..4312
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
ORIGIN
        1 caggctggat cggtcccggt gtcttctatg gaggtcaaaa cagcgtggat ggcgtctcca
       61 ggcgatctga cggttcacta aacgagctct gcttataggg tacccggagt actgtcctcc
      121 gcggagtact gtcctccgtc cctatcagtg atagagatcc ctatcagtga tagagaaccg
      181 gtgtgagttt ggggaccctt gattgttctt tctttttcgc tattgaaaaa ttcatgttat
      241 atggaggggg caaagttttc agggtgttgt ttagaatggg aagatgtccc ttgtatcacc
      301 atggaccctc atgataattt tgtttctttc actttctact ctgttgacaa ccattgtctc
      361 ctcttatttt cttttcattt tctgtaactt ttttcgttaa actttagctt gcatttgtaa
      421 cgaattttta aattcacttt cgtttatttg tcagattgta agtactttct ctaatcactt
      481 ttttttcaag gcaatcaggg taattatatt gtacttcagc acagttttag agaacaattg
      541 ttataattaa atgataaggt agaatatttc tgcatataaa ttctggctgg cgtggaaata
      601 ttcttattgg tagaaacaac tacatcctgg taatcatcct gcctttctct ttatggttac
      661 aatgatatac actgtttgag atgaggataa aatactctga gtccaaaccg ggcccctctg
      721 ctaaccatgt tcatgccttc ttctttttcc tacagctcct gggcaacgtg ctggttgttg
      781 tgctgtctca tcattttggc aaagaattca ctcctcaggt gcaggctgcc tatcagaagg
      841 tggtggctgg tgtggccaat gccctggctc acaaatacca ctgagatcga tctttttccc
      901 tctgccaaaa attatgggga catcatgaag ccccttgagc atctgacttc tgggtaataa
      961 aggaaattta ttttcattgc aatagtgtgt gggaattttt tgtgtctctc actcggaagg
     1021 acatatggga gggcaaatca tttaaaacat cagaatgagt atttggttta gagtttggca
     1081 acatatgcca tatgctggct gccatgaaca aaggtggcta taaagaggtc atcagtatat
     1141 gaaacagccc cctgctgtcc attccttatt ccatagaaaa gccttgactt gaggttagat
     1201 tttttttata ttttgttttg tgttattttt ttctttaaca tccctaaaat tttccttaca
     1261 tgttttacta gccagatttt tcctcctctc ctgactactc ccagtcatag ctgtccctct
     1321 tctcttatga actcgactgc attaatgaat cggccaacgc gcggggagag gcggtttgcg
     1381 tattgggcgc tcttccgctt cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg
     1441 gcgagcggta tcagctcact caaaggcggt aatacggtta tccacagaat caggggataa
     1501 cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc
     1561 gttgctggcg tttttccata ggctccgccc ccctgacgag catcacaaaa atcgacgctc
     1621 aagtcagagg tggcgaaacc cgacaggact ataaagatac caggcgtttc cccctggaag
     1681 ctccctcgtg cgctctcctg ttccgaccct gccgcttacc ggatacctgt ccgcctttct
     1741 cccttcggga agcgtggcgc tttctcaatg ctcacgctgt aggtatctca gttcggtgta
     1801 ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc gttcagcccg accgctgcgc
     1861 cttatccggt aactatcgtc ttgagtccaa cccggtaaga cacgacttat cgccactggc
     1921 agcagccact ggtaacagga ttagcagagc gaggtatgta ggcggtgcta cagagttctt
     1981 gaagtggtgg cctaactacg gctacactag aaggacagta tttggtatct gcgctctgct
     2041 gaagccagtt accttcggaa aaagagttgg tagctcttga tccggcaaac aaaccaccgc
     2101 tggtagcggt ggtttttttg tttgcaagca gcagattacg cgcagaaaaa aaggatctca
     2161 agaagatcct ttgatctttt ctacggggtc tgacgctcag tggaacgaaa actcacgtta
     2221 agggattttg gtcatgagat tatcaaaaag gatcttcacc tagatccttt taaattaaaa
     2281 atgaagtttt aaatcaatct aaagtatata tgagtaaact tggtctgaca gttaccaatg
     2341 cttaatcagt gaggcaccta tctcagcgat ctgtctattt cgttcatcca tagttgcctg
     2401 actccccgtc gtgtagataa ctacgatacg ggagggctta ccatctggcc ccagtgctgc
     2461 aatgataccg cgagacccac gctcaccggc tccagattta tcagcaataa accagccagc
     2521 cggaagggcc gagcgcagaa gtggtcctgc aactttatcc gcctccatcc agtctattaa
     2581 ttgttgccgg gaagctagag taagtagttc gccagttaat agtttgcgca acgttgttgc
     2641 cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat tcagctccgg
     2701 ttcccaacga tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag cggttagctc
     2761 cttcggtcct ccgatcgttg tcagaagtaa gttggccgca gtgttatcac tcatggttat
     2821 ggcagcactg cataattctc ttactgtcat gccatccgta agatgctttt ctgtgactgg
     2881 tgagtactca accaagtcat tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc
     2941 ggcgtcaata cgggataata ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg
     3001 aaaacgttct tcggggcgaa aactctcaag gatcttaccg ctgttgagat ccagttcgat
     3061 gtaacccact cgtgcaccca actgatcttc agcatctttt actttcacca gcgtttctgg
     3121 gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga ataagggcga cacggaaatg
     3181 ttgaatactc atactcttcc tttttcaata ttattgaagc atttatcagg gttattgtct
     3241 catgagcgga tacatatttg aatgtattta gaaaaataaa caaatagggg ttccgcgcac
     3301 atttccccga aaagtgccac ctgacgtcta agaaaccatt attatcatga cattaaccta
     3361 taaaaatagg cgtatcacga ggccctttcg tcttcactcg acgctggtcg agctgatact
     3421 tcccgtccgc caggggacat gccggcgatg ctgaaggtcg cgcgcattcc cgatgaagag
     3481 gccggttacc gcctgttgac ctggtgggac gggcagggcg ccgcccgagt cttgcctcgg
     3541 cggcgggcgc tctgctcatg gagcgcgcgt ccggggacct tgcacagata gcgtggtccg
     3601 gccagacgac gaggcttgca ggatcataat cagccatacc acatttgtag aggttttact
     3661 tgctttaaaa aacctcccac acctccccct gaacctgaaa cataaaatga atgcaattgt
     3721 tgttgttaac ttgtttattg cagcttataa tggttacaaa taaagcaata gcatcacaaa
     3781 tttcacaaat aaagcatttt tttcactgca ttctagttgt ggtttgtcca aactcatcaa
     3841 tgtatcttat catgtctgga tcctctagaa ctaggtcgac aagcttgata tcgaattctt
     3901 acttgtacag ctcgtccatg ccgagagtga tcccggcggc ggtcacgaac tccagcagga
     3961 ccatgtgatc gcgcttctcg ttggggtctt tgctcagggc ggactggtag ctcaggtagt
     4021 ggttgtcggg cagcagcacg gggccgtcgc cgatgggggt gttctgctgg tagtggtcgg
     4081 cgagctgcac gctgccgtcc tcgatgttgt ggcggatctt gaagttcacc ttgatgccgt
     4141 tcttctgctt gtcggccatg atatagacgt tgtggctgtt gtagttgtac tccagcttgt
     4201 gccccaggat gttgccgtcc tccttgaagt cgatgccctt cagctcgatg cggttcacca
     4261 gggtgtcgcc ctcgaacttc acctcggcgc gggtcttgta gttgccgtcg tccttgaaga
     4321 agatggtgcg ctcctggacg tagccttcgg gcatggcgga cttgaagaag tcgtgctgct
     4381 tcatgtggtc ggggtagcgg gcgaagcact gcaggccgta gccgaaggtg gtcacgaggg
     4441 tgggccaggg cacgggcagc ttgccggtgg tgcagatgaa cttcagggtc agcttgccgt
     4501 aggtggcatc gccctcgccc tcgccggaca cgctgaactt gtggccgttt acgtcgccgt
     4561 ccagctcgac caggatgggc accaccccgg tgaacagctc ctcgcccttg ctcaccatac
     4621 tagcgcggcc tcgacgatat ctctagagct gagaacttca gg
//
