LOCUS       sequence-169170-        4661 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Hybrid reporter construct with 4 transcription factor binding sites 
            (2 UAS, 2 TetO) driving EYFP.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4661)
  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 4661)
  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: pU2T2-YFP
FEATURES             Location/Qualifiers
     source          1..4661
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(64..88)
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     protein_bind    104..122
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    123..141
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     intron          182..754
                     /label=beta-globin intron
                     /note="intron from rabbit beta-globin gene"
     primer_bind     complement(890..909)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    955..1010
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(1009..1028)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(1401..1418)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1572..2160)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1652..1671)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2331..3122,3123..3191))
                     /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     2954..2973
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3192..3296)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     3364..3382
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     polyA_signal    3727..3861
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3764..3783)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3818..3837
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     CDS             complement(join(3898..4611,4612..4614,4615..4617))
                     /codon_start=1
                     /product="enhanced YFP"
                     /label=EYFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTFGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(3943..3964)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     4292..4311
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
ORIGIN
        1 caggctggat cggtcccggt gtcttctatg gaggtcaaaa cagcgtggat ggcgtctcca
       61 ggcgatctga cggttcacta aacgagctct gcttataggt acctccctat cagtgataga
      121 gatccctatc agtgatagag acggagtact gtcctccgcg gagtactgtc ctccgaccgg
      181 tgtgagtttg gggacccttg attgttcttt ctttttcgct attgaaaaat tcatgttata
      241 tggagggggc aaagttttca gggtgttgtt tagaatggga agatgtccct tgtatcacca
      301 tggaccctca tgataatttt gtttctttca ctttctactc tgttgacaac cattgtctcc
      361 tcttattttc ttttcatttt ctgtaacttt tttcgttaaa ctttagcttg catttgtaac
      421 gaatttttaa attcactttc gtttatttgt cagattgtaa gtactttctc taatcacttt
      481 tttttcaagg caatcagggt aattatattg tacttcagca cagttttaga gaacaattgt
      541 tataattaaa tgataaggta gaatatttct gcatataaat tctggctggc gtggaaatat
      601 tcttattggt agaaacaact acatcctggt aatcatcctg cctttctctt tatggttaca
      661 atgatataca ctgtttgaga tgaggataaa atactctgag tccaaaccgg gcccctctgc
      721 taaccatgtt catgccttct tctttttcct acagctcctg ggcaacgtgc tggttgttgt
      781 gctgtctcat cattttggca aagaattcac tcctcaggtg caggctgcct atcagaaggt
      841 ggtggctggt gtggccaatg ccctggctca caaataccac tgagatcgat ctttttccct
      901 ctgccaaaaa ttatggggac atcatgaagc cccttgagca tctgacttct gggtaataaa
      961 ggaaatttat tttcattgca atagtgtgtg ggaatttttt gtgtctctca ctcggaagga
     1021 catatgggag ggcaaatcat ttaaaacatc agaatgagta tttggtttag agtttggcaa
     1081 catatgccat atgctggctg ccatgaacaa aggtggctat aaagaggtca tcagtatatg
     1141 aaacagcccc ctgctgtcca ttccttattc catagaaaag ccttgacttg aggttagatt
     1201 ttttttatat tttgttttgt gttatttttt tctttaacat ccctaaaatt ttccttacat
     1261 gttttactag ccagattttt cctcctctcc tgactactcc cagtcatagc tgtccctctt
     1321 ctcttatgaa ctcgactgca ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt
     1381 attgggcgct cttccgcttc ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg
     1441 cgagcggtat cagctcactc aaaggcggta atacggttat ccacagaatc aggggataac
     1501 gcaggaaaga acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg
     1561 ttgctggcgt ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca
     1621 agtcagaggt ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc
     1681 tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc
     1741 ccttcgggaa gcgtggcgct ttctcaatgc tcacgctgta ggtatctcag ttcggtgtag
     1801 gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc
     1861 ttatccggta actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca
     1921 gcagccactg gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg
     1981 aagtggtggc ctaactacgg ctacactaga aggacagtat ttggtatctg cgctctgctg
     2041 aagccagtta ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct
     2101 ggtagcggtg gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa
     2161 gaagatcctt tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa
     2221 gggattttgg tcatgagatt atcaaaaagg atcttcacct agatcctttt aaattaaaaa
     2281 tgaagtttta aatcaatcta aagtatatat gagtaaactt ggtctgacag ttaccaatgc
     2341 ttaatcagtg aggcacctat ctcagcgatc tgtctatttc gttcatccat agttgcctga
     2401 ctccccgtcg tgtagataac tacgatacgg gagggcttac catctggccc cagtgctgca
     2461 atgataccgc gagacccacg ctcaccggct ccagatttat cagcaataaa ccagccagcc
     2521 ggaagggccg agcgcagaag tggtcctgca actttatccg cctccatcca gtctattaat
     2581 tgttgccggg aagctagagt aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc
     2641 attgctacag gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt cagctccggt
     2701 tcccaacgat caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc
     2761 ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag tgttatcact catggttatg
     2821 gcagcactgc ataattctct tactgtcatg ccatccgtaa gatgcttttc tgtgactggt
     2881 gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg
     2941 gcgtcaatac gggataatac cgcgccacat agcagaactt taaaagtgct catcattgga
     3001 aaacgttctt cggggcgaaa actctcaagg atcttaccgc tgttgagatc cagttcgatg
     3061 taacccactc gtgcacccaa ctgatcttca gcatctttta ctttcaccag cgtttctggg
     3121 tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa taagggcgac acggaaatgt
     3181 tgaatactca tactcttcct ttttcaatat tattgaagca tttatcaggg ttattgtctc
     3241 atgagcggat acatatttga atgtatttag aaaaataaac aaataggggt tccgcgcaca
     3301 tttccccgaa aagtgccacc tgacgtctaa gaaaccatta ttatcatgac attaacctat
     3361 aaaaataggc gtatcacgag gccctttcgt cttcactcga cgctggtcga gctgatactt
     3421 cccgtccgcc aggggacatg ccggcgatgc tgaaggtcgc gcgcattccc gatgaagagg
     3481 ccggttaccg cctgttgacc tggtgggacg ggcagggcgc cgcccgagtc ttgcctcggc
     3541 ggcgggcgct ctgctcatgg agcgcgcgtc cggggacctt gcacagatag cgtggtccgg
     3601 ccagacgacg aggcttgcag gatcataatc agccatacca catttgtaga ggttttactt
     3661 gctttaaaaa acctcccaca cctccccctg aacctgaaac ataaaatgaa tgcaattgtt
     3721 gttgttaact tgtttattgc agcttataat ggttacaaat aaagcaatag catcacaaat
     3781 ttcacaaata aagcattttt ttcactgcat tctagttgtg gtttgtccaa actcatcaat
     3841 gtatcttatc atgtctggat cctctagaac taggtcgaca agcttgatat cgaattctta
     3901 cttgtacagc tcgtccatgc cgagagtgat cccggcggcg gtcacgaact ccagcaggac
     3961 catgtgatcg cgcttctcgt tggggtcttt gctcagggcg gactggtagc tcaggtagtg
     4021 gttgtcgggc agcagcacgg ggccgtcgcc gatgggggtg ttctgctggt agtggtcggc
     4081 gagctgcacg ctgccgtcct cgatgttgtg gcggatcttg aagttcacct tgatgccgtt
     4141 cttctgcttg tcggccatga tatagacgtt gtggctgttg tagttgtact ccagcttgtg
     4201 ccccaggatg ttgccgtcct ccttgaagtc gatgcccttc agctcgatgc ggttcaccag
     4261 ggtgtcgccc tcgaacttca cctcggcgcg ggtcttgtag ttgccgtcgt ccttgaagaa
     4321 gatggtgcgc tcctggacgt agccttcggg catggcggac ttgaagaagt cgtgctgctt
     4381 catgtggtcg gggtagcggg cgaagcactg caggccgtag ccgaaggtgg tcacgagggt
     4441 gggccagggc acgggcagct tgccggtggt gcagatgaac ttcagggtca gcttgccgta
     4501 ggtggcatcg ccctcgccct cgccggacac gctgaacttg tggccgttta cgtcgccgtc
     4561 cagctcgacc aggatgggca ccaccccggt gaacagctcc tcgcccttgc tcaccatact
     4621 agcgcggcct cgacgatatc tctagagctg agaacttcag g
//
