LOCUS       sequence-169177-        4679 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Hybrid reporter construct with 5 transcription factor binding sites 
            (2 TetO, 3 UAS) driving EYFP.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4679)
  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 4679)
  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: pT2U3-YFP
FEATURES             Location/Qualifiers
     source          1..4679
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(64..88)
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     protein_bind    156..174
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    175..193
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     intron          200..772
                     /label=beta-globin intron
                     /note="intron from rabbit beta-globin gene"
     primer_bind     complement(908..927)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    973..1028
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(1027..1046)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(1419..1436)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1590..2178)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1670..1689)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2349..3140,3141..3209))
                     /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     2972..2991
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3210..3314)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     3382..3400
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     polyA_signal    3745..3879
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3782..3801)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3836..3855
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     CDS             complement(join(3916..4629,4630..4632,4633..4635))
                     /codon_start=1
                     /product="enhanced YFP"
                     /label=EYFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTFGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(3961..3982)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     4310..4329
                     /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 gtcctccgcg gagtactgtc ctccgtccct atcagtgata gagatcccta
      181 tcagtgatag agaaccggtg tgagtttggg gacccttgat tgttctttct ttttcgctat
      241 tgaaaaattc atgttatatg gagggggcaa agttttcagg gtgttgttta gaatgggaag
      301 atgtcccttg tatcaccatg gaccctcatg ataattttgt ttctttcact ttctactctg
      361 ttgacaacca ttgtctcctc ttattttctt ttcattttct gtaacttttt tcgttaaact
      421 ttagcttgca tttgtaacga atttttaaat tcactttcgt ttatttgtca gattgtaagt
      481 actttctcta atcacttttt tttcaaggca atcagggtaa ttatattgta cttcagcaca
      541 gttttagaga acaattgtta taattaaatg ataaggtaga atatttctgc atataaattc
      601 tggctggcgt ggaaatattc ttattggtag aaacaactac atcctggtaa tcatcctgcc
      661 tttctcttta tggttacaat gatatacact gtttgagatg aggataaaat actctgagtc
      721 caaaccgggc ccctctgcta accatgttca tgccttcttc tttttcctac agctcctggg
      781 caacgtgctg gttgttgtgc tgtctcatca ttttggcaaa gaattcactc ctcaggtgca
      841 ggctgcctat cagaaggtgg tggctggtgt ggccaatgcc ctggctcaca aataccactg
      901 agatcgatct ttttccctct gccaaaaatt atggggacat catgaagccc cttgagcatc
      961 tgacttctgg gtaataaagg aaatttattt tcattgcaat agtgtgtggg aattttttgt
     1021 gtctctcact cggaaggaca tatgggaggg caaatcattt aaaacatcag aatgagtatt
     1081 tggtttagag tttggcaaca tatgccatat gctggctgcc atgaacaaag gtggctataa
     1141 agaggtcatc agtatatgaa acagccccct gctgtccatt ccttattcca tagaaaagcc
     1201 ttgacttgag gttagatttt ttttatattt tgttttgtgt tatttttttc tttaacatcc
     1261 ctaaaatttt ccttacatgt tttactagcc agatttttcc tcctctcctg actactccca
     1321 gtcatagctg tccctcttct cttatgaact cgactgcatt aatgaatcgg ccaacgcgcg
     1381 gggagaggcg gtttgcgtat tgggcgctct tccgcttcct cgctcactga ctcgctgcgc
     1441 tcggtcgttc ggctgcggcg agcggtatca gctcactcaa aggcggtaat acggttatcc
     1501 acagaatcag gggataacgc aggaaagaac atgtgagcaa aaggccagca aaaggccagg
     1561 aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc tccgcccccc tgacgagcat
     1621 cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga caggactata aagataccag
     1681 gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga
     1741 tacctgtccg cctttctccc ttcgggaagc gtggcgcttt ctcaatgctc acgctgtagg
     1801 tatctcagtt cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga accccccgtt
     1861 cagcccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc ggtaagacac
     1921 gacttatcgc cactggcagc agccactggt aacaggatta gcagagcgag gtatgtaggc
     1981 ggtgctacag agttcttgaa gtggtggcct aactacggct acactagaag gacagtattt
     2041 ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa gagttggtag ctcttgatcc
     2101 ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt gcaagcagca gattacgcgc
     2161 agaaaaaaag gatctcaaga agatcctttg atcttttcta cggggtctga cgctcagtgg
     2221 aacgaaaact cacgttaagg gattttggtc atgagattat caaaaaggat cttcacctag
     2281 atccttttaa attaaaaatg aagttttaaa tcaatctaaa gtatatatga gtaaacttgg
     2341 tctgacagtt accaatgctt aatcagtgag gcacctatct cagcgatctg tctatttcgt
     2401 tcatccatag ttgcctgact ccccgtcgtg tagataacta cgatacggga gggcttacca
     2461 tctggcccca gtgctgcaat gataccgcga gacccacgct caccggctcc agatttatca
     2521 gcaataaacc agccagccgg aagggccgag cgcagaagtg gtcctgcaac tttatccgcc
     2581 tccatccagt ctattaattg ttgccgggaa gctagagtaa gtagttcgcc agttaatagt
     2641 ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg
     2701 gcttcattca gctccggttc ccaacgatca aggcgagtta catgatcccc catgttgtgc
     2761 aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg
     2821 ttatcactca tggttatggc agcactgcat aattctctta ctgtcatgcc atccgtaaga
     2881 tgcttttctg tgactggtga gtactcaacc aagtcattct gagaatagtg tatgcggcga
     2941 ccgagttgct cttgcccggc gtcaatacgg gataataccg cgccacatag cagaacttta
     3001 aaagtgctca tcattggaaa acgttcttcg gggcgaaaac tctcaaggat cttaccgctg
     3061 ttgagatcca gttcgatgta acccactcgt gcacccaact gatcttcagc atcttttact
     3121 ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata
     3181 agggcgacac ggaaatgttg aatactcata ctcttccttt ttcaatatta ttgaagcatt
     3241 tatcagggtt attgtctcat gagcggatac atatttgaat gtatttagaa aaataaacaa
     3301 ataggggttc cgcgcacatt tccccgaaaa gtgccacctg acgtctaaga aaccattatt
     3361 atcatgacat taacctataa aaataggcgt atcacgaggc cctttcgtct tcactcgacg
     3421 ctggtcgagc tgatacttcc cgtccgccag gggacatgcc ggcgatgctg aaggtcgcgc
     3481 gcattcccga tgaagaggcc ggttaccgcc tgttgacctg gtgggacggg cagggcgccg
     3541 cccgagtctt gcctcggcgg cgggcgctct gctcatggag cgcgcgtccg gggaccttgc
     3601 acagatagcg tggtccggcc agacgacgag gcttgcagga tcataatcag ccataccaca
     3661 tttgtagagg ttttacttgc tttaaaaaac ctcccacacc tccccctgaa cctgaaacat
     3721 aaaatgaatg caattgttgt tgttaacttg tttattgcag cttataatgg ttacaaataa
     3781 agcaatagca tcacaaattt cacaaataaa gcattttttt cactgcattc tagttgtggt
     3841 ttgtccaaac tcatcaatgt atcttatcat gtctggatcc tctagaacta ggtcgacaag
     3901 cttgatatcg aattcttact tgtacagctc gtccatgccg agagtgatcc cggcggcggt
     3961 cacgaactcc agcaggacca tgtgatcgcg cttctcgttg gggtctttgc tcagggcgga
     4021 ctggtagctc aggtagtggt tgtcgggcag cagcacgggg ccgtcgccga tgggggtgtt
     4081 ctgctggtag tggtcggcga gctgcacgct gccgtcctcg atgttgtggc ggatcttgaa
     4141 gttcaccttg atgccgttct tctgcttgtc ggccatgata tagacgttgt ggctgttgta
     4201 gttgtactcc agcttgtgcc ccaggatgtt gccgtcctcc ttgaagtcga tgcccttcag
     4261 ctcgatgcgg ttcaccaggg tgtcgccctc gaacttcacc tcggcgcggg tcttgtagtt
     4321 gccgtcgtcc ttgaagaaga tggtgcgctc ctggacgtag ccttcgggca tggcggactt
     4381 gaagaagtcg tgctgcttca tgtggtcggg gtagcgggcg aagcactgca ggccgtagcc
     4441 gaaggtggtc acgagggtgg gccagggcac gggcagcttg ccggtggtgc agatgaactt
     4501 cagggtcagc ttgccgtagg tggcatcgcc ctcgccctcg ccggacacgc tgaacttgtg
     4561 gccgtttacg tcgccgtcca gctcgaccag gatgggcacc accccggtga acagctcctc
     4621 gcccttgctc accatactag cgcggcctcg acgatatctc tagagctgag aacttcagg
//
