LOCUS       sequence-359750-        6136 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Fluorescent reporter for AhR activity.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6136)
  AUTHORS   Degrelle SA, Ferecatu I, Fournier T
  TITLE     Novel fluorescent and secreted transcriptional reporters for 
            quantifying activity of the xenobiotic sensor aryl hydrocarbon 
            receptor (AHR).
  JOURNAL   Environ Int. 2022 Sep 24;169:107545. doi: 
            10.1016/j.envint.2022.107545.
  PUBMED    36179647
REFERENCE   2  (bases 1 to 6136)
  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"; journalName: "Environ 
            Int."; date: "2022-09-24"; volume: "169:107545. doi"; pages: " 
            10.1016/j.envint.2022.107545"
COMMENT     Sequence Label: XRE-H2B-mScarlet-I
FEATURES             Location/Qualifiers
     source          1..6136
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      complement(1728..2316)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1808..1827)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     polyA_signal    2645..2692
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     primer_bind     2717..2736
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     CDS             complement(2924..3718)
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     CDS             complement(2924..3718)
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(3035..3054)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     3645..3664
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     promoter        complement(3753..4110)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3767..3902
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     enhancer        3919..4110
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     4067..4087
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        complement(4112..4216)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      4243..4698
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4330..4349)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4540..4561
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     polyA_signal    4705..4826
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4716..4735)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     4770..4789
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     CDS             complement(4952..5647)
                     /codon_start=1
                     /product="bright monomeric red fluorescent protein evolved 
                     from a synthetic template (Bindels et al., 2016)"
                     /label=mScarlet-I
                     /note="matures faster than mScarlet"
                     /translation="MVSKGEAVIKEFMRFKVHMEGSMNGHEFEIEGEGEGRPYEGTQTA
                     KLKVTKGGPLPFSWDILSPQFMYGSRAFIKHPADIPDYYKQSFPEGFKWERVMNFEDGG
                     AVTVTQDTSLEDGTLIYKVKLRGTNFPPDGPVMQKKTMGWEASTERLYPEDGVLKGDIK
                     MALRLKDGGRYLADFKTTYKAKKPVQMPGAYNVDRKLDITSHNEDYTVVEQYERSEGRH
                     STGGMDELYK"
     primer_bind     complement(5227..5246)
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     primer_bind     5496..5514
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     regulatory      5644..5653
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             complement(5666..6043)
                     /codon_start=1
                     /gene="HIST1H2BJ"
                     /product="human histone H2B"
                     /label=H2B
                     /translation="MPEPAKSAPAPKKGSKKAVTKAQKKGGKKRKRSRKESYSIYVYKV
                     LKQVHPDTGISSKAMGIMNSFVNDIFERIAGEASRLAHYNKRSTITSREIQTAVRLLLP
                     GELAKHAVSEGTKAITKYTSAK"
ORIGIN
        1 gatcagggaa ggttccaagg aactgtcacc ttcagggtga gggtgaaggc actgccacct
       61 ttataggcgg gcttgtacgt gtggccacgc ctccttccct ggagcgagct gagatggagg
      121 gagaagcagc ctgaaccggg ctggtctctc tgggattgga gagaaaggtg gcggagggcg
      181 gcgggggtgg ggggtcgggt gggcaagaac ccactatggg acacatctcg gtgcctgtac
      241 atagaggggt gagggctggg aacacctgga agtcccaatt ccaaggcgtc ccaagggcag
      301 tgcagaaccc agccgaggag ggggcttgag ggagaagtcc caggacagcc cgaagagagg
      361 gtacgggaag ctccatcctg gggcgcgggg actcctcttc gtcatttttg cacccactgg
      421 aacgctgggc gtgcagatgc ctccccagcg ctacagccta ccaggactcg gcaggcgggg
      481 gcggggctgc cccgtggtga ccttcttccc ggggttactg agtcccggct cgcgtgagaa
      541 gcgctgcgac cccagccctg aggtcacggg ggccggaagg ccagaggcgc cgcggggctg
      601 gacctgtccc ccagagcccg ggcgactagc tgaggttcgc gcctcttgat tgaaggatcg
      661 gaatggatgg cgaaggggca ggctcgctga gcgctcacta gcggctcctc gcagcctcta
      721 ggatcgcctt agtgctgatt gcaacgcgcg gtccctccag ccaggaccca ccggcccttg
      781 agggtccctc ttggctcccg gggtggctag tgctttgatt ggcagagcac agaaatccgg
      841 cggcgggggg ccacggaaag actcgggccc agggaccacg gagggggcac ctggcgcgga
      901 gcccccaccc tacccccggc tagcttgcgt gcgccggcga catccctcta gggggcagag
      961 gtcaggcggc ccgtccccgc cccacctggc ccggaggcgc ggtgcccagg cgttgcgtga
     1021 gaaggaccgg aggcccgcgc agccacccag ccgacccatt ccccggcccg gcgcgggggc
     1081 tcgcagtcac gcgagggggg agggagtcgg ggcggggctc ttaaagggcc agcctcgcgc
     1141 gcggcgtggg gttggggtgg agggagagga actgtcgaag ggtggcgggt gcgcgattga
     1201 ataaggggat gcggtgcaaa gggtctaggt ctgcgtgtgg cttctgcctg cgaggagagg
     1261 cagcctgcat gtgtccgcat tttcggtcca cgcctgtggc acgacacgaa gccccagtgc
     1321 catttggcat ggcctagctg cctgcctccg acgctgtccc gccctccgga accttcctgt
     1381 tacagggttt ccaggaaaaa aaaagttgta tttgcgtgcc tagctcaacc tggccccagc
     1441 taacatcgtt acgcgccatc cccgacgtgc tccccaaccc ggtggcacct ccatccgtcg
     1501 ctggtccagg ccagaaaata atcccctcac ctcccattcc ggtcatatgc ggcctcgtgc
     1561 attgcacaaa tatttaccaa gcaactacta tgtgccagaa gcattaataa ctaatgcatg
     1621 gcggtaatac ggttatccac agaatcaggg gataacgcag gaaagaacat gtgagcaaaa
     1681 ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc
     1741 cgcccccctg acgagcatca caaaaatcga cgctcaagtc agaggtggcg aaacccgaca
     1801 ggactataaa gataccaggc gtttccccct ggaagctccc tcgtgcgctc tcctgttccg
     1861 accctgccgc ttaccggata cctgtccgcc tttctccctt cgggaagcgt ggcgctttct
     1921 catagctcac gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt
     1981 gtgcacgaac cccccgttca gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag
     2041 tccaacccgg taagacacga cttatcgcca ctggcagcag ccactggtaa caggattagc
     2101 agagcgaggt atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa ctacggctac
     2161 actagaagaa cagtatttgg tatctgcgct ctgctgaagc cagttacctt cggaaaaaga
     2221 gttggtagct cttgatccgg caaacaaacc accgctggta gcggtggttt ttttgtttgc
     2281 aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag atcctttgat cttttctacg
     2341 gggtctgacg ctcagtggaa cgaaaactca cgttaaggga ttttggtcat gagattatca
     2401 aaaaggatct tcacctagat ccttttaaat taaaaatgaa gttttaaatc aatctaaagt
     2461 atatatgagt aacctgaggc tatggcaggg cctgccgccc cgacgttggc tgcgagccct
     2521 gggccttcac ccgaacttgg ggggtggggt ggggaaaagg aagaaacgcg ggcgtattgg
     2581 ccccaatggg gtctcggtgg ggtatcgaca gagtgccagc cctgggaccg aaccccgcgt
     2641 ttatgaacaa acgacccaac accgtgcgtt ttattctgtc tttttattgc cgtcatagcg
     2701 cgggttcctt ccggtattgt ctccttccgt gtttcagtta gcctccccct agggtgggcg
     2761 aagaactcca gcatgagatc cccgcgctgg aggatcatcc agccggcgtc ccggaaaacg
     2821 attccgaagc ccaacctttc atagaaggcg gcggtggaat cgaaatctcg tgatggcagg
     2881 ttgggcgtcg cttggtcggt catttcgaac cccagagtcc cgctcagaag aactcgtcaa
     2941 gaaggcgata gaaggcgatg cgctgcgaat cgggagcggc gataccgtaa agcacgagga
     3001 agcggtcagc ccattcgccg ccaagctctt cagcaatatc acgggtagcc aacgctatgt
     3061 cctgatagcg gtccgccaca cccagccggc cacagtcgat gaatccagaa aagcggccat
     3121 tttccaccat gatattcggc aagcaggcat cgccatgggt cacgacgaga tcctcgccgt
     3181 cgggcatgct cgccttgagc ctggcgaaca gttcggctgg cgcgagcccc tgatgctctt
     3241 cgtccagatc atcctgatcg acaagaccgg cttccatccg agtacgtgct cgctcgatgc
     3301 gatgtttcgc ttggtggtcg aatgggcagg tagccggatc aagcgtatgc agccgccgca
     3361 ttgcatcagc catgatggat actttctcgg caggagcaag gtgagatgac aggagatcct
     3421 gccccggcac ttcgcccaat agcagccagt cccttcccgc ttcagtgaca acgtcgagca
     3481 cagctgcgca aggaacgccc gtcgtggcca gccacgatag ccgcgctgcc tcgtcttgca
     3541 gttcattcag ggcaccggac aggtcggtct tgacaaaaag aaccgggcgc ccctgcgctg
     3601 acagccggaa cacggcggca tcagagcagc cgattgtctg ttgtgcccag tcatagccga
     3661 atagcctctc cacccaagcg gccggagaac ctgcgtgcaa tccatcttgt tcaatcatgc
     3721 gaaacgatcc tcatcctgtc tcttgatcga tctttgcaaa agcctaggcc tccaaaaaag
     3781 cctcctcact acttctggaa tagctcagag gccgaggcgg cctcggcctc tgcataaata
     3841 aaaaaaatta gtcagccatg gggcggagaa tgggcggaac tgggcggagt taggggcggg
     3901 atgggcggag ttaggggcgg gactatggtt gctgactaat tgagatgcat gctttgcata
     3961 cttctgcctg ctggggagcc tggggacttt ccacacctgg ttgctgacta attgagatgc
     4021 atgctttgca tacttctgcc tgctggggag cctggggact ttccacaccc taactgacac
     4081 acattccaca gctggttctt tccgcctcag gactcttcct ttttcaatat tattgaagca
     4141 tttatcaggg ttattgtctc atgagcggat acatatttga atgtatttag aaaaataaac
     4201 aaataggggt tccgcgcaca tttccccgaa aagtgccacc tgacgcgccc tgtagcggcg
     4261 cattaagcgc ggcgggtgtg gtggttacgc gcagcgtgac cgctacactt gccagcgccc
     4321 tagcgcccgc tcctttcgct ttcttccctt cctttctcgc cacgttcgcc ggctttcccc
     4381 gtcaagctct aaatcggggg ctccctttag ggttccgatt tagtgcttta cggcacctcg
     4441 accccaaaaa acttgattag ggtgatggtt cacgtagtgg gccatcgccc tgatagacgg
     4501 tttttcgccc tttgacgttg gagtccacgt tctttaatag tggactcttg ttccaaactg
     4561 gaacaacact caaccctatc tcggtctatt cttttgattt ataagggatt ttgccgattt
     4621 cggcctattg gttaaaaaat gagctgattt aacaaaaatt taacgcgaat tttaacaaaa
     4681 tattaacgct tacaatttac gccttaagat acattgatga gtttggacaa accacaacta
     4741 gaatgcagtg aaaaaaatgc tttatttgtg aaatttgtga tgctattgct ttatttgtaa
     4801 ccattataag ctgcaataaa caagttaaca acaacaattg cattcatttt atgtttcagg
     4861 ttcaggggga ggtgtgggag gttttttaaa gcaagtaaaa cctctacaaa tgtggtatgg
     4921 ctgattatga tctagagtcg cggccgcttt acttgtacag ctcgtccatg ccgccggtgg
     4981 agtggcggcc ctcggagcgt tcgtactgtt ccaccacggt gtagtcctcg ttgtgggagg
     5041 tgatgtccaa cttgcggtcg acgttgtagg cgccgggcat ctgcacgggc ttcttggcct
     5101 tgtaggtggt cttgaagtcc gccaggtagc ggccgccgtc cttcaggcgc agggccatct
     5161 taatgtcgcc cttcagcacg ccgtcctcgg ggtacaaccg ctcggtggac gcttcccagc
     5221 ccattgtctt cttctgcatt acggggccgt caggagggaa gttggtgccg cggagcttca
     5281 ccttgtagat cagggtgccg tcctccaggg aggtgtcctg ggtcacggtc acggcgccgc
     5341 cgtcctcgaa gttcatcacg cgctcccact tgaagccctc ggggaaggac tgcttatagt
     5401 agtcggggat gtcggcgggg tgcttgatga aggccctgga gccgtacatg aactgagggg
     5461 acaggatgtc ccaggagaag ggcagggggc cacccttggt caccttcagc ttggcggtct
     5521 gggtgccctc gtaggggcgg ccctcgccct cgccctcgat ctcgaactcg tggccgttca
     5581 tggagccctc catgtgcacc ttgaaccgca tgaactcctt gatcactgcc tcgcccttgc
     5641 tcaccatggt ggcgaccggt ggatccttag cgctggtgta cttggtgatg gccttagtac
     5701 cctcggacac ggcgtgcttg gccaactccc caggcagcag caggcgcacg gccgtctgga
     5761 tctccctgga ggtgatggtc gagcgcttgt tgtaatgcgc caggcgggaa gcctcacctg
     5821 cgatgcgctc gaaaatgtcg ttcacaaacg aattcatgat gcccatggcc ttggacgaaa
     5881 tgccggtgtc agggtggacc tgcttcagaa ccttgtacac atagatggaa tagctctcct
     5941 tgcggctgcg cttgcgcttc ttgccgcctt tcttctgcgc cttagtcacc gccttcttgg
     6001 agcccttttt cggggcggga gcagacttcg ctggctctgg catggtggcg gtaccgtcga
     6061 ctgcagaatt cgaagcttga gctcgagatc tgagctcagc tgggaagggt ggactcttgg
     6121 agcctgggat cacaag
//
