LOCUS       sequence-488912-        7375 bp    DNA     circular SYN 09-OCT-2025
DEFINITION  AHR GFP reporter plasmid in a lentiviral backbone.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7375)
  AUTHORS   Verma M, Desai K, Ding Y, Wang X, Bae M, Rakoff-Nahoum S, Balskus 
            EP, Fischbach MA, Jost M
  TITLE     Systematic identification and characterization of regulators of aryl
            hydrocarbon receptor signaling
  JOURNAL   bioRxiv 2025.09.15.676391
REFERENCE   2  (bases 1 to 7375)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Oct 9, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "bioRxiv 
            2025.09.15.676391"
COMMENT     Sequence Label: pMV0021 [15ARE23bp-GFP]
FEATURES             Location/Qualifiers
     source          1..7375
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(119..137)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        205..309
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(310..378,379..1170)
                     /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(528..547)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      1341..1929
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1830..1849
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2083..2100
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    2340..2361
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(2376..2381,2382..2399,2400..2406)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2414..2430
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     primer_bind     2419..2441
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     2438..2454
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     2438..2454
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     2504..2523
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     LTR             2715..3348
                     /label=3' LTR
                     /note="3' long terminal repeat (LTR) from HIV-1"
     misc_feature    3395..3520
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    4017..4250
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             4435..4479
                     /codon_start=1
                     /product="antigenic peptide corresponding to amino acids 
                     655 to 669 of the HIV envelope protein gp41 (Lutje Hulsik 
                     et al., 2013)"
                     /label=gp41 peptide
                     /note="recognized by the 2H10 single-chain llama nanobody"
                     /translation="KNEQELLELDKWASL"
     misc_feature    4772..4889
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     regulatory      5539..5548
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(5545..5547,5548..5550,5551..6261)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     ANFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(5590..5611)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(5851..5870)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     6198..6219
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     CDS             6265..6291
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     misc_feature    6479..7067
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(6532..6552)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(6950..6961)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             7142..7375
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
ORIGIN
        1 gtagtagttc atgtcatctt attattcagt atttataact tgcaaagaaa tgaatatcag
       61 agagtgagag gcccgggtta attaaggaaa gggctagatc attcttgaag acgaaagggc
      121 ctcgtgatac gcctattttt ataggttaat gtcatgataa taatggtttc ttagacgtca
      181 ggtggcactt ttcggggaaa tgtgcgcgga acccctattt gtttattttt ctaaatacat
      241 tcaaatatgt atccgctcat gagacaataa ccctgataaa tgcttcaata atattgaaaa
      301 aggaagagta tgagtattca acatttccgt gtcgccctta ttcccttttt tgcggcattt
      361 tgccttcctg tttttgctca cccagaaacg ctggtgaaag taaaagatgc tgaagatcag
      421 ttgggtgcac gagtgggtta catcgaactg gatctcaaca gcggtaagat ccttgagagt
      481 tttcgccccg aagaacgttt tccaatgatg agcactttta aagttctgct atgtggcgcg
      541 gtattatccc gtgttgacgc cgggcaagag caactcggtc gccgcataca ctattctcag
      601 aatgacttgg ttgagtactc accagtcaca gaaaagcatc ttacggatgg catgacagta
      661 agagaattat gcagtgctgc cataaccatg agtgataaca ctgcggccaa cttacttctg
      721 acaacgatcg gaggaccgaa ggagctaacc gcttttttgc acaacatggg ggatcatgta
      781 actcgccttg atcgttggga accggagctg aatgaagcca taccaaacga cgagcgtgac
      841 accacgatgc ctgtagcaat ggcaacaacg ttgcgcaaac tattaactgg cgaactactt
      901 actctagctt cccggcaaca attaatagac tggatggagg cggataaagt tgcaggacca
      961 cttctgcgct cggcccttcc ggctggctgg tttattgctg ataaatctgg agccggtgag
     1021 cgtgggtctc gcggtatcat tgcagcactg gggccagatg gtaagccctc ccgtatcgta
     1081 gttatctaca cgacggggag tcaggcaact atggatgaac gaaatagaca gatcgctgag
     1141 ataggtgcct cactgattaa gcattggtaa ctgtcagacc aagtttactc atatatactt
     1201 tagattgatt taaaacttca tttttaattt aaaaggatct aggtgaagat cctttttgat
     1261 aatctcatga ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc agaccccgta
     1321 gaaaagatca aaggatcttc ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa
     1381 acaaaaaaac caccgctacc agcggtggtt tgtttgccgg atcaagagct accaactctt
     1441 tttccgaagg taactggctt cagcagagcg cagataccaa atactgttct tctagtgtag
     1501 ccgtagttag gccaccactt caagaactct gtagcaccgc ctacatacct cgctctgcta
     1561 atcctgttac cagtggctgc tgccagtggc gataagtcgt gtcttaccgg gttggactca
     1621 agacgatagt taccggataa ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag
     1681 cccagcttgg agcgaacgac ctacaccgaa ctgagatacc tacagcgtga gctatgagaa
     1741 agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga
     1801 acaggagagc gcacgaggga gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc
     1861 gggtttcgcc acctctgact tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc
     1921 ctatggaaaa acgccagcaa cgcggccttt ttacggttcc tggccttttg ctggcctttt
     1981 gctcacatgt tctttcctgc gttatcccct gattctgtgg ataaccgtat taccgccttt
     2041 gagtgagctg ataccgctcg ccgcagccga acgaccgagc gcagcgagtc agtgagcgag
     2101 gaagcggaag agcgcccaat acgcaaaccg cctctccccg cgcgttggcc gattcattaa
     2161 tgcagcaagc tcatggctga ctaatttttt ttatttatgc agaggccgag gccgcctcgg
     2221 cctctgagct attccagaag tagtgaggag gcttttttgg aggcctaggc ttttgcaaaa
     2281 agctccccgt ggcacgacag gtttcccgac tggaaagcgg gcagtgagcg caacgcaatt
     2341 aatgtgagtt agctcactca ttaggcaccc caggctttac actttatgct tccggctcgt
     2401 atgttgtgtg gaattgtgag cggataacaa tttcacacag gaaacagcta tgacatgatt
     2461 acgaatttca caaataaagc atttttttca ctgcattcta gttgtggttt gtccaaactc
     2521 atcaatgtat cttatcatgt ctggatcaac tggataactc aagctaacca aaatcatccc
     2581 aaacttccca ccccataccc tattaccact gccaattacc tgtggtttca tttactctaa
     2641 acctgtgatt cctctgaatt attttcattt taaagaaatt gtatttgtta aatatgtact
     2701 acaaacttag tagttggaag ggctaattca ctcccaaaga agacaagata tccttgatct
     2761 gtggatctac cacacacaag gctacttccc tgattagcag aactacacac cagggccagg
     2821 ggtcagatat ccactgacct ttggatggtg ctacaagcta gtaccagttg agccagataa
     2881 ggtagaagag gccaataaag gagagaacac cagcttgtta caccctgtga gcctgcatgg
     2941 gatggatgac ccggagagag aagtgttaga gtggaggttt gacagccgcc tagcatttca
     3001 tcacgtggcc cgagagctgc atccggagta cttcaagaac tgctgatatc gagcttgcta
     3061 caagggactt tccgctgggg actttccagg gaggcgtggc ctgggcggga ctggggagtg
     3121 gcgagccctc agatcctgca tataagcagc tgctttttgc ctgtactggg tctctctggt
     3181 tagaccagat ctgagcctgg gagctctctg gctaactagg gaacccactg cttaagcctc
     3241 aataaagctt gccttgagtg cttcaagtag tgtgtgcccg tctgttgtgt gactctggta
     3301 actagagatc cctcagaccc ttttagtcag tgtggaaaat ctctagcagt ggcgcccgaa
     3361 cagggacttg aaagcgaaag ggaaaccaga ggagctctct cgacgcagga ctcggcttgc
     3421 tgaagcgcgc acggcaagag gcgaggggcg gcgactggtg agtacgccaa aaattttgac
     3481 tagcggaggc tagaaggaga gagatgggtg cgagagcgtc agtattaagc gggggagaat
     3541 tagatcgcga tgggaaaaaa ttcggttaag gccaggggga aagaaaaaat ataaattaaa
     3601 acatatagta tgggcaagca gggagctaga acgattcgca gttaatcctg gcctgttaga
     3661 aacatcagaa ggctgtagac aaatactggg acagctacaa ccatcccttc agacaggatc
     3721 agaagaactt agatcattat ataatacagt agcaaccctc tattgtgtgc atcaaaggat
     3781 agagataaaa gacaccaagg aagctttaga caagatagag gaagagcaaa acaaaagtaa
     3841 gaccaccgca cagcaagcgg ccggccgctg atcttcagac ctggaggagg agatatgagg
     3901 gacaattgga gaagtgaatt atataaatat aaagtagtaa aaattgaacc attaggagta
     3961 gcacccacca aggcaaagag aagagtggtg cagagagaaa aaagagcagt gggaatagga
     4021 gctttgttcc ttgggttctt gggagcagca ggaagcacta tgggcgcagc gtcaatgacg
     4081 ctgacggtac aggccagaca attattgtct ggtatagtgc agcagcagaa caatttgctg
     4141 agggctattg aggcgcaaca gcatctgttg caactcacag tctggggcat caagcagctc
     4201 caggcaagaa tcctggctgt ggaaagatac ctaaaggatc aacagctcct ggggatttgg
     4261 ggttgctctg gaaaactcat ttgcaccact gctgtgcctt ggaatgctag ttggagtaat
     4321 aaatctctgg aacagatttg gaatcacacg acctggatgg agtgggacag agaaattaac
     4381 aattacacaa gcttaataca ctccttaatt gaagaatcgc aaaaccagca agaaaagaat
     4441 gaacaagaat tattggaatt agataaatgg gcaagtttgt ggaattggtt taacataaca
     4501 aattggctgt ggtatataaa attattcata atgatagtag gaggcttggt aggtttaaga
     4561 atagtttttg ctgtactttc tatagtgaat agagttaggc agggatattc accattatcg
     4621 tttcagaccc acctcccaac cccgagggga cccgacaggc ccgaaggaat agaagaagaa
     4681 ggtggagaga gagacagaga cagatccatt cgattagtga acggatctcg acggtatcgc
     4741 cgaattcaca aatggcagta ttcatccaca attttaaaag aaaagggggg attggggggt
     4801 acagtgcagg ggaaagaata gtagacataa tagcaacaga catacaaact aaagaattac
     4861 aaaaacaaat tacaaaaatt caaaattttc gggtttatta cagggacagc agagatccag
     4921 tttggactag gatccggtac cgagctctta cgcgtgctag ctggcgtgag gggctccccc
     4981 tgtgactaac ttggcgtggc gccaaaacat cttcggatcc ctagcgtgct tgtccaatct
     5041 aactcaccga attgcgtgtt cttacatttt agaccctaat atcgcgtgaa cagtaagcgc
     5101 tttttgtagg cagctggcgt gtcttattaa tacatttgct aggctggcgt gcctggcgtt
     5161 ccacgccgcc acgttagcgt ggttcgttca actgttgatt ggtgttgcgt gtctgcgtgt
     5221 ccaaccttag aatttcgcgt gatcctccac aaggtctagt ggtatggcgt gtgaaattcg
     5281 gctcagttac ttcgtagcgt ggacctggct tacgtcttag tacgttgcgt gcgattccta
     5341 gagacaggtg ctcgcgtgta ggtggtgttc agttcggtaa ctggcgtgcg attcctcgag
     5401 atctgcactc attcataaaa cgcttgttat aaaagcagtg gctgcggcgc ctcgtactcc
     5461 aaccgcatct gcagcgagca actgagaagc caagactgag ccggcggccg atatcaagct
     5521 tgcggtaccg cgggcccggc caccatggtg agcaagggcg aggagctgtt caccggggtg
     5581 gtgcccatcc tggtcgagct ggacggcgac gtaaacggcc acaagttcag cgtgtctggc
     5641 gagggcgagg gcgatgccac ctacggcaag ctgaccctga agttcatctg caccaccggc
     5701 aagctgcccg tgccctggcc caccctcgtg accaccctga cctacggcgt gcagtgcttc
     5761 agccgctacc ccgaccacat gaagcagcac gacttcttca agtccgccat gcccgaaggc
     5821 tacgtccagg agcgcaccat cttcttcaag gacgacggca actacaagac ccgcgccgag
     5881 gtgaagttcg agggcgacac cctggtgaac cgcatcgagc tgaagggcat cgacttcaag
     5941 gaggacggca acatcctggg gcacaagctg gagtacaact acaacagcca caacgtctat
     6001 atcatggccg acaagcagaa gaacggcatc aaggcgaact tcaagatccg ccacaacatc
     6061 gaggacggca gcgtgcagct cgccgaccac taccagcaga acacccccat cggcgacggc
     6121 cccgtgctgc tgcccgacaa ccactacctg agcacccagt ccgccctgag caaagacccc
     6181 aacgagaagc gcgatcacat ggtcctgctg gagttcgtga ccgccgccgg gatcactctc
     6241 ggcatggacg agctgtacaa ggcctatccc tatgacgtgc ccgattatgc cggcagctaa
     6301 taagttaaca ataagcggcc gcgactctag atcataatca gccataccac atttgtagag
     6361 gttttacttg ctttaaaaaa cctcccacac ctccccctga acctgaaaca taaaatgaat
     6421 gcaattgttg ttgtttagtc cctcccaatt cgatatcaag cttatcgcga tagatcctaa
     6481 tcaacctctg gattacaaaa tttgtgaaag attgactggt attcttaact atgttgctcc
     6541 ttttacgcta tgtggatacg ctgctttaat gcctttgtat catgctattg cttcccgtat
     6601 ggctttcatt ttctcctcct tgtataaatc ctggttgctg tctctttatg aggagttgtg
     6661 gcccgttgtc aggcaacgtg gcgtggtgtg cactgtgttt gctgacgcaa cccccactgg
     6721 ttggggcatt gccaccacct gtcagctcct ttccgggact ttcgctttcc ccctccctat
     6781 tgccacggcg gaactcatcg ccgcctgcct tgcccgctgc tggacagggg ctcggctgtt
     6841 gggcactgac aattccgtgg tgttgtcggg gaaatcatcg tcctttcctt ggctgctcgc
     6901 ctgtgttgcc acctggattc tgcgcgggac gtccttctgc tacgtccctt cggccctcaa
     6961 tccagcggac cttccttccc gcggcctgct gccggctctg cggcctcttc cgcgtcttcg
     7021 ccttcgccct cagacgagtc ggatctccct ttgggccgcc tccccgcctg agatccttta
     7081 agaccaatga cttacaaggc agctgtagat cttagccact ttttaaaaga aaagggggga
     7141 ctggaagggc taattcactc ccaacgaaga caagatctgc tttttgcttg tactgggtct
     7201 ctctggttag accagatctg agcctgggag ctctctggct aactagggaa cccactgctt
     7261 aagcctcaat aaagcttgcc ttgagtgctt caagtagtgt gtgcccgtct gttgtgtgac
     7321 tctggtaact agagatccct cagacccttt tagtcagtgt ggaaaatctc tagca
//
