LOCUS       sequence-268046-        6383 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  NRF2 reporter plasmid with GFP under control of 8 ARE elements and 
            constitutively transcribed BFP.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6383)
  AUTHORS   Wyler E, Franke V, Menegatti J, Kocks C, Boltengagen A, Praktiknjo 
            S, Walch-Ruckheim B, Bosse J, Rajewsky N, Grasser F, Akalin A, 
            Landthaler M
  TITLE     Single-cell RNA-sequencing of herpes simplex virus 1-infected cells 
            connects NRF2 activation to an antiviral program.
  JOURNAL   Nat Commun. 2019 Oct 25;10(1):4878. doi: 10.1038/s41467-019-12894-z.
  PUBMED    31653857
REFERENCE   2  (bases 1 to 6383)
  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: "Nat 
            Commun."; date: "2019-10-25"; volume: "10(1):4878. doi"; pages: " 
            10.1038/s41467-019-12894-z"
COMMENT     Sequence Label: pREP-8xARE-GFP-SV40-BFP
FEATURES             Location/Qualifiers
     source          1..6383
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    1..92
                     /label=pause site
                     /note="RNA polymerase II transcriptional pause signal from 
                     the human alpha-2 globin gene"
     primer_bind     41..60
                     /label=RVprimer3
                     /note="pGL3 vector, forward primer"
     promoter        221..252
                     /label=minP
                     /note="minimal TATA-box promoter with low basal activity"
     regulatory      285..294
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(291..293,294..296,297..1010)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(291..293,294..296,297..1007)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(336..357)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(597..616)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     944..965
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(1029..1048)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     1148..1170
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     promoter        1210..1526
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      1377..1512
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             1623..2222
                     /codon_start=1
                     /gene="pac from Streptomyces alboniger"
                     /product="puromycin N-acetyltransferase"
                     /label=PuroR
                     /note="confers resistance to puromycin"
                     /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
                     VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
                     AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
                     APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
     primer_bind     complement(1623..1642)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     2119..2139
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     polyA_signal    2452..2533
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     promoter        2786..3143
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        2786..2977
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(2809..2829)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      2994..3129
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3165..3866
                     /codon_start=1
                     /product="monomeric blue fluorescent protein"
                     /label=TagBFP
                     /note="mammalian codon-optimized"
                     /translation="MSELIKENMHMKLYMEGTVDNHHFKCTSEGEGKPYEGTQTMRIKV
                     VEGGPLPFAFDILATSFLYGSKTFINHTQGIPDFFKQSFPEGFTWERVTTYEDGGVLTA
                     TQDTSLQDGCLIYNVKIRGVNFTSNGPVMQKKTLGWEAFTETLYPADGGLEGRNDMALK
                     LVGGSHLIANIKTTYRSKKPAKNLKMPGVYYVDYRLERIKEANNETYVEQHEVAVARYC
                     DLPSKLGHKLN"
     polyA_signal    3890..4011
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3901..3920)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     CDS             join(4172..4240,4241..5032)
                     /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     complement(4390..4409)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5203..5791
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5692..5711
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5945..5962
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    6079..6100
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(6115..6120,6121..6138,6139..6145)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6153..6169
                     /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     6158..6180
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     6177..6193
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     6177..6193
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     promoter        6213..6231
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     6213..6230
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     6243..6262
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        6243..6261
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     polyA_signal    6322..6370
                     /note="synthetic polyadenylation signal"
ORIGIN
        1 aacatacgct ctccatcaaa acaaaacgaa acaaaacaaa ctagcaaaat aggctgtccc
       61 cagtgcaagt gcaggtgcca gaacatttct ctggcctaac tggccgtgac actgcagcag
      121 tgactacgca tgcgtgacga tgcagacgtg acactgcact ggtgacaaag cacccgtgac
      181 taggcaagcg tgacgtagca tcggtgacac agcaagacac tagagggtat ataatggaag
      241 ctcgacttcc agcttggcaa tccggtactg ttggtaaact cgaggccacc atggtgagca
      301 agggcgagga gctgttcacc ggggtggtgc ccatcctggt cgagctggac ggcgacgtaa
      361 acggccacaa gttcagcgtg tccggcgagg gcgagggcga tgccacctac ggcaagctga
      421 ccctgaagtt catctgcacc accggcaagc tgcccgtgcc ctggcccacc ctcgtgacca
      481 ccctgaccta cggcgtgcag tgcttcagcc gctaccccga ccacatgaag cagcacgact
      541 tcttcaagtc cgccatgccc gaaggctacg tccaggagcg caccatcttc ttcaaggacg
      601 acggcaacta caagacccgc gccgaggtga agttcgaggg cgacaccctg gtgaaccgca
      661 tcgagctgaa gggcatcgac ttcaaggagg acggcaacat cctggggcac aagctggagt
      721 acaactacaa cagccacaac gtctatatca tggccgacaa gcagaagaac ggcatcaagg
      781 tgaacttcaa gatccgccac aacatcgagg acggcagcgt gcagctcgcc gaccactacc
      841 agcagaacac ccccatcggc gacggccccg tgctgctgcc cgacaaccac tacctgagca
      901 cccagtccgc cctgagcaaa gaccccaacg agaagcgcga tcacatggtc ctgctggagt
      961 tcgtgaccgc cgccgggatc actctcggca tggacgagct gtacaagtaa catatggtgc
     1021 actctcagta caatctgctc tgatgccgca tagttaagcc agccccgaca cccgccaaca
     1081 cccgctgacg cgccctgacg ggcttgtctg ctcccggcat ccgcttacag acaagctgtg
     1141 accgtctccg ggagctgcat gtgtcagagg ttttcaccgt catcaccgaa acgcgcgaga
     1201 cgaaagattg gtgtggaaag tccccaggct ccccagcagg cagaagtatg caaagcatgc
     1261 atctcaatta gtcagcaacc aggtgtggaa agtccccagg ctccccagca ggcagaagta
     1321 tgcaaagcat gcatctcaat tagtcagcaa ccatagtccc gcccctaact ccgcccatcc
     1381 cgcccctaac tccgcccagt tccgcccatt ctccgcccca tggctgacta atttttttta
     1441 tttatgcaga ggccgaggcc gcctcggcct ctgagctatt ccagaagtag tgaggaggct
     1501 tttttggagg cctaggcttt tgcaaaaagc tagcttgcat gcctgcaggt cggccgccac
     1561 gaccggtgcc gccaccatcc cctgacccac gcccctgacc cctcacaagg agacgacctt
     1621 ccatgaccga gtacaagccc acggtgcgcc tcgccacccg cgacgacgtc ccccgggccg
     1681 tacgcaccct cgccgccgcg ttcgccgact accccgccac gcgccacacc gtcgacccgg
     1741 accgccacat cgagcgggtc accgagctgc aagaactctt cctcacgcgc gtcgggctcg
     1801 acatcggcaa ggtgtgggtc gcggacgacg gcgccgcggt ggcggtctgg accacgccgg
     1861 agagcgtcga agcgggggcg gtgttcgccg agatcggccc gcgcatggcc gagttgagcg
     1921 gttcccggct ggccgcgcag caacagatgg aaggcctcct ggcgccgcac cggcccaagg
     1981 agcccgcgtg gttcctggcc accgtcggcg tctcgcccga ccaccagggc aagggtctgg
     2041 gcagcgccgt cgtgctcccc ggagtggagg cggccgagcg cgccggggtg cccgccttcc
     2101 tggagacctc cgcgccccgc aacctcccct tctacgagcg gctcggcttc accgtcaccg
     2161 ccgacgtcga ggtgcccgaa ggaccgcgca cctggtgcat gacccgcaag cccggtgcct
     2221 gacgcccgcc ccacgacccg cagcgcccga ccgaaaggag cgcacgaccc catggctccg
     2281 accgaagcca cccggggcgg ccccgccgac cccgcacccg cccccgaggc ccaccgactc
     2341 tagaggatca taatcagcca taccacattt gtagaggttt tacttgcttt aaaaaacctc
     2401 ccacacctcc ccctgaacct gaaacataaa atgaatgcaa ttgttgttgt taacttgttt
     2461 attgcagctt ataatggtta caaataaagc aatagcatca caaatttcac aaataaagca
     2521 tttttttcac tgcaatctcg tgatacgcct atttttatag gttaatgtca tgataataat
     2581 ggtttcttag acgtcaggtg gcacttttcg gggaaatgtg cgcggaaccc ctatttgttt
     2641 atttttctaa atacattcaa atatgtatcc gctcatgaga caataaccct gataaatgct
     2701 tcaataatat ctcgtcatgt gggcccatct gcgcagcacc atggcctgaa ataacctctg
     2761 aaagaggaac ttggttaggt accttctgag gcggaaagaa ccagctgtgg aatgtgtgtc
     2821 agttagggtg tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa agcatgcatc
     2881 tcaattagtc agcaaccagg tgtggaaagt ccccaggctc cccagcaggc agaagtatgc
     2941 aaagcatgca tctcaattag tcagcaacca tagtcccgcc cctaactccg cccatcccgc
     3001 ccctaactcc gcccagttcc gcccattctc cgccccatgg ctgactaatt ttttttattt
     3061 atgcagaggc cgaggccgcc tcggcctctg agctattcca gaagtagtga ggaggctttt
     3121 ttggaggcct aggcttttgc aaaaagcttg attcttctga cacgatgagc gagctgatta
     3181 aggagaacat gcacatgaag ctgtacatgg agggcaccgt ggacaaccat cacttcaagt
     3241 gcacatccga gggcgaaggc aagccctacg agggcaccca gaccatgaga atcaaggtgg
     3301 tcgagggcgg ccctctcccc ttcgccttcg acatcctggc tactagcttc ctctacggca
     3361 gcaagacctt catcaaccac acccagggca tccccgactt cttcaagcag tccttccctg
     3421 agggcttcac atgggagaga gtcaccacat acgaagacgg gggcgtgctg accgctaccc
     3481 aggacaccag cctccaggac ggctgcctca tctacaacgt caagatcaga ggggtgaact
     3541 tcacatccaa cggccctgtg atgcagaaga aaacactcgg ctgggaggcc ttcaccgaga
     3601 cgctgtaccc cgctgacggc ggcctggaag gcagaaacga catggccctg aagctcgtgg
     3661 gcgggagcca tctgatcgca aacatcaaga ccacatatag atccaagaaa cccgctaaga
     3721 acctcaagat gcctggcgtc tactatgtgg actacagact ggaaagaatc aaggaggcca
     3781 acaacgagac ctacgtcgag cagcacgagg tggcagtggc cagatactgc gacctcccta
     3841 gcaaactggg gcacaagctt aattaagcgg ccgcttcgag cagacatgat aagatacatt
     3901 gatgagtttg gacaaaccac aactagaatg cagtgaaaaa aatgctttat ttgtgaaatt
     3961 tgtgatgcta ttgctttatt tgtaaccatt ataagctgca ataaacaagt taacaacaac
     4021 aattgcattc attttatgtt tcaggttcag ggggaggtgt gggaggtttt ttaaagcaag
     4081 taaaacctct acaaatgtgg taaaatcgat aaggatcgat ccaggtggca ctttgggccc
     4141 atctgctgtc atatattgaa aaaggaagag tatgagtatt caacatttcc gtgtcgccct
     4201 tattcccttt tttgcggcat tttgccttcc tgtttttgct cacccagaaa cgctggtgaa
     4261 agtaaaagat gctgaagatc agttgggtgc acgagtgggt tacatcgaac tggatctcaa
     4321 cagcggtaag atccttgaga gttttcgccc cgaagaacgt tttccaatga tgagcacttt
     4381 taaagttctg ctatgtggcg cggtattatc ccgtattgac gccgggcaag agcaactcgg
     4441 tcgccgcata cactattctc agaatgactt ggttgagtac tcaccagtca cagaaaagca
     4501 tcttacggat ggcatgacag taagagaatt atgcagtgct gccataacca tgagtgataa
     4561 cactgcggcc aacttacttc tgacaacgat cggaggaccg aaggagctaa ccgctttttt
     4621 gcacaacatg ggggatcatg taactcgcct tgatcgttgg gaaccggagc tgaatgaagc
     4681 cataccaaac gacgagcgtg acaccacgat gcctgtagca atggcaacaa cgttgcgcaa
     4741 actattaact ggcgaactac ttactctagc ttcccggcaa caattaatag actggatgga
     4801 ggcggataaa gttgcaggac cacttctgcg ctcggccctt ccggctggct ggtttattgc
     4861 tgataaatct ggagccggtg agcgtgggtc tcgcggtatc attgcagcac tggggccaga
     4921 tggtaagccc tcccgtatcg tagttatcta cacgacgggg agtcaggcaa ctatggatga
     4981 acgaaataga cagatcgctg agataggtgc ctcactgatt aagcattggt aactgtcaga
     5041 ccaagtttac tcatatatac tttagattga tttaaaactt catttttaat ttaaaaggat
     5101 ctaggtgaag atcctttttg ataatctcat gaccaaaatc ccttaacgtg agttttcgtt
     5161 ccactgagcg tcagaccccg tagaaaagat caaaggatct tcttgagatc ctttttttct
     5221 gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg tttttttgcc
     5281 ggatcaagag ctaccaactc tttttccgaa ggtaactggc ttcagcagag cgcagatacc
     5341 aaatactgtt cttctagtgt agccgtagtt aggccaccac ttcaagaact ctgtagcacc
     5401 gcctacatac ctcgctctgc taatcctgtt accagtggct gctgccagtg gcgataagtc
     5461 gtgtcttacc gggttggact caagacgata gttaccggat aaggcgcagc ggtcgggctg
     5521 aacggggggt tcgtgcacac agcccagctt ggagcgaacg acctacaccg aactgagata
     5581 cctacagcgt gagctatgag aaagcgccac gcttcccgaa gggagaaagg cggacaggta
     5641 tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg gagcttccag ggggaaacgc
     5701 ctggtatctt tatagtcctg tcgggtttcg ccacctctga cttgagcgtc gatttttgtg
     5761 atgctcgtca ggggggcgga gcctatggaa aaacgccagc aacgcggcct ttttacggtt
     5821 cctggccttt tgctggcctt ttgctcacat gttctttcct gcgttatccc ctgattctgt
     5881 ggataaccgt attaccgcct ttgagtgagc tgataccgct cgccgcagcc gaacgaccga
     5941 gcgcagcgag tcagtgagcg aggaagcgga agagcgccca atacgcaaac cgcctctccc
     6001 cgcgcgttgg ccgattcatt aatgcagctg gcacgacagg tttcccgact ggaaagcggg
     6061 cagtgagcgc aacgcaatta atgtgagtta gctcactcat taggcacccc aggctttaca
     6121 ctttatgctt ccggctcgta tgttgtgtgg aattgtgagc ggataacaat ttcacacagg
     6181 aaacagctat gacatgatta cgaattgcaa cgatttaggt gacactatag aagagaagga
     6241 attaatacga ctcactatag ggagagagag agaattaccc tcactaaagg gaggagaagc
     6301 atgaattcgg cctaactggc caataaaata tctttatttt cattacatct gtgtgttggt
     6361 tttttgtgtg aatcgatagt act
//
