LOCUS       sequence-272429-        7712 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  mammalian expression of mRFP1-tagged human NRF2.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7712)
  AUTHORS   Pereira EJ, Burns JS, Lee CY, Marohl T, Calderon D, Wang L, Atkins 
            KA, Wang CC, Janes KA
  TITLE     Sporadic activation of an oxidative stress-dependent NRF2-p53 
            signaling network in breast epithelial spheroids and 
            premalignancies.
  JOURNAL   Sci Signal. 2020 Apr 14;13(627). pii: 13/627/eaba4200. doi: 
            10.1126/scisignal.aba4200.
  PUBMED    32291314
REFERENCE   2  (bases 1 to 7712)
  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: "Sci 
            Signal."; date: "2020-04-14"; volume: "13(627). pii: 
            13/627/eaba4200. doi"; pages: " 10.1126/scisignal.aba4200"
COMMENT     Sequence Label: pBABE_hygro_mRFP1_NRF2
FEATURES             Location/Qualifiers
     source          1..7712
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     LTR             1..470
                     /note="long terminal repeat from Moloney murine leukemia 
                     virus"
     misc_feature    535..734
                     /label=MMLV Psi
                     /note="packaging signal of Moloney murine leukemia virus 
                     (MMLV)"
     CDS             935..1351
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag 
                     gene lacking the start codon"
     primer_bind     1281..1303
                     /label=pLXSN 5'
                     /note="Murine stem cell virus, forward primer. Also called 
                     MSCV"
     primer_bind     1319..1335
                     /label=pBABE 5'
                     /note="Psi packaging signal, forward primer for pBABE 
                     vectors"
     CDS             1362..2036
                     /codon_start=1
                     /product="monomeric derivative of DsRed (Campbell et al., 
                     2002)"
                     /label=mRFP1
                     /translation="MASSEDVIKEFMRFKVRMEGSVNGHEFEIEGEGEGRPYEGTQTAK
                     LKVTKGGPLPFAWDILSPQFQYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNFEDGGV
                     VTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASTERMYPEDGALKGEIKM
                     RLKLKDGGHYDAEVKTTYMAKKPVQLPGAYKTDIKLDITSHNEDYTIVEQYERAEGRHS
                     TGA"
     CDS             1362..2036
                     /codon_start=1
                     /product="monomeric derivative of DsRed (Campbell et al., 
                     2002)"
                     /label=mRFP1
                     /note="mammalian codon-optimized"
                     /translation="MASSEDVIKEFMRFKVRMEGSVNGHEFEIEGEGEGRPYEGTQTAK
                     LKVTKGGPLPFAWDILSPQFQYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNFEDGGV
                     VTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASTERMYPEDGALKGEIKM
                     RLKLKDGGHYDAEVKTTYMAKKPVQLPGAYKTDIKLDITSHNEDYTIVEQYERAEGRHS
                     TGA"
     primer_bind     complement(1492..1510)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     1760..1779
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     primer_bind     1954..1977
                     /label=DsRed1-C
                     /note="DsRed1, forward primer"
     CDS             2052..2075
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     primer_bind     complement(3902..3922)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3907..4236
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      4087..4222
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4310..5335
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
                     YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP
                     ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
                     TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
                     GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
                     NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK
                     E"
     CDS             4310..5335
                     /codon_start=1
                     /gene="aph(4)-Ia"
                     /product="aminoglycoside phosphotransferase from E. coli"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
                     YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP
                     ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
                     TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
                     GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
                     NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK
                     E"
     primer_bind     5852..5871
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     LTR             6133..6602
                     /note="long terminal repeat from Moloney murine leukemia 
                     virus"
     CDS             complement(join(6717..7508,7509..7577))
                     /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     7340..7359
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7578..7682)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 ctgaatatgg gccaaacagg atatctgtgg taagcagttc ctgccccggc tcagggccaa
       61 gaacagatgg aacagctgaa tatgggccaa acaggatatc tgtggtaagc agttcctgcc
      121 ccggctcagg gccaagaaca gatggtcccc agatgcggtc cagccctcag cagtttctag
      181 agaaccatca gatgtttcca gggtgcccca aggacctgaa atgaccctgt gccttatttg
      241 aactaaccaa tcagttcgct tctcgcttct gttcgcgcgc ttctgctccc cgagctcaat
      301 aaaagagccc acaacccctc actcggggcg ccagtcctcc gattgactga gtcgcccggg
      361 tacccgtgta tccaataaac cctcttgcag ttgcatccga cttgtggtct cgctgttcct
      421 tgggagggtc tcctctgagt gattgactac ccgtcagcgg gggtctttca tttgggggct
      481 cgtccgggat cgggagaccc ctgcccaggg accaccgacc caccaccggg aggcaagctg
      541 gccagcaact tatctgtgtc tgtccgattg tctagtgtct atgactgatt ttatgcgcct
      601 gcgtcggtac tagttagcta actagctctg tatctggcgg acccgtggtg gaactgacga
      661 gttctgaaca cccggccgca accctgggag acgtcccagg gactttgggg gccgtttttg
      721 tggcccgacc tgaggaaggg agtcgatgtg gaatccgacc ccgtcaggat atgtggttct
      781 ggtaggagac gagaacctaa aacagttccc gcctccgtct gaatttttgc tttcggtttg
      841 gaaccgaagc cgcgcgtctt gtctgctgca gcgctgcagc atcgttctgt gttgtctctg
      901 tctgactgtg tttctgtatt tgtctgaaaa ttagggccag actgttacca ctcccttaag
      961 tttgacctta ggtcactgga aagatgtcga gcggatcgct cacaaccagt cggtagatgt
     1021 caagaagaga cgttgggtta ccttctgctc tgcagaatgg ccaaccttta acgtcggatg
     1081 gccgcgagac ggcaccttta accgagacct catcacccag gttaagatca aggtcttttc
     1141 acctggcccg catggacacc cagaccaggt cccctacatc gtgacctggg aagccttggc
     1201 ttttgacccc cctccctggg tcaagccctt tgtacaccct aagcctccgc ctcctcttcc
     1261 tccatccgcc ccgtctctcc cccttgaacc tcctcgttcg accccgcctc gatcctccct
     1321 ttatccagcc ctcactcctt ctctaggcgc cggccggatc tatggcctcc tccgaggacg
     1381 tcatcaagga gttcatgcgc ttcaaggtgc gcatggaggg ctccgtgaac ggccacgagt
     1441 tcgagatcga gggcgagggc gagggccgcc cctacgaggg cacccagacc gccaagctga
     1501 aggtgaccaa gggcggcccc ctgcccttcg cctgggacat cctgtcccct cagttccagt
     1561 acggctccaa ggcctacgtg aagcaccccg ccgacatccc cgactacttg aagctgtcct
     1621 tccccgaggg cttcaagtgg gagcgcgtga tgaacttcga ggacggcggc gtggtgaccg
     1681 tgacccagga ctcctccctg caggacggcg agttcatcta caaggtgaag ctgcgcggca
     1741 ccaacttccc ctccgacggc cccgtaatgc agaagaagac catgggctgg gaggcctcca
     1801 ccgagcggat gtaccccgag gacggcgccc tgaagggcga gatcaagatg aggctgaagc
     1861 tgaaggacgg cggccactac gacgccgagg tcaagaccac ctacatggcc aagaagcccg
     1921 tgcagctgcc cggcgcctac aagaccgaca tcaagctgga catcacctcc cacaacgagg
     1981 actacaccat cgtggaacag tacgagcgcg ccgagggccg ccactccacc ggcgccctcg
     2041 agggatccat ggactacaag gatgacgatg acaagatgga cttggagctg ccgccgccgg
     2101 gactcccgtc ccagcaggac atggatttga ttgacatact ttggaggcaa gatatagatc
     2161 ttggagtaag tcgagaagta tttgacttca gtcagcgacg gaaagagtat gagctggaaa
     2221 aacagaaaaa acttgaaaag gaaagacaag aacaactcca aaaggagcaa gagaaagcct
     2281 ttttcgctca gttacaacta gatgaagaga caggtgaatt tctcccaatt cagccagccc
     2341 agcacatcca gtcagaaacc agtggatctg ccaactactc ccaggttgcc cacattccca
     2401 aatcagatgc tttgtacttt gatgactgca tgcagctttt ggcgcagaca ttcccgtttg
     2461 tagatgacaa tgaggtttct tcggctacgt ttcagtcact tgttcctgat attcccggtc
     2521 acatcgagag cccagtcttc attgctacta atcaggctca gtcacctgaa acttctgttg
     2581 ctcaggtagc ccctgttgat ttagacggta tgcaacagga cattgagcaa gtttgggagg
     2641 agctattatc cattcctgag ttacagtgtc ttaatattga aaatgacaag ctggttgaga
     2701 ctaccatggt tccaagtcca gaagccaaac tgacagaagt tgacaattat catttttact
     2761 catctatacc ctcaatggaa aaagaagtag gtaactgtag tccacatttt cttaatgctt
     2821 ttgaggattc cttcagcagc atcctctcca cagaagaccc caaccagttg acagtgaact
     2881 cattaaattc agatgccaca gtcaacacag attttggtga tgaattttat tctgctttca
     2941 tagctgagcc cagtatcagc aacagcatgc cctcacctgc tactttaagc cattcactct
     3001 ctgaacttct aaatgggccc attgatgttt ctgatctatc actttgcaaa gctttcaacc
     3061 aaaaccaccc tgaaagcaca gcagaattca atgattctga ctccggcatt tcactaaaca
     3121 caagtcccag tgtggcatca ccagaacact cagtggaatc ttccagctat ggagacacac
     3181 tacttggcct cagtgattct gaagtggaag agctagatag tgcccctgga agtgtcaaac
     3241 agaatggtcc taaaacacca gtacattctt ctggggatat ggtacaaccc ttgtcaccat
     3301 ctcaggggca gagcactcac gtgcatgatg cccaatgtga gaacacacca gagaaagaat
     3361 tgcctgtaag tcctggtcat cggaaaaccc cattcacaaa agacaaacat tcaagccgct
     3421 tggaggctca tctcacaaga gatgaactta gggcaaaagc tctccatatc ccattccctg
     3481 tagaaaaaat cattaacctc cctgttgttg acttcaacga aatgatgtcc aaagagcagt
     3541 tcaatgaagc tcaacttgca ttaattcggg atatacgtag gaggggtaag aataaagtgg
     3601 ctgctcagaa ttgcagaaaa agaaaactgg aaaatatagt agaactagag caagatttag
     3661 atcatttgaa agatgaaaaa gaaaaattgc tcaaagaaaa aggagaaaat gacaaaagcc
     3721 ttcacctact gaaaaaacaa ctcagcacct tatatctcga agttttcagc atgctacgtg
     3781 atgaagatgg aaaaccttat tctcctagtg aatactccct gcagcaaaca agagatggca
     3841 atgttttcct tgttcccaaa agtaagaagc cagatgttaa gaaaaactag gtcgaccctg
     3901 tggaatgtgt gtcagttagg gtgtggaaag tccccaggct ccccagcagg cagaagtatg
     3961 caaagcatgc atctcaatta gtcagcaacc aggtgtggaa agtccccagg ctccccagca
     4021 ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa ccatagtccc gcccctaact
     4081 ccgcccatcc cgcccctaac tccgcccagt tccgcccatt ctccgcccca tggctgacta
     4141 atttttttta tttatgcaga ggccgaggcc gcctcggcct ctgagctatt ccagaagtag
     4201 tgaggaggct tttttggagg cctaggcttt tgcaaaaagc ttgccagctg gggcgccctc
     4261 tggtaaggtt gggaagccct gcagatctga tcccgggggg caatgagata tgaaaaagcc
     4321 tgaactcacc gcgacgtctg tcgagaagtt tctgatcgaa aagttcgaca gcgtctccga
     4381 cctgatgcag ctctcggagg gcgaagaatc tcgtgctttc agcttcgatg taggagggcg
     4441 tggatatgtc ctgcgggtaa atagctgcgc cgatggtttc tacaaagatc gttatgttta
     4501 tcggcacttt gcatcggccg cgctcccgat tccggaagtg cttgacattg gggaatttag
     4561 cgagagcctg acctattgca tctcccgccg tgcacagggt gtcacgttgc aagacctgcc
     4621 tgaaaccgaa ctgcccgctg ttctgcagcc ggtcgcggag gccatggatg cgatcgctgc
     4681 ggccgatctt agccagacga gcgggttcgg cccattcgga ccgcaaggaa tcggtcaata
     4741 cactacatgg cgtgatttca tatgcgcgat tgctgatccc catgtgtatc actggcaaac
     4801 tgtgatggac gacaccgtca gtgcgtccgt cgcgcaggct ctcgatgagc tgatgctttg
     4861 ggccgaggac tgccccgaag tccggcacct cgtgcacgcg gatttcggct ccaacaatgt
     4921 cctgacggac aatggccgca taacagcggt cattgactgg agcgaggcga tgttcgggga
     4981 ttcccaatac gaggtcgcca acatcttctt ctggaggccg tggttggctt gtatggagca
     5041 gcagacgcgc tacttcgagc ggaggcatcc ggagcttgca ggatcgccgc ggctccgggc
     5101 gtatatgctc cgcattggtc ttgaccaact ctatcagagc ttggttgacg gcaatttcga
     5161 tgatgcagct tgggcgcagg gtcgatgcga cgcaatcgtc cgatccggag ccgggactgt
     5221 cgggcgtaca caaatcgccc gcagaagcgc ggccgtctgg accgatggct gtgtagaagt
     5281 actcgccgat agtggaaacc gacgccccag cactcgtccg agggcaaagg aatagagtag
     5341 atgccgaccg ggatctatcg atgcggcggg tgtggtggtt acgcgtaatc tgctgcttgc
     5401 aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc
     5461 tttttccgaa ggtaactggc ttcagcagag cgcagatacc aaatactgtt cttctagtgt
     5521 agccgtagtt aggccaccac ttcaagaact ctgtagcacc gcctacatac ctcgctctgc
     5581 taatcctgtt accagtggct gctgccagtg gcgataagtc gtgtcttacc gggttggact
     5641 caagacgata gttaccggat aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac
     5701 agcccagctt ggagcgaacg acctacaccg aactgagata cctacagcgt gagctatgag
     5761 aaagcgccac gcttcccgaa gggagaaagg cggacaggta tccggtaagc ggcagggtcg
     5821 gaacaggaga gcgcacgagg gagcttccag ggggaaacgc ctggtatctt tatagtcctg
     5881 tcgggtttcg ccacctctga cttgagcgtc gatttttgtg atgctcgtca ggggggcgga
     5941 gcctatggaa aaacgccagc aacgcatcga taaaataaaa gattttattt agtctccaga
     6001 aaaagggggg aatgaaagac cccacctgta ggtttggcaa gctagcttaa gtaacgccat
     6061 tttgcaaggc atggaaaaat acataactga gaatagagaa gttcagatca aggtcaggaa
     6121 cagatggaac agctgaatat gggccaaaca ggatatctgt ggtaagcagt tcctgccccg
     6181 gctcagggcc aagaacagat ggaacagctg aatatgggcc aaacaggata tctgtggtaa
     6241 gcagttcctg ccccggctca gggccaagaa cagatggtcc ccagatgcgg tccagccctc
     6301 agcagtttct agagaaccat cagatgtttc cagggtgccc caaggacctg aaatgaccct
     6361 gtgccttatt tgaactaacc aatcagttcg cttctcgctt ctgttcgcgc gcttctgctc
     6421 cccgagctca ataaaagagc ccacaacccc tcactcgggg cgccagtcct ccgattgact
     6481 gagtcgcccg ggtacccgtg tatccaataa accctcttgc agttgcatcc gacttgtggt
     6541 ctcgctgttc cttgggaggg tctcctctga gtgattgact acccgtcagc gggggtcttt
     6601 cacatgcagc atgtatcaaa attaatttgg ttttttttct taagtattta cattaaatgg
     6661 ccatagtgcg gccgcaaatc aatctaaagt atatatgagt aaacttggtc tgacagttac
     6721 caatgcttaa tcagtgaggc acctatctca gcgatctgtc tatttcgttc atccatagtt
     6781 gcctgactcc ccgtcgtgta gataactacg atacgggagg gcttaccatc tggccccagt
     6841 gctgcaatga taccgcgaga cccacgctca ccggctccag atttatcagc aataaaccag
     6901 ccagccggaa gggccgagcg cagaagtggt cctgcaactt tatccgcctc catccagtct
     6961 attaattgtt gccgggaagc tagagtaagt agttcgccag ttaatagttt gcgcaacgtt
     7021 gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc
     7081 tccggttccc aacgatcaag gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt
     7141 agctccttcg gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg
     7201 gttatggcag cactgcataa ttctcttact gtcatgccat ccgtaagatg cttttctgtg
     7261 actggtgagt actcaaccaa gtcattctga gaatagtgta tgcggcgacc gagttgctct
     7321 tgcccggcgt caatacggga taataccgcg ccacatagca gaactttaaa agtgctcatc
     7381 attggaaaac gttcttcggg gcgaaaactc tcaaggatct taccgctgtt gagatccagt
     7441 tcgatgtaac ccactcgtgc acccaactga tcttcagcat cttttacttt caccagcgtt
     7501 tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg
     7561 aaatgttgaa tactcatact cttccttttt caatattatt gaagcattta tcagggttat
     7621 tgtctcatga gcggatacat atttgaatgt atttagaaaa ataaacaaat aggggttccg
     7681 cgcacatttc cccgaaaagt gccacctgca gc
//
