LOCUS       sequence-271531-        7712 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  mammalian expression of mRFP1-tagged human NRF2(C506S, Leuzip mut).
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(C506S, L4A)
FEATURES             Location/Qualifiers
     source          1..7712
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    342..541
                     /label=MMLV Psi
                     /note="packaging signal of Moloney murine leukemia virus 
                     (MMLV)"
     CDS             742..1158
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag 
                     gene lacking the start codon"
     primer_bind     1088..1110
                     /label=pLXSN 5'
                     /note="Murine stem cell virus, forward primer. Also called 
                     MSCV"
     primer_bind     1126..1142
                     /label=pBABE 5'
                     /note="Psi packaging signal, forward primer for pBABE 
                     vectors"
     CDS             1169..1843
                     /codon_start=1
                     /product="monomeric derivative of DsRed (Campbell et al., 
                     2002)"
                     /label=mRFP1
                     /translation="MASSEDVIKEFMRFKVRMEGSVNGHEFEIEGEGEGRPYEGTQTAK
                     LKVTKGGPLPFAWDILSPQFQYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNFEDGGV
                     VTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASTERMYPEDGALKGEIKM
                     RLKLKDGGHYDAEVKTTYMAKKPVQLPGAYKTDIKLDITSHNEDYTIVEQYERAEGRHS
                     TGA"
     CDS             1169..1843
                     /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(1299..1317)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     1567..1586
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     primer_bind     1761..1784
                     /label=DsRed1-C
                     /note="DsRed1, forward primer"
     CDS             1859..1882
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     primer_bind     complement(3709..3729)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3714..4043
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3894..4029
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4117..5142
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
                     YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP
                     ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
                     TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
                     GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
                     NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK
                     E"
     CDS             4117..5142
                     /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     5659..5678
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     LTR             5940..6409
                     /note="long terminal repeat from Moloney murine leukemia 
                     virus"
     CDS             complement(join(6524..7315,7316..7384))
                     /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     7147..7166
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7385..7489)
                     /gene="bla"
                     /label=AmpR promoter
     LTR             join(7520..7712,1..277)
                     /note="long terminal repeat from Moloney murine leukemia 
                     virus"
ORIGIN
        1 gtttccaggg tgccccaagg acctgaaatg accctgtgcc ttatttgaac taaccaatca
       61 gttcgcttct cgcttctgtt cgcgcgcttc tgctccccga gctcaataaa agagcccaca
      121 acccctcact cggggcgcca gtcctccgat tgactgagtc gcccgggtac ccgtgtatcc
      181 aataaaccct cttgcagttg catccgactt gtggtctcgc tgttccttgg gagggtctcc
      241 tctgagtgat tgactacccg tcagcggggg tctttcattt gggggctcgt ccgggatcgg
      301 gagacccctg cccagggacc accgacccac caccgggagg caagctggcc agcaacttat
      361 ctgtgtctgt ccgattgtct agtgtctatg actgatttta tgcgcctgcg tcggtactag
      421 ttagctaact agctctgtat ctggcggacc cgtggtggaa ctgacgagtt ctgaacaccc
      481 ggccgcaacc ctgggagacg tcccagggac tttgggggcc gtttttgtgg cccgacctga
      541 ggaagggagt cgatgtggaa tccgaccccg tcaggatatg tggttctggt aggagacgag
      601 aacctaaaac agttcccgcc tccgtctgaa tttttgcttt cggtttggaa ccgaagccgc
      661 gcgtcttgtc tgctgcagcg ctgcagcatc gttctgtgtt gtctctgtct gactgtgttt
      721 ctgtatttgt ctgaaaatta gggccagact gttaccactc ccttaagttt gaccttaggt
      781 cactggaaag atgtcgagcg gatcgctcac aaccagtcgg tagatgtcaa gaagagacgt
      841 tgggttacct tctgctctgc agaatggcca acctttaacg tcggatggcc gcgagacggc
      901 acctttaacc gagacctcat cacccaggtt aagatcaagg tcttttcacc tggcccgcat
      961 ggacacccag accaggtccc ctacatcgtg acctgggaag ccttggcttt tgacccccct
     1021 ccctgggtca agccctttgt acaccctaag cctccgcctc ctcttcctcc atccgccccg
     1081 tctctccccc ttgaacctcc tcgttcgacc ccgcctcgat cctcccttta tccagccctc
     1141 actccttctc taggcgccgg ccggatctat ggcctcctcc gaggacgtca tcaaggagtt
     1201 catgcgcttc aaggtgcgca tggagggctc cgtgaacggc cacgagttcg agatcgaggg
     1261 cgagggcgag ggccgcccct acgagggcac ccagaccgcc aagctgaagg tgaccaaggg
     1321 cggccccctg cccttcgcct gggacatcct gtcccctcag ttccagtacg gctccaaggc
     1381 ctacgtgaag caccccgccg acatccccga ctacttgaag ctgtccttcc ccgagggctt
     1441 caagtgggag cgcgtgatga acttcgagga cggcggcgtg gtgaccgtga cccaggactc
     1501 ctccctgcag gacggcgagt tcatctacaa ggtgaagctg cgcggcacca acttcccctc
     1561 cgacggcccc gtaatgcaga agaagaccat gggctgggag gcctccaccg agcggatgta
     1621 ccccgaggac ggcgccctga agggcgagat caagatgagg ctgaagctga aggacggcgg
     1681 ccactacgac gccgaggtca agaccaccta catggccaag aagcccgtgc agctgcccgg
     1741 cgcctacaag accgacatca agctggacat cacctcccac aacgaggact acaccatcgt
     1801 ggaacagtac gagcgcgccg agggccgcca ctccaccggc gccctcgagg gatccatgga
     1861 ctacaaggat gacgatgaca agatggactt ggagctgccg ccgccgggac tcccgtccca
     1921 gcaggacatg gatttgattg acatactttg gaggcaagat atagatcttg gagtaagtcg
     1981 agaagtattt gacttcagtc agcgacggaa agagtatgag ctggaaaaac agaaaaaact
     2041 tgaaaaggaa agacaagaac aactccaaaa ggagcaagag aaagcctttt tcgctcagtt
     2101 acaactagat gaagagacag gtgaatttct cccaattcag ccagcccagc acatccagtc
     2161 agaaaccagt ggatctgcca actactccca ggttgcccac attcccaaat cagatgcttt
     2221 gtactttgat gactgcatgc agcttttggc gcagacattc ccgtttgtag atgacaatga
     2281 ggtttcttcg gctacgtttc agtcacttgt tcctgatatt cccggtcaca tcgagagccc
     2341 agtcttcatt gctactaatc aggctcagtc acctgaaact tctgttgctc aggtagcccc
     2401 tgttgattta gacggtatgc aacaggacat tgagcaagtt tgggaggagc tattatccat
     2461 tcctgagtta cagtgtctta atattgaaaa tgacaagctg gttgagacta ccatggttcc
     2521 aagtccagaa gccaaactga cagaagttga caattatcat ttttactcat ctataccctc
     2581 aatggaaaaa gaagtaggta actgtagtcc acattttctt aatgcttttg aggattcctt
     2641 cagcagcatc ctctccacag aagaccccaa ccagttgaca gtgaactcat taaattcaga
     2701 tgccacagtc aacacagatt ttggtgatga attttattct gctttcatag ctgagcccag
     2761 tatcagcaac agcatgccct cacctgctac tttaagccat tcactctctg aacttctaaa
     2821 tgggcccatt gatgtttctg atctatcact ttgcaaagct ttcaaccaaa accaccctga
     2881 aagcacagca gaattcaatg attctgactc cggcatttca ctaaacacaa gtcccagtgt
     2941 ggcatcacca gaacactcag tggaatcttc cagctatgga gacacactac ttggcctcag
     3001 tgattctgaa gtggaagagc tagatagtgc ccctggaagt gtcaaacaga atggtcctaa
     3061 aacaccagta cattcttctg gggatatggt acaacccttg tcaccatctc aggggcagag
     3121 cactcacgtg catgatgccc aatgtgagaa cacaccagag aaagaattgc ctgtaagtcc
     3181 tggtcatcgg aaaaccccat tcacaaaaga caaacattca agccgcttgg aggctcatct
     3241 cacaagagat gaacttaggg caaaagctct ccatatccca ttccctgtag aaaaaatcat
     3301 taacctccct gttgttgact tcaacgaaat gatgtccaaa gagcagttca atgaagctca
     3361 acttgcatta attcgggata tacgtaggag gggtaagaat aaagtggctg ctcagaatag
     3421 cagaaaaaga aaactggaaa atatagtaga actagagcaa gatttagatc atttgaaaga
     3481 tgaaaaagaa aaattgctca aagaaaaagg agaaaatgac aaaagccttc acctagcgaa
     3541 aaaacaagcc agcaccgcat atgccgaagt tttcagcatg ctacgtgatg aagatggaaa
     3601 accttattct cctagtgaat actccctgca gcaaacaaga gatggcaatg ttttccttgt
     3661 tcccaaaagt aagaagccag atgttaagaa aaactaggtc gaccctgtgg aatgtgtgtc
     3721 agttagggtg tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa agcatgcatc
     3781 tcaattagtc agcaaccagg tgtggaaagt ccccaggctc cccagcaggc agaagtatgc
     3841 aaagcatgca tctcaattag tcagcaacca tagtcccgcc cctaactccg cccatcccgc
     3901 ccctaactcc gcccagttcc gcccattctc cgccccatgg ctgactaatt ttttttattt
     3961 atgcagaggc cgaggccgcc tcggcctctg agctattcca gaagtagtga ggaggctttt
     4021 ttggaggcct aggcttttgc aaaaagcttg ccagctgggg cgccctctgg taaggttggg
     4081 aagccctgca gatctgatcc cggggggcaa tgagatatga aaaagcctga actcaccgcg
     4141 acgtctgtcg agaagtttct gatcgaaaag ttcgacagcg tctccgacct gatgcagctc
     4201 tcggagggcg aagaatctcg tgctttcagc ttcgatgtag gagggcgtgg atatgtcctg
     4261 cgggtaaata gctgcgccga tggtttctac aaagatcgtt atgtttatcg gcactttgca
     4321 tcggccgcgc tcccgattcc ggaagtgctt gacattgggg aatttagcga gagcctgacc
     4381 tattgcatct cccgccgtgc acagggtgtc acgttgcaag acctgcctga aaccgaactg
     4441 cccgctgttc tgcagccggt cgcggaggcc atggatgcga tcgctgcggc cgatcttagc
     4501 cagacgagcg ggttcggccc attcggaccg caaggaatcg gtcaatacac tacatggcgt
     4561 gatttcatat gcgcgattgc tgatccccat gtgtatcact ggcaaactgt gatggacgac
     4621 accgtcagtg cgtccgtcgc gcaggctctc gatgagctga tgctttgggc cgaggactgc
     4681 cccgaagtcc ggcacctcgt gcacgcggat ttcggctcca acaatgtcct gacggacaat
     4741 ggccgcataa cagcggtcat tgactggagc gaggcgatgt tcggggattc ccaatacgag
     4801 gtcgccaaca tcttcttctg gaggccgtgg ttggcttgta tggagcagca gacgcgctac
     4861 ttcgagcgga ggcatccgga gcttgcagga tcgccgcggc tccgggcgta tatgctccgc
     4921 attggtcttg accaactcta tcagagcttg gttgacggca atttcgatga tgcagcttgg
     4981 gcgcagggtc gatgcgacgc aatcgtccga tccggagccg ggactgtcgg gcgtacacaa
     5041 atcgcccgca gaagcgcggc cgtctggacc gatggctgtg tagaagtact cgccgatagt
     5101 ggaaaccgac gccccagcac tcgtccgagg gcaaaggaat agagtagatg ccgaccggga
     5161 tctatcgatg cggcgggtgt ggtggttacg cgtaatctgc tgcttgcaaa caaaaaaacc
     5221 accgctacca gcggtggttt gtttgccgga tcaagagcta ccaactcttt ttccgaaggt
     5281 aactggcttc agcagagcgc agataccaaa tactgttctt ctagtgtagc cgtagttagg
     5341 ccaccacttc aagaactctg tagcaccgcc tacatacctc gctctgctaa tcctgttacc
     5401 agtggctgct gccagtggcg ataagtcgtg tcttaccggg ttggactcaa gacgatagtt
     5461 accggataag gcgcagcggt cgggctgaac ggggggttcg tgcacacagc ccagcttgga
     5521 gcgaacgacc tacaccgaac tgagatacct acagcgtgag ctatgagaaa gcgccacgct
     5581 tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa caggagagcg
     5641 cacgagggag cttccagggg gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca
     5701 cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa
     5761 cgccagcaac gcatcgataa aataaaagat tttatttagt ctccagaaaa aggggggaat
     5821 gaaagacccc acctgtaggt ttggcaagct agcttaagta acgccatttt gcaaggcatg
     5881 gaaaaataca taactgagaa tagagaagtt cagatcaagg tcaggaacag atggaacagc
     5941 tgaatatggg ccaaacagga tatctgtggt aagcagttcc tgccccggct cagggccaag
     6001 aacagatgga acagctgaat atgggccaaa caggatatct gtggtaagca gttcctgccc
     6061 cggctcaggg ccaagaacag atggtcccca gatgcggtcc agccctcagc agtttctaga
     6121 gaaccatcag atgtttccag ggtgccccaa ggacctgaaa tgaccctgtg ccttatttga
     6181 actaaccaat cagttcgctt ctcgcttctg ttcgcgcgct tctgctcccc gagctcaata
     6241 aaagagccca caacccctca ctcggggcgc cagtcctccg attgactgag tcgcccgggt
     6301 acccgtgtat ccaataaacc ctcttgcagt tgcatccgac ttgtggtctc gctgttcctt
     6361 gggagggtct cctctgagtg attgactacc cgtcagcggg ggtctttcac atgcagcatg
     6421 tatcaaaatt aatttggttt tttttcttaa gtatttacat taaatggcca tagtgcggcc
     6481 gcaaatcaat ctaaagtata tatgagtaaa cttggtctga cagttaccaa tgcttaatca
     6541 gtgaggcacc tatctcagcg atctgtctat ttcgttcatc catagttgcc tgactccccg
     6601 tcgtgtagat aactacgata cgggagggct taccatctgg ccccagtgct gcaatgatac
     6661 cgcgagaccc acgctcaccg gctccagatt tatcagcaat aaaccagcca gccggaaggg
     6721 ccgagcgcag aagtggtcct gcaactttat ccgcctccat ccagtctatt aattgttgcc
     6781 gggaagctag agtaagtagt tcgccagtta atagtttgcg caacgttgtt gccattgcta
     6841 caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc ggttcccaac
     6901 gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc
     6961 ctccgatcgt tgtcagaagt aagttggccg cagtgttatc actcatggtt atggcagcac
     7021 tgcataattc tcttactgtc atgccatccg taagatgctt ttctgtgact ggtgagtact
     7081 caaccaagtc attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa
     7141 tacgggataa taccgcgcca catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt
     7201 cttcggggcg aaaactctca aggatcttac cgctgttgag atccagttcg atgtaaccca
     7261 ctcgtgcacc caactgatct tcagcatctt ttactttcac cagcgtttct gggtgagcaa
     7321 aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa tgttgaatac
     7381 tcatactctt cctttttcaa tattattgaa gcatttatca gggttattgt ctcatgagcg
     7441 gatacatatt tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc acatttcccc
     7501 gaaaagtgcc acctgcagcc tgaatatggg ccaaacagga tatctgtggt aagcagttcc
     7561 tgccccggct cagggccaag aacagatgga acagctgaat atgggccaaa caggatatct
     7621 gtggtaagca gttcctgccc cggctcaggg ccaagaacag atggtcccca gatgcggtcc
     7681 agccctcagc agtttctaga gaaccatcag at
//
