LOCUS       sequence-380109-        6706 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression of human NRF2 (NFE2L2) fused to EGFP at the 
            N-terminus. Parton lab clone KTH.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6706)
  AUTHORS   Wu Y, Lim YW, Stroud DA, Martel N, Hall TE, Lo HP, Ferguson C, Ryan 
            MT, McMahon KA, Parton RG
  TITLE     Caveolae sense oxidative stress through membrane lipid peroxidation 
            and cytosolic release of CAVIN1 to regulate NRF2.
  JOURNAL   Dev Cell. 2023 Mar 13;58(5):376-397.e4. doi: 
            10.1016/j.devcel.2023.02.004. Epub 2023 Feb 28.
  PUBMED    36858041
REFERENCE   2  (bases 1 to 6706)
  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"; doi: 
            "10.1016/j.devcel.2023.02.004"; journalName: "Dev Cell"; date: 
            "2023-03-13- 13"; volume: "58"; issue: "5"; pages: "376-397.e4"
COMMENT     Sequence Label: EGFP-HsNRF2 (NFE2L2)
FEATURES             Location/Qualifiers
     source          1..6706
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        81..99
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     81..98
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     protein_bind    154..178
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     regulatory      179..188
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(185..187,188..190,191..901)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(185..187,188..190,191..901)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(230..251)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(491..510)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     838..859
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     protein_bind    complement(903..927)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     protein_bind    complement(2744..2764)
                     /gene="mutant version of attB"
                     /label=attB3
                     /bound_moiety="BP Clonase(TM)"
                     /note="core recombination site for the Gateway(R) BP 
                     reaction"
     polyA_signal    2804..2938
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2828..2847)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2882..2901
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(3051..3071)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(3051..3069)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(3090..3106)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3090..3106)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3103..3125)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3114..3130
                     /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)."
     promoter        complement(join(3138..3144,3145..3162,3163..3168))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3183..3204
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3321..3338)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3492..4080)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3572..3591)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4251..5042,5043..5111))
                     /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     4874..4893
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5112..5216)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(5242..5697)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5379..5400)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     5591..5610
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        join(5768..6706,1..46)
                     /label=CMV IE94 promoter
                     /note="enhancer/promoter region of simian cytomegalovirus 
                     major immediate early transcription unit IE94"
ORIGIN
        1 gcgtgcctaa tgggaggtct atataagcaa tgctcgttta gggaaccgcc attctgcctg
       61 gggacgtcgg agcaagcttg atttaggtga cactatagaa tacaagctac ttgttctttt
      121 tgcaggatcc catcgattcg aattcaagga tcaacaagtt tgtacaaaaa agcaggctgc
      181 caccatggtg agcaagggcg aggagctgtt caccggggtg gtgcccatcc tggtcgagct
      241 ggacggcgac gtaaacggcc acaagttcag cgtgtccggc gagggcgagg gcgatgccac
      301 ctacggcaag ctgaccctga agttcatctg caccaccggc aagctgcccg tgccctggcc
      361 caccctcgtg accaccctga cctacggcgt gcagtgcttc agccgctacc ccgaccacat
      421 gaagcagcac gacttcttca agtccgccat gcccgaaggc tacgtccagg agcgcaccat
      481 cttcttcaag gacgacggca actacaagac ccgcgccgag gtgaagttcg agggcgacac
      541 cctggtgaac cgcatcgagc tgaagggcat cgacttcaag gaggacggca acatcctggg
      601 gcacaagctg gagtacaact acaacagcca caacgtctat atcatggccg acaagcagaa
      661 gaacggcatc aaggtgaact tcaagatccg ccacaacatc gaggacggca gcgtgcagct
      721 cgccgaccac taccagcaga acacccccat cggcgacggc cccgtgctgc tgcccgacaa
      781 ccactacctg agcacccagt ccgccctgag caaagacccc aacgagaagc gcgatcacat
      841 ggtcctgctg gagttcgtga ccgccgccgg gatcactctc ggcatggacg agctgtacaa
      901 gcacccagct ttcttgtaca aagtggtgat ggacttggag ctgccgccgc cgggactccc
      961 gtcccagcag gacatggatt tgattgacat actttggagg caagatatag atcttggagt
     1021 aagtcgagaa gtatttgact tcagtcagcg acggaaagag tatgagctgg aaaaacagaa
     1081 aaaacttgaa aaggaaagac aagaacaact ccaaaaggag caagagaaag cctttttcgc
     1141 tcagttacaa ctagatgaag agacaggtga atttctccca attcagccag cccagcacat
     1201 ccagtcagaa accagtggat ctgccaacta ctcccaggtt gcccacattc ccaaatcaga
     1261 tgctttgtac tttgatgact gcatgcagct tttggcgcag acattcccgt ttgtagatga
     1321 caatgaggtt tcttcggcta cgtttcagtc acttgttcct gatattcccg gtcacatcga
     1381 gagcccagtc ttcattgcta ctaatcaggc tcagtcacct gaaacttctg ttgctcaggt
     1441 agcccctgtt gatttagacg gtatgcaaca ggacattgag caagtttggg aggagctatt
     1501 atccattcct gagttacagt gtcttaatat tgaaaatgac aagctggttg agactaccat
     1561 ggttccaagt ccagaagcca aactgacaga agttgacaat tatcattttt actcatctat
     1621 accctcaatg gaaaaagaag taggtaactg tagtccacat tttcttaatg cttttgagga
     1681 ttccttcagc agcatcctct ccacagaaga ccccaaccag ttgacagtga actcattaaa
     1741 ttcagatgcc acagtcaaca cagattttgg tgatgaattt tattctgctt tcatagctga
     1801 gcccagtatc agcaacagca tgccctcacc tgctacttta agccattcac tctctgaact
     1861 tctaaatggg cccattgatg tttctgatct atcactttgc aaagctttca accaaaacca
     1921 ccctgaaagc acagcagaat tcaatgattc tgactccggc atttcactaa acacaagtcc
     1981 cagtgtggca tcaccagaac actcagtgga atcttccagc tatggagaca cactacttgg
     2041 cctcagtgat tctgaagtgg aagagctaga tagtgcccct ggaagtgtca aacagaatgg
     2101 tcctaaaaca ccagtacatt cttctgggga tatggtacaa cccttgtcac catctcaggg
     2161 gcagagcact cacgtgcatg atgcccaatg tgagaacaca ccagagaaag aattgcctgt
     2221 aagtcctggt catcggaaaa ccccattcac aaaagacaaa cattcaagcc gcttggaggc
     2281 tcatctcaca agagatgaac ttagggcaaa agctctccat atcccattcc ctgtagaaaa
     2341 aatcattaac ctccctgttg ttgacttcaa cgaaatgatg tccaaagagc agttcaatga
     2401 agctcaactt gcattaattc gggatatacg taggaggggt aagaataaag tggctgctca
     2461 gaattgcaga aaaagaaaac tggaaaatat agtagaacta gagcaagatt tagatcattt
     2521 gaaagatgaa aaagaaaaat tgctcaaaga aaaaggagaa aatgacaaaa gccttcacct
     2581 actgaaaaaa caactcagca ccttatatct cgaagttttc agcatgctac gtgatgaaga
     2641 tggaaaacct tattctccta gtgaatactc cctgcagcaa acaagagatg gcaatgtttt
     2701 ccttgttccc aaaagtaaga agccagatgt taagaaaaac tagcaacttt attatacata
     2761 gttgcactcg agcctctaga actatagtga gtcgtattac gtagatccag acatgataag
     2821 atacattgat gagtttggac aaaccacaac tagaatgcag tgaaaaaaat gctttatttg
     2881 tgaaatttgt gatgctattg ctttatttgt aaccattata agctgcaata aacaagttaa
     2941 caacaacaat tgcattcatt ttatgtttca ggttcagggg gaggtgtggg aggtttttta
     3001 attcgcggcc gcggcgccaa tgcattgggc ccggtaccca gcttttgttc cctttagtga
     3061 gggttaattg cgcgcttggc gtaatcatgg tcatagctgt ttcctgtgtg aaattgttat
     3121 ccgctcacaa ttccacacaa catacgagcc gggagcataa agtgtaaagc ctggggtgcc
     3181 taatgagtga gctaactcac attaattgcg ttgcgctcac tgcccgcttt ccagtcggga
     3241 aacctgtcgt gccagctgca ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt
     3301 attgggcgct cttccgcttc ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg
     3361 cgagcggtat cagctcactc aaaggcggta atacggttat ccacagaatc aggggataac
     3421 gcaggaaaga acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg
     3481 ttgctggcgt ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca
     3541 agtcagaggt ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc
     3601 tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc
     3661 ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta ggtatctcag ttcggtgtag
     3721 gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc
     3781 ttatccggta actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca
     3841 gcagccactg gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg
     3901 aagtggtggc ctaactacgg ctacactaga agaacagtat ttggtatctg cgctctgctg
     3961 aagccagtta ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct
     4021 ggtagcggtg gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa
     4081 gaagatcctt tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa
     4141 gggattttgg tcatgagatt atcaaaaagg atcttcacct agatcctttt aaattaaaaa
     4201 tgaagtttta aatcaatcta aagtatatat gagtaaactt ggtctgacag ttaccaatgc
     4261 ttaatcagtg aggcacctat ctcagcgatc tgtctatttc gttcatccat agttgcctga
     4321 ctccccgtcg tgtagataac tacgatacgg gagggcttac catctggccc cagtgctgca
     4381 atgataccgc gagacccacg ctcaccggct ccagatttat cagcaataaa ccagccagcc
     4441 ggaagggccg agcgcagaag tggtcctgca actttatccg cctccatcca gtctattaat
     4501 tgttgccggg aagctagagt aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc
     4561 attgctacag gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt cagctccggt
     4621 tcccaacgat caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc
     4681 ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag tgttatcact catggttatg
     4741 gcagcactgc ataattctct tactgtcatg ccatccgtaa gatgcttttc tgtgactggt
     4801 gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg
     4861 gcgtcaatac gggataatac cgcgccacat agcagaactt taaaagtgct catcattgga
     4921 aaacgttctt cggggcgaaa actctcaagg atcttaccgc tgttgagatc cagttcgatg
     4981 taacccactc gtgcacccaa ctgatcttca gcatctttta ctttcaccag cgtttctggg
     5041 tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa taagggcgac acggaaatgt
     5101 tgaatactca tactcttcct ttttcaatat tattgaagca tttatcaggg ttattgtctc
     5161 atgagcggat acatatttga atgtatttag aaaaataaac aaataggggt tccgcgcaca
     5221 tttccccgaa aagtgccacc taaattgtaa gcgttaatat tttgttaaaa ttcgcgttaa
     5281 atttttgtta aatcagctca ttttttaacc aataggccga aatcggcaaa atcccttata
     5341 aatcaaaaga atagaccgag atagggttga gtgttgttcc agtttggaac aagagtccac
     5401 tattaaagaa cgtggactcc aacgtcaaag ggcgaaaaac cgtctatcag ggcgatggcc
     5461 cactacgtga accatcaccc taatcaagtt ttttggggtc gaggtgccgt aaagcactaa
     5521 atcggaaccc taaagggagc ccccgattta gagcttgacg gggaaagccg gcgaacgtgg
     5581 cgagaaagga agggaagaaa gcgaaaggag cgggcgctag ggcgctggca agtgtagcgg
     5641 tcacgctgcg cgtaaccacc acacccgccg cgcttaatgc gccgctacag ggcgcgtccc
     5701 attcgccatt caggctgcgc aactgttggg aagggcgatc ggtgcgggcc tcttcgctat
     5761 tacgccagtc gaccgccaat tcaatatggc gtatatggac tcatgccaat tcaatatggt
     5821 ggatctggac ctgtgccaat tcaatatggc gtatatggac tcgtgccaat tcaatatggt
     5881 ggatctggac cccagccaat tcaatatggc ggacttggca ccatgccaat tcaatatggc
     5941 ggacctggca ctgtgccaac tggggagggg tctacttggc acggtgccaa gtttgaggag
     6001 gggtcttggc cctgtgccaa gtccgccata ttgaattggc atggtgccaa taatggcggc
     6061 catattggct atatgccagg atcaatatat aggcaatatc caatatggcc ctatgccaat
     6121 atggctattg gccaggttca atactatgta ttggccctat gccatatagt attccatata
     6181 tgggttttcc tattgacgta gatagcccct cccaatgggc ggtcccatat accatatatg
     6241 gggcttccta ataccgccca tagccactcc cccattgacg tcaatggtct ctatatatgg
     6301 tctttcctat tgacgtcata tgggcggtcc tattgacgta tatggcgcct cccccattga
     6361 cgtcaattac ggtaaatggc ccgcctggct caatgcccat tgacgtcaat aggaccaccc
     6421 accattgacg tcaatgggat ggctcattgc ccattcatat ccgttctcac gccccctatt
     6481 gacgtcaatg acggtaaatg gcccacttgg cagtacatca atatctatta atagtaactt
     6541 ggcaagtaca ttactattgg aagtacgcca gggtacattg gcagtactcc cattgacgtc
     6601 aatggcggta aatggcccgc gatggctgcc aagtacatcc ccattgacgt caatggggag
     6661 gggcaatgac gcaaatgggc gttccattga cgtaaatggg cggtag
//
