LOCUS       sequence-213583-        5888 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  expression of mutant p47phox.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5888)
  AUTHORS   Hervera A, De Virgiliis F, Palmisano I, Zhou L, Tantardini E, Kong 
            G, Hutson T, Danzi MC, Perry RB, Santos CXC, Kapustin AN, Fleck RA, 
            Del Rio JA, Carroll T, Lemmon V, Bixby JL, Shah AM, Fainzilber M, Di
            Giovanni S
  TITLE     Reactive oxygen species regulate axonal regeneration through the 
            release of exosomal NADPH oxidase 2 complexes into injured axons.
  JOURNAL   Nat Cell Biol. 2018 Mar;20(3):307-319. doi: 
            10.1038/s41556-018-0039-x. Epub 2018 Feb 12.
  PUBMED    29434374
REFERENCE   2  (bases 1 to 5888)
  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: 
            "10.1038/s41556-018-0039-x"; journalName: "Nat Cell Biol"; date: 
            "2018-03"; volume: "20"; issue: "3"; pages: "307-319"
COMMENT     Sequence Label: pEGFPc1 hp47phox (S303/304/328D)
FEATURES             Location/Qualifiers
     source          1..5888
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        130..433
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        434..637
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     588..608
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      676..685
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(682..684,685..687,688..1398)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(682..684,685..687,688..1398)
                     /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(727..748)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(988..1007)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1335..1356
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    2745..2866
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2782..2801)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2836..2855
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(2873..3328)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3010..3031)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3222..3241
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3355..3459
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3461..3818
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3461..3652
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3484..3504)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3669..3804
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3853..4647
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     CDS             3853..4647
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(3907..3926)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4517..4536
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(4835..4854)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    4879..4926
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5255..5843
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5744..5763
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 ccttttgctc acatgttctt tcctgcgtta tcccctgatt ctgtggataa ccgtattacc
       61 gccatgcatt agttattaat agtaatcaat tacggggtca ttagttcata gcccatatat
      121 ggagttccgc gttacataac ttacggtaaa tggcccgcct ggctgaccgc ccaacgaccc
      181 ccgcccattg acgtcaataa tgacgtatgt tcccatagta acgccaatag ggactttcca
      241 ttgacgtcaa tgggtggagt atttacggta aactgcccac ttggcagtac atcaagtgta
      301 tcatatgcca agtacgcccc ctattgacgt caatgacggt aaatggcccg cctggcatta
      361 tgcccagtac atgaccttat gggactttcc tacttggcag tacatctacg tattagtcat
      421 cgctattacc atggtgatgc ggttttggca gtacatcaat gggcgtggat agcggtttga
      481 ctcacgggga tttccaagtc tccaccccat tgacgtcaat gggagtttgt tttggcacca
      541 aaatcaacgg gactttccaa aatgtcgtaa caactccgcc ccattgacgc aaatgggcgg
      601 taggcgtgta cggtgggagg tctatataag cagagctggt ttagtgaacc gtcagatccg
      661 ctagcgctac cggtcgccac catggtgagc aagggcgagg agctgttcac cggggtggtg
      721 cccatcctgg tcgagctgga cggcgacgta aacggccaca agttcagcgt gtccggcgag
      781 ggcgagggcg atgccaccta cggcaagctg accctgaagt tcatctgcac caccggcaag
      841 ctgcccgtgc cctggcccac cctcgtgacc accctgacct acggcgtgca gtgcttcagc
      901 cgctaccccg accacatgaa gcagcacgac ttcttcaagt ccgccatgcc cgaaggctac
      961 gtccaggagc gcaccatctt cttcaaggac gacggcaact acaagacccg cgccgaggtg
     1021 aagttcgagg gcgacaccct ggtgaaccgc atcgagctga agggcatcga cttcaaggag
     1081 gacggcaaca tcctggggca caagctggag tacaactaca acagccacaa cgtctatatc
     1141 atggccgaca agcagaagaa cggcatcaag gtgaacttca agatccgcca caacatcgag
     1201 gacggcagcg tgcagctcgc cgaccactac cagcagaaca cccccatcgg cgacggcccc
     1261 gtgctgctgc ccgacaacca ctacctgagc acccagtccg ccctgagcaa agaccccaac
     1321 gagaagcgcg atcacatggt cctgctggag ttcgtgaccg ccgccgggat cactctcggc
     1381 atggacgagc tgtacaagtc cggactcaga tccatggggg acaccttcat ccgtcacatc
     1441 gccctgctgg gctttgagaa gcgcttcgta cccagccagc actatgtgta catgttcctg
     1501 gtgaaatggc aggacctgtc ggagaaggtg gtctaccggc gcttcaccga gatctacgag
     1561 ttccataaaa ccttaaaaga aatgttccct attgaggcag gggcgatcaa tccagagaac
     1621 aggatcatcc cccacctccc agctcccaag tggtttgacg ggcagcgggc cgccgagaac
     1681 cgccagggca cacttaccga gtactgcagc acgctcatga gcctgcccac caagatctcc
     1741 cgctgtcccc acctcctcga cttcttcaag gtgcgccctg atgacctcaa gctccccacg
     1801 gacaaccaga caaaaaagcc agagacatac ttgatgccca aagatggcaa gagtaccgcg
     1861 acagacatca ccggccccat catcctgcag acgtaccgcg ccattgccaa ctacgagaag
     1921 acctcgggct ccgagatggc tctgtccacg ggggacgtgg tggaggtcgt agagaagagc
     1981 gagagcggtt ggtggttctg tcagatgaaa gcaaagcgag gctggatacc agcatccttc
     2041 ctcgagcccc tggacagtcc tgacgagacg gaagaccctg agcccaacta tgcaggtgag
     2101 ccatacgtcg ccatcaaggc ctacactgct gtggaggggg acgaggtgtc cctgctcgag
     2161 ggtgaagctg ttgaggtcat tcacaagctc ctggacggct ggtgggtcat caggaaagac
     2221 gacgtcacag gctacttccc gtccatgtac ctgcaaaagt cagggcaaga cgtgtcccag
     2281 gcccaacgcc agatcaagcg gggggcgccg ccccgcaggg acgacatccg caacgcgcac
     2341 agcatccatc agcggtcgcg gaagcgcctc agccaggacg cctatcgccg caacgacgtc
     2401 cgttttctgc agcagcgacg ccgccaggcg cggccgggac cgcagagccc cgggagcccg
     2461 ctcgaggagg agcggcagac gcagcgctct aaaccgcagc cggcggtgcc cccgcggccg
     2521 agcgccgacc tcatcctgaa ccgctgcagc gagagcacca agcggaagct ggcgtctgcc
     2581 gtctgaattc tgcagtcgac ggtaccgcgg gcccgggatc caccggatct agataactga
     2641 tcataatcag ccataccaca tttgtagagg ttttacttgc tttaaaaaac ctcccacacc
     2701 tccccctgaa cctgaaacat aaaatgaatg caattgttgt tgttaacttg tttattgcag
     2761 cttataatgg ttacaaataa agcaatagca tcacaaattt cacaaataaa gcattttttt
     2821 cactgcattc tagttgtggt ttgtccaaac tcatcaatgt atcttaacgc gtaaattgta
     2881 agcgttaata ttttgttaaa attcgcgtta aatttttgtt aaatcagctc attttttaac
     2941 caataggccg aaatcggcaa aatcccttat aaatcaaaag aatagaccga gatagggttg
     3001 agtgttgttc cagtttggaa caagagtcca ctattaaaga acgtggactc caacgtcaaa
     3061 gggcgaaaaa ccgtctatca gggcgatggc ccactacgtg aaccatcacc ctaatcaagt
     3121 tttttggggt cgaggtgccg taaagcacta aatcggaacc ctaaagggag cccccgattt
     3181 agagcttgac ggggaaagcc ggcgaacgtg gcgagaaagg aagggaagaa agcgaaagga
     3241 gcgggcgcta gggcgctggc aagtgtagcg gtcacgctgc gcgtaaccac cacacccgcc
     3301 gcgcttaatg cgccgctaca gggcgcgtca ggtggcactt ttcggggaaa tgtgcgcgga
     3361 acccctattt gtttattttt ctaaatacat tcaaatatgt atccgctcat gagacaataa
     3421 ccctgataaa tgcttcaata atattgaaaa aggaagagtc ctgaggcgga aagaaccagc
     3481 tgtggaatgt gtgtcagtta gggtgtggaa agtccccagg ctccccagca ggcagaagta
     3541 tgcaaagcat gcatctcaat tagtcagcaa ccaggtgtgg aaagtcccca ggctccccag
     3601 caggcagaag tatgcaaagc atgcatctca attagtcagc aaccatagtc ccgcccctaa
     3661 ctccgcccat cccgccccta actccgccca gttccgccca ttctccgccc catggctgac
     3721 taattttttt tatttatgca gaggccgagg ccgcctcggc ctctgagcta ttccagaagt
     3781 agtgaggagg cttttttgga ggcctaggct tttgcaaaga tcgatcaaga gacaggatga
     3841 ggatcgtttc gcatgattga acaagatgga ttgcacgcag gttctccggc cgcttgggtg
     3901 gagaggctat tcggctatga ctgggcacaa cagacaatcg gctgctctga tgccgccgtg
     3961 ttccggctgt cagcgcaggg gcgcccggtt ctttttgtca agaccgacct gtccggtgcc
     4021 ctgaatgaac tgcaagacga ggcagcgcgg ctatcgtggc tggccacgac gggcgttcct
     4081 tgcgcagctg tgctcgacgt tgtcactgaa gcgggaaggg actggctgct attgggcgaa
     4141 gtgccggggc aggatctcct gtcatctcac cttgctcctg ccgagaaagt atccatcatg
     4201 gctgatgcaa tgcggcggct gcatacgctt gatccggcta cctgcccatt cgaccaccaa
     4261 gcgaaacatc gcatcgagcg agcacgtact cggatggaag ccggtcttgt cgatcaggat
     4321 gatctggacg aagagcatca ggggctcgcg ccagccgaac tgttcgccag gctcaaggcg
     4381 agcatgcccg acggcgagga tctcgtcgtg acccatggcg atgcctgctt gccgaatatc
     4441 atggtggaaa atggccgctt ttctggattc atcgactgtg gccggctggg tgtggcggac
     4501 cgctatcagg acatagcgtt ggctacccgt gatattgctg aagagcttgg cggcgaatgg
     4561 gctgaccgct tcctcgtgct ttacggtatc gccgctcccg attcgcagcg catcgccttc
     4621 tatcgccttc ttgacgagtt cttctgagcg ggactctggg gttcgaaatg accgaccaag
     4681 cgacgcccaa cctgccatca cgagatttcg attccaccgc cgccttctat gaaaggttgg
     4741 gcttcggaat cgttttccgg gacgccggct ggatgatcct ccagcgcggg gatctcatgc
     4801 tggagttctt cgcccaccct agggggaggc taactgaaac acggaaggag acaataccgg
     4861 aaggaacccg cgctatgacg gcaataaaaa gacagaataa aacgcacggt gttgggtcgt
     4921 ttgttcataa acgcggggtt cggtcccagg gctggcactc tgtcgatacc ccaccgagac
     4981 cccattgggg ccaatacgcc cgcgtttctt ccttttcccc accccacccc ccaagttcgg
     5041 gtgaaggccc agggctcgca gccaacgtcg gggcggcagg ccctgccata gcctcaggtt
     5101 actcatatat actttagatt gatttaaaac ttcattttta atttaaaagg atctaggtga
     5161 agatcctttt tgataatctc atgaccaaaa tcccttaacg tgagttttcg ttccactgag
     5221 cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa
     5281 tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg ccggatcaag
     5341 agctaccaac tctttttccg aaggtaactg gcttcagcag agcgcagata ccaaatactg
     5401 ttcttctagt gtagccgtag ttaggccacc acttcaagaa ctctgtagca ccgcctacat
     5461 acctcgctct gctaatcctg ttaccagtgg ctgctgccag tggcgataag tcgtgtctta
     5521 ccgggttgga ctcaagacga tagttaccgg ataaggcgca gcggtcgggc tgaacggggg
     5581 gttcgtgcac acagcccagc ttggagcgaa cgacctacac cgaactgaga tacctacagc
     5641 gtgagctatg agaaagcgcc acgcttcccg aagggagaaa ggcggacagg tatccggtaa
     5701 gcggcagggt cggaacagga gagcgcacga gggagcttcc agggggaaac gcctggtatc
     5761 tttatagtcc tgtcgggttt cgccacctct gacttgagcg tcgatttttg tgatgctcgt
     5821 caggggggcg gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct
     5881 tttgctgg
//
