LOCUS       sequence-213636-        6620 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  mutant p47phox.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6620)
  TITLE     Di Giovanni Lab neuroscience plasmids
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 6620)
  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: 
            "Unpublished"
COMMENT     Sequence Label: p87AAVEGFP hp47phox (S303/304/328D)
FEATURES             Location/Qualifiers
     source          1..6620
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        69..372
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        373..576
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     527..547
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     573..597
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     intron          725..1103
                     /label=chimeric intron
                     /note="fusion between human cytomegalovirus intron A and 
                     human beta-globin intron 2"
     promoter        1174..1192
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     regulatory      1252..1261
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(1258..1260,1261..1263,1264..1974)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(1258..1260,1261..1263,1264..1974)
                     /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(1303..1324)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1564..1583)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1911..1932
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(3203..3222)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(3204..3222)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     polyA_signal    3230..3706
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(3429..3448)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     repeat_region   3746..3886
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   3746..3875
                     /label=AAV2 ITR
     rep_origin      3961..4416
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4048..4067)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4258..4279
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4433..4452)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4552..4574
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(4612..4630)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4698..4802
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4803..4871,4872..5663)
                     /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(5021..5040)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5834..6422
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6323..6342
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     repeat_region   6484..6613
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
ORIGIN
        1 cacgcgtcta gttattaata gtaatcaatt acggggtcat tagttcatag cccatatatg
       61 gagttccgcg ttacataact tacggtaaat ggcccgcctg gctgaccgcc caacgacccc
      121 cgcccattga cgtcaataat gacgtatgtt cccatagtaa cgccaatagg gactttccat
      181 tgacgtcaat gggtggagta tttacggtaa actgcccact tggcagtaca tcaagtgtat
      241 catatgccaa gtacgccccc tattgacgtc aatgacggta aatggcccgc ctggcattat
      301 gcccagtaca tgaccttatg ggactttcct acttggcagt acatctacgt attagtcatc
      361 gctattacca tggtgatgcg gttttggcag tacatcaatg ggcgtggata gcggtttgac
      421 tcacggggat ttccaagtct ccaccccatt gacgtcaatg ggagtttgtt ttggcaccaa
      481 aatcaacggg actttccaaa atgtcgtaac aactccgccc cattgacgca aatgggcggt
      541 aggcgtgtac ggtgggaggt ctatataagc agagctcgtt tagtgaaccg tcagatcgcc
      601 tggagacgcc atccacgctg ttttgacctc catagaagac accgggaccg atccagcctc
      661 cgcggattcg aatcccggcc gggaacggtg cattggaacg cggattcccc gtgccaagag
      721 tgacgtaagt accgcctata gagtctatag gcccacaaaa aatgctttct tcttttaata
      781 tacttttttg tttatcttat ttctaatact ttccctaatc tctttctttc agggcaataa
      841 tgatacaatg tatcatgcct ctttgcacca ttctaaagaa taacagtgat aatttctggg
      901 ttaaggcaat agcaatattt ctgcatataa atatttctgc atataaattg taactgatgt
      961 aagaggtttc atattgctaa tagcagctac aatccagcta ccattctgct tttattttat
     1021 ggttgggata aggctggatt attctgagtc caagctaggc ccttttgcta atcatgttca
     1081 tacctcttat cttcctccca cagctcctgg gcaacgtgct ggtctgtgtg ctggcccatc
     1141 actttggcaa agaattggga ttcgaacatc gataattaac cctcactaaa gggaacaaaa
     1201 gctggagctc caccgcggtg gcggccgctc tagcccgggc ggatcccggt cgccaccatg
     1261 gtgagcaagg gcgaggagct gttcaccggg gtggtgccca tcctggtcga gctggacggc
     1321 gacgtaaacg gccacaagtt cagcgtgtcc ggcgagggcg agggcgatgc cacctacggc
     1381 aagctgaccc tgaagttcat ctgcaccacc ggcaagctgc ccgtgccctg gcccaccctc
     1441 gtgaccaccc tgacctacgg cgtgcagtgc ttcagccgct accccgacca catgaagcag
     1501 cacgacttct tcaagtccgc catgcccgaa ggctacgtcc aggagcgcac catcttcttc
     1561 aaggacgacg gcaactacaa gacccgcgcc gaggtgaagt tcgagggcga caccctggtg
     1621 aaccgcatcg agctgaaggg catcgacttc aaggaggacg gcaacatcct ggggcacaag
     1681 ctggagtaca actacaacag ccacaacgtc tatatcatgg ccgacaagca gaagaacggc
     1741 atcaaggtga acttcaagat ccgccacaac atcgaggacg gcagcgtgca gctcgccgac
     1801 cactaccagc agaacacccc catcggcgac ggccccgtgc tgctgcccga caaccactac
     1861 ctgagcaccc agtccgccct gagcaaagac cccaacgaga agcgcgatca catggtcctg
     1921 ctggagttcg tgaccgccgc cgggatcact ctcggcatgg acgagctgta caagtccgga
     1981 ctcagatcca tgggggacac cttcatccgt cacatcgccc tgctgggctt tgagaagcgc
     2041 ttcgtaccca gccagcacta tgtgtacatg ttcctggtga aatggcagga cctgtcggag
     2101 aaggtggtct accggcgctt caccgagatc tacgagttcc ataaaacctt aaaagaaatg
     2161 ttccctattg aggcaggggc gatcaatcca gagaacagga tcatccccca cctcccagct
     2221 cccaagtggt ttgacgggca gcgggccgcc gagaaccgcc agggcacact taccgagtac
     2281 tgcagcacgc tcatgagcct gcccaccaag atctcccgct gtccccacct cctcgacttc
     2341 ttcaaggtgc gccctgatga cctcaagctc cccacggaca accagacaaa aaagccagag
     2401 acatacttga tgcccaaaga tggcaagagt accgcgacag acatcaccgg ccccatcatc
     2461 ctgcagacgt accgcgccat tgccaactac gagaagacct cgggctccga gatggctctg
     2521 tccacggggg acgtggtgga ggtcgtagag aagagcgaga gcggttggtg gttctgtcag
     2581 atgaaagcaa agcgaggctg gataccagca tccttcctcg agcccctgga cagtcctgac
     2641 gagacggaag accctgagcc caactatgca ggtgagccat acgtcgccat caaggcctac
     2701 actgctgtgg agggggacga ggtgtccctg ctcgagggtg aagctgttga ggtcattcac
     2761 aagctcctgg acggctggtg ggtcatcagg aaagacgacg tcacaggcta cttcccgtcc
     2821 atgtacctgc aaaagtcagg gcaagacgtg tcccaggccc aacgccagat caagcggggg
     2881 gcgccgcccc gcagggacga catccgcaac gcgcacagca tccatcagcg gtcgcggaag
     2941 cgcctcagcc aggacgccta tcgccgcaac gacgtccgtt ttctgcagca gcgacgccgc
     3001 caggcgcggc cgggaccgca gagccccggg agcccgctcg aggaggagcg gcagacgcag
     3061 cgctctaaac cgcagccggc ggtgcccccg cggccgagcg ccgacctcat cctgaaccgc
     3121 tgcagcgaga gcaccaagcg gaagctggcg tctgccgtct gaattctgca gtcgacggta
     3181 ccaggtaagt gtacccaatt cgccctatag tgagtcgtat tagatctacg ggtggcatcc
     3241 ctgtgacccc tccccagtgc ctctcctggc cctggaagtt gccactccag tgcccaccag
     3301 ccttgtccta ataaaattaa gttgcatcat tttgtctgac taggtgtcct tctataatat
     3361 tatggggtgg aggggggtgg tatggagcaa ggggcaagtt gggaagacaa cctgtagggc
     3421 ctgcggggtc tattgggaac caagctggag tgcagtggca caatcttggc tcactgcaat
     3481 ctccgcctcc tgggttcaag cgattctcct gcctcagcct cccgagttgt tgggattcca
     3541 ggcatgcatg accaggctca gctaattttt gtttttttgg tagagacggg gtttcaccat
     3601 attggccagg ctggtctcca actcctaatc tcaggtgatc tacccacctt ggcctcccaa
     3661 attgctggga ttacaggcgt gaaccactgc tcccttccct gtccttctga ttttgtaggt
     3721 aaccacgtgc ggaccgagcg gccgcaggaa cccctagtga tggagttggc cactccctct
     3781 ctgcgcgctc gctcgctcac tgaggccggg cgaccaaagg tcgcccgacg cccgggcttt
     3841 gcccgggcgg cctcagtgag cgagcgagcg cgcagctgcc tgcaggggcg cctgatgcgg
     3901 tattttctcc ttacgcatct gtgcggtatt tcacaccgca tacgtcaaag caaccatagt
     3961 acgcgccctg tagcggcgca ttaagcgcgg cgggtgtggt ggttacgcgc agcgtgaccg
     4021 ctacacttgc cagcgcctta gcgcccgctc ctttcgcttt cttcccttcc tttctcgcca
     4081 cgttcgccgg ctttccccgt caagctctaa atcgggggct ccctttaggg ttccgattta
     4141 gtgctttacg gcacctcgac cccaaaaaac ttgatttggg tgatggttca cgtagtgggc
     4201 catcgccctg atagacggtt tttcgccctt tgacgttgga gtccacgttc tttaatagtg
     4261 gactcttgtt ccaaactgga acaacactca actctatctc gggctattct tttgatttat
     4321 aagggatttt gccgatttcg gtctattggt taaaaaatga gctgatttaa caaaaattta
     4381 acgcgaattt taacaaaata ttaacgttta caattttatg gtgcactctc agtacaatct
     4441 gctctgatgc cgcatagtta agccagcccc gacacccgcc aacacccgct gacgcgccct
     4501 gacgggcttg tctgctcccg gcatccgctt acagacaagc tgtgaccgtc tccgggagct
     4561 gcatgtgtca gaggttttca ccgtcatcac cgaaacgcgc gagacgaaag ggcctcgtga
     4621 tacgcctatt tttataggtt aatgtcatga taataatggt ttcttagacg tcaggtggca
     4681 cttttcgggg aaatgtgcgc ggaaccccta tttgtttatt tttctaaata cattcaaata
     4741 tgtatccgct catgagacaa taaccctgat aaatgcttca ataatattga aaaaggaaga
     4801 gtatgagtat tcaacatttc cgtgtcgccc ttattccctt ttttgcggca ttttgccttc
     4861 ctgtttttgc tcacccagaa acgctggtga aagtaaaaga tgctgaagat cagttgggtg
     4921 cacgagtggg ttacatcgaa ctggatctca acagcggtaa gatccttgag agttttcgcc
     4981 ccgaagaacg ttttccaatg atgagcactt ttaaagttct gctatgtggc gcggtattat
     5041 cccgtattga cgccgggcaa gagcaactcg gtcgccgcat acactattct cagaatgact
     5101 tggttgagta ctcaccagtc acagaaaagc atcttacgga tggcatgaca gtaagagaat
     5161 tatgcagtgc tgccataacc atgagtgata acactgcggc caacttactt ctgacaacga
     5221 tcggaggacc gaaggagcta accgcttttt tgcacaacat gggggatcat gtaactcgcc
     5281 ttgatcgttg ggaaccggag ctgaatgaag ccataccaaa cgacgagcgt gacaccacga
     5341 tgcctgtagc aatggcaaca acgttgcgca aactattaac tggcgaacta cttactctag
     5401 cttcccggca acaattaata gactggatgg aggcggataa agttgcagga ccacttctgc
     5461 gctcggccct tccggctggc tggtttattg ctgataaatc tggagccggt gagcgtgggt
     5521 ctcgcggtat cattgcagca ctggggccag atggtaagcc ctcccgtatc gtagttatct
     5581 acacgacggg gagtcaggca actatggatg aacgaaatag acagatcgct gagataggtg
     5641 cctcactgat taagcattgg taactgtcag accaagttta ctcatatata ctttagattg
     5701 atttaaaact tcatttttaa tttaaaagga tctaggtgaa gatccttttt gataatctca
     5761 tgaccaaaat cccttaacgt gagttttcgt tccactgagc gtcagacccc gtagaaaaga
     5821 tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa
     5881 aaccaccgct accagcggtg gtttgtttgc cggatcaaga gctaccaact ctttttccga
     5941 aggtaactgg cttcagcaga gcgcagatac caaatactgt tcttctagtg tagccgtagt
     6001 taggccacca cttcaagaac tctgtagcac cgcctacata cctcgctctg ctaatcctgt
     6061 taccagtggc tgctgccagt ggcgataagt cgtgtcttac cgggttggac tcaagacgat
     6121 agttaccgga taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca cagcccagct
     6181 tggagcgaac gacctacacc gaactgagat acctacagcg tgagctatga gaaagcgcca
     6241 cgcttcccga agggagaaag gcggacaggt atccggtaag cggcagggtc ggaacaggag
     6301 agcgcacgag ggagcttcca gggggaaacg cctggtatct ttatagtcct gtcgggtttc
     6361 gccacctctg acttgagcgt cgatttttgt gatgctcgtc aggggggcgg agcctatgga
     6421 aaaacgccag caacgcggcc tttttacggt tcctggcctt ttgctggcct tttgctcaca
     6481 tgtcctgcag gcagctgcgc gctcgctcgc tcactgaggc cgcccgggcg tcgggcgacc
     6541 tttggtcgcc cggcctcagt gagcgagcga gcgcgcagag agggagtggc caactccatc
     6601 actaggggtt cctgcggccg
//
