LOCUS       sequence-235989-        5662 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AAV vector, NFkappaB-dependent eGFP  expression.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5662)
  AUTHORS   Chtarto A, Bockstael O, Gebara E, Vermoesen K, Melas C, Pythoud C, 
            Levivier M, De Witte O, Luthi-Carter R, Clinkers R, Tenenbaum L
  TITLE     An adeno-associated virus-based intracellular sensor of pathological
            nuclear factor-kappaB activation for disease-inducible gene 
            transfer.
  JOURNAL   PLoS One. 2013;8(1):e53156. doi: 10.1371/journal.pone.0053156. Epub 
            2013 Jan 3.
  PUBMED    23301037
REFERENCE   2  (bases 1 to 5662)
  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: "PLoS One.";
            date: "2013"; volume: "8(1):e53156. doi"; pages: " 
            10.1371/journal.pone.0053156. Epub 2013 Jan 3"
COMMENT     Sequence Label: pSC-NF8-d1-eGFP
FEATURES             Location/Qualifiers
     source          1..5662
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     polyA_signal    31..79
                     /note="synthetic polyadenylation signal"
     misc_feature    93..184
                     /label=pause site
                     /note="RNA polymerase II transcriptional pause signal from 
                     the human alpha-2 globin gene"
     primer_bind     133..152
                     /label=RVprimer3
                     /note="pGL3 vector, forward primer"
     primer_bind     complement(326..342)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     356..380
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     primer_bind     complement(421..437)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     intron          466..562
                     /label=SV40 intron
                     /note="modified SV40 intron with splice donor and acceptor 
                     sites"
     regulatory      633..642
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(639..641,642..644,645..1358)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(639..641,642..644,645..1355)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(684..705)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(945..964)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1292..1313
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    1371..1505
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1395..1414)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     1449..1468
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(1591..1608)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    1597..1708
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      complement(2140..2728)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2220..2239)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2899..3690,3691..3759))
                     /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     3522..3541
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3760..3864)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     3932..3950
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(3988..4010)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     4110..4129
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     rep_origin      complement(4611..5124)
                     /direction=LEFT
                     /label=M13 ori
                     /note="M13 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4806..4827)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     5018..5037
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     repeat_region   5495..5624
                     /label=AAV2 ITR
ORIGIN
        1 tagccatgct ctaggatctg aattggccgc aataaaatat ctttattttc attacatctg
       61 tgtgttggtt ttttgtgtga atcgatagta ctaacatacg ctctccatca aaacaaaacg
      121 aaacaaaaca aactagcaaa ataggctgtc cccagtgcaa gtgcaggtgc cagaacattt
      181 ctctatcgat aggtaccgag ctcttacgcg tgctagcggg aatttccggg aatttccggg
      241 aatttccggg aatttccaga tccgcgtgct agcgggaatt tccgggaatt tccgggaatt
      301 tccgggaatt tccagatcag cttatcgata ccgtcgacct cgcctatata agcagagctc
      361 gtttagtgaa ccgtcagatc gcctggagaa ttcgagctcg gtacccgggg atcagcttat
      421 cgataccgtc gacctcgagg aactgaaaaa ccagaaagtt aactggtaag tttagtcttt
      481 ttgtctttta tttcaggtcc cggatccggt ggtggtgcaa atcaaagaac tgctcctcag
      541 tggatgttgc ctttacttct aggcctgtac ggaagtgtta cttctgctct aaaagctgcg
      601 gaattgtacc cgcggcccgg gatccaccgg tcgccaccat ggtgagcaag ggcgaggagc
      661 tgttcaccgg ggtggtgccc atcctggtcg agctggacgg cgacgtaaac ggccacaagt
      721 tcagcgtgtc cggcgagggc gagggcgatg ccacctacgg caagctgacc ctgaagttca
      781 tctgcaccac cggcaagctg cccgtgccct ggcccaccct cgtgaccacc ctgacctacg
      841 gcgtgcagtg cttcagccgc taccccgacc acatgaagca gcacgacttc ttcaagtccg
      901 ccatgcccga aggctacgtc caggagcgca ccatcttctt caaggacgac ggcaactaca
      961 agacccgcgc cgaggtgaag ttcgagggcg acaccctggt gaaccgcatc gagctgaagg
     1021 gcatcgactt caaggaggac ggcaacatcc tggggcacaa gctggagtac aactacaaca
     1081 gccacaacgt ctatatcatg gccgacaagc agaagaacgg catcaaggtg aacttcaaga
     1141 tccgccacaa catcgaggac ggcagcgtgc agctcgccga ccactaccag cagaacaccc
     1201 ccatcggcga cggccccgtg ctgctgcccg acaaccacta cctgagcacc cagtccgccc
     1261 tgagcaaaga ccccaacgag aagcgcgatc acatggtcct gctggagttc gtgaccgccg
     1321 ccgggatcac tctcggcatg gacgagctgt acaagtaaag cggccgcgcg gatccagaca
     1381 tgataagata cattgatgag tttggacaaa ccacaactag aatgcagtga aaaaaatgct
     1441 ttatttgtga aatttgtgat gctattgctt tatttgtaac cattataagc tgcaataaac
     1501 aagttaacaa caacaattgc attcatttta tgtttcaggt tcagggggag gtgtgggagg
     1561 ttttttagtc gactagagct cgctgatcag cctcgactgt gccttctagt tgccagccat
     1621 ctgttgtttg cccctccccc gtgccttcct tgaccctgga aggtgccact cccactgtcc
     1681 tttcctaata aaatgaggaa attgcatcgc attgtctgag taggtgtcat tctattctgg
     1741 ggggtggggt ggggcaggac agcaaggggg aggattggga agacaatagc aggaacccca
     1801 ctccctctct gcgcgctcgc tcgctcactg aggccgggcg accaaaggtc gcccgacgcc
     1861 cgggctttgc ccgggcggcc tcagtgagcg agcgagcgcg cagctgcatt aatgaatcgg
     1921 ccaacgcgcg gggagaggcg gtttgcgtat tgggcgctct tccgcttcct cgctcactga
     1981 ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca gctcactcaa aggcggtaat
     2041 acggttatcc acagaatcag gggataacgc aggaaagaac atgtgagcaa aaggccagca
     2101 aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc tccgcccccc
     2161 tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga caggactata
     2221 aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc
     2281 gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt ctcatagctc
     2341 acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga
     2401 accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc
     2461 ggtaagacac gacttatcgc cactggcagc agccactggt aacaggatta gcagagcgag
     2521 gtatgtaggc ggtgctacag agttcttgaa gtggtggcct aactacggct acactagaag
     2581 aacagtattt ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa gagttggtag
     2641 ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt gcaagcagca
     2701 gattacgcgc agaaaaaaag gatctcaaga agatcctttg atcttttcta cggggtctga
     2761 cgctcagtgg aacgaaaact cacgttaagg gattttggtc atgagattat caaaaaggat
     2821 cttcacctag atccttttaa attaaaaatg aagttttaaa tcaatctaaa gtatatatga
     2881 gtaaacttgg tctgacagtt accaatgctt aatcagtgag gcacctatct cagcgatctg
     2941 tctatttcgt tcatccatag ttgcctgact ccccgtcgtg tagataacta cgatacggga
     3001 gggcttacca tctggcccca gtgctgcaat gataccgcga gacccacgct caccggctcc
     3061 agatttatca gcaataaacc agccagccgg aagggccgag cgcagaagtg gtcctgcaac
     3121 tttatccgcc tccatccagt ctattaattg ttgccgggaa gctagagtaa gtagttcgcc
     3181 agttaatagt ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc
     3241 gtttggtatg gcttcattca gctccggttc ccaacgatca aggcgagtta catgatcccc
     3301 catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca gaagtaagtt
     3361 ggccgcagtg ttatcactca tggttatggc agcactgcat aattctctta ctgtcatgcc
     3421 atccgtaaga tgcttttctg tgactggtga gtactcaacc aagtcattct gagaatagtg
     3481 tatgcggcga ccgagttgct cttgcccggc gtcaatacgg gataataccg cgccacatag
     3541 cagaacttta aaagtgctca tcattggaaa acgttcttcg gggcgaaaac tctcaaggat
     3601 cttaccgctg ttgagatcca gttcgatgta acccactcgt gcacccaact gatcttcagc
     3661 atcttttact ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa
     3721 aaagggaata agggcgacac ggaaatgttg aatactcata ctcttccttt ttcaatatta
     3781 ttgaagcatt tatcagggtt attgtctcat gagcggatac atatttgaat gtatttagaa
     3841 aaataaacaa ataggggttc cgcgcacatt tccccgaaaa gtgccacctg acgtctaaga
     3901 aaccattatt atcatgacat taacctataa aaataggcgt atcacgaggc cctttcgtct
     3961 cgcgcgtttc ggtgatgacg gtgaaaacct ctgacacatg cagctcccgg agacggtcac
     4021 agcttgtctg taagcggatg ccgggagcag acaagcccgt cagggcgcgt cagcgggtgt
     4081 tggcgggtgt cggggctggc ttaactatgc ggcatcagag cagattgtac tgagagtgca
     4141 ccatatgcgg tgtgaaatac cgcacagatg cgtaaggaga aaataccgca tcaggcgatt
     4201 ccaacatcca ataaatcata caggcaaggc aaagaattag caaaattaag caataaagcc
     4261 tcagagcata aagctaaatc ggttgtacca aaaacattat gaccctgtaa tacttttgcg
     4321 ggagaagcct ttatttcaac gcaaggataa aaatttttag aaccctcata tattttaaat
     4381 gcaatgcctg agtaatgtgt aggtaaagat tcaaacgggt gagaaaggcc ggagacagtc
     4441 aaatcaccat caatatgata ttcaaccgtt ctagctgata aattcatgcc ggagagggta
     4501 gctatttttg agaggtctct acaaaggcta tcaggtcatt gcctgagagt ctggagcaaa
     4561 caagagaatc gatgaacggt aatcgtaaaa ctagcatgtc aatcatatgt accccggttg
     4621 ataatcagaa aagccccaaa aacaggaaga ttgtataagc aaatatttaa attgtaagcg
     4681 ttaatatttt gttaaaattc gcgttaaatt tttgttaaat cagctcattt tttaaccaat
     4741 aggccgaaat cggcaaaatc ccttataaat caaaagaata gaccgagata gggttgagtg
     4801 ttgttccagt ttggaacaag agtccactat taaagaacgt ggactccaac gtcaaagggc
     4861 gaaaaaccgt ctatcagggc gatggcccac tacgtgaacc atcaccctaa tcaagttttt
     4921 tggggtcgag gtgccgtaaa gcactaaatc ggaaccctaa agggagcccc cgatttagag
     4981 cttgacgggg aaagccggcg aacgtggcga gaaaggaagg gaagaaagcg aaaggagcgg
     5041 gcgctagggc gctggcaagt gtagcggtca cgctgcgcgt aaccaccaca cccgccgcgc
     5101 ttaatgcgcc gctacagggc gcgtactatg gttgctttga cgagcacgta taacgtgctt
     5161 tcctcgttag aatcagagcg ggagctaaac aggaggccga ttaaagggat tttagacagg
     5221 aacggtacgc cagaatcctg agaagtgttt ttataatcag tgaggccacc gagtaaaaga
     5281 gtctgtccat cacgcaaatt aaccgttgtc gcaatacttc tttgattagt aataacatca
     5341 cttgcctgag tagaagaact caaactatcg gccttgctgg taatatccag aacaatatta
     5401 ccgccagcca ttgcaacgga atcgccattc gccattcagg ctgcgcaact gttgggaagg
     5461 gcgatcggtg cgggcctctt cgctattacg ccagctgcgc gctcgctcgc tcactgaggc
     5521 cgcccgggca aagcccgggc gtcgggcgac ctttggtcgc ccggcctcag tgagcgagcg
     5581 agcgcgcaga gagggagtgg ccaactccat cactaggggt tccttgtagt taatgattaa
     5641 cccgccatgc tacttatcta cg
//
