LOCUS       sequence-488699-        7171 bp    DNA     circular SYN 08-OCT-2025
DEFINITION  AAV plasmid expressing GFP under the CAG promoter..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7171)
  AUTHORS   Nader WO, Brown KS, Boyle NR, Kaplelach AK, Abdelaziz SM, Davis SE, 
            Aljabi Q, Hakim AR, Davidson AG, Vollmer GA, Wright LC, Echols JB, 
            Saad J, Pena NS, Arrant AE
  TITLE     TFEB overexpression alleviates autophagy-lysosomal deficits caused 
            by progranulin insufficiency.
  JOURNAL   Sci Rep. 2025 Jul 19;15(1):26217. doi: 10.1038/s41598-025-12268-0.
  PUBMED    40683940
REFERENCE   2  (bases 1 to 7171)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Oct 8, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Sci Rep."; 
            date: "2025-07-19"; volume: "15(1):26217. doi"; pages: " 
            10.1038/s41598-025-12268-0"
COMMENT     Sequence Label: AAV-CAG-GFP
FEATURES             Location/Qualifiers
     source          1..7171
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   4..133
                     /label=AAV2 ITR
     primer_bind     complement(189..208)
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     enhancer        275..654
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        656..930
                     /label=chicken beta-actin promoter
     primer_bind     1343..1365
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     1427..1446
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     regulatory      1504..1513
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(1510..1512,1513..1515,1516..2229)
                     /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(1510..1512,1513..1515,1516..2226)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(1555..1576)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1816..1835)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2163..2184
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     misc_feature    2254..2842
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(2307..2327)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(2725..2736)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(2934..2953)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    2999..3054
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(3053..3072)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     repeat_region   3454..3583
                     /label=AAV2 ITR
     rep_origin      3954..4467
                     /direction=RIGHT
                     /label=M13 ori
                     /note="M13 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4041..4060)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4251..4272
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4949..4968)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5068..5090
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(5128..5146)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5214..5318
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5319..5387,5388..6179)
                     /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(5537..5556)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6350..6938
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6839..6858
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 cagctgcgcg ctcgctcgct cactgaggcc gcccgggcaa agcccgggcg tcgggcgacc
       61 tttggtcgcc cggcctcagt gagcgagcga gcgcgcagag agggagtggc caactccatc
      121 actaggggtt ccttgtagtt aatgattaac ccgccatgct acttatctac gtagccatgc
      181 tctagcggcc tcggcctctg cataaataaa aaaaattagt cagccatgag cttggcccat
      241 tgcatacgtt gtatccatat cataatatgt actcgacatt gattattgac tagttattaa
      301 tagtaatcaa ttacggggtc attagttcat agcccatata tggagttccg cgttacataa
      361 cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt gacgtcaata
      421 atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca atgggtggag
      481 tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc aagtacgccc
      541 cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta catgacctta
      601 tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac catggtcgag
      661 gtgagcccca cgttctgctt cactctcccc atctcccccc cctccccacc cccaattttg
      721 tatttattta ttttttaatt attttgtgca gcgatggggg cggggggggg ggggcgcgcg
      781 ccaggcgggg cggggcgggg cgaggggcgg ggcggggcga ggcggagagg tgcggcggca
      841 gccaatcaga gcggcgcgct ccgaaagttt ccttttatgg cgaggcggcg gcggcggcgg
      901 ccctataaaa agcgaagcgc gcggcgggcg ggagtcgctg cgcgctgcct tcgccccgtg
      961 ccccgctccg ccgccgcctc gcgccgcccg ccccggctct gactgaccgc gttactccca
     1021 caggtgagcg ggcgggacgg cccttctcct ccgggctgta attagcgctt ggtttaatga
     1081 cggcttgttt cttttctgtg gctgcgtgaa agccttgagg ggctccggga gggccctttg
     1141 tgcgggggga gcggctcggg gggtgcgtgc gtgtgtgtgt gcgtggggag cgccgcgtgc
     1201 ggctccgcgc tgcccggcgg ctgtgagcgc tgcgggcgcg gcgcggggct ttgtgcgctc
     1261 cgcagtgtgc gcgaggggag cgcggccggg ggcggtgccc cgcgggggga cggctgcctt
     1321 cgggggggac ggggcagggc ggggttcggc ttctggcgtg tgaccggcgg ctctagagcc
     1381 tctgctaacc atgttcatgc cttcttcttt ttcctacagc tcctgggcaa cgtgctggtt
     1441 attgtgctgt ctcatcattt tggcaaagaa tctcgagctc aagcttcgaa ttctgcagtc
     1501 gacgccacca tggtgagcaa gggcgaggag ctgttcaccg gggtggtgcc catcctggtc
     1561 gagctggacg gcgacgtaaa cggccacaag ttcagcgtgt ccggcgaggg cgagggcgat
     1621 gccacctacg gcaagctgac cctgaagttc atctgcacca ccggcaagct gcccgtgccc
     1681 tggcccaccc tcgtgaccac cctgacctac ggcgtgcagt gcttcagccg ctaccccgac
     1741 cacatgaagc agcacgactt cttcaagtcc gccatgcccg aaggctacgt ccaggagcgc
     1801 accatcttct tcaaggacga cggcaactac aagacccgcg ccgaggtgaa gttcgagggc
     1861 gacaccctgg tgaaccgcat cgagctgaag ggcatcgact tcaaggagga cggcaacatc
     1921 ctggggcaca agctggagta caactacaac agccacaacg tctatatcat ggccgacaag
     1981 cagaagaacg gcatcaaggt gaacttcaag atccgccaca acatcgagga cggcagcgtg
     2041 cagctcgccg accactacca gcagaacacc cccatcggcg acggccccgt gctgctgccc
     2101 gacaaccact acctgagcac ccagtccgcc ctgagcaaag accccaacga gaagcgcgat
     2161 cacatggtcc tgctggagtt cgtgaccgcc gccgggatca ctctcggcat ggacgagctg
     2221 tacaagtaat gagatatcgg taccggatcc gataatcaac ctctggatta caaaatttgt
     2281 gaaagattga ctggtattct taactatgtt gctcctttta cgctatgtgg atacgctgct
     2341 ttaatgcctt tgtatcatgc tattgcttcc cgtatggctt tcattttctc ctccttgtat
     2401 aaatcctggt tgctgtctct ttatgaggag ttgtggcccg ttgtcaggca acgtggcgtg
     2461 gtgtgcactg tgtttgctga cgcaaccccc actggttggg gcattgccac cacctgtcag
     2521 ctcctttccg ggactttcgc tttccccctc cctattgcca cggcggaact catcgccgcc
     2581 tgccttgccc gctgctggac aggggctcgg ctgttgggca ctgacaattc cgtggtgttg
     2641 tcggggaaat catcgtcctt tccttggctg ctcgcctgtg ttgccacctg gattctgcgc
     2701 gggacgtcct tctgctacgt cccttcggcc ctcaatccag cggaccttcc ttcccgcggc
     2761 ctgctgccgg ctctgcggcc tcttccgcgt cttcgccttc gccctcagac gagtcggatc
     2821 tccctttggg ccgcctcccc gcatcggatc caattcactc ctcaggtgca ggctgcctat
     2881 cagaaggtgg tggctggtgt ggccaatgcc ctggctcaca aataccactg agatcttttt
     2941 ccctctgcca aaaattatgg ggacatcatg aagccccttg agcatctgac ttctggctaa
     3001 taaaggaaat ttattttcat tgcaatagtg tgttggaatt ttttgtgtct ctcactcgga
     3061 aggacatatg ggagggcaaa tcatttaaaa catcagaatg agtatttggt ttagagtttg
     3121 gcaacatatg cccatatgct ggctgccatg aacaaaggtt ggctataaag aggtcatcag
     3181 tatatgaaac agccccctgc tgtccattcc ttattccata gaaaagcctt gacttgaggt
     3241 tagatttttt ttatattttg ttttgtgtta tttttttctt taacatccct aaaattttcc
     3301 ttacatgttt tactagccag atttttcctc ctctcctgac tactcccagt catagctgtc
     3361 cctcttctct tatggagatc cctcgacctg cagcccaagc tctagagcat ggctacgtag
     3421 ataagtagca tggcgggtta atcattaact acaaggaacc cctagtgatg gagttggcca
     3481 ctccctctct gcgcgctcgc tcgctcactg aggccgggcg accaaaggtc gcccgacgcc
     3541 cgggctttgc ccgggcggcc tcagtgagcg agcgagcgcg cagctggcgt aatagcgaag
     3601 aggcccgcac cgatcgccct tcccaacagt tgcgcagcct gaatggcgaa tggcgattcc
     3661 gttgcaatgg ctggcggtaa tattgttctg gatattacca gcaaggccga tagtttgagt
     3721 tcttctactc aggcaagtga tgttattact aatcaaagaa gtattgcgac aacggttaat
     3781 ttgcgtgatg gacagactct tttactcggt ggcctcactg attataaaaa cacttctcag
     3841 gattctggcg taccgttcct gtctaaaatc cctttaatcg gcctcctgtt tagctcccgc
     3901 tctgattcta acgaggaaag cacgttatac gtgctcgtca aagcaaccat agtacgcgcc
     3961 ctgtagcggc gcattaagcg cggcgggtgt ggtggttacg cgcagcgtga ccgctacact
     4021 tgccagcgcc ctagcgcccg ctcctttcgc tttcttccct tcctttctcg ccacgttcgc
     4081 cggctttccc cgtcaagctc taaatcgggg gctcccttta gggttccgat ttagtgcttt
     4141 acggcacctc gaccccaaaa aacttgatta gggtgatggt tcacgtagtg ggccatcgcc
     4201 ctgatagacg gtttttcgcc ctttgacgtt ggagtccacg ttctttaata gtggactctt
     4261 gttccaaact ggaacaacac tcaaccctat ctcggtctat tcttttgatt tataagggat
     4321 tttgccgatt tcggcctatt ggttaaaaaa tgagctgatt taacaaaaat ttaacgcgaa
     4381 ttttaacaaa atattaacgc ttacaattta aatatttgct tatacaatct tcctgttttt
     4441 ggggcttttc tgattatcaa ccggggtaca tatgattgac atgctagttt tacgattacc
     4501 gttcatcgat tctcttgttt gctccagact ctcaggcaat gacctgatag cctttgtaga
     4561 gacctctcaa aaatagctac cctctccggc atgaatttat cagctagaac ggttgaatat
     4621 catattgatg gtgatttgac tgtctccggc ctttctcacc cgtttgaatc tttacctaca
     4681 cattactcag gcattgcatt taaaatatat gagggttcta aaaattttta tccttgcgtt
     4741 gaaataaagg cttctcccgc aaaagtatta cagggtcata atgtttttgg tacaaccgat
     4801 ttagctttat gctctgaggc tttattgctt aattttgcta attctttgcc ttgcctgtat
     4861 gatttattgg atgttggaat cgcctgatgc ggtattttct ccttacgcat ctgtgcggta
     4921 tttcacaccg catatggtgc actctcagta caatctgctc tgatgccgca tagttaagcc
     4981 agccccgaca cccgccaaca cccgctgacg cgccctgacg ggcttgtctg ctcccggcat
     5041 ccgcttacag acaagctgtg accgtctccg ggagctgcat gtgtcagagg ttttcaccgt
     5101 catcaccgaa acgcgcgaga cgaaagggcc tcgtgatacg cctattttta taggttaatg
     5161 tcatgataat aatggtttct tagacgtcag gtggcacttt tcggggaaat gtgcgcggaa
     5221 cccctatttg tttatttttc taaatacatt caaatatgta tccgctcatg agacaataac
     5281 cctgataaat gcttcaataa tattgaaaaa ggaagagtat gagtattcaa catttccgtg
     5341 tcgcccttat tccctttttt gcggcatttt gccttcctgt ttttgctcac ccagaaacgc
     5401 tggtgaaagt aaaagatgct gaagatcagt tgggtgcacg agtgggttac atcgaactgg
     5461 atctcaacag cggtaagatc cttgagagtt ttcgccccga agaacgtttt ccaatgatga
     5521 gcacttttaa agttctgcta tgtggcgcgg tattatcccg tattgacgcc gggcaagagc
     5581 aactcggtcg ccgcatacac tattctcaga atgacttggt tgagtactca ccagtcacag
     5641 aaaagcatct tacggatggc atgacagtaa gagaattatg cagtgctgcc ataaccatga
     5701 gtgataacac tgcggccaac ttacttctga caacgatcgg aggaccgaag gagctaaccg
     5761 cttttttgca caacatgggg gatcatgtaa ctcgccttga tcgttgggaa ccggagctga
     5821 atgaagccat accaaacgac gagcgtgaca ccacgatgcc tgtagcaatg gcaacaacgt
     5881 tgcgcaaact attaactggc gaactactta ctctagcttc ccggcaacaa ttaatagact
     5941 ggatggaggc ggataaagtt gcaggaccac ttctgcgctc ggcccttccg gctggctggt
     6001 ttattgctga taaatctgga gccggtgagc gtgggtctcg cggtatcatt gcagcactgg
     6061 ggccagatgg taagccctcc cgtatcgtag ttatctacac gacggggagt caggcaacta
     6121 tggatgaacg aaatagacag atcgctgaga taggtgcctc actgattaag cattggtaac
     6181 tgtcagacca agtttactca tatatacttt agattgattt aaaacttcat ttttaattta
     6241 aaaggatcta ggtgaagatc ctttttgata atctcatgac caaaatccct taacgtgagt
     6301 tttcgttcca ctgagcgtca gaccccgtag aaaagatcaa aggatcttct tgagatcctt
     6361 tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca gcggtggttt
     6421 gtttgccgga tcaagagcta ccaactcttt ttccgaaggt aactggcttc agcagagcgc
     6481 agataccaaa tactgttctt ctagtgtagc cgtagttagg ccaccacttc aagaactctg
     6541 tagcaccgcc tacatacctc gctctgctaa tcctgttacc agtggctgct gccagtggcg
     6601 ataagtcgtg tcttaccggg ttggactcaa gacgatagtt accggataag gcgcagcggt
     6661 cgggctgaac ggggggttcg tgcacacagc ccagcttgga gcgaacgacc tacaccgaac
     6721 tgagatacct acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg
     6781 acaggtatcc ggtaagcggc agggtcggaa caggagagcg cacgagggag cttccagggg
     6841 gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat
     6901 ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac gcggcctttt
     6961 tacggttcct ggccttttgc tggccttttg ctcacatgtt ctttcctgcg ttatcccctg
     7021 attctgtgga taaccgtatt accgcctttg agtgagctga taccgctcgc cgcagccgaa
     7081 cgaccgagcg cagcgagtca gtgagcgagg aagcggaaga gcgcccaata cgcaaaccgc
     7141 ctctccccgc gcgttggccg attcattaat g
//
