LOCUS       sequence-487083-        7884 bp    DNA     circular SYN 24-SEP-2025
DEFINITION  AAV plasmid expressing HA-tagged codon-optimized mouse TFEB under 
            the CAG promoter..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7884)
  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 7884)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Sep 24, 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-HA-mTFEB
FEATURES             Location/Qualifiers
     source          1..7884
                     /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..931
                     /label=chicken beta-actin promoter
     primer_bind     1344..1366
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     1428..1447
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     CDS             1488..1514
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     misc_feature    2967..3555
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(3020..3040)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3438..3449)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(3647..3666)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    3712..3767
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(3766..3785)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     repeat_region   4167..4296
                     /label=AAV2 ITR
     rep_origin      4667..5180
                     /direction=RIGHT
                     /label=M13 ori
                     /note="M13 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4754..4773)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4964..4985
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(5662..5681)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5781..5803
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(5841..5859)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5927..6031
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6032..6100,6101..6892)
                     /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(6250..6269)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      7063..7651
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7552..7571
                     /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 gggggggcgc
      781 gccaggcggg gcggggcggg gcgaggggcg gggcggggcg aggcggagag gtgcggcggc
      841 agccaatcag agcggcgcgc tccgaaagtt tccttttatg gcgaggcggc ggcggcggcg
      901 gccctataaa aagcgaagcg cgcggcgggc gggagtcgct gcgcgctgcc ttcgccccgt
      961 gccccgctcc gccgccgcct cgcgccgccc gccccggctc tgactgaccg cgttactccc
     1021 acaggtgagc gggcgggacg gcccttctcc tccgggctgt aattagcgct tggtttaatg
     1081 acggcttgtt tcttttctgt ggctgcgtga aagccttgag gggctccggg agggcccttt
     1141 gtgcgggggg agcggctcgg ggggtgcgtg cgtgtgtgtg tgcgtgggga gcgccgcgtg
     1201 cggctccgcg ctgcccggcg gctgtgagcg ctgcgggcgc ggcgcggggc tttgtgcgct
     1261 ccgcagtgtg cgcgagggga gcgcggccgg gggcggtgcc ccgcgggggg acggctgcct
     1321 tcggggggga cggggcaggg cggggttcgg cttctggcgt gtgaccggcg gctctagagc
     1381 ctctgctaac catgttcatg ccttcttctt tttcctacag ctcctgggca acgtgctggt
     1441 tattgtgctg tctcatcatt ttggcaaaga attctcgagc cacaatgtac ccttacgacg
     1501 tgccagacta cgccggaatg gctagccgaa taggactgag gatgcagctt atgagagagc
     1561 aggcccaaca agaggagcag agagagagaa tgcaacagca agctgtgatg cactacatgc
     1621 aacagcaaca gcaacaacaa cagcagctgg ggggccctcc aaccccagcc attaatactc
     1681 ctgtccactt tcagtcccct ccccctgtcc ctggagaagt tctcaaagtt cagagttatc
     1741 tcgagaatcc tacatcctat cacctccagc aaagtcaaca ccagaaggtg cgagagtatc
     1801 tctctgagac ttatggcaac aagtttgctg ctcatgtaag tcccgcccaa ggctctccaa
     1861 aacctgcccc cgctgctagt cccggagtac gcgctggcca tgttctttca acttccgccg
     1921 gtaactctgc acctaatagt ccaatggcta tgcttcacat cagcagcaac ccagaaaagg
     1981 aatttgacga tgttatagat aatatcatga ggcttgattc cgtactcggg tacattaacc
     2041 cagaaatgca aatgccaaac acccttcccc tcagcagctc acatctgaat gtatactccg
     2101 gcgatccaca agtgaccgcc tccatggtag gggtgaccag ctctagttgt cctgctgatt
     2161 tgactcagaa aagggagttg actgatgcag agagtcgagc cctcgcaaaa gagcgacaga
     2221 aaaaagacaa tcataacctt atagagcgac ggcgcaggtt caatattaat gaccgcatca
     2281 aggagttggg aatgttgatc cctaaagcta atgatctcga tgtgcggtgg aacaaaggta
     2341 ccatactcaa agccagtgtt gattacataa ggcgcatgca aaaagacctc caaaaatcca
     2401 gagaactgga aaatcattcc cgcaggctgg aaatgacaaa taaacaactg tggctgcgga
     2461 tccaagaact tgagatgcaa gcacgagtgc acggtttgcc cactacaagt cccagtggcg
     2521 ttaatatggc cgagcttgcc caacaggtgg taaagcaaga gctgccctcc gaggatgggc
     2581 caggggaagc ccttatgctt ggtccagagg tgccagagcc agagcagatg cctgctctcc
     2641 ctcctcaagc acctctgccc tctgctgctc agccccaatc accttttcac cacctcgatt
     2701 tctctcacgg gctgagcttt gggggcgggg gcgatgaagg accaactggt tatccagata
     2761 ccctcggaac agagcatggc tctccattcc ctaaccttag caagaaagac ctggatctga
     2821 tgttgctcga tgacagcctt ctgcctctcg ccagcgaccc actctttagc acaatgtctc
     2881 cagaggcctc aaaggcaagc tctaggagaa gttcattctc tatggaagaa ggagacgtgt
     2941 tgtagtgaat cggtaccgga tccgataatc aacctctgga ttacaaaatt tgtgaaagat
     3001 tgactggtat tcttaactat gttgctcctt ttacgctatg tggatacgct gctttaatgc
     3061 ctttgtatca tgctattgct tcccgtatgg ctttcatttt ctcctccttg tataaatcct
     3121 ggttgctgtc tctttatgag gagttgtggc ccgttgtcag gcaacgtggc gtggtgtgca
     3181 ctgtgtttgc tgacgcaacc cccactggtt ggggcattgc caccacctgt cagctccttt
     3241 ccgggacttt cgctttcccc ctccctattg ccacggcgga actcatcgcc gcctgccttg
     3301 cccgctgctg gacaggggct cggctgttgg gcactgacaa ttccgtggtg ttgtcgggga
     3361 aatcatcgtc ctttccttgg ctgctcgcct gtgttgccac ctggattctg cgcgggacgt
     3421 ccttctgcta cgtcccttcg gccctcaatc cagcggacct tccttcccgc ggcctgctgc
     3481 cggctctgcg gcctcttccg cgtcttcgcc ttcgccctca gacgagtcgg atctcccttt
     3541 gggccgcctc cccgcatcgg atccaattca ctcctcaggt gcaggctgcc tatcagaagg
     3601 tggtggctgg tgtggccaat gccctggctc acaaatacca ctgagatctt tttccctctg
     3661 ccaaaaatta tggggacatc atgaagcccc ttgagcatct gacttctggc taataaagga
     3721 aatttatttt cattgcaata gtgtgttgga attttttgtg tctctcactc ggaaggacat
     3781 atgggagggc aaatcattta aaacatcaga atgagtattt ggtttagagt ttggcaacat
     3841 atgcccatat gctggctgcc atgaacaaag gttggctata aagaggtcat cagtatatga
     3901 aacagccccc tgctgtccat tccttattcc atagaaaagc cttgacttga ggttagattt
     3961 tttttatatt ttgttttgtg ttattttttt ctttaacatc cctaaaattt tccttacatg
     4021 ttttactagc cagatttttc ctcctctcct gactactccc agtcatagct gtccctcttc
     4081 tcttatggag atccctcgac ctgcagccca agctctagag catggctacg tagataagta
     4141 gcatggcggg ttaatcatta actacaagga acccctagtg atggagttgg ccactccctc
     4201 tctgcgcgct cgctcgctca ctgaggccgg gcgaccaaag gtcgcccgac gcccgggctt
     4261 tgcccgggcg gcctcagtga gcgagcgagc gcgcagctgg cgtaatagcg aagaggcccg
     4321 caccgatcgc ccttcccaac agttgcgcag cctgaatggc gaatggcgat tccgttgcaa
     4381 tggctggcgg taatattgtt ctggatatta ccagcaaggc cgatagtttg agttcttcta
     4441 ctcaggcaag tgatgttatt actaatcaaa gaagtattgc gacaacggtt aatttgcgtg
     4501 atggacagac tcttttactc ggtggcctca ctgattataa aaacacttct caggattctg
     4561 gcgtaccgtt cctgtctaaa atccctttaa tcggcctcct gtttagctcc cgctctgatt
     4621 ctaacgagga aagcacgtta tacgtgctcg tcaaagcaac catagtacgc gccctgtagc
     4681 ggcgcattaa gcgcggcggg tgtggtggtt acgcgcagcg tgaccgctac acttgccagc
     4741 gccctagcgc ccgctccttt cgctttcttc ccttcctttc tcgccacgtt cgccggcttt
     4801 ccccgtcaag ctctaaatcg ggggctccct ttagggttcc gatttagtgc tttacggcac
     4861 ctcgacccca aaaaacttga ttagggtgat ggttcacgta gtgggccatc gccctgatag
     4921 acggtttttc gccctttgac gttggagtcc acgttcttta atagtggact cttgttccaa
     4981 actggaacaa cactcaaccc tatctcggtc tattcttttg atttataagg gattttgccg
     5041 atttcggcct attggttaaa aaatgagctg atttaacaaa aatttaacgc gaattttaac
     5101 aaaatattaa cgcttacaat ttaaatattt gcttatacaa tcttcctgtt tttggggctt
     5161 ttctgattat caaccggggt acatatgatt gacatgctag ttttacgatt accgttcatc
     5221 gattctcttg tttgctccag actctcaggc aatgacctga tagcctttgt agagacctct
     5281 caaaaatagc taccctctcc ggcatgaatt tatcagctag aacggttgaa tatcatattg
     5341 atggtgattt gactgtctcc ggcctttctc acccgtttga atctttacct acacattact
     5401 caggcattgc atttaaaata tatgagggtt ctaaaaattt ttatccttgc gttgaaataa
     5461 aggcttctcc cgcaaaagta ttacagggtc ataatgtttt tggtacaacc gatttagctt
     5521 tatgctctga ggctttattg cttaattttg ctaattcttt gccttgcctg tatgatttat
     5581 tggatgttgg aatcgcctga tgcggtattt tctccttacg catctgtgcg gtatttcaca
     5641 ccgcatatgg tgcactctca gtacaatctg ctctgatgcc gcatagttaa gccagccccg
     5701 acacccgcca acacccgctg acgcgccctg acgggcttgt ctgctcccgg catccgctta
     5761 cagacaagct gtgaccgtct ccgggagctg catgtgtcag aggttttcac cgtcatcacc
     5821 gaaacgcgcg agacgaaagg gcctcgtgat acgcctattt ttataggtta atgtcatgat
     5881 aataatggtt tcttagacgt caggtggcac ttttcgggga aatgtgcgcg gaacccctat
     5941 ttgtttattt ttctaaatac attcaaatat gtatccgctc atgagacaat aaccctgata
     6001 aatgcttcaa taatattgaa aaaggaagag tatgagtatt caacatttcc gtgtcgccct
     6061 tattcccttt tttgcggcat tttgccttcc tgtttttgct cacccagaaa cgctggtgaa
     6121 agtaaaagat gctgaagatc agttgggtgc acgagtgggt tacatcgaac tggatctcaa
     6181 cagcggtaag atccttgaga gttttcgccc cgaagaacgt tttccaatga tgagcacttt
     6241 taaagttctg ctatgtggcg cggtattatc ccgtattgac gccgggcaag agcaactcgg
     6301 tcgccgcata cactattctc agaatgactt ggttgagtac tcaccagtca cagaaaagca
     6361 tcttacggat ggcatgacag taagagaatt atgcagtgct gccataacca tgagtgataa
     6421 cactgcggcc aacttacttc tgacaacgat cggaggaccg aaggagctaa ccgctttttt
     6481 gcacaacatg ggggatcatg taactcgcct tgatcgttgg gaaccggagc tgaatgaagc
     6541 cataccaaac gacgagcgtg acaccacgat gcctgtagca atggcaacaa cgttgcgcaa
     6601 actattaact ggcgaactac ttactctagc ttcccggcaa caattaatag actggatgga
     6661 ggcggataaa gttgcaggac cacttctgcg ctcggccctt ccggctggct ggtttattgc
     6721 tgataaatct ggagccggtg agcgtgggtc tcgcggtatc attgcagcac tggggccaga
     6781 tggtaagccc tcccgtatcg tagttatcta cacgacgggg agtcaggcaa ctatggatga
     6841 acgaaataga cagatcgctg agataggtgc ctcactgatt aagcattggt aactgtcaga
     6901 ccaagtttac tcatatatac tttagattga tttaaaactt catttttaat ttaaaaggat
     6961 ctaggtgaag atcctttttg ataatctcat gaccaaaatc ccttaacgtg agttttcgtt
     7021 ccactgagcg tcagaccccg tagaaaagat caaaggatct tcttgagatc ctttttttct
     7081 gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc
     7141 ggatcaagag ctaccaactc tttttccgaa ggtaactggc ttcagcagag cgcagatacc
     7201 aaatactgtt cttctagtgt agccgtagtt aggccaccac ttcaagaact ctgtagcacc
     7261 gcctacatac ctcgctctgc taatcctgtt accagtggct gctgccagtg gcgataagtc
     7321 gtgtcttacc gggttggact caagacgata gttaccggat aaggcgcagc ggtcgggctg
     7381 aacggggggt tcgtgcacac agcccagctt ggagcgaacg acctacaccg aactgagata
     7441 cctacagcgt gagctatgag aaagcgccac gcttcccgaa gggagaaagg cggacaggta
     7501 tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg gagcttccag ggggaaacgc
     7561 ctggtatctt tatagtcctg tcgggtttcg ccacctctga cttgagcgtc gatttttgtg
     7621 atgctcgtca ggggggcgga gcctatggaa aaacgccagc aacgcggcct ttttacggtt
     7681 cctggccttt tgctggcctt ttgctcacat gttctttcct gcgttatccc ctgattctgt
     7741 ggataaccgt attaccgcct ttgagtgagc tgataccgct cgccgcagcc gaacgaccga
     7801 gcgcagcgag tcagtgagcg aggaagcgga agagcgccca atacgcaaac cgcctctccc
     7861 cgcgcgttgg ccgattcatt aatg
//
