LOCUS       sequence-488713-        8316 bp    DNA     circular SYN 08-OCT-2025
DEFINITION  AAV plasmid expressing human GRN with a C-terminal HA tag under the 
            CAG promoter..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8316)
  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 8316)
  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-GRN-HA
FEATURES             Location/Qualifiers
     source          1..8316
                     /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             3345..3371
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     misc_feature    3399..3987
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(3452..3472)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3870..3881)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(4079..4098)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    4144..4199
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(4198..4217)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     repeat_region   4599..4728
                     /label=AAV2 ITR
     rep_origin      5099..5612
                     /direction=RIGHT
                     /label=M13 ori
                     /note="M13 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5186..5205)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5396..5417
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(6094..6113)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     6213..6235
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(6273..6291)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        6359..6463
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6464..6532,6533..7324)
                     /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(6682..6701)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      7495..8083
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7984..8003
                     /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 gggggsgcgc
      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 atctcgagct caagcttcga attctgcagt
     1501 cgagtcgact ggatccggta ccgaggagat ctgccgccgc gatcgccatg tggaccctgg
     1561 tgagctgggt ggccttaaca gcagggctgg tggctggaac gcggtgccca gatggtcagt
     1621 tctgccctgt ggcctgctgc ctggaccccg gaggagccag ctacagctgc tgccgtcccc
     1681 ttctggacaa atggcccaca acactgagca ggcatctggg tggcccctgc caggttgatg
     1741 cccactgctc tgccggccac tcctgcatct ttaccgtctc agggacttcc agttgctgcc
     1801 ccttcccaga ggccgtggca tgcggggatg gccatcactg ctgcccacgg ggcttccact
     1861 gcagtgcaga cgggcgatcc tgcttccaaa gatcaggtaa caactccgtg ggtgccatcc
     1921 agtgccctga tagtcagttc gaatgcccgg acttctccac gtgctgtgtt atggtcgatg
     1981 gctcctgggg gtgctgcccc atgccccagg cttcctgctg tgaagacagg gtgcactgct
     2041 gtccgcacgg tgccttctgc gacctggttc acacccgctg catcacaccc acgggcaccc
     2101 accccctggc aaagaagctc cctgcccaga ggactaacag ggcagtggcc ttgtccagct
     2161 cggtcatgtg tccggacgca cggtcccggt gccctgatgg ttctacctgc tgtgagctgc
     2221 ccagtgggaa gtatggctgc tgcccaatgc ccaacgccac ctgctgctcc gatcacctgc
     2281 actgctgccc ccaagacact gtgtgtgacc tgatccagag taagtgcctc tccaaggaga
     2341 acgctaccac ggacctcctc actaagctgc ctgcgcacac agtgggggat gtgaaatgtg
     2401 acatggaggt gagctgccca gatggctata cctgctgccg tctacagtcg ggggcctggg
     2461 gctgctgccc ttttacccag gctgtgtgct gtgaggacca catacactgc tgtcccgcgg
     2521 ggtttacgtg tgacacgcag aagggtacct gtgaacaggg gccccaccag gtgccctgga
     2581 tggagaaggc cccagctcac ctcagcctgc cagacccaca agccttgaag agagatgtcc
     2641 cctgtgataa tgtcagcagc tgtccctcct ccgatacctg ctgccaactc acgtctgggg
     2701 agtggggctg ctgtccaatc ccagaggctg tctgctgctc ggaccaccag cactgctgcc
     2761 cccagggcta cacgtgtgta gctgaggggc agtgtcagcg aggaagcgag atcgtggctg
     2821 gactggagaa gatgcctgcc cgccgggctt ccttatccca ccccagagac atcggctgtg
     2881 accagcacac cagctgcccg gtggggcaga cctgctgccc gagcctgggt gggagctggg
     2941 cctgctgcca gttgccccat gctgtgtgct gcgaggatcg ccagcactgc tgcccggctg
     3001 gctacacctg caacgtgaag gctcgatcct gcgagaagga agtggtctct gcccagcctg
     3061 ccaccttcct ggcccgtagc cctcacgtgg gtgtgaagga cgtggagtgt ggggaaggac
     3121 acttctgcca tgataaccag acctgctgcc gagacaaccg acagggctgg gcctgctgtc
     3181 cctaccgcca gggcgtctgt tgtgctgatc ggcgccactg ctgtcctgct ggcttccgct
     3241 gcgcagccag gggtaccaag tgtttgcgca gggaggcccc gcgctgggac gcccctttga
     3301 gggacccagc cttgagacag ctgctgacgc gtacgcggcc gctctaccca tacgatgttc
     3361 cagattacgc ttaagatatc atcggtaccg gatccgataa tcaacctctg gattacaaaa
     3421 tttgtgaaag attgactggt attcttaact atgttgctcc ttttacgcta tgtggatacg
     3481 ctgctttaat gcctttgtat catgctattg cttcccgtat ggctttcatt ttctcctcct
     3541 tgtataaatc ctggttgctg tctctttatg aggagttgtg gcccgttgtc aggcaacgtg
     3601 gcgtggtgtg cactgtgttt gctgacgcaa cccccactgg ttggggcatt gccaccacct
     3661 gtcagctcct ttccgggact ttcgctttcc ccctccctat tgccacggcg gaactcatcg
     3721 ccgcctgcct tgcccgctgc tggacagggg ctcggctgtt gggcactgac aattccgtgg
     3781 tgttgtcggg gaaatcatcg tcctttcctt ggctgctcgc ctgtgttgcc acctggattc
     3841 tgcgcgggac gtccttctgc tacgtccctt cggccctcaa tccagcggac cttccttccc
     3901 gcggcctgct gccggctctg cggcctcttc cgcgtcttcg ccttcgccct cagacgagtc
     3961 ggatctccct ttgggccgcc tccccgcatc ggatccaatt cactcctcag gtgcaggctg
     4021 cctatcagaa ggtggtggct ggtgtggcca atgccctggc tcacaaatac cactgagatc
     4081 tttttccctc tgccaaaaat tatggggaca tcatgaagcc ccttgagcat ctgacttctg
     4141 gctaataaag gaaatttatt ttcattgcaa tagtgtgttg gaattttttg tgtctctcac
     4201 tcggaaggac atatgggagg gcaaatcatt taaaacatca gaatgagtat ttggtttaga
     4261 gtttggcaac atatgcccat atgctggctg ccatgaacaa aggttggcta taaagaggtc
     4321 atcagtatat gaaacagccc cctgctgtcc attccttatt ccatagaaaa gccttgactt
     4381 gaggttagat tttttttata ttttgttttg tgttattttt ttctttaaca tccctaaaat
     4441 tttccttaca tgttttacta gccagatttt tcctcctctc ctgactactc ccagtcatag
     4501 ctgtccctct tctcttatgg agatccctcg acctgcagcc caagctctag agcatggcta
     4561 cgtagataag tagcatggcg ggttaatcat taactacaag gaacccctag tgatggagtt
     4621 ggccactccc tctctgcgcg ctcgctcgct cactgaggcc gggcgaccaa aggtcgcccg
     4681 acgcccgggc tttgcccggg cggcctcagt gagcgagcga gcgcgcagct ggcgtaatag
     4741 cgaagaggcc cgcaccgatc gcccttccca acagttgcgc agcctgaatg gcgaatggcg
     4801 attccgttgc aatggctggc ggtaatattg ttctggatat taccagcaag gccgatagtt
     4861 tgagttcttc tactcaggca agtgatgtta ttactaatca aagaagtatt gcgacaacgg
     4921 ttaatttgcg tgatggacag actcttttac tcggtggcct cactgattat aaaaacactt
     4981 ctcaggattc tggcgtaccg ttcctgtcta aaatcccttt aatcggcctc ctgtttagct
     5041 cccgctctga ttctaacgag gaaagcacgt tatacgtgct cgtcaaagca accatagtac
     5101 gcgccctgta gcggcgcatt aagcgcggcg ggtgtggtgg ttacgcgcag cgtgaccgct
     5161 acacttgcca gcgccctagc gcccgctcct ttcgctttct tcccttcctt tctcgccacg
     5221 ttcgccggct ttccccgtca agctctaaat cgggggctcc ctttagggtt ccgatttagt
     5281 gctttacggc acctcgaccc caaaaaactt gattagggtg atggttcacg tagtgggcca
     5341 tcgccctgat agacggtttt tcgccctttg acgttggagt ccacgttctt taatagtgga
     5401 ctcttgttcc aaactggaac aacactcaac cctatctcgg tctattcttt tgatttataa
     5461 gggattttgc cgatttcggc ctattggtta aaaaatgagc tgatttaaca aaaatttaac
     5521 gcgaatttta acaaaatatt aacgcttaca atttaaatat ttgcttatac aatcttcctg
     5581 tttttggggc ttttctgatt atcaaccggg gtacatatga ttgacatgct agttttacga
     5641 ttaccgttca tcgattctct tgtttgctcc agactctcag gcaatgacct gatagccttt
     5701 gtagagacct ctcaaaaata gctaccctct ccggcatgaa tttatcagct agaacggttg
     5761 aatatcatat tgatggtgat ttgactgtct ccggcctttc tcacccgttt gaatctttac
     5821 ctacacatta ctcaggcatt gcatttaaaa tatatgaggg ttctaaaaat ttttatcctt
     5881 gcgttgaaat aaaggcttct cccgcaaaag tattacaggg tcataatgtt tttggtacaa
     5941 ccgatttagc tttatgctct gaggctttat tgcttaattt tgctaattct ttgccttgcc
     6001 tgtatgattt attggatgtt ggaatcgcct gatgcggtat tttctcctta cgcatctgtg
     6061 cggtatttca caccgcatat ggtgcactct cagtacaatc tgctctgatg ccgcatagtt
     6121 aagccagccc cgacacccgc caacacccgc tgacgcgccc tgacgggctt gtctgctccc
     6181 ggcatccgct tacagacaag ctgtgaccgt ctccgggagc tgcatgtgtc agaggttttc
     6241 accgtcatca ccgaaacgcg cgagacgaaa gggcctcgtg atacgcctat ttttataggt
     6301 taatgtcatg ataataatgg tttcttagac gtcaggtggc acttttcggg gaaatgtgcg
     6361 cggaacccct atttgtttat ttttctaaat acattcaaat atgtatccgc tcatgagaca
     6421 ataaccctga taaatgcttc aataatattg aaaaaggaag agtatgagta ttcaacattt
     6481 ccgtgtcgcc cttattccct tttttgcggc attttgcctt cctgtttttg ctcacccaga
     6541 aacgctggtg aaagtaaaag atgctgaaga tcagttgggt gcacgagtgg gttacatcga
     6601 actggatctc aacagcggta agatccttga gagttttcgc cccgaagaac gttttccaat
     6661 gatgagcact tttaaagttc tgctatgtgg cgcggtatta tcccgtattg acgccgggca
     6721 agagcaactc ggtcgccgca tacactattc tcagaatgac ttggttgagt actcaccagt
     6781 cacagaaaag catcttacgg atggcatgac agtaagagaa ttatgcagtg ctgccataac
     6841 catgagtgat aacactgcgg ccaacttact tctgacaacg atcggaggac cgaaggagct
     6901 aaccgctttt ttgcacaaca tgggggatca tgtaactcgc cttgatcgtt gggaaccgga
     6961 gctgaatgaa gccataccaa acgacgagcg tgacaccacg atgcctgtag caatggcaac
     7021 aacgttgcgc aaactattaa ctggcgaact acttactcta gcttcccggc aacaattaat
     7081 agactggatg gaggcggata aagttgcagg accacttctg cgctcggccc ttccggctgg
     7141 ctggtttatt gctgataaat ctggagccgg tgagcgtggg tctcgcggta tcattgcagc
     7201 actggggcca gatggtaagc cctcccgtat cgtagttatc tacacgacgg ggagtcaggc
     7261 aactatggat gaacgaaata gacagatcgc tgagataggt gcctcactga ttaagcattg
     7321 gtaactgtca gaccaagttt actcatatat actttagatt gatttaaaac ttcattttta
     7381 atttaaaagg atctaggtga agatcctttt tgataatctc atgaccaaaa tcccttaacg
     7441 tgagttttcg ttccactgag cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga
     7501 tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt
     7561 ggtttgtttg ccggatcaag agctaccaac tctttttccg aaggtaactg gcttcagcag
     7621 agcgcagata ccaaatactg ttcttctagt gtagccgtag ttaggccacc acttcaagaa
     7681 ctctgtagca ccgcctacat acctcgctct gctaatcctg ttaccagtgg ctgctgccag
     7741 tggcgataag tcgtgtctta ccgggttgga ctcaagacga tagttaccgg ataaggcgca
     7801 gcggtcgggc tgaacggggg gttcgtgcac acagcccagc ttggagcgaa cgacctacac
     7861 cgaactgaga tacctacagc gtgagctatg agaaagcgcc acgcttcccg aagggagaaa
     7921 ggcggacagg tatccggtaa gcggcagggt cggaacagga gagcgcacga gggagcttcc
     7981 agggggaaac gcctggtatc tttatagtcc tgtcgggttt cgccacctct gacttgagcg
     8041 tcgatttttg tgatgctcgt caggggggcg gagcctatgg aaaaacgcca gcaacgcggc
     8101 ctttttacgg ttcctggcct tttgctggcc ttttgctcac atgttctttc ctgcgttatc
     8161 ccctgattct gtggataacc gtattaccgc ctttgagtga gctgataccg ctcgccgcag
     8221 ccgaacgacc gagcgcagcg agtcagtgag cgaggaagcg gaagagcgcc caatacgcaa
     8281 accgcctctc cccgcgcgtt ggccgattca ttaatg
//
