LOCUS       sequence-492354-        6684 bp    DNA     circular SYN 10-NOV-2025
DEFINITION  AAV transgene plasmid encoding TdTomato with miR-122 target site.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6684)
  AUTHORS   Lin C, Chen X, Hoang JD, Ristic F, Fan Y, Jang S, Chung JHA, 
            Sullivan EE, Gawda T, Kavvathas B, Tran I, Li Y, Steele AD, Shay TF,
            Gradinaru V
  TITLE     AAVs Targeting Human Carbonic Anhydrase IV Enhance Gene Delivery to 
            the Brain
  JOURNAL   bioRxiv 2025.04.21.649868
REFERENCE   2  (bases 1 to 6684)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Nov 10, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "bioRxiv 
            2025.04.21.649868"
COMMENT     Sequence Label: pAAV-CAG-TdTomato-miR122
FEATURES             Location/Qualifiers
     source          1..6684
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   1..130
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     enhancer        210..513
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        515..790
                     /label=chicken beta-actin promoter
     primer_bind     1120..1142
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     1204..1223
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     regulatory      1266..1275
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     misc_feature    2811..3399
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(2864..2884)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3282..3293)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    3431..3907
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(3630..3649)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     repeat_region   3947..4087
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   3947..4076
                     /label=AAV2 ITR
     rep_origin      4162..4617
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4249..4268)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4459..4480
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4634..4653)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4753..4775
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(4813..4831)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4899..5003
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5004..5072,5073..5864)
                     /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(5222..5241)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6035..6623
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6524..6543
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 cctgcaggca gctgcgcgct cgctcgctca ctgaggccgc ccgggcgtcg ggcgaccttt
       61 ggtcgcccgg cctcagtgag cgagcgagcg cgcagagagg gagtggccaa ctccatcact
      121 aggggttcct gcggccgcac gcgtaacact agttattaat agtaatcaat tacggggtca
      181 ttagttcata gcccatatat ggagttccgc gttacataac ttacggtaaa tggcccgcct
      241 ggctgaccgc ccaacgaccc ccgcccattg acgtcaataa tgacgtatgt tcccatagta
      301 acgccaatag ggactttcca ttgacgtcaa tgggtggagt atttacggta aactgcccac
      361 ttggcagtac atcaagtgta tcatatgcca agtacgcccc ctattgacgt caatgacggt
      421 aaatggcccg cctggcatta tgcccagtac atgaccttat gggactttcc tacttggcag
      481 tacatctacg tattagtcat cgctattacc atggtcgagg tgagccccac gttctgcttc
      541 actctcccca tctccccccc ctccccaccc ccaattttgt atttatttat tttttaatta
      601 tttttggcag cgatgggggg gggggggggg ggggcgcgcg ccaggcgggg cggggcgggg
      661 cgaggggcgg ggcggggcga ggcggagagg tgcggcggca gccaatcaga gcggcgcgct
      721 ccgaaagttt ccttttatgg cgaggcggcg gcggcggcgg ccctataaaa agcgaagcgc
      781 gcggcgggcg ggagtcgctg cgcgctgcct tcgccccgtg ccccgctccg ccgccgcctc
      841 gcgccgcccg ccccggctct gactgaccgc gttactccca caggtgagcg ggcgggacgg
      901 cccttctcct ccgggctgta attagcgctt ggtttaatga cggcttgttt cttttctgtg
      961 gctgcgtgaa agccttgagg ggctccggga gcgccggcag gaaggaaatg ggcggggagg
     1021 gccttcgtgc gtcgccgcgc cgccgtcccc ttctccctct ccagcctcgg ggctgtccgc
     1081 ggggggacgg ctgccttcgg gggggacggg gcagggcggg gttcggcttc tggcgtgtga
     1141 ccggcggctc tagagcctct gctaaccatg ttcatgcctt cttctttttc ctacagctcc
     1201 tgggcaacgt gctggttatt gtgctgtctc atcattttgg caaagaattg cggcccaacg
     1261 gtaccgccac catggtgagt aagggcgagg aagtgatcaa agagttcatg cggtttaagg
     1321 tgagaatgga aggaagcatg aacggccacg agttcgaaat tgagggagaa ggagagggac
     1381 ggccctacga gggcacccag acagccaagc tgaaagtgac aaagggcggg cctctgccat
     1441 tcgcttggga catcctgagc ccacagttta tgtacggctc caaggcctat gtgaaacatc
     1501 cagctgacat tcccgattat aagaaactga gcttccccga ggggtttaag tgggaaagag
     1561 tgatgaactt cgaggacgga ggcctggtga ctgtgaccca ggacagctcc ctgcaggatg
     1621 ggaccctgat ctacaaggtg aaaatgagag ggacaaattt tccccctgat ggacctgtga
     1681 tgcagaagaa aactatggga tgggaggcct ccaccgaaag gctgtatcca cgcgacgggg
     1741 tgctgaaagg agaaatccac caggctctga agctgaaaga tgggggacat tacctggtgg
     1801 agttcaagac aatctacatg gccaagaaac ctgtgcagct gccaggctac tattacgtgg
     1861 acacaaaact ggatatcact tcacacaacg aggactacac tattgtggag cagtatgaac
     1921 ggagcgaggg gagacaccat ctgttcctgg gccatgggac tggaagtacc ggctcagggt
     1981 ctagtggaac cgcctcaagc gaggataaca atatggctgt gatcaaagag ttcatgaggt
     2041 ttaaggtgcg catggagggc agcatgaatg ggcacgaatt tgagattgaa ggagagggcg
     2101 aagggaggcc ttacgagggc acacagactg ccaagctgaa agtgaccaag ggaggaccac
     2161 tgcctttcgc ttgggatatc ctgtctcctc agtttatgta cggaagtaag gcctatgtca
     2221 agcatcccgc tgacattcct gattacaaga aactgtcttt cccagagggc tttaagtggg
     2281 agagagtgat gaattttgaa gatggaggcc tggtgaccgt gacacaggac tcctctctgc
     2341 aggatggcac tctgatctac aaagtcaaaa tgcgcggcac caattttcca cccgatgggc
     2401 ccgtgatgca gaagaaaaca atggggtggg aggccagcac tgaacggctg tatcctagag
     2461 acggagtgct gaagggcgaa atccaccagg ccctgaagct gaaagacggc ggccactacc
     2521 tggtggagtt caaaaccatc tacatggcca agaaaccagt gcagctgccc ggctattact
     2581 atgtggacac caagctggat atcacatccc acaatgaaga ctacaccatt gtggaacagt
     2641 atgagaggtc tgaaggacgc caccatctgt ttctgtacgg catggatgag ctgtataagt
     2701 gaattcaccg gtcaaacacc attgtcacac tccatactat caaacaccat tgtcacactc
     2761 cagatcaaca aacaccattg tcacactcca cgccggcgaa gcttatcgat aatcaacctc
     2821 tggattacaa aatttgtgaa agattgactg gtattcttaa ctatgttgct ccttttacgc
     2881 tatgtggata cgctgcttta atgcctttgt atcatgctat tgcttcccgt atggctttca
     2941 ttttctcctc cttgtataaa tcctggttgc tgtctcttta tgaggagttg tggcccgttg
     3001 tcaggcaacg tggcgtggtg tgcactgtgt ttgctgacgc aacccccact ggttggggca
     3061 ttgccaccac ctgtcagctc ctttccggga ctttcgcttt ccccctccct attgccacgg
     3121 cggaactcat cgccgcctgc cttgcccgct gctggacagg ggctcggctg ttgggcactg
     3181 acaattccgt ggtgttgtcg gggaaatcat cgtcctttcc ttggctgctc gcctatgttg
     3241 ccacctggat tctgcgcggg acgtccttct gctacgtccc ttcggccctc aatccagcgg
     3301 accttccttc ccgcggcctg ctgccggctc tgcggcctct tccgcgtctt cgccttcgcc
     3361 ctcagacgag tcggatctcc ctttgggccg cctccccgca tcgataccga gcgctgctcg
     3421 agagatctac gggtggcatc cctgtgaccc ctccccagtg cctctcctgg ccctggaagt
     3481 tgccactcca gtgcccacca gccttgtcct aataaaatta agttgcatca ttttgtctga
     3541 ctaggtgtcc ttctataata ttatggggtg gaggggggtg gtatggagca aggggcaagt
     3601 tgggaagaca acctgtaggg cctgcggggt ctattgggaa ccaagctgga gtgcagtggc
     3661 acaatcttgg ctcactgcaa tctccgcctc ctgggttcaa gcgattctcc tgcctcagcc
     3721 tcccgagttg ttgggattcc aggcatgcat gaccaggctc agctaatttt tgtttttttg
     3781 gtagagacgg ggtttcacca tattggccag gctggtctcc aactcctaat ctcaggtgat
     3841 ctacccacct tggcctccca aattgctggg attacaggcg tgaaccactg ctcccttccc
     3901 tgtccttctg attttgtagg taaccacgtg cggaccgagc ggccgcagga acccctagtg
     3961 atggagttgg ccactccctc tctgcgcgct cgctcgctca ctgaggccgg gcgaccaaag
     4021 gtcgcccgac gcccgggctt tgcccgggcg gcctcagtga gcgagcgagc gcgcagctgc
     4081 ctgcaggggc gcctgatgcg gtattttctc cttacgcatc tgtgcggtat ttcacaccgc
     4141 atacgtcaaa gcaaccatag tacgcgccct gtagcggcgc attaagcgcg gcgggtgtgg
     4201 tggttacgcg cagcgtgacc gctacacttg ccagcgcctt agcgcccgct cctttcgctt
     4261 tcttcccttc ctttctcgcc acgttcgccg gctttccccg tcaagctcta aatcgggggc
     4321 tccctttagg gttccgattt agtgctttac ggcacctcga ccccaaaaaa cttgatttgg
     4381 gtgatggttc acgtagtggg ccatcgccct gatagacggt ttttcgccct ttgacgttgg
     4441 agtccacgtt ctttaatagt ggactcttgt tccaaactgg aacaacactc aactctatct
     4501 cgggctattc ttttgattta taagggattt tgccgatttc ggtctattgg ttaaaaaatg
     4561 agctgattta acaaaaattt aacgcgaatt ttaacaaaat attaacgttt acaattttat
     4621 ggtgcactct cagtacaatc tgctctgatg ccgcatagtt aagccagccc cgacacccgc
     4681 caacacccgc tgacgcgccc tgacgggctt gtctgctccc ggcatccgct tacagacaag
     4741 ctgtgaccgt ctccgggagc tgcatgtgtc agaggttttc accgtcatca ccgaaacgcg
     4801 cgagacgaaa gggcctcgtg atacgcctat ttttataggt taatgtcatg ataataatgg
     4861 tttcttagac gtcaggtggc acttttcggg gaaatgtgcg cggaacccct atttgtttat
     4921 ttttctaaat acattcaaat atgtatccgc tcatgagaca ataaccctga taaatgcttc
     4981 aataatattg aaaaaggaag agtatgagta ttcaacattt ccgtgtcgcc cttattccct
     5041 tttttgcggc attttgcctt cctgtttttg ctcacccaga aacgctggtg aaagtaaaag
     5101 atgctgaaga tcagttgggt gcacgagtgg gttacatcga actggatctc aacagcggta
     5161 agatccttga gagttttcgc cccgaagaac gttttccaat gatgagcact tttaaagttc
     5221 tgctatgtgg cgcggtatta tcccgtattg acgccgggca agagcaactc ggtcgccgca
     5281 tacactattc tcagaatgac ttggttgagt actcaccagt cacagaaaag catcttacgg
     5341 atggcatgac agtaagagaa ttatgcagtg ctgccataac catgagtgat aacactgcgg
     5401 ccaacttact tctgacaacg atcggaggac cgaaggagct aaccgctttt ttgcacaaca
     5461 tgggggatca tgtaactcgc cttgatcgtt gggaaccgga gctgaatgaa gccataccaa
     5521 acgacgagcg tgacaccacg atgcctgtag caatggcaac aacgttgcgc aaactattaa
     5581 ctggcgaact acttactcta gcttcccggc aacaattaat agactggatg gaggcggata
     5641 aagttgcagg accacttctg cgctcggccc ttccggctgg ctggtttatt gctgataaat
     5701 ctggagccgg tgagcgtggg tctcgcggta tcattgcagc actggggcca gatggtaagc
     5761 cctcccgtat cgtagttatc tacacgacgg ggagtcaggc aactatggat gaacgaaata
     5821 gacagatcgc tgagataggt gcctcactga ttaagcattg gtaactgtca gaccaagttt
     5881 actcatatat actttagatt gatttaaaac ttcattttta atttaaaagg atctaggtga
     5941 agatcctttt tgataatctc atgaccaaaa tcccttaacg tgagttttcg ttccactgag
     6001 cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa
     6061 tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg ccggatcaag
     6121 agctaccaac tctttttccg aaggtaactg gcttcagcag agcgcagata ccaaatactg
     6181 ttcttctagt gtagccgtag ttaggccacc acttcaagaa ctctgtagca ccgcctacat
     6241 acctcgctct gctaatcctg ttaccagtgg ctgctgccag tggcgataag tcgtgtctta
     6301 ccgggttgga ctcaagacga tagttaccgg ataaggcgca gcggtcgggc tgaacggggg
     6361 gttcgtgcac acagcccagc ttggagcgaa cgacctacac cgaactgaga tacctacagc
     6421 gtgagctatg agaaagcgcc acgcttcccg aagggagaaa ggcggacagg tatccggtaa
     6481 gcggcagggt cggaacagga gagcgcacga gggagcttcc agggggaaac gcctggtatc
     6541 tttatagtcc tgtcgggttt cgccacctct gacttgagcg tcgatttttg tgatgctcgt
     6601 caggggggcg gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct
     6661 tttgctggcc ttttgctcac atgt
//
