LOCUS       sequence-492353-        6207 bp    DNA     circular SYN 10-NOV-2025
DEFINITION  AAV transgene plasmid encoding human CA4 with miR-122 target site.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6207)
  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 6207)
  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-hCA4-miR122
FEATURES             Location/Qualifiers
     source          1..6207
                     /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"
     primer_bind     1133..1155
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     1217..1236
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     misc_feature    2334..2922
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(2387..2407)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(2805..2816)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    2954..3430
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(3153..3172)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     repeat_region   3470..3610
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   3470..3599
                     /label=AAV2 ITR
     rep_origin      3685..4140
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3772..3791)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3982..4003
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4157..4176)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4276..4298
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(4336..4354)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4422..4526
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4527..4595,4596..5387)
                     /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(4745..4764)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5558..6146
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6047..6066
                     /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 ttttgtgcag cgatgggggc gggggggggg gggggggggg gggggggcgc gcgccaggcg
      661 gggcggggcg gggcgagggg cggggcgggg cgaggcggag aggtgcggcg gcagccaatc
      721 agagcggcgc gctccgaaag tttcctttta tggcgaggcg gcggcggcgg cggccctata
      781 aaaagcgaag cgcgcggcgg gcgggagtcg ctgcgcgctg ccttcgcccc gtgccccgct
      841 ccgccgccgc ctcgcgccgc ccgccccggc tctgactgac cgcgttactc ccacaggtga
      901 gcgggcggga cggcccttct cctccgggct gtaattagcg cttggtttaa tgacggcttg
      961 tttcttttct gtggctgcgt gaaagccttg aggggctccg ggagcgccgg caggaaggaa
     1021 atgggcgggg agggccttcg tgcgtcgccg cgccgccgtc cccttctccc tctccagcct
     1081 cggggctgtc cgcgggggga cggctgcctt cgggggggac ggggcagggc ggggttcggc
     1141 ttctggcgtg tgaccggcgg ctctagagcc tctgctaacc atgttcatgc cttcttcttt
     1201 ttcctacagc tcctgggcaa cgtgctggtt attgtgctgt ctcatcattt tggcaaagaa
     1261 ttgcggccca acggtaccgc caccatgcag ctccttcttg ctctgctggc gctggcatac
     1321 gtggccccct ctactgaaga cgagtcacac tggtgctacg aggttcaagc cgagtcctcc
     1381 aactacccct gcttggtgcc agtcaagtgg ggtggaaact gccagaagga ccgccagtcc
     1441 cccatcaaca tcgtcaccac caaggcaaag gtggacaaaa aactgggacg cttcttcttc
     1501 tctggctacg ataagaagca aacgtggact gtccaaaata acgggcactc agtgatgatg
     1561 ttgctggaga acaaggccag catttctgga ggaggactgc ctgccccata ccaggccaaa
     1621 cagttgcacc tgcactggtc cgacttgcca tataagggct cggagcacag cctcgatggg
     1681 gagcactttg ccatggagat gcacatagta catgagaaag agaaggggac atcgaggaat
     1741 gtgaaagagg cccaggaccc tgaagacgaa attgcggtgc tggcctttct ggtggaggct
     1801 ggaacccagg tgaacgaggg cttccagcca ctggtggagg cactgtctaa tatccccaaa
     1861 cctgagatga gcactacgat ggcagagagc agcctgttgg acctgctccc caaggaggag
     1921 aaactgaggc actacttccg ctacctgggc tcactcacca caccgacctg cgatgagaag
     1981 gtcgtctgga ctgtgttccg ggagcccatt cagcttcaca gagaacagat cctggcattc
     2041 tctcagaagc tgtactacga caaggaacag acagtgagca tgaaggacaa tgtcaggccc
     2101 ctgcagcagc tggggcagcg cacggtgata aagtcccatg ccccaggaca gctgctgtct
     2161 ttgcccctgc ccactctgtt ggtccccaca ctcacctgcc tggtagccaa cttcctccag
     2221 taagaattca ccggtcaaac accattgtca cactccatac tatcaaacac cattgtcaca
     2281 ctccagatca acaaacacca ttgtcacact ccacgccggc gaagcttatc gataatcaac
     2341 ctctggatta caaaatttgt gaaagattga ctggtattct taactatgtt gctcctttta
     2401 cgctatgtgg atacgctgct ttaatgcctt tgtatcatgc tattgcttcc cgtatggctt
     2461 tcattttctc ctccttgtat aaatcctggt tgctgtctct ttatgaggag ttgtggcccg
     2521 ttgtcaggca acgtggcgtg gtgtgcactg tgtttgctga cgcaaccccc actggttggg
     2581 gcattgccac cacctgtcag ctcctttccg ggactttcgc tttccccctc cctattgcca
     2641 cggcggaact catcgccgcc tgccttgccc gctgctggac aggggctcgg ctgttgggca
     2701 ctgacaattc cgtggtgttg tcggggaaat catcgtcctt tccttggctg ctcgcctatg
     2761 ttgccacctg gattctgcgc gggacgtcct tctgctacgt cccttcggcc ctcaatccag
     2821 cggaccttcc ttcccgcggc ctgctgccgg ctctgcggcc tcttccgcgt cttcgccttc
     2881 gccctcagac gagtcggatc tccctttggg ccgcctcccc gcatcgatac cgagcgctgc
     2941 tcgagagatc tacgggtggc atccctgtga cccctcccca gtgcctctcc tggccctgga
     3001 agttgccact ccagtgccca ccagccttgt cctaataaaa ttaagttgca tcattttgtc
     3061 tgactaggtg tccttctata atattatggg gtggaggggg gtggtatgga gcaaggggca
     3121 agttgggaag acaacctgta gggcctgcgg ggtctattgg gaaccaagct ggagtgcagt
     3181 ggcacaatct tggctcactg caatctccgc ctcctgggtt caagcgattc tcctgcctca
     3241 gcctcccgag ttgttgggat tccaggcatg catgaccagg ctcagctaat ttttgttttt
     3301 ttggtagaga cggggtttca ccatattggc caggctggtc tccaactcct aatctcaggt
     3361 gatctaccca ccttggcctc ccaaattgct gggattacag gcgtgaacca ctgctccctt
     3421 ccctgtcctt ctgattttgt aggtaaccac gtgcggaccg agcggccgca ggaaccccta
     3481 gtgatggagt tggccactcc ctctctgcgc gctcgctcgc tcactgaggc cgggcgacca
     3541 aaggtcgccc gacgcccggg ctttgcccgg gcggcctcag tgagcgagcg agcgcgcagc
     3601 tgcctgcagg ggcgcctgat gcggtatttt ctccttacgc atctgtgcgg tatttcacac
     3661 cgcatacgtc aaagcaacca tagtacgcgc cctgtagcgg cgcattaagc gcggcgggtg
     3721 tggtggttac gcgcagcgtg accgctacac ttgccagcgc cttagcgccc gctcctttcg
     3781 ctttcttccc ttcctttctc gccacgttcg ccggctttcc ccgtcaagct ctaaatcggg
     3841 ggctcccttt agggttccga tttagtgctt tacggcacct cgaccccaaa aaacttgatt
     3901 tgggtgatgg ttcacgtagt gggccatcgc cctgatagac ggtttttcgc cctttgacgt
     3961 tggagtccac gttctttaat agtggactct tgttccaaac tggaacaaca ctcaactcta
     4021 tctcgggcta ttcttttgat ttataaggga ttttgccgat ttcggtctat tggttaaaaa
     4081 atgagctgat ttaacaaaaa tttaacgcga attttaacaa aatattaacg tttacaattt
     4141 tatggtgcac tctcagtaca atctgctctg atgccgcata gttaagccag ccccgacacc
     4201 cgccaacacc cgctgacgcg ccctgacggg cttgtctgct cccggcatcc gcttacagac
     4261 aagctgtgac cgtctccggg agctgcatgt gtcagaggtt ttcaccgtca tcaccgaaac
     4321 gcgcgagacg aaagggcctc gtgatacgcc tatttttata ggttaatgtc atgataataa
     4381 tggtttctta gacgtcaggt ggcacttttc ggggaaatgt gcgcggaacc cctatttgtt
     4441 tatttttcta aatacattca aatatgtatc cgctcatgag acaataaccc tgataaatgc
     4501 ttcaataata ttgaaaaagg aagagtatga gtattcaaca tttccgtgtc gcccttattc
     4561 ccttttttgc ggcattttgc cttcctgttt ttgctcaccc agaaacgctg gtgaaagtaa
     4621 aagatgctga agatcagttg ggtgcacgag tgggttacat cgaactggat ctcaacagcg
     4681 gtaagatcct tgagagtttt cgccccgaag aacgttttcc aatgatgagc acttttaaag
     4741 ttctgctatg tggcgcggta ttatcccgta ttgacgccgg gcaagagcaa ctcggtcgcc
     4801 gcatacacta ttctcagaat gacttggttg agtactcacc agtcacagaa aagcatctta
     4861 cggatggcat gacagtaaga gaattatgca gtgctgccat aaccatgagt gataacactg
     4921 cggccaactt acttctgaca acgatcggag gaccgaagga gctaaccgct tttttgcaca
     4981 acatggggga tcatgtaact cgccttgatc gttgggaacc ggagctgaat gaagccatac
     5041 caaacgacga gcgtgacacc acgatgcctg tagcaatggc aacaacgttg cgcaaactat
     5101 taactggcga actacttact ctagcttccc ggcaacaatt aatagactgg atggaggcgg
     5161 ataaagttgc aggaccactt ctgcgctcgg cccttccggc tggctggttt attgctgata
     5221 aatctggagc cggtgagcgt gggtctcgcg gtatcattgc agcactgggg ccagatggta
     5281 agccctcccg tatcgtagtt atctacacga cggggagtca ggcaactatg gatgaacgaa
     5341 atagacagat cgctgagata ggtgcctcac tgattaagca ttggtaactg tcagaccaag
     5401 tttactcata tatactttag attgatttaa aacttcattt ttaatttaaa aggatctagg
     5461 tgaagatcct ttttgataat ctcatgacca aaatccctta acgtgagttt tcgttccact
     5521 gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg agatcctttt tttctgcgcg
     5581 taatctgctg cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc
     5641 aagagctacc aactcttttt ccgaaggtaa ctggcttcag cagagcgcag ataccaaata
     5701 ctgttcttct agtgtagccg tagttaggcc accacttcaa gaactctgta gcaccgccta
     5761 catacctcgc tctgctaatc ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc
     5821 ttaccgggtt ggactcaaga cgatagttac cggataaggc gcagcggtcg ggctgaacgg
     5881 ggggttcgtg cacacagccc agcttggagc gaacgaccta caccgaactg agatacctac
     5941 agcgtgagct atgagaaagc gccacgcttc ccgaagggag aaaggcggac aggtatccgg
     6001 taagcggcag ggtcggaaca ggagagcgca cgagggagct tccaggggga aacgcctggt
     6061 atctttatag tcctgtcggg tttcgccacc tctgacttga gcgtcgattt ttgtgatgct
     6121 cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc ggccttttta cggttcctgg
     6181 ccttttgctg gccttttgct cacatgt
//
