LOCUS       sequence-492356-        8395 bp    DNA     circular SYN 10-NOV-2025
DEFINITION  Rep/Cap plasmid for producing variant AAV-hCA4-Alpha 49.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8395)
  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 8395)
  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: pUCmini-iCAP-Alpha 49
FEATURES             Location/Qualifiers
     source          1..8395
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             2576..2686
                     /codon_start=1
                     /product="3 tandem repeats of the minimal activation domain
                     of herpes simplex virus VP16 (Cheng et al., 2013)"
                     /label=VP48
                     /translation="DALDDFDLDMLPADALDDFDLDMLPADALDDFDLDML"
     promoter        complement(2725..2743)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(2726..2743)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(2763..2780)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2769..2993
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     protein_bind    3055..3073
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    3092..3110
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    3129..3147
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    3166..3184
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     CDS             5901..5912
                     /codon_start=1
                     /product="WELQut protease recognition and cleavage site"
                     /label=WELQut site
                     /translation="WELQ"
     rep_origin      complement(6468..7056)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6548..6567)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7227..8018,8019..8087))
                     /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     7850..7869
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(8088..8192)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     8322..8344
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     8336..8353
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     8337..8353
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     8363..8382
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        8363..8381
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
ORIGIN
        1 tcgacggtat cgggggagct cgcagggtct ccattttgaa gcgggaggtt tgaacgcgca
       61 gccgccatgc cggggtttta cgagattgtg attaaggtcc ccagcgacct tgacgagcat
      121 ctgcccggca tttctgacag ctttgtgaac tgggtggccg agaaggaatg ggagttgccg
      181 ccagattctg acatggatct gaatctgatt gagcaggcac ccctgaccgt ggccgagaag
      241 ctgcagcgcg actttctgac ggaatggcgc cgtgtgagta aggccccgga ggctcttttc
      301 tttgtgcaat ttgagaaggg agagagctac ttccacatgc acgtgctcgt ggaaaccacc
      361 ggggtgaaat ccatggtttt gggacgtttc ctgagtcaga ttcgcgaaaa actgattcag
      421 agaatttacc gcgggatcga gccgactttg ccaaactggt tcgcggtcac aaagaccaga
      481 aatggcgccg gaggcgggaa caaggtggtg gatgagtgct acatccccaa ttacttgctc
      541 cccaaaaccc agcctgagct ccagtgggcg tggactaata tggaacagta tttaagcgcc
      601 tgtttgaatc tcacggagcg taaacggttg gtggcgcagc atctgacgca cgtgtcgcag
      661 acgcaggagc agaacaaaga gaatcagaat cccaattctg atgcgccggt gatcagatca
      721 aaaacttcag ccaggtacat ggagctggtc gggtggctcg tggacaaggg gattacctcg
      781 gagaagcagt ggatccagga ggaccaggcc tcatacatct ccttcaatgc ggcctccaac
      841 tcgcggtccc aaatcaaggc tgccttggac aatgcgggaa agattatgag cctgactaaa
      901 accgcccccg actacctggt gggccagcag cccgtggagg acatttccag caatcggatt
      961 tataaaattt tggaactaaa cgggtacgat ccccaatatg cggcttccgt ctttctggga
     1021 tgggccacga aaaagttcgg caagaggaac accatctggc tgtttgggcc tgcaactacc
     1081 gggaagacca acatcgcgga ggccatagcc cacactgtgc ccttctacgg gtgcgtaaac
     1141 tggaccaatg agaactttcc cttcaacgac tgtgtggaca agatggtgat ctggtgggag
     1201 gaggggaaga tgaccgccaa ggtcgtggag tcggccaaag ccattctcgg aggaagcaag
     1261 gtgcgcgtgg accagaaatg caagtcctcg gcccagatag acccgactcc cgtgatcgtc
     1321 acctccaaca ccaatatgtg cgccgtgatt gacgggaact caacgacctt cgaacaccag
     1381 cagccgttgc aagaccggat gttcaaattt gaactcaccc gccgtctgga tcatgacttt
     1441 gggaaggtca ccaagcagga agtcaaagac tttttccggt gggcaaagga tcacgtggtt
     1501 gaggtggagc atgaattcta cgtcaaaaag ggtggagcca agaaaagacc cgcccccagt
     1561 gacgcagata taagtgagcc caaacgggtg cgcgagtcag ttgcgcagcc atcgacgtca
     1621 gacgcggaag cttcgatcaa ctacgcggac aggtaccaaa acaaatgttc tcgtcacgtg
     1681 ggcatgaatc tgatgctgtt tccctgcaga caatgcgaga gactgaatca gaattcaaat
     1741 atctgcttca ctcacggtgt caaagactgt ttagagtgct ttcccgtgtc agaatctcaa
     1801 cccgtttctg tcgtcaaaaa ggcgtatcag aaactgtgct acattcatca catcatggga
     1861 aaggtgccag acgcttgcac tgcttgcgac ctggtcaatg tggacttgga tgactgtgtt
     1921 tctgaacaat aaatgactta aaccaggtat gagtcggctg gataaatcta aagtcataaa
     1981 cggcgctctg gaattactca atgaagtcgg tatcgaaggc ctgacgacaa ggaaactcgc
     2041 tcaaaagctg ggagttgagc agcctaccct gtactggcac gtgaagaaca agcgggccct
     2101 gctcgatgcc ctggccatcg agatgctgga caggcatcat acccacttct gccccctgga
     2161 aggcgagtca tggcaagact ttctgcggaa caacgccaag tcattccgct gtgctctcct
     2221 ctcacatcgc gacggggcta aagtgcatct cggcacccgc ccaacagaga aacagtacga
     2281 aaccctggaa aatcagctcg cgttcctgtg tcagcaaggc ttctccctgg agaacgcact
     2341 gtacgctctg tccgccgtgg gccactttac actgggctgc gtattggagg aacaggagca
     2401 tcaagtagca aaagaggaaa gagagacacc taccaccgat tctatgcccc cacttctgag
     2461 acaagcaatt gagctgttcg accggcaggg agccgaacct gccttccttt tcggcctgga
     2521 actaatcata tgtggcctgg agaaacagct aaagtgcgaa agcggcgggc cggccgacgc
     2581 ccttgacgat tttgacttag acatgctccc agccgatgcc cttgacgact ttgaccttga
     2641 tatgctgcct gctgacgctc ttgacgattt tgaccttgac atgctccccg ggtaaatgca
     2701 tgaattcgat ctagagggcc ctattctata gtgtcaccta aatgctagag ctcgctgatc
     2761 agcctcgact gtgccttcta gttgccagcc atctgttgtt tgcccctccc ccgtgccttc
     2821 cttgaccctg gaaggtgcca ctcccactgt cctttcctaa taaaatgagg aaattgcatc
     2881 gcattgtctg agtaggtgtc attctattct ggggggtggg gtggggcagg acagcaaggg
     2941 ggaggattgg gaagacaata gcaggcatgc tggggatgcg gtgggctcta tggcttctga
     3001 ggcggaaaga accagctggg gctcgaatca agctatcaag tgccacctga cgtctcccta
     3061 tcagtgatag agaagtcgac acgtctcgag ctccctatca gtgatagaga aggtacgtct
     3121 agaacgtctc cctatcagtg atagagaagt cgacacgtct cgagctccct atcagtgata
     3181 gagaaggtac cccctatata agcagagaga tctgttcaaa tttgaactga ctaagcggct
     3241 cccgccagat tttggcaaga ttactaagca ggaagtcaag gacttttttg cttgggcaaa
     3301 ggtcaatcag gtgccggtga ctcacgagtt taaagttccc agggaattgg cgggaactaa
     3361 aggggcggag aaatctctaa aacgcccact gggtgacgtc accaatacta gctataaaag
     3421 tctggagaag cgggccaggc tctcatttgt tcccgagacg cctcgcagtt cagacgtgac
     3481 tgttgatccc gctcctctgc gaccgctagc ttcgatcaac tacgcagaca ggtaccaaaa
     3541 caagtgttct cgtcacgtgg gcattaatct gattctgttt ccctgcagac aatgcgagag
     3601 aatgaatcag aactcaaata tctgcttcac tcacggacag aaagactgtt tagagtgctt
     3661 tcccgtgtca gaatctcaac ccgtttctgt cgtcaaaaag gcgtatcaga aactgtgcta
     3721 cattcatcat atcatgggaa aggtgccaga cgcttgcact gcctgcgatc tggtcaatgt
     3781 ggatttggat gactgcatct ttgaacaata aatgacttaa gccaggtatg gctgccgatg
     3841 gttatcttcc agattggctc gaggacaacc ttagtgaagg aattcgcgag tggtgggctt
     3901 tgaaacctgg agcccctcaa cccaaggcaa atcaacaaca tcaagacaac gctagaggtc
     3961 ttgtgcttcc gggttacaaa taccttggac ccggcaacgg actcgacaag ggggagccgg
     4021 tcaacgcagc agacgcggcg gccctcgagc acgacaaagc ctacgaccag cagctcaagg
     4081 ccggagacaa cccgtacctc aagtacaacc acgccgacgc cgagttccag gagcggctca
     4141 aagaagatac gtcttttggg ggcaacctcg ggcgagcagt cttccaggcc aaaaagaggc
     4201 ttcttgaacc tcttggtctg gttgaggaag cggctaagac ggctcctgga aagaagaggc
     4261 ctgtagagca gtctcctcag gaaccggact cctccgcggg tattggcaaa tcgggtgcac
     4321 agcccgctaa aaagagactc aatttcggtc agactggcga cacagagtca gtcccagacc
     4381 ctcaaccaat cggagaacct cccgcagccc cctcaggtgt gggatctctt acaatggctt
     4441 caggtggtgg cgcaccagtg gcagacaata acgaaggtgc cgatggagtg ggtagttcct
     4501 cgggaaattg gcattgcgat tcccaatggc tgggggacag agtcatcacc accagcaccc
     4561 gaacctgggc cctgcccacc tacaacaatc acctctacaa gcaaatctcc aacagcacat
     4621 ctggaggatc ttcaaatgac aacgcctact tcggctacag caccccctgg gggtattttg
     4681 acttcaacag attccactgc cacttctcac cacgtgactg gcagcgactc atcaacaaca
     4741 actggggatt ccggcctaag cgactcaact tcaagctctt taacattcag gtcaaagagg
     4801 ttacggacaa caatggagtc aagaccatcg ccaataacct taccagcacg gtccaggtct
     4861 tcacggactc agactatcag ctcccgtacg tgctcgggtc ggctcacgag ggctgcctcc
     4921 cgccgttccc agcggacgtt ttcatgattc ctcagtacgg gtatctgacg cttaatgatg
     4981 gaagccaggc cgtgggtcgt tcgtcctttt actgcctgga atatttcccg tcgcaaatgc
     5041 taagaacggg taacaacttc cagttcagct acgagtttga gaacgtacct ttccatagca
     5101 gctacgctca cagccaaagc ctggaccgac taatgaatcc actcatcgac caatacttgt
     5161 actatctctc aaagactatt aacggttctg gacagaatca acaaacgcta aaattcagtg
     5221 tggccggacc cagcaacatg gctgtccagg gaagaaacta catacctgga cccagctacc
     5281 gacaacaacg tgtctcaacc actgtgactc aaaacaacaa cagcgaattt gcttggcctg
     5341 gagcttcttc ttgggctctc aatggacgta atagcttgat gaatcctgga cctgctatgg
     5401 cctctcacaa agaaggagag gaccgtttct ttcctttgtc tggatcttta atttttggca
     5461 aacaaggtac tggcagagac aacgtggatg cggacaaagt catgataacc aacgaagaag
     5521 aaattaaaac tactaacccg gtagcaacgg agtcctatgg acaagtggcc acaaaccacc
     5581 agagtgacgg aattgtgggg tcgactattc gggcacaggc gcagaccggt tgggttcaaa
     5641 accaaggaat acttccgggt atggtttggc aggacagaga tgtgtacctg caaggaccca
     5701 tttgggccaa aattcctcac acggacggca actttcaccc ttctccgctg atgggagggt
     5761 ttggaatgaa gcacccgcct cctcagatcc tcatcaaaaa cacacctgta cctgcggatc
     5821 ctccaacggc cttcaacaag gacaagctga actctttcat cacccagtat tctactggtc
     5881 aagtcagcgt ggagatcgag tgggagctgc agaaggaaaa cagcaagcgc tggaacccgg
     5941 agatccagta cacttccaac tattacaagt ctaataatgt tgaatttgct gttaatactg
     6001 aaggtgtata tagtgaaccc cgccccattg gcaccagata cctgactcgt aatctgtaag
     6061 tcgacttgct tgttaatcaa taaaccgttt aattcgtttc agttgaactt tggtctctgc
     6121 gaagggcaat tcgtttaaac ctgcaggact agaggtcctg tattagaggt cacgtgagtg
     6181 ttttgcgaca ttttgcgaca ccatgtggtc acgctgggta tttaagcccg agtgagcacg
     6241 cagggtctcc attttgaagc gggaggtttg aacgcgcagc cgccaagccg aattctgcag
     6301 atatcacatg tcctaggaac tatcgatcca tcacactggc ggccgctcga ctagagcggc
     6361 cgccaccgcg gtggagctcc agcttttgcg gaccgaatcg gaaagaacat gtgagcaaaa
     6421 ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc
     6481 cgcccccctg acgagcatca caaaaatcga cgctcaagtc agaggtggcg aaacccgaca
     6541 ggactataaa gataccaggc gtttccccct ggaagctccc tcgtgcgctc tcctgttccg
     6601 accctgccgc ttaccggata cctgtccgcc tttctccctt cgggaagcgt ggcgctttct
     6661 catagctcac gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt
     6721 gtgcacgaac cccccgttca gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag
     6781 tccaacccgg taagacacga cttatcgcca ctggcagcag ccactggtaa caggattagc
     6841 agagcgaggt atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa ctacggctac
     6901 actagaagaa cagtatttgg tatctgcgct ctgctgaagc cagttacctt cggaaaaaga
     6961 gttggtagct cttgatccgg caaacaaacc accgctggta gcggtggttt ttttgtttgc
     7021 aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag atcctttgat cttttctacg
     7081 gggtctgacg ctcagtggaa cgaaaactca cgttaaggga ttttggtcat gagattatca
     7141 aaaaggatct tcacctagat ccttttaaat taaaaatgaa gttttaaatc aatctaaagt
     7201 atatatgagt aaacttggtc tgacagttac caatgcttaa tcagtgaggc acctatctca
     7261 gcgatctgtc tatttcgttc atccatagtt gcctgactcc ccgtcgtgta gataactacg
     7321 atacgggagg gcttaccatc tggccccagt gctgcaatga taccgcgaga cccacgctca
     7381 ccggctccag atttatcagc aataaaccag ccagccggaa gggccgagcg cagaagtggt
     7441 cctgcaactt tatccgcctc catccagtct attaattgtt gccgggaagc tagagtaagt
     7501 agttcgccag ttaatagttt gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca
     7561 cgctcgtcgt ttggtatggc ttcattcagc tccggttccc aacgatcaag gcgagttaca
     7621 tgatccccca tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat cgttgtcaga
     7681 agtaagttgg ccgcagtgtt atcactcatg gttatggcag cactgcataa ttctcttact
     7741 gtcatgccat ccgtaagatg cttttctgtg actggtgagt actcaaccaa gtcattctga
     7801 gaatagtgta tgcggcgacc gagttgctct tgcccggcgt caatacggga taataccgcg
     7861 ccacatagca gaactttaaa agtgctcatc attggaaaac gttcttcggg gcgaaaactc
     7921 tcaaggatct taccgctgtt gagatccagt tcgatgtaac ccactcgtgc acccaactga
     7981 tcttcagcat cttttacttt caccagcgtt tctgggtgag caaaaacagg aaggcaaaat
     8041 gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa tactcatact cttccttttt
     8101 caatattatt gaagcattta tcagggttat tgtctcatga gcggatacat atttgaatgt
     8161 atttagaaaa ataaacaaat aggggttccg cgcacatttc cccgaaaagt gccacctgac
     8221 gtcccaatga tacgcgtcgg tgcgggcctc ttcgctatta cgccagctgg cgaaaggggg
     8281 atgtgctgca aggcgattaa gttgggtaac gccagggttt tcccagtcac gacgttgtaa
     8341 aacgacggcc agtgagcgcg cgtaatacga ctcactatag ggcgaattgg gtaca
//
