LOCUS       sequence-455914-        5335 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AAV vector for expressing CMV-driven mCherry reporter gene..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5335)
  AUTHORS   Kumar S, Hsiao YW, Wong VHY, Aubin D, Wang JH, Lisowski L, Rakoczy 
            EP, Li F, Alarcon-Martinez L, Gonzalez-Cordero A, Bui BV, Liu GS
  TITLE     Characterization of RNA editing and gene therapy with a compact 
            CRISPR-Cas13 in the retina.
  JOURNAL   Proc Natl Acad Sci U S A. 2024 Nov 5;121(45):e2408345121. doi: 
            10.1073/pnas.2408345121. Epub 2024 Oct 30.
  PUBMED    39475642
REFERENCE   2  (bases 1 to 5335)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Proc Natl 
            Acad Sci U S A."; date: "2024-11-5"; volume: "121(45):e2408345121. 
            doi"; pages: " 10.1073/pnas.2408345121. Epub 2024 Oct 30"
COMMENT     Sequence Label: pC081: pAAV.CMV.mCherry
FEATURES             Location/Qualifiers
     source          1..5335
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(join(107..898,899..967))
                     /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     730..749
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(968..1072)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     1140..1158
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(1196..1218)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     1318..1337
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     rep_origin      complement(1354..1809)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1491..1512)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     1703..1722
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     repeat_region   1884..2024
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   1895..2024
                     /label=AAV2 ITR
     polyA_signal    2064..2540
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     2322..2341
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     CDS             complement(2577..3287)
                     /codon_start=1
                     /product="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
                     /label=mCherry
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEG
                     TQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNF
                     EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK
                     GEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERA
                     EGRHSTGGMDELYK"
     primer_bind     complement(2855..2874)
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     primer_bind     3124..3142
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     regulatory      3284..3293
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     intron          3382..3760
                     /label=chimeric intron
                     /note="fusion between human cytomegalovirus intron A and 
                     human beta-globin intron 2"
     primer_bind     complement(3888..3912)
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     promoter        complement(3909..4112)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(3938..3958)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        4113..4416
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     repeat_region   4492..4621
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     rep_origin      complement(4683..5271)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4763..4782)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 ttttggtcat gagattatca aaaaggatct tcacctagat ccttttaaat taaaaatgaa
       61 gttttaaatc aatctaaagt atatatgagt aaacttggtc tgacagttac caatgcttaa
      121 tcagtgaggc acctatctca gcgatctgtc tatttcgttc atccatagtt gcctgactcc
      181 ccgtcgtgta gataactacg atacgggagg gcttaccatc tggccccagt gctgcaatga
      241 taccgcgaga cccacgctca ccggctccag atttatcagc aataaaccag ccagccggaa
      301 gggccgagcg cagaagtggt cctgcaactt tatccgcctc catccagtct attaattgtt
      361 gccgggaagc tagagtaagt agttcgccag ttaatagttt gcgcaacgtt gttgccattg
      421 ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc tccggttccc
      481 aacgatcaag gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt agctccttcg
      541 gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg gttatggcag
      601 cactgcataa ttctcttact gtcatgccat ccgtaagatg cttttctgtg actggtgagt
      661 actcaaccaa gtcattctga gaatagtgta tgcggcgacc gagttgctct tgcccggcgt
      721 caatacggga taataccgcg ccacatagca gaactttaaa agtgctcatc attggaaaac
      781 gttcttcggg gcgaaaactc tcaaggatct taccgctgtt gagatccagt tcgatgtaac
      841 ccactcgtgc acccaactga tcttcagcat cttttacttt caccagcgtt tctgggtgag
      901 caaaaacagg aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa
      961 tactcatact cttccttttt caatattatt gaagcattta tcagggttat tgtctcatga
     1021 gcggatacat atttgaatgt atttagaaaa ataaacaaat aggggttccg cgcacatttc
     1081 cccgaaaagt gccacctgac gtctaagaaa ccattattat catgacatta acctataaaa
     1141 ataggcgtat cacgaggccc tttcgtctcg cgcgtttcgg tgatgacggt gaaaacctct
     1201 gacacatgca gctcccggag acggtcacag cttgtctgta agcggatgcc gggagcagac
     1261 aagcccgtca gggcgcgtca gcgggtgttg gcgggtgtcg gggctggctt aactatgcgg
     1321 catcagagca gattgtactg agagtgcacc ataaaattgt aaacgttaat attttgttaa
     1381 aattcgcgtt aaatttttgt taaatcagct cattttttaa ccaatagacc gaaatcggca
     1441 aaatccctta taaatcaaaa gaatagcccg agatagagtt gagtgttgtt ccagtttgga
     1501 acaagagtcc actattaaag aacgtggact ccaacgtcaa agggcgaaaa accgtctatc
     1561 agggcgatgg cccactacgt gaaccatcac ccaaatcaag ttttttgggg tcgaggtgcc
     1621 gtaaagcact aaatcggaac cctaaaggga gcccccgatt tagagcttga cggggaaagc
     1681 cggcgaacgt ggcgagaaag gaagggaaga aagcgaaagg agcgggcgct aaggcgctgg
     1741 caagtgtagc ggtcacgctg cgcgtaacca ccacacccgc cgcgcttaat gcgccgctac
     1801 agggcgcgta ctatggttgc tttgacgtat gcggtgtgaa ataccgcaca gatgcgtaag
     1861 gagaaaatac cgcatcaggc gcccctgcag gcagctgcgc gctcgctcgc tcactgaggc
     1921 cgcccgggca aagcccgggc gtcgggcgac ctttggtcgc ccggcctcag tgagcgagcg
     1981 agcgcgcaga gagggagtgg ccaactccat cactaggggt tcctgcggcc gctcggtccg
     2041 cacgtggtta cctacaaaat cagaaggaca gggaagggag cagtggttca cgcctgtaat
     2101 cccagcaatt tgggaggcca aggtgggtag atcacctgag attaggagtt ggagaccagc
     2161 ctggccaata tggtgaaacc ccgtctctac caaaaaaaca aaaattagct gagcctggtc
     2221 atgcatgcct ggaatcccaa caactcggga ggctgaggca ggagaatcgc ttgaacccag
     2281 gaggcggaga ttgcagtgag ccaagattgt gccactgcac tccagcttgg ttcccaatag
     2341 accccgcagg ccctacaggt tgtcttccca acttgcccct tgctccatac cacccccctc
     2401 caccccataa tattatagaa ggacacctag tcagacaaaa tgatgcaact taattttatt
     2461 aggacaaggc tggtgggcac tggagtggca acttccaggg ccaggagagg cactggggag
     2521 gggtcacagg gatgccaccc gtagatctct cgagcagcgc tgctcgaggc aagcttctac
     2581 ttgtacagct cgtccatgcc gccggtggag tggcggccct cggcgcgttc gtactgttcc
     2641 acgatggtgt agtcctcgtt gtgggaggtg atgtccaact tgatgttgac gttgtaggcg
     2701 ccgggcagct gcacgggctt cttggccttg taggtggtct tgacctcagc gtcgtagtgg
     2761 ccgccgtcct tcagcttcag cctctgcttg atctcgccct tcagggcgcc gtcctcgggg
     2821 tacatccgct cggaggaggc ctcccagccc atggtcttct tctgcattac ggggccgtcg
     2881 gaggggaagt tggtgccgcg cagcttcacc ttgtagatga actcgccgtc ctgcagggag
     2941 gagtcctggg tcacggtcac cacgccgccg tcctcgaagt tcatcacgcg ctcccacttg
     3001 aagccctcgg ggaaggacag cttcaagtag tcggggatgt cggcggggtg cttcacgtag
     3061 gccttggagc cgtacatgaa ctgaggggac aggatgtccc aggcgaaggg cagggggcca
     3121 cccttggtca ccttcagctt ggcggtctgg gtgccctcgt aggggcggcc ctcgccctcg
     3181 ccctcgatct cgaactcgtg gccgttcacg gagccctcca tgtgcacctt gaagcgcatg
     3241 aactccttga tgatggccat gttatcctcc tcgcccttgc tcaccatggt ggcggatccc
     3301 cggggaattc aatcgatgtt cgaatcccaa ttctttgcca aagtgatggg ccagcacaca
     3361 gaccagcacg ttgcccagga gctgtgggag gaagataaga ggtatgaaca tgattagcaa
     3421 aagggcctag cttggactca gaataatcca gccttatccc aaccataaaa taaaagcaga
     3481 atggtagctg gattgtagct gctattagca atatgaaacc tcttacatca gttacaattt
     3541 atatgcagaa atatttatat gcagaaatat tgctattgcc ttaacccaga aattatcact
     3601 gttattcttt agaatggtgc aaagaggcat gatacattgt atcattattg ccctgaaaga
     3661 aagagattag ggaaagtatt agaaataaga taaacaaaaa agtatattaa aagaagaaag
     3721 cattttttgt gggcctatag actctatagg cggtacttac gtcactcttg gcacggggaa
     3781 tccgcgttcc aatgcaccgt tcccggccgg gattcgaatc cgcggaggct ggatcggtcc
     3841 cggtgtcttc tatggaggtc aaaacagcgt ggatggcgtc tccaggcgat ctgacggttc
     3901 actaaacgag ctctgcttat atagacctcc caccgtacac gcctaccgcc catttgcgtc
     3961 aatggggcgg agttgttacg acattttgga aagtcccgtt gattttggtg ccaaaacaaa
     4021 ctcccattga cgtcaatggg gtggagactt ggaaatcccc gtgagtcaaa ccgctatcca
     4081 cgcccattga tgtactgcca aaaccgcatc accatggtaa tagcgatgac taatacgtag
     4141 atgtactgcc aagtaggaaa gtcccataag gtcatgtact gggcataatg ccaggcgggc
     4201 catttaccgt cattgacgtc aatagggggc gtacttggca tatgatacac ttgatgtact
     4261 gccaagtggg cagtttaccg taaatactcc acccattgac gtcaatggaa agtccctatt
     4321 ggcgttacta tgggaacata cgtcattatt gacgtcaatg ggcgggggtc gttgggcggt
     4381 cagccaggcg ggccatttac cgtaagttat gtaacgcgga actccatata tgggctatga
     4441 actaatgacc ccgtaattga ttactattaa taactagacg cgtgcggccg caggaacccc
     4501 tagtgatgga gttggccact ccctctctgc gcgctcgctc gctcactgag gccgggcgac
     4561 caaaggtcgc ccgacgcccg ggcggcctca gtgagcgagc gagcgcgcag ctgcctgcag
     4621 gacatgtgag caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg
     4681 tttttccata ggctccgccc ccctgacgag catcacaaaa atcgacgctc aagtcagagg
     4741 tggcgaaacc cgacaggact ataaagatac caggcgtttc cccctggaag ctccctcgtg
     4801 cgctctcctg ttccgaccct gccgcttacc ggatacctgt ccgcctttct cccttcggga
     4861 agcgtggcgc tttctcatag ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc
     4921 tccaagctgg gctgtgtgca cgaacccccc gttcagcccg accgctgcgc cttatccggt
     4981 aactatcgtc ttgagtccaa cccggtaaga cacgacttat cgccactggc agcagccact
     5041 ggtaacagga ttagcagagc gaggtatgta ggcggtgcta cagagttctt gaagtggtgg
     5101 cctaactacg gctacactag aagaacagta tttggtatct gcgctctgct gaagccagtt
     5161 accttcggaa aaagagttgg tagctcttga tccggcaaac aaaccaccgc tggtagcggt
     5221 ggtttttttg tttgcaagca gcagattacg cgcagaaaaa aaggatctca agaagatcct
     5281 ttgatctttt ctacggggtc tgacgctcag tggaacgaaa actcacgtta aggga
//
