LOCUS       sequence-258218-        4465 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AAV vector with CaMKIIa promoter, EGFP, and W3SL(shorten expression 
            cassette with relatively high expression to maximize expression of 
            larger genes) .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4465)
  AUTHORS   Choi JH, Yu NK, Baek GC, Bakes J, Seo D, Nam HJ, Baek SH, Lim CS, 
            Lee YS, Kaang BK
  TITLE     Optimization of AAV expression cassettes to improve packaging 
            capacity and transgene expression in neurons.
  JOURNAL   Mol Brain. 2014 Mar 11;7:17. doi: 10.1186/1756-6606-7-17.
  PUBMED    24618276
REFERENCE   2  (bases 1 to 4465)
  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: "Mol 
            Brain."; date: "2014-03-11"; volume: "7:17. doi"; pages: " 
            10.1186/1756-6606-7-17"
COMMENT     Sequence Label: pAAV-CW3SL-EGFP
FEATURES             Location/Qualifiers
     source          1..4465
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   1..130
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     regulatory      550..559
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(556..558,559..561,562..1275)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(556..558,559..561,562..1272)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(601..622)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(862..881)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1209..1230
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(1348..1368)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     repeat_region   1728..1868
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   1728..1857
                     /label=AAV2 ITR
     rep_origin      1943..2398
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2030..2049)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2240..2261
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(2415..2434)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     2534..2556
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(2594..2612)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        2680..2784
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2785..2853,2854..3645)
                     /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(3003..3022)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3816..4404
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4305..4324
                     /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 gcggccgcag atgctgcaca cttgtggact aagtttgttc gcatcccctt
      181 ctccaacccc ctcagtacat caccctgggg gaacagggtc cacttgctcc tgggcccaca
      241 cagtcctgca gtattgtgta tataaggcca gggcaaagag gagcaggttt taaagtgaaa
      301 ggcaggcagg tgttggggag gcagttaccg gggcaacggg aacagggcgt ttcggaggtg
      361 gttgccatgg ggacctggat gctgacgaag gctcgcgagg ctgtgagcag ccacagtgcc
      421 ctgctcagaa gccccaagct cgtcagtcaa gccggttctc cgtttgcact caggagcacg
      481 ggcaggcgag tggcccctag ttctgggggc agcgtgaaaa gacggtatcg ataagcttga
      541 ttcgagctag ccaccatggt gagcaagggc gaggagctgt tcaccggggt ggtgcccatc
      601 ctggtcgagc tggacggcga cgtaaacggc cacaagttca gcgtgtccgg cgagggcgag
      661 ggcgatgcca cctacggcaa gctgaccctg aagttcatct gcaccaccgg caagctgccc
      721 gtgccctggc ccaccctcgt gaccaccctg acctacggcg tgcagtgctt cagccgctac
      781 cccgaccaca tgaagcagca cgacttcttc aagtccgcca tgcccgaagg ctacgtccag
      841 gagcgcacca tcttcttcaa ggacgacggc aactacaaga cccgcgccga ggtgaagttc
      901 gagggcgaca ccctggtgaa ccgcatcgag ctgaagggca tcgacttcaa ggaggacggc
      961 aacatcctgg ggcacaagct ggagtacaac tacaacagcc acaacgtcta tatcatggcc
     1021 gacaagcaga agaacggcat caaggtgaac ttcaagatcc gccacaacat cgaggacggc
     1081 agcgtgcagc tcgccgacca ctaccagcag aacaccccca tcggcgacgg ccccgtgctg
     1141 ctgcccgaca accactacct gagcacccag tccgccctga gcaaagaccc caacgagaag
     1201 cgcgatcaca tggtcctgct ggagttcgtg accgccgccg ggatcactct cggcatggac
     1261 gagctgtaca agtaaatcga attccgctcg agataatcaa cctctggatt acaaaatttg
     1321 tgaaagattg actggtattc ttaactatgt tgctcctttt acgctatgtg gatacgctgc
     1381 tttaatgcct ttgtatcatg ctattgcttc ccgtatggct ttcattttct cctccttgta
     1441 taaatcctgg ttagttcttg ccacggcgga actcatcgcc gcctgccttg cccgctgctg
     1501 gacaggggct cggctgttgg gcactgacaa ttccgtggtg tttatttgtg aaatttgtga
     1561 tgctattgct ttatttgtaa ccatctagct ttatttgtga aatttgtgat gctattgctt
     1621 tatttgtaac cattataagc tgcaataaac aagttaacaa caacaattgc attcatttta
     1681 tgtttcaggt tcagggggag atgtgggagg ttttttaaag cggccgcagg aacccctagt
     1741 gatggagttg gccactccct ctctgcgcgc tcgctcgctc actgaggccg ggcgaccaaa
     1801 ggtcgcccga cgcccgggct ttgcccgggc ggcctcagtg agcgagcgag cgcgcagctg
     1861 cctgcagggg cgcctgatgc ggtattttct ccttacgcat ctgtgcggta tttcacaccg
     1921 catacgtcaa agcaaccata gtacgcgccc tgtagcggcg cattaagcgc ggcgggtgtg
     1981 gtggttacgc gcagcgtgac cgctacactt gccagcgcct tagcgcccgc tcctttcgct
     2041 ttcttccctt cctttctcgc cacgttcgcc ggctttcccc gtcaagctct aaatcggggg
     2101 ctccctttag ggttccgatt tagtgcttta cggcacctcg accccaaaaa acttgatttg
     2161 ggtgatggtt cacgtagtgg gccatcgccc tgatagacgg tttttcgccc tttgacgttg
     2221 gagtccacgt tctttaatag tggactcttg ttccaaactg gaacaacact caactctatc
     2281 tcgggctatt cttttgattt ataagggatt ttgccgattt cggtctattg gttaaaaaat
     2341 gagctgattt aacaaaaatt taacgcgaat tttaacaaaa tattaacgtt tacaatttta
     2401 tggtgcactc tcagtacaat ctgctctgat gccgcatagt taagccagcc ccgacacccg
     2461 ccaacacccg ctgacgcgcc ctgacgggct tgtctgctcc cggcatccgc ttacagacaa
     2521 gctgtgaccg tctccgggag ctgcatgtgt cagaggtttt caccgtcatc accgaaacgc
     2581 gcgagacgaa agggcctcgt gatacgccta tttttatagg ttaatgtcat gataataatg
     2641 gtttcttaga cgtcaggtgg cacttttcgg ggaaatgtgc gcggaacccc tatttgttta
     2701 tttttctaaa tacattcaaa tatgtatccg ctcatgagac aataaccctg ataaatgctt
     2761 caataatatt gaaaaaggaa gagtatgagt attcaacatt tccgtgtcgc ccttattccc
     2821 ttttttgcgg cattttgcct tcctgttttt gctcacccag aaacgctggt gaaagtaaaa
     2881 gatgctgaag atcagttggg tgcacgagtg ggttacatcg aactggatct caacagcggt
     2941 aagatccttg agagttttcg ccccgaagaa cgttttccaa tgatgagcac ttttaaagtt
     3001 ctgctatgtg gcgcggtatt atcccgtatt gacgccgggc aagagcaact cggtcgccgc
     3061 atacactatt ctcagaatga cttggttgag tactcaccag tcacagaaaa gcatcttacg
     3121 gatggcatga cagtaagaga attatgcagt gctgccataa ccatgagtga taacactgcg
     3181 gccaacttac ttctgacaac gatcggagga ccgaaggagc taaccgcttt tttgcacaac
     3241 atgggggatc atgtaactcg ccttgatcgt tgggaaccgg agctgaatga agccatacca
     3301 aacgacgagc gtgacaccac gatgcctgta gcaatggcaa caacgttgcg caaactatta
     3361 actggcgaac tacttactct agcttcccgg caacaattaa tagactggat ggaggcggat
     3421 aaagttgcag gaccacttct gcgctcggcc cttccggctg gctggtttat tgctgataaa
     3481 tctggagccg gtgagcgtgg gtctcgcggt atcattgcag cactggggcc agatggtaag
     3541 ccctcccgta tcgtagttat ctacacgacg gggagtcagg caactatgga tgaacgaaat
     3601 agacagatcg ctgagatagg tgcctcactg attaagcatt ggtaactgtc agaccaagtt
     3661 tactcatata tactttagat tgatttaaaa cttcattttt aatttaaaag gatctaggtg
     3721 aagatccttt ttgataatct catgaccaaa atcccttaac gtgagttttc gttccactga
     3781 gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag atcctttttt tctgcgcgta
     3841 atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa
     3901 gagctaccaa ctctttttcc gaaggtaact ggcttcagca gagcgcagat accaaatact
     3961 gttcttctag tgtagccgta gttaggccac cacttcaaga actctgtagc accgcctaca
     4021 tacctcgctc tgctaatcct gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt
     4081 accgggttgg actcaagacg atagttaccg gataaggcgc agcggtcggg ctgaacgggg
     4141 ggttcgtgca cacagcccag cttggagcga acgacctaca ccgaactgag atacctacag
     4201 cgtgagctat gagaaagcgc cacgcttccc gaagggagaa aggcggacag gtatccggta
     4261 agcggcaggg tcggaacagg agagcgcacg agggagcttc cagggggaaa cgcctggtat
     4321 ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt gtgatgctcg
     4381 tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc
     4441 ttttgctggc cttttgctca catgt
//
