LOCUS       sequence-100033-        4476 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 4476)
  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 4476)
  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..4476
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   1..141
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   12..141
                     /label=AAV2 ITR
     regulatory      561..570
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(567..569,570..572,573..1286)
                     /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(567..569,570..572,573..1283)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(612..633)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(873..892)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1220..1241
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(1359..1379)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     repeat_region   1739..1879
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   1739..1868
                     /label=AAV2 ITR
     rep_origin      1954..2409
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2041..2060)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2251..2272
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(2426..2445)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     2545..2567
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(2605..2623)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        2691..2795
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2796..2864,2865..3656)
                     /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(3014..3033)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3827..4415
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4316..4335
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 cctgcaggca gctgcgcgct cgctcgctca ctgaggccgc ccgggcaaag cccgggcgtc
       61 gggcgacctt tggtcgcccg gcctcagtga gcgagcgagc gcgcagagag ggagtggcca
      121 actccatcac taggggttcc tgcggccgca gatgctgcac acttgtggac taagtttgtt
      181 cgcatcccct tctccaaccc cctcagtaca tcaccctggg ggaacagggt ccacttgctc
      241 ctgggcccac acagtcctgc agtattgtgt atataaggcc agggcaaaga ggagcaggtt
      301 ttaaagtgaa aggcaggcag gtgttgggga ggcagttacc ggggcaacgg gaacagggcg
      361 tttcggaggt ggttgccatg gggacctgga tgctgacgaa ggctcgcgag gctgtgagca
      421 gccacagtgc cctgctcaga agccccaagc tcgtcagtca agccggttct ccgtttgcac
      481 tcaggagcac gggcaggcga gtggccccta gttctggggg cagcgtgaaa agacggtatc
      541 gataagcttg attcgagcta gccaccatgg tgagcaaggg cgaggagctg ttcaccgggg
      601 tggtgcccat cctggtcgag ctggacggcg acgtaaacgg ccacaagttc agcgtgtccg
      661 gcgagggcga gggcgatgcc acctacggca agctgaccct gaagttcatc tgcaccaccg
      721 gcaagctgcc cgtgccctgg cccaccctcg tgaccaccct gacctacggc gtgcagtgct
      781 tcagccgcta ccccgaccac atgaagcagc acgacttctt caagtccgcc atgcccgaag
      841 gctacgtcca ggagcgcacc atcttcttca aggacgacgg caactacaag acccgcgccg
      901 aggtgaagtt cgagggcgac accctggtga accgcatcga gctgaagggc atcgacttca
      961 aggaggacgg caacatcctg gggcacaagc tggagtacaa ctacaacagc cacaacgtct
     1021 atatcatggc cgacaagcag aagaacggca tcaaggtgaa cttcaagatc cgccacaaca
     1081 tcgaggacgg cagcgtgcag ctcgccgacc actaccagca gaacaccccc atcggcgacg
     1141 gccccgtgct gctgcccgac aaccactacc tgagcaccca gtccgccctg agcaaagacc
     1201 ccaacgagaa gcgcgatcac atggtcctgc tggagttcgt gaccgccgcc gggatcactc
     1261 tcggcatgga cgagctgtac aagtaaatcg aattccgctc gagataatca acctctggat
     1321 tacaaaattt gtgaaagatt gactggtatt cttaactatg ttgctccttt tacgctatgt
     1381 ggatacgctg ctttaatgcc tttgtatcat gctattgctt cccgtatggc tttcattttc
     1441 tcctccttgt ataaatcctg gttagttctt gccacggcgg aactcatcgc cgcctgcctt
     1501 gcccgctgct ggacaggggc tcggctgttg ggcactgaca attccgtggt gtttatttgt
     1561 gaaatttgtg atgctattgc tttatttgta accatctagc tttatttgtg aaatttgtga
     1621 tgctattgct ttatttgtaa ccattataag ctgcaataaa caagttaaca acaacaattg
     1681 cattcatttt atgtttcagg ttcaggggga gatgtgggag gttttttaaa gcggccgcag
     1741 gaacccctag tgatggagtt ggccactccc tctctgcgcg ctcgctcgct cactgaggcc
     1801 gggcgaccaa aggtcgcccg acgcccgggc tttgcccggg cggcctcagt gagcgagcga
     1861 gcgcgcagct gcctgcaggg gcgcctgatg cggtattttc tccttacgca tctgtgcggt
     1921 atttcacacc gcatacgtca aagcaaccat agtacgcgcc ctgtagcggc gcattaagcg
     1981 cggcgggtgt ggtggttacg cgcagcgtga ccgctacact tgccagcgcc ctagcgcccg
     2041 ctcctttcgc tttcttccct tcctttctcg ccacgttcgc cggctttccc cgtcaagctc
     2101 taaatcgggg gctcccttta gggttccgat ttagtgcttt acggcacctc gaccccaaaa
     2161 aacttgattt gggtgatggt tcacgtagtg ggccatcgcc ctgatagacg gtttttcgcc
     2221 ctttgacgtt ggagtccacg ttctttaata gtggactctt gttccaaact ggaacaacac
     2281 tcaaccctat ctcgggctat tcttttgatt tataagggat tttgccgatt tcggcctatt
     2341 ggttaaaaaa tgagctgatt taacaaaaat ttaacgcgaa ttttaacaaa atattaacgt
     2401 ttacaatttt atggtgcact ctcagtacaa tctgctctga tgccgcatag ttaagccagc
     2461 cccgacaccc gccaacaccc gctgacgcgc cctgacgggc ttgtctgctc ccggcatccg
     2521 cttacagaca agctgtgacc gtctccggga gctgcatgtg tcagaggttt tcaccgtcat
     2581 caccgaaacg cgcgagacga aagggcctcg tgatacgcct atttttatag gttaatgtca
     2641 tgataataat ggtttcttag acgtcaggtg gcacttttcg gggaaatgtg cgcggaaccc
     2701 ctatttgttt atttttctaa atacattcaa atatgtatcc gctcatgaga caataaccct
     2761 gataaatgct tcaataatat tgaaaaagga agagtatgag tattcaacat ttccgtgtcg
     2821 cccttattcc cttttttgcg gcattttgcc ttcctgtttt tgctcaccca gaaacgctgg
     2881 tgaaagtaaa agatgctgaa gatcagttgg gtgcacgagt gggttacatc gaactggatc
     2941 tcaacagcgg taagatcctt gagagttttc gccccgaaga acgttttcca atgatgagca
     3001 cttttaaagt tctgctatgt ggcgcggtat tatcccgtat tgacgccggg caagagcaac
     3061 tcggtcgccg catacactat tctcagaatg acttggttga gtactcacca gtcacagaaa
     3121 agcatcttac ggatggcatg acagtaagag aattatgcag tgctgccata accatgagtg
     3181 ataacactgc ggccaactta cttctgacaa cgatcggagg accgaaggag ctaaccgctt
     3241 ttttgcacaa catgggggat catgtaactc gccttgatcg ttgggaaccg gagctgaatg
     3301 aagccatacc aaacgacgag cgtgacacca cgatgcctgt agcaatggca acaacgttgc
     3361 gcaaactatt aactggcgaa ctacttactc tagcttcccg gcaacaatta atagactgga
     3421 tggaggcgga taaagttgca ggaccacttc tgcgctcggc ccttccggct ggctggttta
     3481 ttgctgataa atctggagcc ggtgagcgtg ggtctcgcgg tatcattgca gcactggggc
     3541 cagatggtaa gccctcccgt atcgtagtta tctacacgac ggggagtcag gcaactatgg
     3601 atgaacgaaa tagacagatc gctgagatag gtgcctcact gattaagcat tggtaactgt
     3661 cagaccaagt ttactcatat atactttaga ttgatttaaa acttcatttt taatttaaaa
     3721 ggatctaggt gaagatcctt tttgataatc tcatgaccaa aatcccttaa cgtgagtttt
     3781 cgttccactg agcgtcagac cccgtagaaa agatcaaagg atcttcttga gatccttttt
     3841 ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt
     3901 tgccggatca agagctacca actctttttc cgaaggtaac tggcttcagc agagcgcaga
     3961 taccaaatac tgtccttcta gtgtagccgt agttaggcca ccacttcaag aactctgtag
     4021 caccgcctac atacctcgct ctgctaatcc tgttaccagt ggctgctgcc agtggcgata
     4081 agtcgtgtct taccgggttg gactcaagac gatagttacc ggataaggcg cagcggtcgg
     4141 gctgaacggg gggttcgtgc acacagccca gcttggagcg aacgacctac accgaactga
     4201 gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca
     4261 ggtatccggt aagcggcagg gtcggaacag gagagcgcac gagggagctt ccagggggaa
     4321 acgcctggta tctttatagt cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt
     4381 tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg gcctttttac
     4441 ggttcctggc cttttgctgg ccttttgctc acatgt
//
