LOCUS       sequence-100032-        4879 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AAV vector with CaMKIIa promoter, EGFP, WPRE, and bGHpA.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4879)
  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 4879)
  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-CWB-EGFP
FEATURES             Location/Qualifiers
     source          1..4879
                     /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"
     misc_feature    1309..1897
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(1362..1382)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(1780..1791)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    1910..2134
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   2142..2282
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   2142..2271
                     /label=AAV2 ITR
     rep_origin      2357..2812
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2444..2463)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2654..2675
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(2829..2848)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     2948..2970
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(3008..3026)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3094..3198
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3199..3267,3268..4059)
                     /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(3417..3436)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4230..4818
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4719..4738
                     /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 aattcctgca gccccgataa tcaacctctg
     1321 gattacaaaa tttgtgaaag attgactggt attcttaact atgttgctcc ttttacgcta
     1381 tgtggatacg ctgctttaat gcctttgtat catgctattg cttcccgtat ggctttcatt
     1441 ttctcctcct tgtataaatc ctggttgctg tctctttatg aggagttgtg gcccgttgtc
     1501 aggcaacgtg gcgtggtgtg cactgtgttt gctgacgcaa cccccactgg ttggggcatt
     1561 gccaccacct gtcagctcct ttccgggact ttcgctttcc ccctccctat tgccacggcg
     1621 gaactcatcg ccgcctgcct tgcccgctgc tggacagggg ctcggctgtt gggcactgac
     1681 aattccgtgg tgttgtcggg gaaatcatcg tcctttcctt ggctgctcgc ctgtgttgcc
     1741 acctggattc tgcgcgggac gtccttctgc tacgtccctt cggccctcaa tccagcggac
     1801 cttccttccc gcggcctgct gccggctctg cggcctcttc cgcgtcttcg ccttcgccct
     1861 cagacgagtc ggatctccct ttgggccgcc tccccgcatc ggggggatcc tgtgccttct
     1921 agttgccagc catctgttgt ttgcccctcc cccgtgcctt ccttgaccct ggaaggtgcc
     1981 actcccactg tcctttccta ataaaatgag gaaattgcat cgcattgtct gagtaggtgt
     2041 cattctattc tggggggtgg ggtggggcag gacagcaagg gggaggattg ggaagacaat
     2101 agcaggcatg ctggggatgc ggtgggctct atggcggccg caggaacccc tagtgatgga
     2161 gttggccact ccctctctgc gcgctcgctc gctcactgag gccgggcgac caaaggtcgc
     2221 ccgacgcccg ggctttgccc gggcggcctc agtgagcgag cgagcgcgca gctgcctgca
     2281 ggggcgcctg atgcggtatt ttctccttac gcatctgtgc ggtatttcac accgcatacg
     2341 tcaaagcaac catagtacgc gccctgtagc ggcgcattaa gcgcggcggg tgtggtggtt
     2401 acgcgcagcg tgaccgctac acttgccagc gccctagcgc ccgctccttt cgctttcttc
     2461 ccttcctttc tcgccacgtt cgccggcttt ccccgtcaag ctctaaatcg ggggctccct
     2521 ttagggttcc gatttagtgc tttacggcac ctcgacccca aaaaacttga tttgggtgat
     2581 ggttcacgta gtgggccatc gccctgatag acggtttttc gccctttgac gttggagtcc
     2641 acgttcttta atagtggact cttgttccaa actggaacaa cactcaaccc tatctcgggc
     2701 tattcttttg atttataagg gattttgccg atttcggcct attggttaaa aaatgagctg
     2761 atttaacaaa aatttaacgc gaattttaac aaaatattaa cgtttacaat tttatggtgc
     2821 actctcagta caatctgctc tgatgccgca tagttaagcc agccccgaca cccgccaaca
     2881 cccgctgacg cgccctgacg ggcttgtctg ctcccggcat ccgcttacag acaagctgtg
     2941 accgtctccg ggagctgcat gtgtcagagg ttttcaccgt catcaccgaa acgcgcgaga
     3001 cgaaagggcc tcgtgatacg cctattttta taggttaatg tcatgataat aatggtttct
     3061 tagacgtcag gtggcacttt tcggggaaat gtgcgcggaa cccctatttg tttatttttc
     3121 taaatacatt caaatatgta tccgctcatg agacaataac cctgataaat gcttcaataa
     3181 tattgaaaaa ggaagagtat gagtattcaa catttccgtg tcgcccttat tccctttttt
     3241 gcggcatttt gccttcctgt ttttgctcac ccagaaacgc tggtgaaagt aaaagatgct
     3301 gaagatcagt tgggtgcacg agtgggttac atcgaactgg atctcaacag cggtaagatc
     3361 cttgagagtt ttcgccccga agaacgtttt ccaatgatga gcacttttaa agttctgcta
     3421 tgtggcgcgg tattatcccg tattgacgcc gggcaagagc aactcggtcg ccgcatacac
     3481 tattctcaga atgacttggt tgagtactca ccagtcacag aaaagcatct tacggatggc
     3541 atgacagtaa gagaattatg cagtgctgcc ataaccatga gtgataacac tgcggccaac
     3601 ttacttctga caacgatcgg aggaccgaag gagctaaccg cttttttgca caacatgggg
     3661 gatcatgtaa ctcgccttga tcgttgggaa ccggagctga atgaagccat accaaacgac
     3721 gagcgtgaca ccacgatgcc tgtagcaatg gcaacaacgt tgcgcaaact attaactggc
     3781 gaactactta ctctagcttc ccggcaacaa ttaatagact ggatggaggc ggataaagtt
     3841 gcaggaccac ttctgcgctc ggcccttccg gctggctggt ttattgctga taaatctgga
     3901 gccggtgagc gtgggtctcg cggtatcatt gcagcactgg ggccagatgg taagccctcc
     3961 cgtatcgtag ttatctacac gacggggagt caggcaacta tggatgaacg aaatagacag
     4021 atcgctgaga taggtgcctc actgattaag cattggtaac tgtcagacca agtttactca
     4081 tatatacttt agattgattt aaaacttcat ttttaattta aaaggatcta ggtgaagatc
     4141 ctttttgata atctcatgac caaaatccct taacgtgagt tttcgttcca ctgagcgtca
     4201 gaccccgtag aaaagatcaa aggatcttct tgagatcctt tttttctgcg cgtaatctgc
     4261 tgcttgcaaa caaaaaaacc accgctacca gcggtggttt gtttgccgga tcaagagcta
     4321 ccaactcttt ttccgaaggt aactggcttc agcagagcgc agataccaaa tactgtcctt
     4381 ctagtgtagc cgtagttagg ccaccacttc aagaactctg tagcaccgcc tacatacctc
     4441 gctctgctaa tcctgttacc agtggctgct gccagtggcg ataagtcgtg tcttaccggg
     4501 ttggactcaa gacgatagtt accggataag gcgcagcggt cgggctgaac ggggggttcg
     4561 tgcacacagc ccagcttgga gcgaacgacc tacaccgaac tgagatacct acagcgtgag
     4621 ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc
     4681 agggtcggaa caggagagcg cacgagggag cttccagggg gaaacgcctg gtatctttat
     4741 agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg
     4801 gggcggagcc tatggaaaaa cgccagcaac gcggcctttt tacggttcct ggccttttgc
     4861 tggccttttg ctcacatgt
//
