LOCUS       sequence-356942-        4340 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AAV basic vector backbone designed to express GFP from a 0.4CaMKIIa 
            promoter. It also contains a poly-Adenylation Signal (pA)..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4340)
  AUTHORS   Holehonnur R, Lella SK, Ho A, Luong JA, Ploski JE
  TITLE     The production of viral vectors designed to express large and 
            difficult to express transgenes within neurons.
  JOURNAL   Mol Brain. 2015 Feb 24;8:12. doi: 10.1186/s13041-015-0100-7.
  PUBMED    25887710
REFERENCE   2  (bases 1 to 4340)
  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: "2015-02-24"; volume: "8:12. doi"; pages: " 
            10.1186/s13041-015-0100-7"
COMMENT     Sequence Label: pJEP224-AAV-Basic-0.4-alpha-CaMKII-GFP-pA
FEATURES             Location/Qualifiers
     source          1..4340
                     /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      587..596
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(593..595,596..598,599..1312)
                     /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(593..595,596..598,599..1309)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(638..659)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(899..918)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1246..1267
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    1407..1528
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1444..1463)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     1498..1517
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     repeat_region   1603..1743
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   1603..1732
                     /label=AAV2 ITR
     rep_origin      1818..2273
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1905..1924)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2115..2136
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(2290..2309)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     2409..2431
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(2469..2487)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        2555..2659
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2660..2728,2729..3520)
                     /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(2878..2897)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3691..4279
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4180..4199
                     /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 gcggcaattc agtcgataac tataacggtc ctaaggtagc gatttaaata
      181 cgcgctctct taaggtagcc ccgggacgcg tacttgtgga ctaagtttgt tcacatcccc
      241 ttctccaacc ccctcagtac atcaccctgg gagaacaagg tccacttgct tctgggccca
      301 cacagtcctg cagtattgtg tatataaggc cagggcaacg gaggagcagg ttttgaagtg
      361 aaaggcaggc aggtgttggg gaggcagtta ccggggcaac gggaacaggg cgtttcggag
      421 gtggttgcca tggggacctg gatgctgacg aaggctcgcg aggctgtgag cagccacagt
      481 gccctgctca gaagccccaa gctcgtcaat caagctggtt ctccatttgc actcaggagc
      541 acgggcaggc gagtggcccc tagttctggg agcagagatc tgcgccgcca ccatggtgag
      601 caagggcgag gagctgttca ccggggtggt gcccatcctg gtcgagctgg acggcgacgt
      661 aaacggccac aagttcagcg tgtccggcga gggcgagggc gatgccacct acggcaagct
      721 gaccctgaag ttcatctgca ccaccggcaa gctgcccgtg ccctggccca ccctcgtgac
      781 caccctgacc tacggcgtgc agtgcttcag ccgctacccc gaccacatga agcagcacga
      841 cttcttcaag tccgccatgc ccgaaggcta cgtccaggag cgcaccatct tcttcaagga
      901 cgacggcaac tacaagaccc gcgccgaggt gaagttcgag ggcgacaccc tggtgaaccg
      961 catcgagctg aagggcatcg acttcaagga ggacggcaac atcctggggc acaagctgga
     1021 gtacaactac aacagccaca acgtctatat catggccgac aagcagaaga acggcatcaa
     1081 ggtgaacttc aagatccgcc acaacatcga ggacggcagc gtgcagctcg ccgaccacta
     1141 ccagcagaac acccccatcg gcgacggccc cgtgctgctg cccgacaacc actacctgag
     1201 cacccagtcc gccctgagca aagaccccaa cgagaagcgc gatcacatgg tcctgctgga
     1261 gttcgtgacc gccgccggga tcactctcgg catggacgag ctgtacaagt aaaagcttca
     1321 catttgtaga ggttttactt gctttaaaaa acctcccaca cctccccctg aacctgaaac
     1381 ataaaatgaa tgcaattgtt gttgttaact tgtttattgc agcttataat ggttacaaat
     1441 aaagcaatag catcacaaat ttcacaaata aagcattttt ttcactgcat tctagttgtg
     1501 gtttgtccaa actcatcaat gtatcttaag gcgtaaagat atcctctctt aaggtagcat
     1561 cgagatttaa attagggata acagggtaat ggcgcgggcc gcaggaaccc ctagtgatgg
     1621 agttggccac tccctctctg cgcgctcgct cgctcactga ggccgggcga ccaaaggtcg
     1681 cccgacgccc gggctttgcc cgggcggcct cagtgagcga gcgagcgcgc agctgcctgc
     1741 aggggcgcct gatgcggtat tttctcctta cgcatctgtg cggtatttca caccgcatac
     1801 gtcaaagcaa ccatagtacg cgccctgtag cggcgcatta agcgcggcgg gtgtggtggt
     1861 tacgcgcagc gtgaccgcta cacttgccag cgccttagcg cccgctcctt tcgctttctt
     1921 cccttccttt ctcgccacgt tcgccggctt tccccgtcaa gctctaaatc gggggctccc
     1981 tttagggttc cgatttagtg ctttacggca cctcgacccc aaaaaacttg atttgggtga
     2041 tggttcacgt agtgggccat cgccctgata gacggttttt cgccctttga cgttggagtc
     2101 cacgttcttt aatagtggac tcttgttcca aactggaaca acactcaact ctatctcggg
     2161 ctattctttt gatttataag ggattttgcc gatttcggtc tattggttaa aaaatgagct
     2221 gatttaacaa aaatttaacg cgaattttaa caaaatatta acgtttacaa ttttatggtg
     2281 cactctcagt acaatctgct ctgatgccgc atagttaagc cagccccgac acccgccaac
     2341 acccgctgac gcgccctgac gggcttgtct gctcccggca tccgcttaca gacaagctgt
     2401 gaccgtctcc gggagctgca tgtgtcagag gttttcaccg tcatcaccga aacgcgcgag
     2461 acgaaagggc ctcgtgatac gcctattttt ataggttaat gtcatgataa taatggtttc
     2521 ttagacgtca ggtggcactt ttcggggaaa tgtgcgcgga acccctattt gtttattttt
     2581 ctaaatacat tcaaatatgt atccgctcat gagacaataa ccctgataaa tgcttcaata
     2641 atattgaaaa aggaagagta tgagtattca acatttccgt gtcgccctta ttcccttttt
     2701 tgcggcattt tgccttcctg tttttgctca cccagaaacg ctggtgaaag taaaagatgc
     2761 tgaagatcag ttgggtgcac gagtgggtta catcgaactg gatctcaaca gcggtaagat
     2821 ccttgagagt tttcgccccg aagaacgttt tccaatgatg agcactttta aagttctgct
     2881 atgtggcgcg gtattatccc gtattgacgc cgggcaagag caactcggtc gccgcataca
     2941 ctattctcag aatgacttgg ttgagtactc accagtcaca gaaaagcatc ttacggatgg
     3001 catgacagta agagaattat gcagtgctgc cataaccatg agtgataaca ctgcggccaa
     3061 cttacttctg acaacgatcg gaggaccgaa ggagctaacc gcttttttgc acaacatggg
     3121 ggatcatgta actcgccttg atcgttggga accggagctg aatgaagcca taccaaacga
     3181 cgagcgtgac accacgatgc ctgtagcaat ggcaacaacg ttgcgcaaac tattaactgg
     3241 cgaactactt actctagctt cccggcaaca attaatagac tggatggagg cggataaagt
     3301 tgcaggacca cttctgcgct cggcccttcc ggctggctgg tttattgctg ataaatctgg
     3361 agccggtgag cgtgggtctc gcggtatcat tgcagcactg gggccagatg gtaagccctc
     3421 ccgtatcgta gttatctaca cgacggggag tcaggcaact atggatgaac gaaatagaca
     3481 gatcgctgag ataggtgcct cactgattaa gcattggtaa ctgtcagacc aagtttactc
     3541 atatatactt tagattgatt taaaacttca tttttaattt aaaaggatct aggtgaagat
     3601 cctttttgat aatctcatga ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc
     3661 agaccccgta gaaaagatca aaggatcttc ttgagatcct ttttttctgc gcgtaatctg
     3721 ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt tgtttgccgg atcaagagct
     3781 accaactctt tttccgaagg taactggctt cagcagagcg cagataccaa atactgttct
     3841 tctagtgtag ccgtagttag gccaccactt caagaactct gtagcaccgc ctacatacct
     3901 cgctctgcta atcctgttac cagtggctgc tgccagtggc gataagtcgt gtcttaccgg
     3961 gttggactca agacgatagt taccggataa ggcgcagcgg tcgggctgaa cggggggttc
     4021 gtgcacacag cccagcttgg agcgaacgac ctacaccgaa ctgagatacc tacagcgtga
     4081 gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg
     4141 cagggtcgga acaggagagc gcacgaggga gcttccaggg ggaaacgcct ggtatcttta
     4201 tagtcctgtc gggtttcgcc acctctgact tgagcgtcga tttttgtgat gctcgtcagg
     4261 ggggcggagc ctatggaaaa acgccagcaa cgcggccttt ttacggttcc tggccttttg
     4321 ctggcctttt gctcacatgt
//
