LOCUS       sequence-356898-        5006 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AAV vector backbone designed to express GFP from a 1.3CaMKIIa 
            promoter..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5006)
  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 5006)
  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: pJEP218-AAV-1.3-alpha-CaMKII-GFP-pA
FEATURES             Location/Qualifiers
     source          1..5006
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   1..130
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     promoter        151..1438
                     /label=CaMKII promoter
                     /note="promoter for the mouse calcium/calmodulin-dependent 
                     kinase II alpha subunit"
     regulatory      1448..1457
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(1454..1456,1457..1459,1460..2173)
                     /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(1454..1456,1457..1459,1460..2170)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(1499..1520)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1760..1779)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2107..2128
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(2211..2230)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     repeat_region   2269..2409
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   2269..2398
                     /label=AAV2 ITR
     rep_origin      2484..2939
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2571..2590)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2781..2802
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(2956..2975)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3075..3097
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(3135..3153)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3221..3325
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3326..3394,3395..4186)
                     /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(3544..3563)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4357..4945
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4846..4865
                     /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 gcggccgcac gcgttcgtaa cattatggcc ttaggtcact tcatctccat
      181 ggggttcttc ttctgatttt ctagaaaatg agatgggggt gcagagagct tcctcagtga
      241 cctgcccagg gtcacatcag aaatgtcaga gctagaactt gaactcagat tactaatctt
      301 aaattccatg ccttgggggc atgcaagtac gatatacaga aggagtgaac tcattagggc
      361 agatgaccaa tgagtttagg aaagaagagt ccagggcagg gtacatctac accacccgcc
      421 cagccctggg tgagtccagc cacgttcacc tcattatagt tgcctctctc cagtcctacc
      481 ttgacgggaa gcacaagcag aaactgggac aggagcccca ggagaccaaa tcttcatggt
      541 ccctctggga ggatgggtgg ggagagctgt ggcagaggcc tcaggagggg ccctgctgct
      601 cagtggtgac agataggggt gagaaagcag acagagtcat tccgtcagca ttctgggtct
      661 gtttggtact tcttctcacg ctaaggtggc ggtgtgatat gcacaatggc taaaaagcag
      721 ggagagctgg aaagaaacaa ggacagagac agaggccaag tcaaccagac caattcccag
      781 aggaagcaaa gaaaccatta cagagactac aagggggaag ggaaggagag atgaattagc
      841 ttcccctgta aaccttagaa cccagctgtt gccagggcaa cggggcaata cctgtctctt
      901 cagaggagat gaagttgcca gggtaactac atcctgtctt tctcaaggac catcccagaa
      961 tgtggcaccc actagccgtt accatagcaa ctgcctcttt gccccactta atcccatccc
     1021 gtctgttaaa agggccctat agttggaggt gggggaggta ggaagagcga tgatcacttg
     1081 tggactaagt ttgttcgcat ccccttctcc aaccccctca gtacatcacc ctgggggaac
     1141 agggtccact tgctcctggg cccacacagt cctgcagtat tgtgtatata aggccagggc
     1201 aaagaggagc aggttttaaa gtgaaaggca ggcaggtgtt ggggaggcag ttaccggggc
     1261 aacgggaaca gggcgtttcg gaggtggttg ccatggggac ctggatgctg acgaaggctc
     1321 gcgaggctgt gagcagccac agtgccctgc tcagaagccc caagctcgtc agtcaagccg
     1381 gttctccgtt tgcactcagg agcacgggca ggcgagtggc ccctagttct gggggcagga
     1441 attcgccgcc accatggtga gcaagggcga ggagctgttc accggggtgg tgcccatcct
     1501 ggtcgagctg gacggcgacg taaacggcca caagttcagc gtgtccggcg agggcgaggg
     1561 cgatgccacc tacggcaagc tgaccctgaa gttcatctgc accaccggca agctgcccgt
     1621 gccctggccc actctcgtga ccaccctgac ctacggcgtg cagtgcttca gccgctaccc
     1681 cgaccacatg aagcagcacg acttcttcaa gtccgccatg cccgaaggct acgtccagga
     1741 gcgcaccatc ttcttcaagg acgacggcaa ctacaagacc cgcgccgagg tgaagttcga
     1801 gggcgacacc ctggtgaacc gcatcgagct gaagggcatc gacttcaagg aggacggcaa
     1861 catcctgggg cacaagctgg agtacaacta caacagccac aacgtctata tcatggccga
     1921 caagcagaag aacggcatca aggtgaactt caagatccgc cacaacatcg aggacggcag
     1981 cgtgcagctc gccgaccact accagcagaa cacccccatc ggcgacggcc ccgtgctgct
     2041 gcccgacaac cactacctga gcacccagtc cgccctgagc aaagacccca acgagaagcg
     2101 cgatcacatg gtcctgctgg agttcgtgac cgccgccggg atcactctcg gcatggacga
     2161 gctgtacaag taactcgagt ttattgcagc ttataatggt tacaaataaa gcaatagcat
     2221 cacaaatttc acaaataaag catttttttc actggaccga gcggccgcag gaacccctag
     2281 tgatggagtt ggccactccc tctctgcgcg ctcgctcgct cactgaggcc gggcgaccaa
     2341 aggtcgcccg acgcccgggc tttgcccggg cggcctcagt gagcgagcga gcgcgcagct
     2401 gcctgcaggg gcgcctgatg cggtattttc tccttacgca tctgtgcggt atttcacacc
     2461 gcatacgtca aagcaaccat agtacgcgcc ctgtagcggc gcattaagcg cggcgggtgt
     2521 ggtggttacg cgcagcgtga ccgctacact tgccagcgcc ttagcgcccg ctcctttcgc
     2581 tttcttccct tcctttctcg ccacgttcgc cggctttccc cgtcaagctc taaatcgggg
     2641 gctcccttta gggttccgat ttagtgcttt acggcacctc gaccccaaaa aacttgattt
     2701 gggtgatggt tcacgtagtg ggccatcgcc ctgatagacg gtttttcgcc ctttgacgtt
     2761 ggagtccacg ttctttaata gtggactctt gttccaaact ggaacaacac tcaactctat
     2821 ctcgggctat tcttttgatt tataagggat tttgccgatt tcggtctatt ggttaaaaaa
     2881 tgagctgatt taacaaaaat ttaacgcgaa ttttaacaaa atattaacgt ttacaatttt
     2941 atggtgcact ctcagtacaa tctgctctga tgccgcatag ttaagccagc cccgacaccc
     3001 gccaacaccc gctgacgcgc cctgacgggc ttgtctgctc ccggcatccg cttacagaca
     3061 agctgtgacc gtctccggga gctgcatgtg tcagaggttt tcaccgtcat caccgaaacg
     3121 cgcgagacga aagggcctcg tgatacgcct atttttatag gttaatgtca tgataataat
     3181 ggtttcttag acgtcaggtg gcacttttcg gggaaatgtg cgcggaaccc ctatttgttt
     3241 atttttctaa atacattcaa atatgtatcc gctcatgaga caataaccct gataaatgct
     3301 tcaataatat tgaaaaagga agagtatgag tattcaacat ttccgtgtcg cccttattcc
     3361 cttttttgcg gcattttgcc ttcctgtttt tgctcaccca gaaacgctgg tgaaagtaaa
     3421 agatgctgaa gatcagttgg gtgcacgagt gggttacatc gaactggatc tcaacagcgg
     3481 taagatcctt gagagttttc gccccgaaga acgttttcca atgatgagca cttttaaagt
     3541 tctgctatgt ggcgcggtat tatcccgtat tgacgccggg caagagcaac tcggtcgccg
     3601 catacactat tctcagaatg acttggttga gtactcacca gtcacagaaa agcatcttac
     3661 ggatggcatg acagtaagag aattatgcag tgctgccata accatgagtg ataacactgc
     3721 ggccaactta cttctgacaa cgatcggagg accgaaggag ctaaccgctt ttttgcacaa
     3781 catgggggat catgtaactc gccttgatcg ttgggaaccg gagctgaatg aagccatacc
     3841 aaacgacgag cgtgacacca cgatgcctgt agcaatggca acaacgttgc gcaaactatt
     3901 aactggcgaa ctacttactc tagcttcccg gcaacaatta atagactgga tggaggcgga
     3961 taaagttgca ggaccacttc tgcgctcggc ccttccggct ggctggttta ttgctgataa
     4021 atctggagcc ggtgagcgtg ggtctcgcgg tatcattgca gcactggggc cagatggtaa
     4081 gccctcccgt atcgtagtta tctacacgac ggggagtcag gcaactatgg atgaacgaaa
     4141 tagacagatc gctgagatag gtgcctcact gattaagcat tggtaactgt cagaccaagt
     4201 ttactcatat atactttaga ttgatttaaa acttcatttt taatttaaaa ggatctaggt
     4261 gaagatcctt tttgataatc tcatgaccaa aatcccttaa cgtgagtttt cgttccactg
     4321 agcgtcagac cccgtagaaa agatcaaagg atcttcttga gatccttttt ttctgcgcgt
     4381 aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt tgccggatca
     4441 agagctacca actctttttc cgaaggtaac tggcttcagc agagcgcaga taccaaatac
     4501 tgttcttcta gtgtagccgt agttaggcca ccacttcaag aactctgtag caccgcctac
     4561 atacctcgct ctgctaatcc tgttaccagt ggctgctgcc agtggcgata agtcgtgtct
     4621 taccgggttg gactcaagac gatagttacc ggataaggcg cagcggtcgg gctgaacggg
     4681 gggttcgtgc acacagccca gcttggagcg aacgacctac accgaactga gatacctaca
     4741 gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca ggtatccggt
     4801 aagcggcagg gtcggaacag gagagcgcac gagggagctt ccagggggaa acgcctggta
     4861 tctttatagt cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc
     4921 gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc
     4981 cttttgctgg ccttttgctc acatgt
//
