LOCUS       sequence-356924-        4255 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AAV vector backbone designed to express GFP from a 0.5Synapsin 
            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 4255)
  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 4255)
  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: pJEP220-AAV-0.5Synapsin-GFP-pA
FEATURES             Location/Qualifiers
     source          1..4255
                     /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      697..706
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(703..705,706..708,709..1422)
                     /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(703..705,706..708,709..1419)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(748..769)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1009..1028)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1356..1377
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(1460..1479)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     repeat_region   1518..1658
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   1518..1647
                     /label=AAV2 ITR
     rep_origin      1733..2188
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1820..1839)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2030..2051
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(2205..2224)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     2324..2346
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(2384..2402)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        2470..2574
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2575..2643,2644..3435)
                     /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(2793..2812)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3606..4194
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4095..4114
                     /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 gcgtacaagt atctgggaag ggtaacaatg caagttagcc
      181 tgaggattta ggaggaggct gaaaaacaga gtaggagcct tactacgggt ccagacccta
      241 cggacaagaa cccccactcc cactccccaa attgcgcatt ccctccccca tcagaggggg
      301 aggggaagag gatgcagcgc ggcgcggcgc gtgcgcactg tcggatttag taccgcggac
      361 agagccttcg cccccgctgc cggcgcgcgc caccacctcc ccagcaccaa aggcgggctg
      421 acgtcactct ccagccctcc ccaaactccc ctacctcacc gccttggtcg cgtccgtgca
      481 gcggtgagtc cagtcgggcc gcaccacaag aggtgcaaga taggggggtg caggcgcgac
      541 catacgctct gcggcggcag agcctcagcg ctgcctcagt ctgcagcggg cagcagagga
      601 gtcgcgtcgt gccagagagc gccgccgtgc tcctgagccc cttgcgctcc gcccccgcgg
      661 cccaccgacc cactgcccct tggatccgaa ttcgccgcca ccatggtgag caagggcgag
      721 gagctgttca ccggggtggt gcccatcctg gtcgagctgg acggcgacgt aaacggccac
      781 aagttcagcg tgtccggcga gggcgagggc gatgccacct acggcaagct gaccctgaag
      841 ttcatctgca ccaccggcaa gctgcccgtg ccctggccca ctctcgtgac caccctgacc
      901 tacggcgtgc agtgcttcag ccgctacccc gaccacatga agcagcacga cttcttcaag
      961 tccgccatgc ccgaaggcta cgtccaggag cgcaccatct tcttcaagga cgacggcaac
     1021 tacaagaccc gcgccgaggt gaagttcgag ggcgacaccc tggtgaaccg catcgagctg
     1081 aagggcatcg acttcaagga ggacggcaac atcctggggc acaagctgga gtacaactac
     1141 aacagccaca acgtctatat catggccgac aagcagaaga acggcatcaa ggtgaacttc
     1201 aagatccgcc acaacatcga ggacggcagc gtgcagctcg ccgaccacta ccagcagaac
     1261 acccccatcg gcgacggccc cgtgctgctg cccgacaacc actacctgag cacccagtcc
     1321 gccctgagca aagaccccaa cgagaagcgc gatcacatgg tcctgctgga gttcgtgacc
     1381 gccgccggga tcactctcgg catggacgag ctgtacaagt aactcgagtt tattgcagct
     1441 tataatggtt acaaataaag caatagcatc acaaatttca caaataaagc atttttttca
     1501 ctggaccgag cggccgcagg aacccctagt gatggagttg gccactccct ctctgcgcgc
     1561 tcgctcgctc actgaggccg ggcgaccaaa ggtcgcccga cgcccgggct ttgcccgggc
     1621 ggcctcagtg agcgagcgag cgcgcagctg cctgcagggg cgcctgatgc ggtattttct
     1681 ccttacgcat ctgtgcggta tttcacaccg catacgtcaa agcaaccata gtacgcgccc
     1741 tgtagcggcg cattaagcgc ggcgggtgtg gtggttacgc gcagcgtgac cgctacactt
     1801 gccagcgcct tagcgcccgc tcctttcgct ttcttccctt cctttctcgc cacgttcgcc
     1861 ggctttcccc gtcaagctct aaatcggggg ctccctttag ggttccgatt tagtgcttta
     1921 cggcacctcg accccaaaaa acttgatttg ggtgatggtt cacgtagtgg gccatcgccc
     1981 tgatagacgg tttttcgccc tttgacgttg gagtccacgt tctttaatag tggactcttg
     2041 ttccaaactg gaacaacact caactctatc tcgggctatt cttttgattt ataagggatt
     2101 ttgccgattt cggtctattg gttaaaaaat gagctgattt aacaaaaatt taacgcgaat
     2161 tttaacaaaa tattaacgtt tacaatttta tggtgcactc tcagtacaat ctgctctgat
     2221 gccgcatagt taagccagcc ccgacacccg ccaacacccg ctgacgcgcc ctgacgggct
     2281 tgtctgctcc cggcatccgc ttacagacaa gctgtgaccg tctccgggag ctgcatgtgt
     2341 cagaggtttt caccgtcatc accgaaacgc gcgagacgaa agggcctcgt gatacgccta
     2401 tttttatagg ttaatgtcat gataataatg gtttcttaga cgtcaggtgg cacttttcgg
     2461 ggaaatgtgc gcggaacccc tatttgttta tttttctaaa tacattcaaa tatgtatccg
     2521 ctcatgagac aataaccctg ataaatgctt caataatatt gaaaaaggaa gagtatgagt
     2581 attcaacatt tccgtgtcgc ccttattccc ttttttgcgg cattttgcct tcctgttttt
     2641 gctcacccag aaacgctggt gaaagtaaaa gatgctgaag atcagttggg tgcacgagtg
     2701 ggttacatcg aactggatct caacagcggt aagatccttg agagttttcg ccccgaagaa
     2761 cgttttccaa tgatgagcac ttttaaagtt ctgctatgtg gcgcggtatt atcccgtatt
     2821 gacgccgggc aagagcaact cggtcgccgc atacactatt ctcagaatga cttggttgag
     2881 tactcaccag tcacagaaaa gcatcttacg gatggcatga cagtaagaga attatgcagt
     2941 gctgccataa ccatgagtga taacactgcg gccaacttac ttctgacaac gatcggagga
     3001 ccgaaggagc taaccgcttt tttgcacaac atgggggatc atgtaactcg ccttgatcgt
     3061 tgggaaccgg agctgaatga agccatacca aacgacgagc gtgacaccac gatgcctgta
     3121 gcaatggcaa caacgttgcg caaactatta actggcgaac tacttactct agcttcccgg
     3181 caacaattaa tagactggat ggaggcggat aaagttgcag gaccacttct gcgctcggcc
     3241 cttccggctg gctggtttat tgctgataaa tctggagccg gtgagcgtgg gtctcgcggt
     3301 atcattgcag cactggggcc agatggtaag ccctcccgta tcgtagttat ctacacgacg
     3361 gggagtcagg caactatgga tgaacgaaat agacagatcg ctgagatagg tgcctcactg
     3421 attaagcatt ggtaactgtc agaccaagtt tactcatata tactttagat tgatttaaaa
     3481 cttcattttt aatttaaaag gatctaggtg aagatccttt ttgataatct catgaccaaa
     3541 atcccttaac gtgagttttc gttccactga gcgtcagacc ccgtagaaaa gatcaaagga
     3601 tcttcttgag atcctttttt tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg
     3661 ctaccagcgg tggtttgttt gccggatcaa gagctaccaa ctctttttcc gaaggtaact
     3721 ggcttcagca gagcgcagat accaaatact gttcttctag tgtagccgta gttaggccac
     3781 cacttcaaga actctgtagc accgcctaca tacctcgctc tgctaatcct gttaccagtg
     3841 gctgctgcca gtggcgataa gtcgtgtctt accgggttgg actcaagacg atagttaccg
     3901 gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca cacagcccag cttggagcga
     3961 acgacctaca ccgaactgag atacctacag cgtgagctat gagaaagcgc cacgcttccc
     4021 gaagggagaa aggcggacag gtatccggta agcggcaggg tcggaacagg agagcgcacg
     4081 agggagcttc cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc
     4141 tgacttgagc gtcgattttt gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc
     4201 agcaacgcgg cctttttacg gttcctggcc ttttgctggc cttttgctca catgt
//
