LOCUS       sequence-433352-        4981 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AAV vector plasmid expressing enhanced green fluorescent protein 
            (eGFP) under the human synapsin (SYN) promoter. It also contains an 
            optimised and truncated version of WPRE as a regulatory element..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4981)
  TITLE     AAV fluorescent transgenes
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 4981)
  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: 
            "Unpublished"
COMMENT     Sequence Label: AAV-SYN-eGFP-oPRE
FEATURES             Location/Qualifiers
     source          1..4981
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   1..130
                     /label=AAV2 ITR
     protein_bind    137..157
                     /gene="mutant version of attB"
                     /label=attB4
                     /bound_moiety="BP Clonase(TM)"
                     /note="core recombination site for the Gateway(R) BP 
                     reaction"
     promoter        179..626
                     /label=hSyn promoter
                     /note="human synapsin I promoter; confers neuron-specific 
                     expression (Kugler et al., 2003)"
     protein_bind    626..650
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     regulatory      651..660
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(657..659,660..662,663..1376)
                     /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(657..659,660..662,663..1373)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(702..723)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(963..982)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1310..1331
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     protein_bind    complement(1377..1401)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     primer_bind     complement(2012..2029)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2018..2225
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   2233..2373
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   2233..2362
                     /label=AAV2 ITR
     rep_origin      2448..2903
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2535..2554)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2745..2766
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(2920..2939)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3039..3061
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(3099..3117)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3185..3289
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3290..3358,3359..4150)
                     /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(3508..3527)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4321..4909
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4810..4829
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     repeat_region   join(4971..4981,1..130)
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
ORIGIN
        1 ctgcgcgctc gctcgctcac tgaggccgcc cgggcaaagc ccgggcgtcg ggcgaccttt
       61 ggtcgcccgg cctcagtgag cgagcgagcg cgcagagagg gagtggccaa ctccatcact
      121 aggggttcct tctagacaac tttgtataga aaagttgctg cagagggccc tgcgtatgag
      181 tgcaagtggg ttttaggacc aggatgaggc ggggtggggg tgcctacctg acgaccgacc
      241 ccgacccact ggacaagcac ccaaccccca ttccccaaat tgcgcatccc ctatcagaga
      301 gggggagggg aaacaggatg cggcgaggcg cgtgcgcact gccagcttca gcaccgcgga
      361 cagtgccttc gcccccgcct ggcggcgcgc gccaccgccg cctcagcact gaaggcgcgc
      421 tgacgtcact cgccggtccc ccgcaaactc cccttcccgg ccaccttggt cgcgtccgcg
      481 ccgccgccgg cccagccgga ccgcaccacg cgaggcgcga gatagggggg cacgggcgcg
      541 accatctgcg ctgcggcgcc ggcgactcag cgctgcctca gtctgcggtg ggcagcggag
      601 gagtcgtgtc gtgcctgaga gcgcagcaag tttgtacaaa aaagcaggct gccaccatgg
      661 tgagcaaggg cgaggagctg ttcaccgggg tggtgcccat cctggtcgag ctggacggcg
      721 acgtaaacgg ccacaagttc agcgtgtccg gcgagggcga gggcgatgcc acctacggca
      781 agctgaccct gaagttcatc tgcaccaccg gcaagctgcc cgtgccctgg cccaccctcg
      841 tgaccaccct gacctacggc gtgcagtgct tcagccgcta ccccgaccac atgaagcagc
      901 acgacttctt caagtccgcc atgcccgaag gctacgtcca ggagcgcacc atcttcttca
      961 aggacgacgg caactacaag acccgcgccg aggtgaagtt cgagggcgac accctggtga
     1021 accgcatcga gctgaagggc atcgacttca aggaggacgg caacatcctg gggcacaagc
     1081 tggagtacaa ctacaacagc cacaacgtct atatcatggc cgacaagcag aagaacggca
     1141 tcaaggtgaa cttcaagatc cgccacaaca tcgaggacgg cagcgtgcag ctcgccgacc
     1201 actaccagca gaacaccccc atcggcgacg gccccgtgct gctgcccgac aaccactacc
     1261 tgagcaccca gtccgccctg agcaaagacc ccaacgagaa gcgcgatcac atggtcctgc
     1321 tggagttcgt gaccgccgcc gggatcactc tcggcatgga cgagctgtac aagtaaaccc
     1381 agctttcttg tacaaagtgg gaattcgagc atcttaccgc catttatacc catatttgtt
     1441 ctgtttttct tgatttgggt atacatttaa atgttaataa aacaaaatgg tggggcaatc
     1501 atttacattt ttagggatat gtaattacta gttcaggtgt attgccacaa gacaaacatg
     1561 ttaagaaact ttcccgttat ttacgctctg ttcctgttaa tcaacctctg gattacaaaa
     1621 tttgtgaaag attgactgat attcttaact atgttgctcc ttttacgctg tgtggatatg
     1681 ctgctttata gcctctgtat ctagctattg cttcccgtac ggctttcgtt ttctcctcct
     1741 tgtataaatc ctggttgctg tctcttttag aggagttgtg gcccgttgtc cgtcaacgtg
     1801 gcgtggtgtg ctctgtgttt gctgacgcaa cccccactgg ctggggcatt gccaccacct
     1861 gtcaactcct ttctgggact ttcgctttcc ccctcccgat cgccacggca gaactcatcg
     1921 ccgcctgcct tgcccgctgc tggacagggg ctaggttgct gggcactgat aattccgtgg
     1981 tgttgtcgaa ttcctagagc tcgctgatca gcctcgactg tgccttctag ttgccagcca
     2041 tctgttgttt gcccctcccc cgtgccttcc ttgaccctgg aaggtgccac tcccactgtc
     2101 ctttcctaat aaaatgagga aattgcatcg cattgtctga gtaggtgtca ttctattctg
     2161 gggggtgggg tggggcagga cagcaagggg gaggattggg aagagaatag caggcatgct
     2221 ggggagggcc gcaggaaccc ctagtgatgg agttggccac tccctctctg cgcgctcgct
     2281 cgctcactga ggccgggcga ccaaaggtcg cccgacgccc gggctttgcc cgggcggcct
     2341 cagtgagcga gcgagcgcgc agctgcctgc aggggcgcct gatgcggtat tttctcctta
     2401 cgcatctgtg cggtatttca caccgcatac gtcaaagcaa ccatagtacg cgccctgtag
     2461 cggcgcatta agcgcggcgg gtgtggtggt tacgcgcagc gtgaccgcta cacttgccag
     2521 cgccttagcg cccgctcctt tcgctttctt cccttccttt ctcgccacgt tcgccggctt
     2581 tccccgtcaa gctctaaatc gggggctccc tttagggttc cgatttagtg ctttacggca
     2641 cctcgacccc aaaaaacttg atttgggtga tggttcacgt agtgggccat cgccctgata
     2701 gacggttttt cgccctttga cgttggagtc cacgttcttt aatagtggac tcttgttcca
     2761 aactggaaca acactcaact ctatctcggg ctattctttt gatttataag ggattttgcc
     2821 gatttcggtc tattggttaa aaaatgagct gatttaacaa aaatttaacg cgaattttaa
     2881 caaaatatta acgtttacaa ttttatggtg cactctcagt acaatctgct ctgatgccgc
     2941 atagttaagc cagccccgac acccgccaac acccgctgac gcgccctgac gggcttgtct
     3001 gctcccggca tccgcttaca gacaagctgt gaccgtctcc gggagctgca tgtgtcagag
     3061 gttttcaccg tcatcaccga aacgcgcgag acgaaagggc ctcgtgatac gcctattttt
     3121 ataggttaat gtcatgataa taatggtttc ttagacgtca ggtggcactt ttcggggaaa
     3181 tgtgcgcgga acccctattt gtttattttt ctaaatacat tcaaatatgt atccgctcat
     3241 gagacaataa ccctgataaa tgcttcaata atattgaaaa aggaagagta tgagtattca
     3301 acatttccgt gtcgccctta ttcccttttt tgcggcattt tgccttcctg tttttgctca
     3361 cccagaaacg ctggtgaaag taaaagatgc tgaagatcag ttgggtgcac gagtgggtta
     3421 catcgaactg gatctcaaca gcggtaagat ccttgagagt tttcgccccg aagaacgttt
     3481 tccaatgatg agcactttta aagttctgct atgtggcgcg gtattatccc gtattgacgc
     3541 cgggcaagag caactcggtc gccgcataca ctattctcag aatgacttgg ttgagtactc
     3601 accagtcaca gaaaagcatc ttacggatgg catgacagta agagaattat gcagtgctgc
     3661 cataaccatg agtgataaca ctgcggccaa cttacttctg acaacgatcg gaggaccgaa
     3721 ggagctaacc gcttttttgc acaacatggg ggatcatgta actcgccttg atcgttggga
     3781 accggagctg aatgaagcca taccaaacga cgagcgtgac accacgatgc ctgtagcaat
     3841 ggcaacaacg ttgcgcaaac tattaactgg cgaactactt actctagctt cccggcaaca
     3901 attaatagac tggatggagg cggataaagt tgcaggacca cttctgcgct cggcccttcc
     3961 ggctggctgg tttattgctg ataaatctgg agccggtgag cgtggaagcc gcggtatcat
     4021 tgcagcactg gggccagatg gtaagccctc ccgtatcgta gttatctaca cgacggggag
     4081 tcaggcaact atggatgaac gaaatagaca gatcgctgag ataggtgcct cactgattaa
     4141 gcattggtaa ctgtcagacc aagtttactc atatatactt tagattgatt taaaacttca
     4201 tttttaattt aaaaggatct aggtgaagat cctttttgat aatctcatga ccaaaatccc
     4261 ttaacgtgag ttttcgttcc actgagcgtc agaccccgta gaaaagatca aaggatcttc
     4321 ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc
     4381 agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt
     4441 cagcagagcg cagataccaa atactgttct tctagtgtag ccgtagttag gccaccactt
     4501 caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc
     4561 tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa
     4621 ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac
     4681 ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg
     4741 gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga
     4801 gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact
     4861 tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa
     4921 cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgt cctgcaggca
     4981 g
//
