LOCUS       sequence-390519-        4496 bp    DNA     circular SYN 14-APR-2025
DEFINITION  AAV vector plasmid expressing enhanced green fluorescent protein 
            (eGFP) under the human synapsin (hSYN) promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4496)
  AUTHORS   Nieuwenhuis B, Haenzi B, Hilton S, Carnicer-Lombarte A, Hobo B, 
            Verhaagen J, Fawcett JW
  TITLE     Optimization of adeno-associated viral vector-mediated transduction 
            of the corticospinal tract: comparison of four promoters.
  JOURNAL   Gene Ther. 2020 Jun 23. pii: 10.1038/s41434-020-0169-1. doi: 
            10.1038/s41434-020-0169-1.
  PUBMED    32576975
REFERENCE   2  (bases 1 to 4496)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 14, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Gene Ther. 
            2020 Jun 23. pii: 10.1038/s41434-020-0169-1. doi: 
            10.1038/s41434-020-0169-1."
COMMENT     Sequence Label: AAV-hSYN-eGFP
FEATURES             Location/Qualifiers
     source          1..4496
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(104..123)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      917..1505
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1406..1425
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     polyA_signal    1811..1945
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1835..1854)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     1889..1908
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(1960..1976)
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1960..1976)
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     CDS             complement(join(1990..2703,2704..2706,2707..2709))
                     /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             complement(join(1993..2703,2704..2706,2707..2709))
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(2035..2056)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     2384..2403
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2643..2664
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     regulatory      2706..2715
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     promoter        complement(2750..3197)
                     /label=hSyn promoter
                     /note="human synapsin I promoter; confers neuron-specific 
                     expression (Kugler et al., 2003)"
     repeat_region   3257..3386
                     /label=AAV2 ITR
     rep_origin      3490..3945
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3577..3596)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3787..3808
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4012..4031)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4131..4153
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(4191..4209)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4277..4381
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4382..4450,4451..4496,1..746)
                     /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"
ORIGIN
        1 gtgcacgagt gggttacatc gaactggatc tcaacagcgg taagatcctt gagagttttc
       61 gccccgaaga acgttttcca atgatgagca cttttaaagt tctgctatgt ggcgcggtat
      121 tatcccgtat tgacgccggg caagagcaac tcggtcgccg catacactat tctcagaatg
      181 acttggttga gtactcacca gtcacagaaa agcatcttac ggatggcatg acagtaagag
      241 aattatgcag tgctgccata accatgagtg ataacactgc ggccaactta cttctgacaa
      301 cgatcggagg accgaaggag ctaaccgctt ttttgcacaa catgggggat catgtaactc
      361 gccttgatcg ttgggaaccg gagctgaatg aagccatacc aaacgacgag cgtgacacca
      421 cgatgcctgt agcaatggca acaacgttgc gcaaactatt aactggcgaa ctacttactc
      481 tagcttcccg gcaacaatta atagactgga tggaggcgga taaagttgca ggaccacttc
      541 tgcgctcggc ccttccggct ggctggttta ttgctgataa atctggagcc ggtgagcgtg
      601 ggtctcgcgg tatcattgca gcactggggc cagatggtaa gccctcccgt atcgtagtta
      661 tctacacgac ggggagtcag gcaactatgg atgaacgaaa tagacagatc gctgagatag
      721 gtgcctcact gattaagcat tggtaactgt cagaccaagt ttactcatat atactttaga
      781 ttgatttaaa acttcatttt taatttaaaa ggatctaggt gaagatcctt tttgataatc
      841 tcatgaccaa aatcccttaa cgtgagtttt cgttccactg agcgtcagac cccgtagaaa
      901 agatcaaagg atcttcttga gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa
      961 aaaaaccacc gctaccagcg gtggtttgtt tgccggatca agagctacca actctttttc
     1021 cgaaggtaac tggcttcagc agagcgcaga taccaaatac tgttcttcta gtgtagccgt
     1081 agttaggcca ccacttcaag aactctgtag caccgcctac atacctcgct ctgctaatcc
     1141 tgttaccagt ggctgctgcc agtggcgata agtcgtgtct taccgggttg gactcaagac
     1201 gatagttacc ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc acacagccca
     1261 gcttggagcg aacgacctac accgaactga gatacctaca gcgtgagcta tgagaaagcg
     1321 ccacgcttcc cgaagggaga aaggcggaca ggtatccggt aagcggcagg gtcggaacag
     1381 gagagcgcac gagggagctt ccagggggaa acgcctggta tctttatagt cctgtcgggt
     1441 ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg cggagcctat
     1501 ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc cttttgctgg ccttttgctc
     1561 acatgttctt tcctgcgtta tcccctgatt ctgtggataa ccgtattacc gcctttgagt
     1621 gagctgatac cgctcgccgc agccgaacga ccgagcgcag cgagtcagtg agcgaggaag
     1681 cggaagagcg cccaatacgc aaaccgcctc tccccgcgcg ttggccgatt cattaatgca
     1741 ggctgcagag agggagtggc caactccatc actaggggtt cctcagatcg ggggatcttc
     1801 gatgctagac gatccagaca tgataagata cattgatgag tttggacaaa ccacaactag
     1861 aatgcagtga aaaaaatgct ttatttgtga aatttgtgat gctattgctt tatttgtaac
     1921 cattataagc tgcaataaac aagttgctcg aagtcgacgg atccactagt tctagagcgg
     1981 ccggccgctt tacttgtaca gctcgtccat gccgagagtg atcccggcgg cggtcacgaa
     2041 ctccagcagg accatgtgat cgcgcttctc gttggggtct ttgctcaggg cggactgggt
     2101 gctcaggtag tggttgtcgg gcagcagcac ggggccgtcg ccgatggggg tgttctgctg
     2161 gtagtggtcg gcgagctgca cgctgccgtc ctcgatgttg tggcggatct tgaagttcac
     2221 cttgatgccg ttcttctgct tgtcggccat gatatagacg ttgtggctgt tgtagttgta
     2281 ctccagcttg tgccccagga tgttgccgtc ctccttgaag tcgatgccct tcagctcgat
     2341 gcggttcacc agggtgtcgc cctcgaactt cacctcggcg cgggtcttgt agttgccgtc
     2401 gtccttgaag aagatggtgc gctcctggac gtagccttcg ggcatggcgg acttgaagaa
     2461 gtcgtgctgc ttcatgtggt cggggtagcg gctgaagcac tgcacgccgt aggtcagggt
     2521 ggtcacgagg gtgggccagg gcacgggcag cttgccggtg gtgcagatga acttcagggt
     2581 cagcttgccg taggtggcat cgccctcgcc ctcgccggac acgctgaact tgtggccgtt
     2641 tacgtcgccg tccagctcga ccaggatggg caccaccccg gtgaacagct cctcgccctt
     2701 gctcaccatg gtggcgaccg gtggatcctt gctagcagct tgaattcgac tgcgctctca
     2761 ggcacgacac gactcctccg ctgcccaccg cagactgagg cagcgctgag tcgccggcgc
     2821 cgcagcgcag atggtcgcgc ccgtgccccc ctatctcgcg cctcgcgtgg tgcggtccgg
     2881 ctgggccggc ggcggcgcgg acgcgaccaa ggtggccggg aaggggagtt tgcgggggac
     2941 cggcgagtga cgtcagcgcg ccttcagtgc tgaggcggcg gtggcgcgcg ccgccaggcg
     3001 ggggcgaagg cactgtccgc ggtgctgaag ctggcagtgc gcacgcgcct cgccgcatcc
     3061 tgtttcccct ccccctctct gataggggat gcgcaatttg gggaatgggg gttgggtgct
     3121 tgtccagtgg gtcggggtcg gtcgtcaggt aggcaccccc accccgcctc atcctggtcc
     3181 taaaacccac ttgcactcat acgcaggtcc ctctgcagct agagtctaga gcggccaatt
     3241 cctgcagccc gatctgagga acccctagtg atggagttgg ccactccctc tctgcgcgct
     3301 cgctcgctca ctgaggccgg gcgaccaaag gtcgcccgac gcccgggctt tgcccgggcg
     3361 gcctcagtga gcgagcgagc gcgcagagag ggagtggccc cccccccccc ccctgcagcc
     3421 tggcgtaata gcgaagaggc ccgcaccgat cgcccttccc aacagttgcg cagcctgaat
     3481 ggcgaatgga cgcgccctgt agcggcgcat taagcgcggc gggtgtggtg gttacgcgca
     3541 gcgtgaccgc tacacttgcc agcgccctag cgcccgctcc tttcgctttc ttcccttcct
     3601 ttctcgccac gttcgccggc tttccccgtc aagctctaaa tcgggggctc cctttagggt
     3661 tccgatttag tgctttacgg cacctcgacc ccaaaaaact tgattagggt gatggttcac
     3721 gtagtgggcc atcgccctga tagacggttt ttcgcccttt gacgttggag tccacgttct
     3781 ttaatagtgg actcttgttc caaactggaa caacactcaa ccctatctcg gtctattctt
     3841 ttgatttata agggattttg ccgatttcgg cctattggtt aaaaaatgag ctgatttaac
     3901 aaaaatttaa cgcgaatttt aacaaaatat taacgcttac aatttcctga tgcggtattt
     3961 tctccttacg catctgtgcg gtatttcaca ccgcatatgg tgcactctca gtacaatctg
     4021 ctctgatgcc gcatagttaa gccagccccg acacccgcca acacccgctg acgcgccctg
     4081 acgggcttgt ctgctcccgg catccgctta cagacaagct gtgaccgtct ccgggagctg
     4141 catgtgtcag aggttttcac cgtcatcacc gaaacgcgcg agacgaaagg gcctcgtgat
     4201 acgcctattt ttataggtta atgtcatgat aataatggtt tcttagacgt caggtggcac
     4261 ttttcgggga aatgtgcgcg gaacccctat ttgtttattt ttctaaatac attcaaatat
     4321 gtatccgctc atgagacaat aaccctgata aatgcttcaa taatattgaa aaaggaagag
     4381 tatgagtatt caacatttcc gtgtcgccct tattcccttt tttgcggcat tttgccttcc
     4441 tgtttttgct cacccagaaa cgctggtgaa agtaaaagat gctgaagatc agttgg
//
