LOCUS       sequence-457689-        5131 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AAV expression of EGFP from hSyn promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5131)
  TITLE     AAV plasmids
REFERENCE   2  (bases 1 to 5131)
  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"
COMMENT     Sequence Label: pAAV.hSyn.eGFP.WPRE.bGH
FEATURES             Location/Qualifiers
     source          1..5131
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   82..211
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     polyA_signal    267..315
                     /note="synthetic polyadenylation signal"
     misc_feature    329..420
                     /label=pause site
                     /note="RNA polymerase II transcriptional pause signal from 
                     the human alpha-2 globin gene"
     primer_bind     369..388
                     /label=RVprimer3
                     /note="pGL3 vector, forward primer"
     promoter        456..903
                     /label=hSyn promoter
                     /note="human synapsin I promoter; confers neuron-specific 
                     expression (Kugler et al., 2003)"
     regulatory      938..947
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(944..946,947..949,950..1663)
                     /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(944..946,947..949,950..1660)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(989..1010)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1250..1269)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1597..1618
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     misc_feature    1685..2273
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(1738..1758)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(2156..2167)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(2306..2323)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2312..2519
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      2640..3095
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2727..2746)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2937..2958
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3162..3181)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3281..3303
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(3341..3359)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3427..3531
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3532..3600,3601..4392)
                     /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(3750..3769)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      join(4563..5131,1..20)
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5052..5071
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 gggggcggag cctatggaaa aacgccagca acgcggcctt tttacggttc ctggcctttt
       61 gctggccttt tgctcacatg tcctgcaggc agctgcgcgc tcgctcgctc actgaggccg
      121 cccgggcgtc gggcgacctt tggtcgcccg gcctcagtga gcgagcgagc gcgcagagag
      181 ggagtggcca actccatcac taggggttcc tgcggccgca cgcgttctag aaatattaag
      241 gtacgggagg tacttggagc ggccgcaata aaatatcttt attttcatta catctgtgtg
      301 ttggtttttt gtgtgaatcg atagtactaa catacgctct ccatcaaaac aaaacgaaac
      361 aaaacaaact agcaaaatag gctgtcccca gtgcaagtgc aggtgccaga acatttctct
      421 attctagact cgagctgcag agggccctgc gtatgagtgc aagtgggttt taggaccagg
      481 atgaggcggg gtgggggtgc ctacctgacg accgaccccg acccactgga caagcaccca
      541 acccccattc cccaaattgc gcatccccta tcagagaggg ggaggggaaa caggatgcgg
      601 cgaggcgcgt gcgcactgcc agcttcagca ccgcggacag tgccttcgcc cccgcctggc
      661 ggcgcgcgcc accgccgcct cagcactgaa ggcgcgctga cgtcactcgc cggtcccccg
      721 caaactcccc ttcccggcca ccttggtcgc gtccgcgccg ccgccggccc agccggaccg
      781 caccacgcga ggcgcgagat aggggggcac gggcgcgacc atctgcgctg cggcgccggc
      841 gactcagcgc tgcctcagtc tgcggtgggc agcggaggag tcgtgtcgtg cctgagagcg
      901 cagtcgaatt caagctgcta gcaaggatcc accggtcgcc accatggtga gcaagggcga
      961 ggagctgttc accggggtgg tgcccatcct ggtcgagctg gacggcgacg taaacggcca
     1021 caagttcagc gtgtccggcg agggcgaggg cgatgccacc tacggcaagc tgaccctgaa
     1081 gttcatctgc accaccggca agctgcccgt gccctggccc accctcgtga ccaccctgac
     1141 ctacggcgtg cagtgcttca gccgctaccc cgaccacatg aagcagcacg acttcttcaa
     1201 gtccgccatg cccgaaggct acgtccagga gcgcaccatc ttcttcaagg acgacggcaa
     1261 ctacaagacc cgcgccgagg tgaagttcga gggcgacacc ctggtgaacc gcatcgagct
     1321 gaagggcatc gacttcaagg aggacggcaa catcctgggg cacaagctgg agtacaacta
     1381 caacagccac aacgtctata tcatggccga caagcagaag aacggcatca aggtgaactt
     1441 caagatccgc cacaacatcg aggacggcag cgtgcagctc gccgaccact accagcagaa
     1501 cacccccatc ggcgacggcc ccgtgctgct gcccgacaac cactacctga gcacccagtc
     1561 cgccctgagc aaagacccca acgagaagcg cgatcacatg gtcctgctgg agttcgtgac
     1621 cgccgccggg atcactctcg gcatggacga gctgtacaag taaagcggcc gcaagcttat
     1681 cgataatcaa cctctggatt acaaaatttg tgaaagattg actggtattc ttaactatgt
     1741 tgctcctttt acgctatgtg gatacgctgc tttaatgcct ttgtatcatg ctattgcttc
     1801 ccgtatggct ttcattttct cctccttgta taaatcctgg ttgctgtctc tttatgagga
     1861 gttgtggccc gttgtcaggc aacgtggcgt ggtgtgcact gtgtttgctg acgcaacccc
     1921 cactggttgg ggcattgcca ccacctgtca gctcctttcc gggactttcg ctttccccct
     1981 ccctattgcc acggcggaac tcatcgccgc ctgccttgcc cgctgctgga caggggctcg
     2041 gctgttgggc actgacaatt ccgtggtgtt gtcggggaaa tcatcgtcct ttccttggct
     2101 gctcgcctgt gttgccacct ggattctgcg cgggacgccc ttctgctacg tcccttcggc
     2161 cctcaatcca gcggaccttc cttcccgcgg cctgctgccg gctctgcggc ctcttccgcg
     2221 tcttcgcctt cgccctcaga cgagtcggat ctccctttgg gccgcctccc cgcatcgata
     2281 ccgtcgacta gagctcgctg atcagcctcg actgtgcctt ctagttgcca gccatctgtt
     2341 gtttgcccct cccccgtgcc ttccttgacc ctggaaggtg ccactcccac tgtcctttcc
     2401 taataaaatg aggaaattgc atcgcattgt ctgagtaggt gtcattctat tctggggggt
     2461 ggggtggggc aggacagcaa gggggaggat tgggaagaca atagcaggca tgctggggag
     2521 agatctgagg aacccctagt gatggagttg gccactccct ctctgcagcc tggcgtaata
     2581 gcgaagaggc ccgcaccgat cgcccttccc aacagttgcg cagcctgaat ggcgaatgga
     2641 cgcgccctgt agcggcgcat taagcgcggc gggtgtggtg gttacgcgca gcgtgaccgc
     2701 tacacttgcc agcgccctag cgcccgctcc tttcgctttc ttcccttcct ttctcgccac
     2761 gttcgccggc tttccccgtc aagctctaaa tcgggggctc cctttagggt tccgatttag
     2821 tgctttacgg cacctcgacc ccaaaaaact tgattagggt gatggttcac gtagtgggcc
     2881 atcgccctga tagacggttt ttcgcccttt gacgttggag tccacgttct ttaatagtgg
     2941 actcttgttc caaactggaa caacactcaa ccctatctcg gtctattctt ttgatttata
     3001 agggattttg ccgatttcgg cctattggtt aaaaaatgag ctgatttaac aaaaatttaa
     3061 cgcgaatttt aacaaaatat taacgcttac aatttcctga tgcggtattt tctccttacg
     3121 catctgtgcg gtatttcaca ccgcatatgg tgcactctca gtacaatctg ctctgatgcc
     3181 gcatagttaa gccagccccg acacccgcca acacccgctg acgcgccctg acgggcttgt
     3241 ctgctcccgg catccgctta cagacaagct gtgaccgtct ccgggagctg catgtgtcag
     3301 aggttttcac cgtcatcacc gaaacgcgcg agacgaaagg gcctcgtgat acgcctattt
     3361 ttataggtta atgtcatgat aataatggtt tcttagacgt caggtggcac ttttcgggga
     3421 aatgtgcgcg gaacccctat ttgtttattt ttctaaatac attcaaatat gtatccgctc
     3481 atgagacaat aaccctgata aatgcttcaa taatattgaa aaaggaagag tatgagtatt
     3541 caacatttcc gtgtcgccct tattcccttt tttgcggcat tttgccttcc tgtttttgct
     3601 cacccagaaa cgctggtgaa agtaaaagat gctgaagatc agttgggtgc acgagtgggt
     3661 tacatcgaac tggatctcaa cagcggtaag atccttgaga gttttcgccc cgaagaacgt
     3721 tttccaatga tgagcacttt taaagttctg ctatgtggcg cggtattatc ccgtattgac
     3781 gccgggcaag agcaactcgg tcgccgcata cactattctc agaatgactt ggttgagtac
     3841 tcaccagtca cagaaaagca tcttacggat ggcatgacag taagagaatt atgcagtgct
     3901 gccataacca tgagtgataa cactgcggcc aacttacttc tgacaacgat cggaggaccg
     3961 aaggagctaa ccgctttttt gcacaacatg ggggatcatg taactcgcct tgatcgttgg
     4021 gaaccggagc tgaatgaagc cataccaaac gacgagcgtg acaccacgat gcctgtagca
     4081 atggcaacaa cgttgcgcaa actattaact ggcgaactac ttactctagc ttcccggcaa
     4141 caattaatag actggatgga ggcggataaa gttgcaggac cacttctgcg ctcggccctt
     4201 ccggctggct ggtttattgc tgataaatct ggagccggtg agcgtgggtc tcgcggtatc
     4261 attgcagcac tggggccaga tggtaagccc tcccgtatcg tagttatcta cacgacgggg
     4321 agtcaggcaa ctatggatga acgaaataga cagatcgctg agataggtgc ctcactgatt
     4381 aagcattggt aactgtcaga ccaagtttac tcatatatac tttagattga tttaaaactt
     4441 catttttaat ttaaaaggat ctaggtgaag atcctttttg ataatctcat gaccaaaatc
     4501 ccttaacgtg agttttcgtt ccactgagcg tcagaccccg tagaaaagat caaaggatct
     4561 tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta
     4621 ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc tttttccgaa ggtaactggc
     4681 ttcagcagag cgcagatacc aaatactgtt cttctagtgt agccgtagtt aggccaccac
     4741 ttcaagaact ctgtagcacc gcctacatac ctcgctctgc taatcctgtt accagtggct
     4801 gctgccagtg gcgataagtc gtgtcttacc gggttggact caagacgata gttaccggat
     4861 aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac agcccagctt ggagcgaacg
     4921 acctacaccg aactgagata cctacagcgt gagctatgag aaagcgccac gcttcccgaa
     4981 gggagaaagg cggacaggta tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg
     5041 gagcttccag ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg ccacctctga
     5101 cttgagcgtc gatttttgtg atgctcgtca g
//
