LOCUS       sequence-492124-        4861 bp    DNA     circular SYN 06-NOV-2025
DEFINITION  AiE2659m is an enhancer sequence designed to drive AAV-mediated 
            transgene expression in specific population sof brain cells..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4861)
  TITLE     Basal Ganglia/CERP Enhancer AAVs
REFERENCE   2  (bases 1 to 4861)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Nov 6, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"
COMMENT     Sequence Label: AiP2132 - pAAV-AiE2659m-minBG-SYFP2-WPRE3-BGHpA
FEATURES             Location/Qualifiers
     source          1..4861
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   293..422
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     rep_origin      complement(484..1072)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(564..583)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(1243..2034,2035..2103))
                     /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     1866..1885
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(2104..2208)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     2276..2294
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(2332..2354)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     2454..2473
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     rep_origin      complement(2490..2945)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2627..2648)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2839..2858
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     repeat_region   3020..3160
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   3031..3160
                     /label=AAV2 ITR
     polyA_signal    3278..3389
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     3576..3596
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(join(3689..4399,4400..4402,4403..4405))
                     /codon_start=1
                     /product="monomeric optimized variant of EYFP, identical to
                     mVenus but with L68V (Kremers et al., 2006)"
                     /label=SYFP2
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KLICTTGKLPVPWPTLVTTLGYGVQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYITADKQKNGIK
                     ANFKIRHNIEDGGVQLADHYQQNTPIGDGPVLLPDNHYLSYQSKLSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(3731..3752)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     4080..4099
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     4339..4360
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     regulatory      4402..4411
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
ORIGIN
        1 agtcagattt tagaatttgc ttgagactga agttttctat ttttaccttg actctagata
       61 agagtattac ttccatgact caaagttaaa attttatagt tcaagctgtg aaatgaagct
      121 caatggtaga acatttacct agaatatgaa gctccagtta aatccagcaa taactttcaa
      181 gtattcatca agtctcactt tacacctctt ctgacctatc ccccaaatta gaaatcagca
      241 ggagttattg ttcagactgt ttttattaga gaaataggac gcgtgcggcc gcaggaaccc
      301 ctagtgatgg agttggccac tccctctctg cgcgctcgct cgctcactga ggccgggcga
      361 ccaaaggtcg cccgacgccc gggcggcctc agtgagcgag cgagcgcgca gctgcctgca
      421 ggacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc
      481 gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag
      541 gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt
      601 gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg
      661 aagcgtggcg ctttctcata gctcacgctg taggtatctc agttcggtgt aggtcgttcg
      721 ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg
      781 taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac
      841 tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg
      901 gcctaactac ggctacacta gaagaacagt atttggtatc tgcgctctgc tgaagccagt
      961 taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg
     1021 tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc
     1081 tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt
     1141 ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa aatgaagttt
     1201 taaatcaatc taaagtatat atgagtaaac ttggtctgac agttaccaat gcttaatcag
     1261 tgaggcacct atctcagcga tctgtctatt tcgttcatcc atagttgcct gactccccgt
     1321 cgtgtagata actacgatac gggagggctt accatctggc cccagtgctg caatgatacc
     1381 gcgagaccca cgctcaccgg ctccagattt atcagcaata aaccagccag ccggaagggc
     1441 cgagcgcaga agtggtcctg caactttatc cgcctccatc cagtctatta attgttgccg
     1501 ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg ccattgctac
     1561 aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg gttcccaacg
     1621 atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc
     1681 tccgatcgtt gtcagaagta agttggccgc agtgttatca ctcatggtta tggcagcact
     1741 gcataattct cttactgtca tgccatccgt aagatgcttt tctgtgactg gtgagtactc
     1801 aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat
     1861 acgggataat accgcgccac atagcagaac tttaaaagtg ctcatcattg gaaaacgttc
     1921 ttcggggcga aaactctcaa ggatcttacc gctgttgaga tccagttcga tgtaacccac
     1981 tcgtgcaccc aactgatctt cagcatcttt tactttcacc agcgtttctg ggtgagcaaa
     2041 aacaggaagg caaaatgccg caaaaaaggg aataagggcg acacggaaat gttgaatact
     2101 catactcttc ctttttcaat attattgaag catttatcag ggttattgtc tcatgagcgg
     2161 atacatattt gaatgtattt agaaaaataa acaaataggg gttccgcgca catttccccg
     2221 aaaagtgcca cctgacgtct aagaaaccat tattatcatg acattaacct ataaaaatag
     2281 gcgtatcacg aggccctttc gtctcgcgcg tttcggtgat gacggtgaaa acctctgaca
     2341 catgcagctc ccggagacgg tcacagcttg tctgtaagcg gatgccggga gcagacaagc
     2401 ccgtcagggc gcgtcagcgg gtgttggcgg gtgtcggggc tggcttaact atgcggcatc
     2461 agagcagatt gtactgagag tgcaccataa aattgtaaac gttaatattt tgttaaaatt
     2521 cgcgttaaat ttttgttaaa tcagctcatt ttttaaccaa tagaccgaaa tcggcaaaat
     2581 cccttataaa tcaaaagaat agcccgagat agagttgagt gttgttccag tttggaacaa
     2641 gagtccacta ttaaagaacg tggactccaa cgtcaaaggg cgaaaaaccg tctatcaggg
     2701 cgatggccca ctacgtgaac catcacccaa atcaagtttt ttggggtcga ggtgccgtaa
     2761 agcactaaat cggaacccta aagggagccc ccgatttaga gcttgacggg gaaagccggc
     2821 gaacgtggcg agaaaggaag ggaagaaagc gaaaggagcg ggcgctaagg cgctggcaag
     2881 tgtagcggtc acgctgcgcg taaccaccac acccgccgcg cttaatgcgc cgctacaggg
     2941 cgcgtactat ggttgctttg acgtatgcgg tgtgaaatac cgcacagatg cgtaaggaga
     3001 aaataccgca tcaggcgccc ctgcaggcag ctgcgcgctc gctcgctcac tgaggccgcc
     3061 cgggcaaagc ccgggcgtcg ggcgaccttt ggtcgcccgg cctcagtgag cgagcgagcg
     3121 cgcagagagg gagtggccaa ctccatcact aggggttcct gcggccgctc ggtccgcacg
     3181 tgagatctca tgcctgctat tgtcttccca atcctccccc ttgctgtcct gccccacccc
     3241 accccccaga atagaatgac acctactcag acaatgcgat gcaatttcct cattttatta
     3301 ggaaaggaca gtgggagtgg caccttccag ggtcaaggaa ggcacggggg aggggcaaac
     3361 aacagatggc tggcaactag aaggcacagt cgaagatctc tcgagccacg gaattgtcag
     3421 tgcccaacag ccgagcccct gtccagcagc gggcaaggca ggcggcgatg agttccgccg
     3481 tggcaagaac taaccaggat ttatacaagg aggagaaaat gaaagccata cgggaagcaa
     3541 tagcatgata caaaggcatt aaagcagcgt atccacatag cgtaaaagga gcaacatagt
     3601 taagaatacc agtcaatctt tcacaaattt tgtaatccag aggttgatta tgatatcgaa
     3661 ttcgcggccg cggcgcgccg tcgacttact tgtacagctc gtccatgccg agagtgatcc
     3721 cggcggcggt cacgaactcc agcaggacca tgtgatcgcg cttctcgttg gggtctttgc
     3781 tcagcttgga ctggtagctc aggtagtggt tgtcgggcag cagcacgggg ccgtcgccga
     3841 tgggggtgtt ctgctggtag tggtcggcga gctgcacgcc gccgtcctcg atgttgtggc
     3901 ggatcttgaa gttggccttg atgccgttct tctgcttgtc ggcggtgata tagacgttgt
     3961 ggctgttgta gttgtactcc agcttgtgcc ccaggatgtt gccgtcctcc ttgaagtcga
     4021 tgcccttcag ctcgatgcgg ttcaccaggg tgtcgccctc gaacttcacc tcggcgcggg
     4081 tcttgtagtt gccgtcgtcc ttgaagaaga tggtgcgctc ctggacgtag ccttcgggca
     4141 tggcggactt gaagaagtcg tgctgcttca tgtggtcggg gtagcgggcg aagcactgca
     4201 cgccgtagcc cagggtggtc acgagggtgg gccagggcac gggcagcttg ccggtggtgc
     4261 agatcagctt cagggtcagc ttgccgtagg tggcatcgcc ctcgccctcg ccggacacgc
     4321 tgaacttgtg gccgtttacg tcgccgtcca gctcgaccag gatgggcacc accccggtga
     4381 acagctcctc gcccttgctc accatggtgg cgaccggtag cgctagcttc gaaagatctg
     4441 gatcccagaa gcaaatgtaa gcaatagatg gctctgccct gacttttatg cccagcccga
     4501 gctccgccgg cgtccacaaa gcccatcagt tctgtctctt ccacttcctc tccgtttagc
     4561 atgagaactc tgtagcagaa taaaaggaag gttcaggtta gcctgggacc agcttcccaa
     4621 ccttgcaccg tcatcccata aggaaggagc aggcctgccc ttaccttgtc tggcccacaa
     4681 aagagagcac caaagtgtca tccgtctccc gaccggggtc agagcgcaga ggccataaac
     4741 ctggaacaag gagtaaagca taaagtgtac catgagccag cacccaagac gtgccctcac
     4801 cctaagccac gaggggggcg gaaatccctt ctagtccttc ctgatattta cactctaaag
     4861 a
//
