LOCUS       sequence-492118-        4757 bp    DNA     circular SYN 06-NOV-2025
DEFINITION  AiE2206m 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 4757)
  TITLE     Basal Ganglia/CERP Enhancer AAVs
REFERENCE   2  (bases 1 to 4757)
  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: AiP1539 - pAAV-AiE2206m-minBG-SYFP2-WPRE3-BGHpA
FEATURES             Location/Qualifiers
     source          1..4757
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   113..242
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     rep_origin      complement(304..892)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(384..403)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(1063..1854,1855..1923))
                     /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     1686..1705
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(1924..2028)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     2096..2114
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(2152..2174)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     2274..2293
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     rep_origin      complement(2310..2765)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2447..2468)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2659..2678
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     repeat_region   2840..2980
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   2851..2980
                     /label=AAV2 ITR
     polyA_signal    3098..3209
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     3396..3416
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(join(3509..4219,4220..4222,4223..4225))
                     /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(3551..3572)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     3900..3919
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     4159..4180
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     regulatory      4222..4231
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
ORIGIN
        1 ctccagagag aaagtgagac aattaaaatg accacgtaca tccgtgggat gagtctcgac
       61 aaaccacaga gatcacgccg gcgattatta ctcttacaac gcgtgcggcc gcaggaaccc
      121 ctagtgatgg agttggccac tccctctctg cgcgctcgct cgctcactga ggccgggcga
      181 ccaaaggtcg cccgacgccc gggcggcctc agtgagcgag cgagcgcgca gctgcctgca
      241 ggacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc
      301 gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag
      361 gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt
      421 gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg
      481 aagcgtggcg ctttctcata gctcacgctg taggtatctc agttcggtgt aggtcgttcg
      541 ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg
      601 taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac
      661 tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg
      721 gcctaactac ggctacacta gaagaacagt atttggtatc tgcgctctgc tgaagccagt
      781 taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg
      841 tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc
      901 tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt
      961 ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa aatgaagttt
     1021 taaatcaatc taaagtatat atgagtaaac ttggtctgac agttaccaat gcttaatcag
     1081 tgaggcacct atctcagcga tctgtctatt tcgttcatcc atagttgcct gactccccgt
     1141 cgtgtagata actacgatac gggagggctt accatctggc cccagtgctg caatgatacc
     1201 gcgagaccca cgctcaccgg ctccagattt atcagcaata aaccagccag ccggaagggc
     1261 cgagcgcaga agtggtcctg caactttatc cgcctccatc cagtctatta attgttgccg
     1321 ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg ccattgctac
     1381 aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg gttcccaacg
     1441 atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc
     1501 tccgatcgtt gtcagaagta agttggccgc agtgttatca ctcatggtta tggcagcact
     1561 gcataattct cttactgtca tgccatccgt aagatgcttt tctgtgactg gtgagtactc
     1621 aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat
     1681 acgggataat accgcgccac atagcagaac tttaaaagtg ctcatcattg gaaaacgttc
     1741 ttcggggcga aaactctcaa ggatcttacc gctgttgaga tccagttcga tgtaacccac
     1801 tcgtgcaccc aactgatctt cagcatcttt tactttcacc agcgtttctg ggtgagcaaa
     1861 aacaggaagg caaaatgccg caaaaaaggg aataagggcg acacggaaat gttgaatact
     1921 catactcttc ctttttcaat attattgaag catttatcag ggttattgtc tcatgagcgg
     1981 atacatattt gaatgtattt agaaaaataa acaaataggg gttccgcgca catttccccg
     2041 aaaagtgcca cctgacgtct aagaaaccat tattatcatg acattaacct ataaaaatag
     2101 gcgtatcacg aggccctttc gtctcgcgcg tttcggtgat gacggtgaaa acctctgaca
     2161 catgcagctc ccggagacgg tcacagcttg tctgtaagcg gatgccggga gcagacaagc
     2221 ccgtcagggc gcgtcagcgg gtgttggcgg gtgtcggggc tggcttaact atgcggcatc
     2281 agagcagatt gtactgagag tgcaccataa aattgtaaac gttaatattt tgttaaaatt
     2341 cgcgttaaat ttttgttaaa tcagctcatt ttttaaccaa tagaccgaaa tcggcaaaat
     2401 cccttataaa tcaaaagaat agcccgagat agagttgagt gttgttccag tttggaacaa
     2461 gagtccacta ttaaagaacg tggactccaa cgtcaaaggg cgaaaaaccg tctatcaggg
     2521 cgatggccca ctacgtgaac catcacccaa atcaagtttt ttggggtcga ggtgccgtaa
     2581 agcactaaat cggaacccta aagggagccc ccgatttaga gcttgacggg gaaagccggc
     2641 gaacgtggcg agaaaggaag ggaagaaagc gaaaggagcg ggcgctaagg cgctggcaag
     2701 tgtagcggtc acgctgcgcg taaccaccac acccgccgcg cttaatgcgc cgctacaggg
     2761 cgcgtactat ggttgctttg acgtatgcgg tgtgaaatac cgcacagatg cgtaaggaga
     2821 aaataccgca tcaggcgccc ctgcaggcag ctgcgcgctc gctcgctcac tgaggccgcc
     2881 cgggcaaagc ccgggcgtcg ggcgaccttt ggtcgcccgg cctcagtgag cgagcgagcg
     2941 cgcagagagg gagtggccaa ctccatcact aggggttcct gcggccgctc ggtccgcacg
     3001 tgagatctca tgcctgctat tgtcttccca atcctccccc ttgctgtcct gccccacccc
     3061 accccccaga atagaatgac acctactcag acaatgcgat gcaatttcct cattttatta
     3121 ggaaaggaca gtgggagtgg caccttccag ggtcaaggaa ggcacggggg aggggcaaac
     3181 aacagatggc tggcaactag aaggcacagt cgaagatctc tcgagccacg gaattgtcag
     3241 tgcccaacag ccgagcccct gtccagcagc gggcaaggca ggcggcgatg agttccgccg
     3301 tggcaagaac taaccaggat ttatacaagg aggagaaaat gaaagccata cgggaagcaa
     3361 tagcatgata caaaggcatt aaagcagcgt atccacatag cgtaaaagga gcaacatagt
     3421 taagaatacc agtcaatctt tcacaaattt tgtaatccag aggttgatta tgatatcgaa
     3481 ttcgcggccg cggcgcgccg tcgacttact tgtacagctc gtccatgccg agagtgatcc
     3541 cggcggcggt cacgaactcc agcaggacca tgtgatcgcg cttctcgttg gggtctttgc
     3601 tcagcttgga ctggtagctc aggtagtggt tgtcgggcag cagcacgggg ccgtcgccga
     3661 tgggggtgtt ctgctggtag tggtcggcga gctgcacgcc gccgtcctcg atgttgtggc
     3721 ggatcttgaa gttggccttg atgccgttct tctgcttgtc ggcggtgata tagacgttgt
     3781 ggctgttgta gttgtactcc agcttgtgcc ccaggatgtt gccgtcctcc ttgaagtcga
     3841 tgcccttcag ctcgatgcgg ttcaccaggg tgtcgccctc gaacttcacc tcggcgcggg
     3901 tcttgtagtt gccgtcgtcc ttgaagaaga tggtgcgctc ctggacgtag ccttcgggca
     3961 tggcggactt gaagaagtcg tgctgcttca tgtggtcggg gtagcgggcg aagcactgca
     4021 cgccgtagcc cagggtggtc acgagggtgg gccagggcac gggcagcttg ccggtggtgc
     4081 agatcagctt cagggtcagc ttgccgtagg tggcatcgcc ctcgccctcg ccggacacgc
     4141 tgaacttgtg gccgtttacg tcgccgtcca gctcgaccag gatgggcacc accccggtga
     4201 acagctcctc gcccttgctc accatggtgg cgaccggtag cgctagcttc gaaagatctg
     4261 gatcccagaa gcaaatgtaa gcaatagatg gctctgccct gacttttatg cccagcccga
     4321 gctccgccgg cgttatcaga accagagcca atgtgcttaa tgaggactga attggcactc
     4381 gtggggacga gatcatgcca gcaagaatgc gcagaagtgg acagacaaag aggatattaa
     4441 aagtacaggg agcgatgagc agatagccag gacaaagaca agacacagaa agggacatcg
     4501 ggcggatatt ggagcagata ggttggtgtg tggagcacac ttccggaaag actttccatt
     4561 gatacgtcgc tgtctccaga gtggagtcaa acaccaggcc gaagggaatg ggagagcagt
     4621 ggggaaggtt aggccacacc tctgatgagg gtacaaggag aaaggcagac ctggaagcag
     4681 cgagggcagc ctagagggca tccagtgcat ctctgcatca ggcactcacc tcaagcctga
     4741 cagtggggta gagactg
//
