LOCUS       sequence-492112-        4754 bp    DNA     circular SYN 06-NOV-2025
DEFINITION  AiE2202m 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 4754)
  TITLE     Basal Ganglia/CERP Enhancer AAVs
REFERENCE   2  (bases 1 to 4754)
  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: AiP1527 - pAAV-AiE2202m-minBG-SYFP2-WPRE3-BGHpA
FEATURES             Location/Qualifiers
     source          1..4754
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   94..234
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   105..234
                     /label=AAV2 ITR
     polyA_signal    352..463
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     650..670
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(join(763..1473,1474..1476,1477..1479))
                     /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(805..826)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     1154..1173
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1413..1434
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     regulatory      1476..1485
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     repeat_region   2121..2250
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     rep_origin      complement(2312..2900)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2392..2411)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3071..3862,3863..3931))
                     /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     3694..3713
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3932..4036)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     4104..4122
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(4160..4182)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     4282..4301
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     rep_origin      complement(join(4318..4754,1..19))
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4455..4476)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4667..4686
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
ORIGIN
        1 gcgccgctac agggcgcgta ctatggttgc tttgacgtat gcggtgtgaa ataccgcaca
       61 gatgcgtaag gagaaaatac cgcatcaggc gcccctgcag gcagctgcgc gctcgctcgc
      121 tcactgaggc cgcccgggca aagcccgggc gtcgggcgac ctttggtcgc ccggcctcag
      181 tgagcgagcg agcgcgcaga gagggagtgg ccaactccat cactaggggt tcctgcggcc
      241 gctcggtccg cacgtgagat ctcatgcctg ctattgtctt cccaatcctc ccccttgctg
      301 tcctgcccca ccccaccccc cagaatagaa tgacacctac tcagacaatg cgatgcaatt
      361 tcctcatttt attaggaaag gacagtggga gtggcacctt ccagggtcaa ggaaggcacg
      421 ggggaggggc aaacaacaga tggctggcaa ctagaaggca cagtcgaaga tctctcgagc
      481 cacggaattg tcagtgccca acagccgagc ccctgtccag cagcgggcaa ggcaggcggc
      541 gatgagttcc gccgtggcaa gaactaacca ggatttatac aaggaggaga aaatgaaagc
      601 catacgggaa gcaatagcat gatacaaagg cattaaagca gcgtatccac atagcgtaaa
      661 aggagcaaca tagttaagaa taccagtcaa tctttcacaa attttgtaat ccagaggttg
      721 attatgatat cgaattcgcg gccgcggcgc gccgtcgact tacttgtaca gctcgtccat
      781 gccgagagtg atcccggcgg cggtcacgaa ctccagcagg accatgtgat cgcgcttctc
      841 gttggggtct ttgctcagct tggactggta gctcaggtag tggttgtcgg gcagcagcac
      901 ggggccgtcg ccgatggggg tgttctgctg gtagtggtcg gcgagctgca cgccgccgtc
      961 ctcgatgttg tggcggatct tgaagttggc cttgatgccg ttcttctgct tgtcggcggt
     1021 gatatagacg ttgtggctgt tgtagttgta ctccagcttg tgccccagga tgttgccgtc
     1081 ctccttgaag tcgatgccct tcagctcgat gcggttcacc agggtgtcgc cctcgaactt
     1141 cacctcggcg cgggtcttgt agttgccgtc gtccttgaag aagatggtgc gctcctggac
     1201 gtagccttcg ggcatggcgg acttgaagaa gtcgtgctgc ttcatgtggt cggggtagcg
     1261 ggcgaagcac tgcacgccgt agcccagggt ggtcacgagg gtgggccagg gcacgggcag
     1321 cttgccggtg gtgcagatca gcttcagggt cagcttgccg taggtggcat cgccctcgcc
     1381 ctcgccggac acgctgaact tgtggccgtt tacgtcgccg tccagctcga ccaggatggg
     1441 caccaccccg gtgaacagct cctcgccctt gctcaccatg gtggcgaccg gtagcgctag
     1501 cttcgaaaga tctggatccc agaagcaaat gtaagcaata gatggctctg ccctgacttt
     1561 tatgcccagc ccgagctccg ccggcgaagg gacttggggc acaggagtcc tgggctcctg
     1621 aatgaaggtc cttgcctgtg ctgggatccc ctgacactgg cagagatctg gattcagacc
     1681 tagcaatagc tctgcagtcc agatacaggg gtgggatgtg ggggcaggga aagccccgag
     1741 gccagtcagt ctggttgttg aaaggaaatg ttcttacagg cacatttcca gcagccactg
     1801 gcagtttaat atccagcgcc caggcaaggg gccttggctg gtgcaggctg caaagagcag
     1861 gcagcaccca cagcagagag gaaatgaaat gcccgccttg ctcccgaaca atgccactga
     1921 tcagttccca gatcaggctg agttccgtga gggccacggt taggcaaggc cagcaggtgc
     1981 acatgaggcc tggggacggg agaaaggtgg ctgcaggctg tagtgctaga cctcctaacg
     2041 gcctgcgggg cttcagtttc accctctgtg gaatagaaat gatcgccggc gattattact
     2101 cttacaacgc gtgcggccgc aggaacccct agtgatggag ttggccactc cctctctgcg
     2161 cgctcgctcg ctcactgagg ccgggcgacc aaaggtcgcc cgacgcccgg gcggcctcag
     2221 tgagcgagcg agcgcgcagc tgcctgcagg acatgtgagc aaaaggccag caaaaggcca
     2281 ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag gctccgcccc cctgacgagc
     2341 atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc gacaggacta taaagatacc
     2401 aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg
     2461 gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta
     2521 ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg
     2581 ttcagcccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggtaagac
     2641 acgacttatc gccactggca gcagccactg gtaacaggat tagcagagcg aggtatgtag
     2701 gcggtgctac agagttcttg aagtggtggc ctaactacgg ctacactaga agaacagtat
     2761 ttggtatctg cgctctgctg aagccagtta ccttcggaaa aagagttggt agctcttgat
     2821 ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag cagattacgc
     2881 gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc tacggggtct gacgctcagt
     2941 ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt atcaaaaagg atcttcacct
     3001 agatcctttt aaattaaaaa tgaagtttta aatcaatcta aagtatatat gagtaaactt
     3061 ggtctgacag ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc tgtctatttc
     3121 gttcatccat agttgcctga ctccccgtcg tgtagataac tacgatacgg gagggcttac
     3181 catctggccc cagtgctgca atgataccgc gagacccacg ctcaccggct ccagatttat
     3241 cagcaataaa ccagccagcc ggaagggccg agcgcagaag tggtcctgca actttatccg
     3301 cctccatcca gtctattaat tgttgccggg aagctagagt aagtagttcg ccagttaata
     3361 gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt gtcacgctcg tcgtttggta
     3421 tggcttcatt cagctccggt tcccaacgat caaggcgagt tacatgatcc cccatgttgt
     3481 gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag
     3541 tgttatcact catggttatg gcagcactgc ataattctct tactgtcatg ccatccgtaa
     3601 gatgcttttc tgtgactggt gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc
     3661 gaccgagttg ctcttgcccg gcgtcaatac gggataatac cgcgccacat agcagaactt
     3721 taaaagtgct catcattgga aaacgttctt cggggcgaaa actctcaagg atcttaccgc
     3781 tgttgagatc cagttcgatg taacccactc gtgcacccaa ctgatcttca gcatctttta
     3841 ctttcaccag cgtttctggg tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa
     3901 taagggcgac acggaaatgt tgaatactca tactcttcct ttttcaatat tattgaagca
     3961 tttatcaggg ttattgtctc atgagcggat acatatttga atgtatttag aaaaataaac
     4021 aaataggggt tccgcgcaca tttccccgaa aagtgccacc tgacgtctaa gaaaccatta
     4081 ttatcatgac attaacctat aaaaataggc gtatcacgag gccctttcgt ctcgcgcgtt
     4141 tcggtgatga cggtgaaaac ctctgacaca tgcagctccc ggagacggtc acagcttgtc
     4201 tgtaagcgga tgccgggagc agacaagccc gtcagggcgc gtcagcgggt gttggcgggt
     4261 gtcggggctg gcttaactat gcggcatcag agcagattgt actgagagtg caccataaaa
     4321 ttgtaaacgt taatattttg ttaaaattcg cgttaaattt ttgttaaatc agctcatttt
     4381 ttaaccaata gaccgaaatc ggcaaaatcc cttataaatc aaaagaatag cccgagatag
     4441 agttgagtgt tgttccagtt tggaacaaga gtccactatt aaagaacgtg gactccaacg
     4501 tcaaagggcg aaaaaccgtc tatcagggcg atggcccact acgtgaacca tcacccaaat
     4561 caagtttttt ggggtcgagg tgccgtaaag cactaaatcg gaaccctaaa gggagccccc
     4621 gatttagagc ttgacgggga aagccggcga acgtggcgag aaaggaaggg aagaaagcga
     4681 aaggagcggg cgctaaggcg ctggcaagtg tagcggtcac gctgcgcgta accaccacac
     4741 ccgccgcgct taat
//
