LOCUS       sequence-492115-        4746 bp    DNA     circular SYN 06-NOV-2025
DEFINITION  AiE2323m 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 4746)
  TITLE     Basal Ganglia/CERP Enhancer AAVs
REFERENCE   2  (bases 1 to 4746)
  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: AiP1684 - pAAV-AiE2323m-minBG-SYFP2-WPRE3-BGHpA
FEATURES             Location/Qualifiers
     source          1..4746
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     165..184
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     repeat_region   326..455
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     regulatory      1083..1092
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(1089..1091,1092..1094,1095..1805)
                     /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(1134..1155)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1395..1414)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1742..1763
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(1898..1918)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     polyA_signal    2105..2216
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   2334..2474
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   2334..2463
                     /label=AAV2 ITR
     rep_origin      2549..3004
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2636..2655)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2846..2867
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3021..3040)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3140..3162
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(3200..3218)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3286..3390
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3391..3459,3460..4251)
                     /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(3609..3628)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      join(4422..4746,1..264)
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
ORIGIN
        1 ggttcgtgca cacagcccag cttggagcga acgacctaca ccgaactgag atacctacag
       61 cgtgagctat gagaaagcgc cacgcttccc gaagggagaa aggcggacag gtatccggta
      121 agcggcaggg tcggaacagg agagcgcacg agggagcttc cagggggaaa cgcctggtat
      181 ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt gtgatgctcg
      241 tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc
      301 ttttgctggc cttttgctca catgtcctgc aggcagctgc gcgctcgctc gctcactgag
      361 gccgcccggg cgtcgggcga cctttggtcg cccggcctca gtgagcgagc gagcgcgcag
      421 agagggagtg gccaactcca tcactagggg ttcctgcggc cgcacgcgtt gtaagagtaa
      481 taatcgtgtg gaagtataag ctgaataaac ccttttctcc ccaatttgct ttttggtcat
      541 gatgtttgtg taggaataga aactcttgac taagacaaat tggtatcagc atagtgaggc
      601 attcctgtga caacctgacc atgttttggg gagaactgtg gaaggacttt ggaactttgg
      661 actagaatag ccattgggca ttaagagctc tgtgagatgt tttgtaggag cttggaagat
      721 aatgttgaga acagtgcaga aggtggaggc ctggcttgtg aaatttcaga gggaagatta
      781 aagattctta tcagggccat tgctgttttt actgtgaaga ttctgtggtt ttggttggct
      841 ggggccaaag aatcagctgt gattaacaag ataccagaac tactaaagca aaacctttgc
      901 attactggga tgattgatgt agtcagctgg agctaagaaa ttagcagtga ttaagaagag
      961 agtgattaag agcatcacta aggtgaacgg agctcgggct gggcataaaa gtcagggcag
     1021 agccatctat tgcttacatt tgcttctggg atccagatct ttcgaagcta gcgctaccgg
     1081 tcgccaccat ggtgagcaag ggcgaggagc tgttcaccgg ggtggtgccc atcctggtcg
     1141 agctggacgg cgacgtaaac ggccacaagt tcagcgtgtc cggcgagggc gagggcgatg
     1201 ccacctacgg caagctgacc ctgaagctga tctgcaccac cggcaagctg cccgtgccct
     1261 ggcccaccct cgtgaccacc ctgggctacg gcgtgcagtg cttcgcccgc taccccgacc
     1321 acatgaagca gcacgacttc ttcaagtccg ccatgcccga aggctacgtc caggagcgca
     1381 ccatcttctt caaggacgac ggcaactaca agacccgcgc cgaggtgaag ttcgagggcg
     1441 acaccctggt gaaccgcatc gagctgaagg gcatcgactt caaggaggac ggcaacatcc
     1501 tggggcacaa gctggagtac aactacaaca gccacaacgt ctatatcacc gccgacaagc
     1561 agaagaacgg catcaaggcc aacttcaaga tccgccacaa catcgaggac ggcggcgtgc
     1621 agctcgccga ccactaccag cagaacaccc ccatcggcga cggccccgtg ctgctgcccg
     1681 acaaccacta cctgagctac cagtccaagc tgagcaaaga ccccaacgag aagcgcgatc
     1741 acatggtcct gctggagttc gtgaccgccg ccgggatcac tctcggcatg gacgagctgt
     1801 acaagtaagt cgacggcgcg ccgcggccgc gaattcgata tcataatcaa cctctggatt
     1861 acaaaatttg tgaaagattg actggtattc ttaactatgt tgctcctttt acgctatgtg
     1921 gatacgctgc tttaatgcct ttgtatcatg ctattgcttc ccgtatggct ttcattttct
     1981 cctccttgta taaatcctgg ttagttcttg ccacggcgga actcatcgcc gcctgccttg
     2041 cccgctgctg gacaggggct cggctgttgg gcactgacaa ttccgtggct cgagagatct
     2101 tcgactgtgc cttctagttg ccagccatct gttgtttgcc cctcccccgt gccttccttg
     2161 accctggaag gtgccactcc cactgtcctt tcctaataaa atgaggaaat tgcatcgcat
     2221 tgtctgagta ggtgtcattc tattctgggg ggtggggtgg ggcaggacag caagggggag
     2281 gattgggaag acaatagcag gcatgagatc tcacgtgcgg accgagcggc cgcaggaacc
     2341 cctagtgatg gagttggcca ctccctctct gcgcgctcgc tcgctcactg aggccgggcg
     2401 accaaaggtc gcccgacgcc cgggctttgc ccgggcggcc tcagtgagcg agcgagcgcg
     2461 cagctgcctg caggggcgcc tgatgcggta ttttctcctt acgcatctgt gcggtatttc
     2521 acaccgcata cgtcaaagca accatagtac gcgccctgta gcggcgcatt aagcgcggcg
     2581 ggtgtggtgg ttacgcgcag cgtgaccgct acacttgcca gcgccttagc gcccgctcct
     2641 ttcgctttct tcccttcctt tctcgccacg ttcgccggct ttccccgtca agctctaaat
     2701 cgggggctcc ctttagggtt ccgatttagt gctttacggc acctcgaccc caaaaaactt
     2761 gatttgggtg atggttcacg tagtgggcca tcgccctgat agacggtttt tcgccctttg
     2821 acgttggagt ccacgttctt taatagtgga ctcttgttcc aaactggaac aacactcaac
     2881 tctatctcgg gctattcttt tgatttataa gggattttgc cgatttcggt ctattggtta
     2941 aaaaatgagc tgatttaaca aaaatttaac gcgaatttta acaaaatatt aacgtttaca
     3001 attttatggt gcactctcag tacaatctgc tctgatgccg catagttaag ccagccccga
     3061 cacccgccaa cacccgctga cgcgccctga cgggcttgtc tgctcccggc atccgcttac
     3121 agacaagctg tgaccgtctc cgggagctgc atgtgtcaga ggttttcacc gtcatcaccg
     3181 aaacgcgcga gacgaaaggg cctcgtgata cgcctatttt tataggttaa tgtcatgata
     3241 ataatggttt cttagacgtc aggtggcact tttcggggaa atgtgcgcgg aacccctatt
     3301 tgtttatttt tctaaataca ttcaaatatg tatccgctca tgagacaata accctgataa
     3361 atgcttcaat aatattgaaa aaggaagagt atgagtattc aacatttccg tgtcgccctt
     3421 attccctttt ttgcggcatt ttgccttcct gtttttgctc acccagaaac gctggtgaaa
     3481 gtaaaagatg ctgaagatca gttgggtgca cgagtgggtt acatcgaact ggatctcaac
     3541 agcggtaaga tccttgagag ttttcgcccc gaagaacgtt ttccaatgat gagcactttt
     3601 aaagttctgc tatgtggcgc ggtattatcc cgtattgacg ccgggcaaga gcaactcggt
     3661 cgccgcatac actattctca gaatgacttg gttgagtact caccagtcac agaaaagcat
     3721 cttacggatg gcatgacagt aagagaatta tgcagtgctg ccataaccat gagtgataac
     3781 actgcggcca acttacttct gacaacgatc ggaggaccga aggagctaac cgcttttttg
     3841 cacaacatgg gggatcatgt aactcgcctt gatcgttggg aaccggagct gaatgaagcc
     3901 ataccaaacg acgagcgtga caccacgatg cctgtagcaa tggcaacaac gttgcgcaaa
     3961 ctattaactg gcgaactact tactctagct tcccggcaac aattaataga ctggatggag
     4021 gcggataaag ttgcaggacc acttctgcgc tcggcccttc cggctggctg gtttattgct
     4081 gataaatctg gagccggtga gcgtgggtct cgcggtatca ttgcagcact ggggccagat
     4141 ggtaagccct cccgtatcgt agttatctac acgacgggga gtcaggcaac tatggatgaa
     4201 cgaaatagac agatcgctga gataggtgcc tcactgatta agcattggta actgtcagac
     4261 caagtttact catatatact ttagattgat ttaaaacttc atttttaatt taaaaggatc
     4321 taggtgaaga tcctttttga taatctcatg accaaaatcc cttaacgtga gttttcgttc
     4381 cactgagcgt cagaccccgt agaaaagatc aaaggatctt cttgagatcc tttttttctg
     4441 cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg
     4501 gatcaagagc taccaactct ttttccgaag gtaactggct tcagcagagc gcagatacca
     4561 aatactgttc ttctagtgta gccgtagtta ggccaccact tcaagaactc tgtagcaccg
     4621 cctacatacc tcgctctgct aatcctgtta ccagtggctg ctgccagtgg cgataagtcg
     4681 tgtcttaccg ggttggactc aagacgatag ttaccggata aggcgcagcg gtcgggctga
     4741 acgggg
//
