LOCUS       sequence-492117-        4709 bp    DNA     circular SYN 06-NOV-2025
DEFINITION  AiE2643m 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 4709)
  TITLE     Basal Ganglia/CERP Enhancer AAVs
REFERENCE   2  (bases 1 to 4709)
  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: AiP20006 - pAAV-AiE2643m-minBG-SYFP2-WPRE3-BGHpA
FEATURES             Location/Qualifiers
     source          1..4709
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     74..93
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     repeat_region   255..395
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   266..395
                     /label=AAV2 ITR
     polyA_signal    513..624
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     811..831
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(join(924..1634,1635..1637,1638..1640))
                     /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(966..987)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     1315..1334
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1574..1595
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     regulatory      1637..1646
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     repeat_region   2237..2366
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     rep_origin      complement(2428..3016)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2508..2527)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3187..3978,3979..4047))
                     /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     3810..3829
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4048..4152)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     4220..4238
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(4276..4298)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     4398..4417
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     rep_origin      complement(join(4434..4709,1..180))
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4571..4592)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
ORIGIN
        1 taaatcggaa ccctaaaggg agcccccgat ttagagcttg acggggaaag ccggcgaacg
       61 tggcgagaaa ggaagggaag aaagcgaaag gagcgggcgc taaggcgctg gcaagtgtag
      121 cggtcacgct gcgcgtaacc accacacccg ccgcgcttaa tgcgccgcta cagggcgcgt
      181 actatggttg ctttgacgta tgcggtgtga aataccgcac agatgcgtaa ggagaaaata
      241 ccgcatcagg cgcccctgca ggcagctgcg cgctcgctcg ctcactgagg ccgcccgggc
      301 aaagcccggg cgtcgggcga cctttggtcg cccggcctca gtgagcgagc gagcgcgcag
      361 agagggagtg gccaactcca tcactagggg ttcctgcggc cgctcggtcc gcacgtgaga
      421 tctcatgcct gctattgtct tcccaatcct cccccttgct gtcctgcccc accccacccc
      481 ccagaataga atgacaccta ctcagacaat gcgatgcaat ttcctcattt tattaggaaa
      541 ggacagtggg agtggcacct tccagggtca aggaaggcac gggggagggg caaacaacag
      601 atggctggca actagaaggc acagtcgaag atctctcgag ccacggaatt gtcagtgccc
      661 aacagccgag cccctgtcca gcagcgggca aggcaggcgg cgatgagttc cgccgtggca
      721 agaactaacc aggatttata caaggaggag aaaatgaaag ccatacggga agcaatagca
      781 tgatacaaag gcattaaagc agcgtatcca catagcgtaa aaggagcaac atagttaaga
      841 ataccagtca atctttcaca aattttgtaa tccagaggtt gattatgata tcgaattcgc
      901 ggccgcggcg cgccgtcgac ttacttgtac agctcgtcca tgccgagagt gatcccggcg
      961 gcggtcacga actccagcag gaccatgtga tcgcgcttct cgttggggtc tttgctcagc
     1021 ttggactggt agctcaggta gtggttgtcg ggcagcagca cggggccgtc gccgatgggg
     1081 gtgttctgct ggtagtggtc ggcgagctgc acgccgccgt cctcgatgtt gtggcggatc
     1141 ttgaagttgg ccttgatgcc gttcttctgc ttgtcggcgg tgatatagac gttgtggctg
     1201 ttgtagttgt actccagctt gtgccccagg atgttgccgt cctccttgaa gtcgatgccc
     1261 ttcagctcga tgcggttcac cagggtgtcg ccctcgaact tcacctcggc gcgggtcttg
     1321 tagttgccgt cgtccttgaa gaagatggtg cgctcctgga cgtagccttc gggcatggcg
     1381 gacttgaaga agtcgtgctg cttcatgtgg tcggggtagc gggcgaagca ctgcacgccg
     1441 tagcccaggg tggtcacgag ggtgggccag ggcacgggca gcttgccggt ggtgcagatc
     1501 agcttcaggg tcagcttgcc gtaggtggca tcgccctcgc cctcgccgga cacgctgaac
     1561 ttgtggccgt ttacgtcgcc gtccagctcg accaggatgg gcaccacccc ggtgaacagc
     1621 tcctcgccct tgctcaccat ggtggcgacc ggtagcgcta gcttcgaaag atctggatcc
     1681 cagaagcaaa tgtaagcaat agatggctct gccctgactt ttatgcccag cccgagctcc
     1741 gccggcgttc ttcttagctt ccaagatgat cgacccaaat ctgagcctca ttacagtgcc
     1801 ttagaggtgc cgaggactat gccctttctc ccttggagaa gtacgatggc tgcgagcaag
     1861 gctcccagga ccaaactgta ttcaaaccct ggctctacca ctttcaaggt acatggcttt
     1921 gtggtggtat ttccattcct ctgcgtccca gctccctgct tcaaactgga gatattaata
     1981 ggacctacgt cacaaggctc ttttcaggat ttagtgagat agcacaggtg aattccttcg
     2041 cacacaggaa gaactcagta aactcagcta taatcactga gcctcactcc agtctcgcaa
     2101 tgtttattta cattttcctt ggtcctggtt ctttatcctt actgatattt taacattgtt
     2161 tatggtttct gcttcggtgg taagccacct cagatccttt gtggattagg gtaggttata
     2221 aaacgcgtgc ggccgcagga acccctagtg atggagttgg ccactccctc tctgcgcgct
     2281 cgctcgctca ctgaggccgg gcgaccaaag gtcgcccgac gcccgggcgg cctcagtgag
     2341 cgagcgagcg cgcagctgcc tgcaggacat gtgagcaaaa ggccagcaaa aggccaggaa
     2401 ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc cgcccccctg acgagcatca
     2461 caaaaatcga cgctcaagtc agaggtggcg aaacccgaca ggactataaa gataccaggc
     2521 gtttccccct ggaagctccc tcgtgcgctc tcctgttccg accctgccgc ttaccggata
     2581 cctgtccgcc tttctccctt cgggaagcgt ggcgctttct catagctcac gctgtaggta
     2641 tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac cccccgttca
     2701 gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag tccaacccgg taagacacga
     2761 cttatcgcca ctggcagcag ccactggtaa caggattagc agagcgaggt atgtaggcgg
     2821 tgctacagag ttcttgaagt ggtggcctaa ctacggctac actagaagaa cagtatttgg
     2881 tatctgcgct ctgctgaagc cagttacctt cggaaaaaga gttggtagct cttgatccgg
     2941 caaacaaacc accgctggta gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag
     3001 aaaaaaagga tctcaagaag atcctttgat cttttctacg gggtctgacg ctcagtggaa
     3061 cgaaaactca cgttaaggga ttttggtcat gagattatca aaaaggatct tcacctagat
     3121 ccttttaaat taaaaatgaa gttttaaatc aatctaaagt atatatgagt aaacttggtc
     3181 tgacagttac caatgcttaa tcagtgaggc acctatctca gcgatctgtc tatttcgttc
     3241 atccatagtt gcctgactcc ccgtcgtgta gataactacg atacgggagg gcttaccatc
     3301 tggccccagt gctgcaatga taccgcgaga cccacgctca ccggctccag atttatcagc
     3361 aataaaccag ccagccggaa gggccgagcg cagaagtggt cctgcaactt tatccgcctc
     3421 catccagtct attaattgtt gccgggaagc tagagtaagt agttcgccag ttaatagttt
     3481 gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc
     3541 ttcattcagc tccggttccc aacgatcaag gcgagttaca tgatccccca tgttgtgcaa
     3601 aaaagcggtt agctccttcg gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt
     3661 atcactcatg gttatggcag cactgcataa ttctcttact gtcatgccat ccgtaagatg
     3721 cttttctgtg actggtgagt actcaaccaa gtcattctga gaatagtgta tgcggcgacc
     3781 gagttgctct tgcccggcgt caatacggga taataccgcg ccacatagca gaactttaaa
     3841 agtgctcatc attggaaaac gttcttcggg gcgaaaactc tcaaggatct taccgctgtt
     3901 gagatccagt tcgatgtaac ccactcgtgc acccaactga tcttcagcat cttttacttt
     3961 caccagcgtt tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa agggaataag
     4021 ggcgacacgg aaatgttgaa tactcatact cttccttttt caatattatt gaagcattta
     4081 tcagggttat tgtctcatga gcggatacat atttgaatgt atttagaaaa ataaacaaat
     4141 aggggttccg cgcacatttc cccgaaaagt gccacctgac gtctaagaaa ccattattat
     4201 catgacatta acctataaaa ataggcgtat cacgaggccc tttcgtctcg cgcgtttcgg
     4261 tgatgacggt gaaaacctct gacacatgca gctcccggag acggtcacag cttgtctgta
     4321 agcggatgcc gggagcagac aagcccgtca gggcgcgtca gcgggtgttg gcgggtgtcg
     4381 gggctggctt aactatgcgg catcagagca gattgtactg agagtgcacc ataaaattgt
     4441 aaacgttaat attttgttaa aattcgcgtt aaatttttgt taaatcagct cattttttaa
     4501 ccaatagacc gaaatcggca aaatccctta taaatcaaaa gaatagcccg agatagagtt
     4561 gagtgttgtt ccagtttgga acaagagtcc actattaaag aacgtggact ccaacgtcaa
     4621 agggcgaaaa accgtctatc agggcgatgg cccactacgt gaaccatcac ccaaatcaag
     4681 ttttttgggg tcgaggtgcc gtaaagcac
//
