LOCUS       sequence-492120-        4858 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AiE2656m 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 4858)
  TITLE     Basal Ganglia/CERP Enhancer AAVs
REFERENCE   2  (bases 1 to 4858)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"
COMMENT     Sequence Label: AiP2129 - pAAV-AiE2656m-minBG-SYFP2-WPRE3-BGHpA
FEATURES             Location/Qualifiers
     source          1..4858
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   489..618
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     rep_origin      complement(680..1268)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(760..779)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(1439..2230,2231..2299))
                     /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     2062..2081
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(2300..2404)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     2472..2490
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(2528..2550)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     2650..2669
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     rep_origin      complement(2686..3141)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2823..2844)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3035..3054
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     repeat_region   3216..3356
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   3227..3356
                     /label=AAV2 ITR
     polyA_signal    3474..3585
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     3772..3792
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(join(3885..4595,4596..4598,4599..4601))
                     /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(3927..3948)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     4276..4295
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     4535..4556
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     regulatory      4598..4607
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
ORIGIN
        1 atcaatattt gtcctattca tcctgcataa atattttctc agacaagtag ctatttacca
       61 tattgagtca acacagactg caagaatata agaaggttgt ctggaaggtt ctctttcccc
      121 aagggtctgg taacacaacc aacaggagga taacagacat tgatcgagtg cccatcatga
      181 gacacaatgc caacatttac atatattaca gatgtcatcc tcataaacac cccagcagag
      241 atgctcttat tcaccttacc ttaaggtgaa agtcctggga ttccgaaagg cccagcaatt
      301 ctcattttta aatgctttgt tttcccatgt caatggacaa agcatttcta tctcctatga
      361 cttctcagtg tccttgagtt tccctctccc cttggccttt ctttatctgt agcatagtgc
      421 tggatactct tttctctaat cctgatgacc tcatttaggg tgatggcttt caatacgcgt
      481 gcggccgcag gaacccctag tgatggagtt ggccactccc tctctgcgcg ctcgctcgct
      541 cactgaggcc gggcgaccaa aggtcgcccg acgcccgggc ggcctcagtg agcgagcgag
      601 cgcgcagctg cctgcaggac atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa
      661 aggccgcgtt gctggcgttt ttccataggc tccgcccccc tgacgagcat cacaaaaatc
      721 gacgctcaag tcagaggtgg cgaaacccga caggactata aagataccag gcgtttcccc
      781 ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga tacctgtccg
      841 cctttctccc ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg tatctcagtt
      901 cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga accccccgtt cagcccgacc
      961 gctgcgcctt atccggtaac tatcgtcttg agtccaaccc ggtaagacac gacttatcgc
     1021 cactggcagc agccactggt aacaggatta gcagagcgag gtatgtaggc ggtgctacag
     1081 agttcttgaa gtggtggcct aactacggct acactagaag aacagtattt ggtatctgcg
     1141 ctctgctgaa gccagttacc ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa
     1201 ccaccgctgg tagcggtggt ttttttgttt gcaagcagca gattacgcgc agaaaaaaag
     1261 gatctcaaga agatcctttg atcttttcta cggggtctga cgctcagtgg aacgaaaact
     1321 cacgttaagg gattttggtc atgagattat caaaaaggat cttcacctag atccttttaa
     1381 attaaaaatg aagttttaaa tcaatctaaa gtatatatga gtaaacttgg tctgacagtt
     1441 accaatgctt aatcagtgag gcacctatct cagcgatctg tctatttcgt tcatccatag
     1501 ttgcctgact ccccgtcgtg tagataacta cgatacggga gggcttacca tctggcccca
     1561 gtgctgcaat gataccgcga gacccacgct caccggctcc agatttatca gcaataaacc
     1621 agccagccgg aagggccgag cgcagaagtg gtcctgcaac tttatccgcc tccatccagt
     1681 ctattaattg ttgccgggaa gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg
     1741 ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg gcttcattca
     1801 gctccggttc ccaacgatca aggcgagtta catgatcccc catgttgtgc aaaaaagcgg
     1861 ttagctcctt cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg ttatcactca
     1921 tggttatggc agcactgcat aattctctta ctgtcatgcc atccgtaaga tgcttttctg
     1981 tgactggtga gtactcaacc aagtcattct gagaatagtg tatgcggcga ccgagttgct
     2041 cttgcccggc gtcaatacgg gataataccg cgccacatag cagaacttta aaagtgctca
     2101 tcattggaaa acgttcttcg gggcgaaaac tctcaaggat cttaccgctg ttgagatcca
     2161 gttcgatgta acccactcgt gcacccaact gatcttcagc atcttttact ttcaccagcg
     2221 tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata agggcgacac
     2281 ggaaatgttg aatactcata ctcttccttt ttcaatatta ttgaagcatt tatcagggtt
     2341 attgtctcat gagcggatac atatttgaat gtatttagaa aaataaacaa ataggggttc
     2401 cgcgcacatt tccccgaaaa gtgccacctg acgtctaaga aaccattatt atcatgacat
     2461 taacctataa aaataggcgt atcacgaggc cctttcgtct cgcgcgtttc ggtgatgacg
     2521 gtgaaaacct ctgacacatg cagctcccgg agacggtcac agcttgtctg taagcggatg
     2581 ccgggagcag acaagcccgt cagggcgcgt cagcgggtgt tggcgggtgt cggggctggc
     2641 ttaactatgc ggcatcagag cagattgtac tgagagtgca ccataaaatt gtaaacgtta
     2701 atattttgtt aaaattcgcg ttaaattttt gttaaatcag ctcatttttt aaccaataga
     2761 ccgaaatcgg caaaatccct tataaatcaa aagaatagcc cgagatagag ttgagtgttg
     2821 ttccagtttg gaacaagagt ccactattaa agaacgtgga ctccaacgtc aaagggcgaa
     2881 aaaccgtcta tcagggcgat ggcccactac gtgaaccatc acccaaatca agttttttgg
     2941 ggtcgaggtg ccgtaaagca ctaaatcgga accctaaagg gagcccccga tttagagctt
     3001 gacggggaaa gccggcgaac gtggcgagaa aggaagggaa gaaagcgaaa ggagcgggcg
     3061 ctaaggcgct ggcaagtgta gcggtcacgc tgcgcgtaac caccacaccc gccgcgctta
     3121 atgcgccgct acagggcgcg tactatggtt gctttgacgt atgcggtgtg aaataccgca
     3181 cagatgcgta aggagaaaat accgcatcag gcgcccctgc aggcagctgc gcgctcgctc
     3241 gctcactgag gccgcccggg caaagcccgg gcgtcgggcg acctttggtc gcccggcctc
     3301 agtgagcgag cgagcgcgca gagagggagt ggccaactcc atcactaggg gttcctgcgg
     3361 ccgctcggtc cgcacgtgag atctcatgcc tgctattgtc ttcccaatcc tcccccttgc
     3421 tgtcctgccc caccccaccc cccagaatag aatgacacct actcagacaa tgcgatgcaa
     3481 tttcctcatt ttattaggaa aggacagtgg gagtggcacc ttccagggtc aaggaaggca
     3541 cgggggaggg gcaaacaaca gatggctggc aactagaagg cacagtcgaa gatctctcga
     3601 gccacggaat tgtcagtgcc caacagccga gcccctgtcc agcagcgggc aaggcaggcg
     3661 gcgatgagtt ccgccgtggc aagaactaac caggatttat acaaggagga gaaaatgaaa
     3721 gccatacggg aagcaatagc atgatacaaa ggcattaaag cagcgtatcc acatagcgta
     3781 aaaggagcaa catagttaag aataccagtc aatctttcac aaattttgta atccagaggt
     3841 tgattatgat atcgaattcg cggccgcggc gcgccgtcga cttacttgta cagctcgtcc
     3901 atgccgagag tgatcccggc ggcggtcacg aactccagca ggaccatgtg atcgcgcttc
     3961 tcgttggggt ctttgctcag cttggactgg tagctcaggt agtggttgtc gggcagcagc
     4021 acggggccgt cgccgatggg ggtgttctgc tggtagtggt cggcgagctg cacgccgccg
     4081 tcctcgatgt tgtggcggat cttgaagttg gccttgatgc cgttcttctg cttgtcggcg
     4141 gtgatataga cgttgtggct gttgtagttg tactccagct tgtgccccag gatgttgccg
     4201 tcctccttga agtcgatgcc cttcagctcg atgcggttca ccagggtgtc gccctcgaac
     4261 ttcacctcgg cgcgggtctt gtagttgccg tcgtccttga agaagatggt gcgctcctgg
     4321 acgtagcctt cgggcatggc ggacttgaag aagtcgtgct gcttcatgtg gtcggggtag
     4381 cgggcgaagc actgcacgcc gtagcccagg gtggtcacga gggtgggcca gggcacgggc
     4441 agcttgccgg tggtgcagat cagcttcagg gtcagcttgc cgtaggtggc atcgccctcg
     4501 ccctcgccgg acacgctgaa cttgtggccg tttacgtcgc cgtccagctc gaccaggatg
     4561 ggcaccaccc cggtgaacag ctcctcgccc ttgctcacca tggtggcgac cggtagcgct
     4621 agcttcgaaa gatctggatc ccagaagcaa atgtaagcaa tagatggctc tgccctgact
     4681 tttatgccca gcccgagctc cgccggcgag tcctttattc atttgttcac agcacaagtg
     4741 ttaacacatt aattactaag acagggttga gagagtgagt agcacagtta ctgtttctaa
     4801 aattcatgtg cggagtctcc cttgtccttg agttatatgg atgccagggt agaacaaa
//
