LOCUS       sequence-502248-        4758 bp    DNA     circular SYN 10-MAR-2026
DEFINITION  AiE2229m is an enhancer sequence, designed to drive AAV-mediated 
            transgene expression in specific populations of brain cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4758)
  TITLE     Allen Institute AAV Enhancer Plasmids
REFERENCE   2  (bases 1 to 4758)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Mar 10, 2026 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"
COMMENT     Sequence Label: AiP1562 - pAAV-AiE2229m-minBG-SYFP2-WPRE3-BGHpA
FEATURES             Location/Qualifiers
     source          1..4758
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      473..1061
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     962..981
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     repeat_region   1123..1252
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     regulatory      1892..1901
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(1898..1900,1901..1903,1904..2614)
                     /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(1943..1964)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2204..2223)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2551..2572
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(2707..2727)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     polyA_signal    2914..3025
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   3143..3283
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   3143..3272
                     /label=AAV2 ITR
     rep_origin      3358..3813
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3445..3464)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3655..3676
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3830..3849)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3949..3971
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(4009..4027)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4095..4199
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4200..4268,4269..4758,1..302)
                     /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(4418..4437)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
ORIGIN
        1 cgttgcgcaa actattaact ggcgaactac ttactctagc ttcccggcaa caattaatag
       61 actggatgga ggcggataaa gttgcaggac cacttctgcg ctcggccctt ccggctggct
      121 ggtttattgc tgataaatct ggagccggtg agcgtgggtc tcgcggtatc attgcagcac
      181 tggggccaga tggtaagccc tcccgtatcg tagttatcta cacgacgggg agtcaggcaa
      241 ctatggatga acgaaataga cagatcgctg agataggtgc ctcactgatt aagcattggt
      301 aactgtcaga ccaagtttac tcatatatac tttagattga tttaaaactt catttttaat
      361 ttaaaaggat ctaggtgaag atcctttttg ataatctcat gaccaaaatc ccttaacgtg
      421 agttttcgtt ccactgagcg tcagaccccg tagaaaagat caaaggatct tcttgagatc
      481 ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg
      541 tttgtttgcc ggatcaagag ctaccaactc tttttccgaa ggtaactggc ttcagcagag
      601 cgcagatacc aaatactgtt cttctagtgt agccgtagtt aggccaccac ttcaagaact
      661 ctgtagcacc gcctacatac ctcgctctgc taatcctgtt accagtggct gctgccagtg
      721 gcgataagtc gtgtcttacc gggttggact caagacgata gttaccggat aaggcgcagc
      781 ggtcgggctg aacggggggt tcgtgcacac agcccagctt ggagcgaacg acctacaccg
      841 aactgagata cctacagcgt gagctatgag aaagcgccac gcttcccgaa gggagaaagg
      901 cggacaggta tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg gagcttccag
      961 ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg ccacctctga cttgagcgtc
     1021 gatttttgtg atgctcgtca ggggggcgga gcctatggaa aaacgccagc aacgcggcct
     1081 ttttacggtt cctggccttt tgctggcctt ttgctcacat gtcctgcagg cagctgcgcg
     1141 ctcgctcgct cactgaggcc gcccgggcgt cgggcgacct ttggtcgccc ggcctcagtg
     1201 agcgagcgag cgcgcagaga gggagtggcc aactccatca ctaggggttc ctgcggccgc
     1261 acgcgttgta agagtaataa tcgccggcgg gaagggttgg aaaacctgag cctgaagtcc
     1321 agagtgggac tgtcccaaag gcaccagaaa gccaccacag ctgcaggctg agcaaaggcc
     1381 ccagatctct ttagcccttg ctgtgtggag cgagtagatg agaagtccct gctaaccccc
     1441 gctgacccct gccacccagc gccttctata aatagtctca ccagatggca ggtgtggaga
     1501 agccagggct ccctcgtagg cggtctcctc tgcccgctct aatcccctgt atcaaaggca
     1561 agaggctcag gtcccctgtg gtcaccacag ccatgacctg tgcctacaga ggtgtggggt
     1621 gatggaggag aggagggctg tagggagaga ggctctatgg gtaagggtga agccagatgg
     1681 cagggcctgg gctgtgaagg gcagcccttc ctatggtcgc cctgctcctt ctacttggac
     1741 ccctgcacca tcacccaagg cagctggttc tcagacccct tcacccctcc cgccggcgga
     1801 gctcgggctg ggcataaaag tcagggcaga gccatctatt gcttacattt gcttctggga
     1861 tccagatctt tcgaagctag cgctaccggt cgccaccatg gtgagcaagg gcgaggagct
     1921 gttcaccggg gtggtgccca tcctggtcga gctggacggc gacgtaaacg gccacaagtt
     1981 cagcgtgtcc ggcgagggcg agggcgatgc cacctacggc aagctgaccc tgaagctgat
     2041 ctgcaccacc ggcaagctgc ccgtgccctg gcccaccctc gtgaccaccc tgggctacgg
     2101 cgtgcagtgc ttcgcccgct accccgacca catgaagcag cacgacttct tcaagtccgc
     2161 catgcccgaa ggctacgtcc aggagcgcac catcttcttc aaggacgacg gcaactacaa
     2221 gacccgcgcc gaggtgaagt tcgagggcga caccctggtg aaccgcatcg agctgaaggg
     2281 catcgacttc aaggaggacg gcaacatcct ggggcacaag ctggagtaca actacaacag
     2341 ccacaacgtc tatatcaccg ccgacaagca gaagaacggc atcaaggcca acttcaagat
     2401 ccgccacaac atcgaggacg gcggcgtgca gctcgccgac cactaccagc agaacacccc
     2461 catcggcgac ggccccgtgc tgctgcccga caaccactac ctgagctacc agtccaagct
     2521 gagcaaagac cccaacgaga agcgcgatca catggtcctg ctggagttcg tgaccgccgc
     2581 cgggatcact ctcggcatgg acgagctgta caagtaagtc gacggcgcgc cgcggccgcg
     2641 aattcgatat cataatcaac ctctggatta caaaatttgt gaaagattga ctggtattct
     2701 taactatgtt gctcctttta cgctatgtgg atacgctgct ttaatgcctt tgtatcatgc
     2761 tattgcttcc cgtatggctt tcattttctc ctccttgtat aaatcctggt tagttcttgc
     2821 cacggcggaa ctcatcgccg cctgccttgc ccgctgctgg acaggggctc ggctgttggg
     2881 cactgacaat tccgtggctc gagagatctt cgactgtgcc ttctagttgc cagccatctg
     2941 ttgtttgccc ctcccccgtg ccttccttga ccctggaagg tgccactccc actgtccttt
     3001 cctaataaaa tgaggaaatt gcatcgcatt gtctgagtag gtgtcattct attctggggg
     3061 gtggggtggg gcaggacagc aagggggagg attgggaaga caatagcagg catgagatct
     3121 cacgtgcgga ccgagcggcc gcaggaaccc ctagtgatgg agttggccac tccctctctg
     3181 cgcgctcgct cgctcactga ggccgggcga ccaaaggtcg cccgacgccc gggctttgcc
     3241 cgggcggcct cagtgagcga gcgagcgcgc agctgcctgc aggggcgcct gatgcggtat
     3301 tttctcctta cgcatctgtg cggtatttca caccgcatac gtcaaagcaa ccatagtacg
     3361 cgccctgtag cggcgcatta agcgcggcgg gtgtggtggt tacgcgcagc gtgaccgcta
     3421 cacttgccag cgccttagcg cccgctcctt tcgctttctt cccttccttt ctcgccacgt
     3481 tcgccggctt tccccgtcaa gctctaaatc gggggctccc tttagggttc cgatttagtg
     3541 ctttacggca cctcgacccc aaaaaacttg atttgggtga tggttcacgt agtgggccat
     3601 cgccctgata gacggttttt cgccctttga cgttggagtc cacgttcttt aatagtggac
     3661 tcttgttcca aactggaaca acactcaact ctatctcggg ctattctttt gatttataag
     3721 ggattttgcc gatttcggtc tattggttaa aaaatgagct gatttaacaa aaatttaacg
     3781 cgaattttaa caaaatatta acgtttacaa ttttatggtg cactctcagt acaatctgct
     3841 ctgatgccgc atagttaagc cagccccgac acccgccaac acccgctgac gcgccctgac
     3901 gggcttgtct gctcccggca tccgcttaca gacaagctgt gaccgtctcc gggagctgca
     3961 tgtgtcagag gttttcaccg tcatcaccga aacgcgcgag acgaaagggc ctcgtgatac
     4021 gcctattttt ataggttaat gtcatgataa taatggtttc ttagacgtca ggtggcactt
     4081 ttcggggaaa tgtgcgcgga acccctattt gtttattttt ctaaatacat tcaaatatgt
     4141 atccgctcat gagacaataa ccctgataaa tgcttcaata atattgaaaa aggaagagta
     4201 tgagtattca acatttccgt gtcgccctta ttcccttttt tgcggcattt tgccttcctg
     4261 tttttgctca cccagaaacg ctggtgaaag taaaagatgc tgaagatcag ttgggtgcac
     4321 gagtgggtta catcgaactg gatctcaaca gcggtaagat ccttgagagt tttcgccccg
     4381 aagaacgttt tccaatgatg agcactttta aagttctgct atgtggcgcg gtattatccc
     4441 gtattgacgc cgggcaagag caactcggtc gccgcataca ctattctcag aatgacttgg
     4501 ttgagtactc accagtcaca gaaaagcatc ttacggatgg catgacagta agagaattat
     4561 gcagtgctgc cataaccatg agtgataaca ctgcggccaa cttacttctg acaacgatcg
     4621 gaggaccgaa ggagctaacc gcttttttgc acaacatggg ggatcatgta actcgccttg
     4681 atcgttggga accggagctg aatgaagcca taccaaacga cgagcgtgac accacgatgc
     4741 ctgtagcaat ggcaacaa
//
