LOCUS       sequence-502286-        4948 bp    DNA     circular SYN 10-MAR-2026
DEFINITION  AiE2294m 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 4948)
  TITLE     Allen Institute AAV Enhancer Plasmids
REFERENCE   2  (bases 1 to 4948)
  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: AiP1627 - pAAV-AiE2294m-minBG-SYFP2-WPRE3-BGHpA
FEATURES             Location/Qualifiers
     source          1..4948
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     260..279
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(498..602)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     670..688
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(726..748)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     848..867
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     rep_origin      complement(884..1339)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1021..1042)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     1233..1252
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     repeat_region   1414..1554
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   1425..1554
                     /label=AAV2 ITR
     polyA_signal    1672..1783
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     1970..1990
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(join(2083..2793,2794..2796,2797..2799))
                     /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(2125..2146)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     2474..2493
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2733..2754
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     regulatory      2796..2805
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     repeat_region   3635..3764
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     rep_origin      complement(3826..4414)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3906..3925)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4585..4948,1..428,429..497))
                     /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"
ORIGIN
        1 tccggttccc aacgatcaag gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt
       61 agctccttcg gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg
      121 gttatggcag cactgcataa ttctcttact gtcatgccat ccgtaagatg cttttctgtg
      181 actggtgagt actcaaccaa gtcattctga gaatagtgta tgcggcgacc gagttgctct
      241 tgcccggcgt caatacggga taataccgcg ccacatagca gaactttaaa agtgctcatc
      301 attggaaaac gttcttcggg gcgaaaactc tcaaggatct taccgctgtt gagatccagt
      361 tcgatgtaac ccactcgtgc acccaactga tcttcagcat cttttacttt caccagcgtt
      421 tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg
      481 aaatgttgaa tactcatact cttccttttt caatattatt gaagcattta tcagggttat
      541 tgtctcatga gcggatacat atttgaatgt atttagaaaa ataaacaaat aggggttccg
      601 cgcacatttc cccgaaaagt gccacctgac gtctaagaaa ccattattat catgacatta
      661 acctataaaa ataggcgtat cacgaggccc tttcgtctcg cgcgtttcgg tgatgacggt
      721 gaaaacctct gacacatgca gctcccggag acggtcacag cttgtctgta agcggatgcc
      781 gggagcagac aagcccgtca gggcgcgtca gcgggtgttg gcgggtgtcg gggctggctt
      841 aactatgcgg catcagagca gattgtactg agagtgcacc ataaaattgt aaacgttaat
      901 attttgttaa aattcgcgtt aaatttttgt taaatcagct cattttttaa ccaatagacc
      961 gaaatcggca aaatccctta taaatcaaaa gaatagcccg agatagagtt gagtgttgtt
     1021 ccagtttgga acaagagtcc actattaaag aacgtggact ccaacgtcaa agggcgaaaa
     1081 accgtctatc agggcgatgg cccactacgt gaaccatcac ccaaatcaag ttttttgggg
     1141 tcgaggtgcc gtaaagcact aaatcggaac cctaaaggga gcccccgatt tagagcttga
     1201 cggggaaagc cggcgaacgt ggcgagaaag gaagggaaga aagcgaaagg agcgggcgct
     1261 aaggcgctgg caagtgtagc ggtcacgctg cgcgtaacca ccacacccgc cgcgcttaat
     1321 gcgccgctac agggcgcgta ctatggttgc tttgacgtat gcggtgtgaa ataccgcaca
     1381 gatgcgtaag gagaaaatac cgcatcaggc gcccctgcag gcagctgcgc gctcgctcgc
     1441 tcactgaggc cgcccgggca aagcccgggc gtcgggcgac ctttggtcgc ccggcctcag
     1501 tgagcgagcg agcgcgcaga gagggagtgg ccaactccat cactaggggt tcctgcggcc
     1561 gctcggtccg cacgtgagat ctcatgcctg ctattgtctt cccaatcctc ccccttgctg
     1621 tcctgcccca ccccaccccc cagaatagaa tgacacctac tcagacaatg cgatgcaatt
     1681 tcctcatttt attaggaaag gacagtggga gtggcacctt ccagggtcaa ggaaggcacg
     1741 ggggaggggc aaacaacaga tggctggcaa ctagaaggca cagtcgaaga tctctcgagc
     1801 cacggaattg tcagtgccca acagccgagc ccctgtccag cagcgggcaa ggcaggcggc
     1861 gatgagttcc gccgtggcaa gaactaacca ggatttatac aaggaggaga aaatgaaagc
     1921 catacgggaa gcaatagcat gatacaaagg cattaaagca gcgtatccac atagcgtaaa
     1981 aggagcaaca tagttaagaa taccagtcaa tctttcacaa attttgtaat ccagaggttg
     2041 attatgatat cgaattcgcg gccgcggcgc gccgtcgact tacttgtaca gctcgtccat
     2101 gccgagagtg atcccggcgg cggtcacgaa ctccagcagg accatgtgat cgcgcttctc
     2161 gttggggtct ttgctcagct tggactggta gctcaggtag tggttgtcgg gcagcagcac
     2221 ggggccgtcg ccgatggggg tgttctgctg gtagtggtcg gcgagctgca cgccgccgtc
     2281 ctcgatgttg tggcggatct tgaagttggc cttgatgccg ttcttctgct tgtcggcggt
     2341 gatatagacg ttgtggctgt tgtagttgta ctccagcttg tgccccagga tgttgccgtc
     2401 ctccttgaag tcgatgccct tcagctcgat gcggttcacc agggtgtcgc cctcgaactt
     2461 cacctcggcg cgggtcttgt agttgccgtc gtccttgaag aagatggtgc gctcctggac
     2521 gtagccttcg ggcatggcgg acttgaagaa gtcgtgctgc ttcatgtggt cggggtagcg
     2581 ggcgaagcac tgcacgccgt agcccagggt ggtcacgagg gtgggccagg gcacgggcag
     2641 cttgccggtg gtgcagatca gcttcagggt cagcttgccg taggtggcat cgccctcgcc
     2701 ctcgccggac acgctgaact tgtggccgtt tacgtcgccg tccagctcga ccaggatggg
     2761 caccaccccg gtgaacagct cctcgccctt gctcaccatg gtggcgaccg gtagcgctag
     2821 cttcgaaaga tctggatccc agaagcaaat gtaagcaata gatggctctg ccctgacttt
     2881 tatgcccagc ccgagctccg ccggcgtggg ctcaggaaag acagtgagat gggggcagag
     2941 gcaattccca caattgtctg aggagcagtg ctgagacaaa ctggctagaa cctctcgccc
     3001 tggagacctc ccagctcctt ctctcgggca ggactggaac cggcaggcct ggcggcgtgg
     3061 ctctgctcaa gccagcagga agaggaatga ggccaggcaa ggcgggccag ctggccagtg
     3121 gggagtcccg ggtgtgggcg caagtagttg gggaagcccc gtcggtgcct cctgggctcc
     3181 attgtgtgtg cagtgctcaa ccccaccccc caacttggcc tcggagtcct cctctataaa
     3241 ttagtcccct gagatgggat ggcctgcgtg ccactggcag gcgaggcgcc acaagagcca
     3301 gagacggcta acagatctgg ggacccaagg acttgcaact caaagaggct gagacgcgct
     3361 ggacccccag ctcacgtggg cggagcttcg aaggggcggg cgcccgtggg gcgggtcctg
     3421 agtgggggcg ggaccggggc cggcacctgg gtgaggtcgc gcagcccctc ccctttgcac
     3481 ttccgtgtgc cgcggcgcca gagcaggagg cggcggccgc ggggatcccg cgagcggccc
     3541 gtggacatct acccttcgag ccgggacccg ggaggtcccc actggcctcc ggacccgcgc
     3601 cggcgattat tactcttaca acgcgtgcgg ccgcaggaac ccctagtgat ggagttggcc
     3661 actccctctc tgcgcgctcg ctcgctcact gaggccgggc gaccaaaggt cgcccgacgc
     3721 ccgggcggcc tcagtgagcg agcgagcgcg cagctgcctg caggacatgt gagcaaaagg
     3781 ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg
     3841 cccccctgac gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg
     3901 actataaaga taccaggcgt ttccccctgg aagctccctc gtgcgctctc ctgttccgac
     3961 cctgccgctt accggatacc tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca
     4021 tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt
     4081 gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc
     4141 caacccggta agacacgact tatcgccact ggcagcagcc actggtaaca ggattagcag
     4201 agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg tggcctaact acggctacac
     4261 tagaagaaca gtatttggta tctgcgctct gctgaagcca gttaccttcg gaaaaagagt
     4321 tggtagctct tgatccggca aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa
     4381 gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat cctttgatct tttctacggg
     4441 gtctgacgct cagtggaacg aaaactcacg ttaagggatt ttggtcatga gattatcaaa
     4501 aaggatcttc acctagatcc ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat
     4561 atatgagtaa acttggtctg acagttacca atgcttaatc agtgaggcac ctatctcagc
     4621 gatctgtcta tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga taactacgat
     4681 acgggagggc ttaccatctg gccccagtgc tgcaatgata ccgcgagacc cacgctcacc
     4741 ggctccagat ttatcagcaa taaaccagcc agccggaagg gccgagcgca gaagtggtcc
     4801 tgcaacttta tccgcctcca tccagtctat taattgttgc cgggaagcta gagtaagtag
     4861 ttcgccagtt aatagtttgc gcaacgttgt tgccattgct acaggcatcg tggtgtcacg
     4921 ctcgtcgttt ggtatggctt cattcagc
//
