LOCUS       sequence-502262-        4758 bp    DNA     circular SYN 10-MAR-2026
DEFINITION  AiE2249m 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: AiP1582 - pAAV-AiE2249m-minBG-SYFP2-WPRE3-BGHpA
FEATURES             Location/Qualifiers
     source          1..4758
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      47..502
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(134..153)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     344..365
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(519..538)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     638..660
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(698..716)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        784..888
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(889..957,958..1749)
                     /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(1107..1126)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      1920..2508
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2409..2428
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     repeat_region   2570..2699
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     regulatory      3339..3348
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(3345..3347,3348..3350,3351..4061)
                     /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(3390..3411)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(3651..3670)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     3998..4019
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(4154..4174)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     polyA_signal    4361..4472
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   4590..4730
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   4590..4719
                     /label=AAV2 ITR
ORIGIN
        1 gcatctgtgc ggtatttcac accgcatacg tcaaagcaac catagtacgc gccctgtagc
       61 ggcgcattaa gcgcggcggg tgtggtggtt acgcgcagcg tgaccgctac acttgccagc
      121 gccttagcgc ccgctccttt cgctttcttc ccttcctttc tcgccacgtt cgccggcttt
      181 ccccgtcaag ctctaaatcg ggggctccct ttagggttcc gatttagtgc tttacggcac
      241 ctcgacccca aaaaacttga tttgggtgat ggttcacgta gtgggccatc gccctgatag
      301 acggtttttc gccctttgac gttggagtcc acgttcttta atagtggact cttgttccaa
      361 actggaacaa cactcaactc tatctcgggc tattcttttg atttataagg gattttgccg
      421 atttcggtct attggttaaa aaatgagctg atttaacaaa aatttaacgc gaattttaac
      481 aaaatattaa cgtttacaat tttatggtgc actctcagta caatctgctc tgatgccgca
      541 tagttaagcc agccccgaca cccgccaaca cccgctgacg cgccctgacg ggcttgtctg
      601 ctcccggcat ccgcttacag acaagctgtg accgtctccg ggagctgcat gtgtcagagg
      661 ttttcaccgt catcaccgaa acgcgcgaga cgaaagggcc tcgtgatacg cctattttta
      721 taggttaatg tcatgataat aatggtttct tagacgtcag gtggcacttt tcggggaaat
      781 gtgcgcggaa cccctatttg tttatttttc taaatacatt caaatatgta tccgctcatg
      841 agacaataac cctgataaat gcttcaataa tattgaaaaa ggaagagtat gagtattcaa
      901 catttccgtg tcgcccttat tccctttttt gcggcatttt gccttcctgt ttttgctcac
      961 ccagaaacgc tggtgaaagt aaaagatgct gaagatcagt tgggtgcacg agtgggttac
     1021 atcgaactgg atctcaacag cggtaagatc cttgagagtt ttcgccccga agaacgtttt
     1081 ccaatgatga gcacttttaa agttctgcta tgtggcgcgg tattatcccg tattgacgcc
     1141 gggcaagagc aactcggtcg ccgcatacac tattctcaga atgacttggt tgagtactca
     1201 ccagtcacag aaaagcatct tacggatggc atgacagtaa gagaattatg cagtgctgcc
     1261 ataaccatga gtgataacac tgcggccaac ttacttctga caacgatcgg aggaccgaag
     1321 gagctaaccg cttttttgca caacatgggg gatcatgtaa ctcgccttga tcgttgggaa
     1381 ccggagctga atgaagccat accaaacgac gagcgtgaca ccacgatgcc tgtagcaatg
     1441 gcaacaacgt tgcgcaaact attaactggc gaactactta ctctagcttc ccggcaacaa
     1501 ttaatagact ggatggaggc ggataaagtt gcaggaccac ttctgcgctc ggcccttccg
     1561 gctggctggt ttattgctga taaatctgga gccggtgagc gtgggtctcg cggtatcatt
     1621 gcagcactgg ggccagatgg taagccctcc cgtatcgtag ttatctacac gacggggagt
     1681 caggcaacta tggatgaacg aaatagacag atcgctgaga taggtgcctc actgattaag
     1741 cattggtaac tgtcagacca agtttactca tatatacttt agattgattt aaaacttcat
     1801 ttttaattta aaaggatcta ggtgaagatc ctttttgata atctcatgac caaaatccct
     1861 taacgtgagt tttcgttcca ctgagcgtca gaccccgtag aaaagatcaa aggatcttct
     1921 tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca
     1981 gcggtggttt gtttgccgga tcaagagcta ccaactcttt ttccgaaggt aactggcttc
     2041 agcagagcgc agataccaaa tactgttctt ctagtgtagc cgtagttagg ccaccacttc
     2101 aagaactctg tagcaccgcc tacatacctc gctctgctaa tcctgttacc agtggctgct
     2161 gccagtggcg ataagtcgtg tcttaccggg ttggactcaa gacgatagtt accggataag
     2221 gcgcagcggt cgggctgaac ggggggttcg tgcacacagc ccagcttgga gcgaacgacc
     2281 tacaccgaac tgagatacct acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg
     2341 agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa caggagagcg cacgagggag
     2401 cttccagggg gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca cctctgactt
     2461 gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac
     2521 gcggcctttt tacggttcct ggccttttgc tggccttttg ctcacatgtc ctgcaggcag
     2581 ctgcgcgctc gctcgctcac tgaggccgcc cgggcgtcgg gcgacctttg gtcgcccggc
     2641 ctcagtgagc gagcgagcgc gcagagaggg agtggccaac tccatcacta ggggttcctg
     2701 cggccgcacg cgttgtaaga gtaataatcg ccggcgctaa gttccccttg cagctacctt
     2761 gggaatggtg gaatacaggt agcaagaaag gcattgcact gcagcgttcc acaccttgca
     2821 ttaagaactg cagagagaaa ggccattctc tgaagactgc ttaactggcc tcatgccacg
     2881 tctccctctg aacttcttac cattctctct gcagttcccc acccccacta taaaatgaat
     2941 gggttagagc agagaagtta ttttgatctt tagttttgag tccttgtcct tggctaggat
     3001 cccaaatatc aaggtgtact ggctccagag gagaggtgaa agcctctctc cgatgcgtca
     3061 gcagtggctc agcacacagc cttagagttc cggctccagg aaggaggatg acaaccaata
     3121 gctgttggca tcctttttaa acatttgtgt ccctcattta agatttatat acaaattact
     3181 catctatggg acgaacaaaa gggaatgctt tgataaaaga tgggatccga tccttaacgc
     3241 cggcggagct cgggctgggc ataaaagtca gggcagagcc atctattgct tacatttgct
     3301 tctgggatcc agatctttcg aagctagcgc taccggtcgc caccatggtg agcaagggcg
     3361 aggagctgtt caccggggtg gtgcccatcc tggtcgagct ggacggcgac gtaaacggcc
     3421 acaagttcag cgtgtccggc gagggcgagg gcgatgccac ctacggcaag ctgaccctga
     3481 agctgatctg caccaccggc aagctgcccg tgccctggcc caccctcgtg accaccctgg
     3541 gctacggcgt gcagtgcttc gcccgctacc ccgaccacat gaagcagcac gacttcttca
     3601 agtccgccat gcccgaaggc tacgtccagg agcgcaccat cttcttcaag gacgacggca
     3661 actacaagac ccgcgccgag gtgaagttcg agggcgacac cctggtgaac cgcatcgagc
     3721 tgaagggcat cgacttcaag gaggacggca acatcctggg gcacaagctg gagtacaact
     3781 acaacagcca caacgtctat atcaccgccg acaagcagaa gaacggcatc aaggccaact
     3841 tcaagatccg ccacaacatc gaggacggcg gcgtgcagct cgccgaccac taccagcaga
     3901 acacccccat cggcgacggc cccgtgctgc tgcccgacaa ccactacctg agctaccagt
     3961 ccaagctgag caaagacccc aacgagaagc gcgatcacat ggtcctgctg gagttcgtga
     4021 ccgccgccgg gatcactctc ggcatggacg agctgtacaa gtaagtcgac ggcgcgccgc
     4081 ggccgcgaat tcgatatcat aatcaacctc tggattacaa aatttgtgaa agattgactg
     4141 gtattcttaa ctatgttgct ccttttacgc tatgtggata cgctgcttta atgcctttgt
     4201 atcatgctat tgcttcccgt atggctttca ttttctcctc cttgtataaa tcctggttag
     4261 ttcttgccac ggcggaactc atcgccgcct gccttgcccg ctgctggaca ggggctcggc
     4321 tgttgggcac tgacaattcc gtggctcgag agatcttcga ctgtgccttc tagttgccag
     4381 ccatctgttg tttgcccctc ccccgtgcct tccttgaccc tggaaggtgc cactcccact
     4441 gtcctttcct aataaaatga ggaaattgca tcgcattgtc tgagtaggtg tcattctatt
     4501 ctggggggtg gggtggggca ggacagcaag ggggaggatt gggaagacaa tagcaggcat
     4561 gagatctcac gtgcggaccg agcggccgca ggaaccccta gtgatggagt tggccactcc
     4621 ctctctgcgc gctcgctcgc tcactgaggc cgggcgacca aaggtcgccc gacgcccggg
     4681 ctttgcccgg gcggcctcag tgagcgagcg agcgcgcagc tgcctgcagg ggcgcctgat
     4741 gcggtatttt ctccttac
//
