LOCUS       sequence-502249-        4973 bp    DNA     circular SYN 10-MAR-2026
DEFINITION  AiE2230m 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 4973)
  TITLE     Allen Institute AAV Enhancer Plasmids
REFERENCE   2  (bases 1 to 4973)
  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: AiP1563 - pAAV-AiE2230m-minBG-SYFP2-WPRE3-BGHpA
FEATURES             Location/Qualifiers
     source          1..4973
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   221..350
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     rep_origin      complement(412..1000)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(492..511)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(1171..1962,1963..2031))
                     /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     1794..1813
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(2032..2136)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     2204..2222
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(2260..2282)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     2382..2401
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     rep_origin      complement(2418..2873)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2555..2576)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2767..2786
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     repeat_region   2948..3088
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   2959..3088
                     /label=AAV2 ITR
     polyA_signal    3206..3317
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     3504..3524
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(join(3617..4327,4328..4330,4331..4333))
                     /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(3659..3680)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     4008..4027
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     4267..4288
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     regulatory      4330..4339
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
ORIGIN
        1 tgaaatgctg aactacagag gatggcttcc tgctaggcta gatcagcgtg aaggggtagg
       61 gatgtaaaag acagaaggat ggtttctgag cttgtggcag tatgtcactg tttggagtgg
      121 gcactgtctg ctttctgctt tggttccaat aggtacaaag gtaccacgtt tctgctgttt
      181 gagcgccggc gattattact cttacaacgc gtgcggccgc aggaacccct agtgatggag
      241 ttggccactc cctctctgcg cgctcgctcg ctcactgagg ccgggcgacc aaaggtcgcc
      301 cgacgcccgg gcggcctcag tgagcgagcg agcgcgcagc tgcctgcagg acatgtgagc
      361 aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag
      421 gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc
      481 gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt
      541 tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct
      601 ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg
      661 ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct
      721 tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat
      781 tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg
      841 ctacactaga agaacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa
      901 aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt
      961 ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc
     1021 tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt
     1081 atcaaaaagg atcttcacct agatcctttt aaattaaaaa tgaagtttta aatcaatcta
     1141 aagtatatat gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg aggcacctat
     1201 ctcagcgatc tgtctatttc gttcatccat agttgcctga ctccccgtcg tgtagataac
     1261 tacgatacgg gagggcttac catctggccc cagtgctgca atgataccgc gagacccacg
     1321 ctcaccggct ccagatttat cagcaataaa ccagccagcc ggaagggccg agcgcagaag
     1381 tggtcctgca actttatccg cctccatcca gtctattaat tgttgccggg aagctagagt
     1441 aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt
     1501 gtcacgctcg tcgtttggta tggcttcatt cagctccggt tcccaacgat caaggcgagt
     1561 tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt
     1621 cagaagtaag ttggccgcag tgttatcact catggttatg gcagcactgc ataattctct
     1681 tactgtcatg ccatccgtaa gatgcttttc tgtgactggt gagtactcaa ccaagtcatt
     1741 ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac gggataatac
     1801 cgcgccacat agcagaactt taaaagtgct catcattgga aaacgttctt cggggcgaaa
     1861 actctcaagg atcttaccgc tgttgagatc cagttcgatg taacccactc gtgcacccaa
     1921 ctgatcttca gcatctttta ctttcaccag cgtttctggg tgagcaaaaa caggaaggca
     1981 aaatgccgca aaaaagggaa taagggcgac acggaaatgt tgaatactca tactcttcct
     2041 ttttcaatat tattgaagca tttatcaggg ttattgtctc atgagcggat acatatttga
     2101 atgtatttag aaaaataaac aaataggggt tccgcgcaca tttccccgaa aagtgccacc
     2161 tgacgtctaa gaaaccatta ttatcatgac attaacctat aaaaataggc gtatcacgag
     2221 gccctttcgt ctcgcgcgtt tcggtgatga cggtgaaaac ctctgacaca tgcagctccc
     2281 ggagacggtc acagcttgtc tgtaagcgga tgccgggagc agacaagccc gtcagggcgc
     2341 gtcagcgggt gttggcgggt gtcggggctg gcttaactat gcggcatcag agcagattgt
     2401 actgagagtg caccataaaa ttgtaaacgt taatattttg ttaaaattcg cgttaaattt
     2461 ttgttaaatc agctcatttt ttaaccaata gaccgaaatc ggcaaaatcc cttataaatc
     2521 aaaagaatag cccgagatag agttgagtgt tgttccagtt tggaacaaga gtccactatt
     2581 aaagaacgtg gactccaacg tcaaagggcg aaaaaccgtc tatcagggcg atggcccact
     2641 acgtgaacca tcacccaaat caagtttttt ggggtcgagg tgccgtaaag cactaaatcg
     2701 gaaccctaaa gggagccccc gatttagagc ttgacgggga aagccggcga acgtggcgag
     2761 aaaggaaggg aagaaagcga aaggagcggg cgctaaggcg ctggcaagtg tagcggtcac
     2821 gctgcgcgta accaccacac ccgccgcgct taatgcgccg ctacagggcg cgtactatgg
     2881 ttgctttgac gtatgcggtg tgaaataccg cacagatgcg taaggagaaa ataccgcatc
     2941 aggcgcccct gcaggcagct gcgcgctcgc tcgctcactg aggccgcccg ggcaaagccc
     3001 gggcgtcggg cgacctttgg tcgcccggcc tcagtgagcg agcgagcgcg cagagaggga
     3061 gtggccaact ccatcactag gggttcctgc ggccgctcgg tccgcacgtg agatctcatg
     3121 cctgctattg tcttcccaat cctccccctt gctgtcctgc cccaccccac cccccagaat
     3181 agaatgacac ctactcagac aatgcgatgc aatttcctca ttttattagg aaaggacagt
     3241 gggagtggca ccttccaggg tcaaggaagg cacgggggag gggcaaacaa cagatggctg
     3301 gcaactagaa ggcacagtcg aagatctctc gagccacgga attgtcagtg cccaacagcc
     3361 gagcccctgt ccagcagcgg gcaaggcagg cggcgatgag ttccgccgtg gcaagaacta
     3421 accaggattt atacaaggag gagaaaatga aagccatacg ggaagcaata gcatgataca
     3481 aaggcattaa agcagcgtat ccacatagcg taaaaggagc aacatagtta agaataccag
     3541 tcaatctttc acaaattttg taatccagag gttgattatg atatcgaatt cgcggccgcg
     3601 gcgcgccgtc gacttacttg tacagctcgt ccatgccgag agtgatcccg gcggcggtca
     3661 cgaactccag caggaccatg tgatcgcgct tctcgttggg gtctttgctc agcttggact
     3721 ggtagctcag gtagtggttg tcgggcagca gcacggggcc gtcgccgatg ggggtgttct
     3781 gctggtagtg gtcggcgagc tgcacgccgc cgtcctcgat gttgtggcgg atcttgaagt
     3841 tggccttgat gccgttcttc tgcttgtcgg cggtgatata gacgttgtgg ctgttgtagt
     3901 tgtactccag cttgtgcccc aggatgttgc cgtcctcctt gaagtcgatg cccttcagct
     3961 cgatgcggtt caccagggtg tcgccctcga acttcacctc ggcgcgggtc ttgtagttgc
     4021 cgtcgtcctt gaagaagatg gtgcgctcct ggacgtagcc ttcgggcatg gcggacttga
     4081 agaagtcgtg ctgcttcatg tggtcggggt agcgggcgaa gcactgcacg ccgtagccca
     4141 gggtggtcac gagggtgggc cagggcacgg gcagcttgcc ggtggtgcag atcagcttca
     4201 gggtcagctt gccgtaggtg gcatcgccct cgccctcgcc ggacacgctg aacttgtggc
     4261 cgtttacgtc gccgtccagc tcgaccagga tgggcaccac cccggtgaac agctcctcgc
     4321 ccttgctcac catggtggcg accggtagcg ctagcttcga aagatctgga tcccagaagc
     4381 aaatgtaagc aatagatggc tctgccctga cttttatgcc cagcccgagc tccgccggcg
     4441 cactgttatg agtaagcttg cagacagaaa gctttggtca cgtggctgcc tcactttcca
     4501 tggttaagga agtgagcatc tagggagacc cagaggcaag tcaagagtct ccaaagcctg
     4561 cccattccct atgcccagga ctgtgactgt cctgctgagt gccagagaca ccacacagcg
     4621 gttccacttg acagcgacca tcaactacca ccatgtctgc tgtaccatgg actgaggcac
     4681 agatgtgagc tcggtcttca ttctccccga ggcctcttgc ttaccaactc aatcaggatt
     4741 tctataactg aacagcccaa caccgccaga gcctggaaca tcaggaaagc ggtctgctct
     4801 gactgccctt atccaggcct ttcatctgac tgtttcaaag tgtcctttcc acattaatta
     4861 agtgatgaat taatcaacaa agtcattggc taattggctt tcatatctgg aagctctttc
     4921 ataaatgtca ggaaaaatga cattttcaac gtttaaagat acactcccca atc
//
