LOCUS       sequence-502277-        5578 bp    DNA     circular SYN 10-MAR-2026
DEFINITION  AiE2266m 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 5578)
  TITLE     Allen Institute AAV Enhancer Plasmids
REFERENCE   2  (bases 1 to 5578)
  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: AiP1599 - pAAV-AiE2266m-minBG-SYFP2-WPRE3-BGHpA
FEATURES             Location/Qualifiers
     source          1..5578
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   651..780
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     rep_origin      complement(842..1430)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(922..941)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(1601..2392,2393..2461))
                     /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     2224..2243
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(2462..2566)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     2634..2652
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(2690..2712)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     2812..2831
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     rep_origin      complement(2848..3303)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2985..3006)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3197..3216
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     repeat_region   3378..3518
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   3389..3518
                     /label=AAV2 ITR
     polyA_signal    3636..3747
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     3934..3954
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(join(4047..4757,4758..4760,4761..4763))
                     /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(4089..4110)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     4438..4457
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     4697..4718
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     regulatory      4760..4769
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
ORIGIN
        1 ctagccaaac tcacgaggag aggagctatt ggttcccgtt ctcccctgcc cgcaccctct
       61 gcttgctcca aatgccaagt tagaccagga agttagaccc tttgtagctg agatgggagg
      121 gatgagacaa ctggtccagg gaaccaaaag gacagcagaa caagatccca cctctaccct
      181 cctacaccca gtttgcttct gggacccaac tgacaaatca gttgacttag actttcaagg
      241 tctctgtctg tcatttgatt ttctttgggg tgagagggta tgttttagag gagacctttg
      301 cccagaaagc tgatagttga ttggcagttc ctaagagcat tgcctgaccc tgctttcaat
      361 ttgtctacag cgccctgacc agggttctag gaggggataa aattagaatc ccttctaaga
      421 gccaggctcc ggggccttaa ggacatccta gcgaactcac agaagatcct ggggtctgat
      481 gtattgggct tttgaaaacc tcataaagct aaaacatact agcaggctcc caaaagccca
      541 aattcccacc ctggggagcc ctcatcagta tccctggtcc tccaccactg gctgcacccc
      601 atcttgttta gaccgccggc gattattact cttacaacgc gtgcggccgc aggaacccct
      661 agtgatggag ttggccactc cctctctgcg cgctcgctcg ctcactgagg ccgggcgacc
      721 aaaggtcgcc cgacgcccgg gcggcctcag tgagcgagcg agcgcgcagc tgcctgcagg
      781 acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt
      841 ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt
      901 ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc
      961 gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa
     1021 gcgtggcgct ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct
     1081 ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta
     1141 actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg
     1201 gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc
     1261 ctaactacgg ctacactaga agaacagtat ttggtatctg cgctctgctg aagccagtta
     1321 ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg
     1381 gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt
     1441 tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg
     1501 tcatgagatt atcaaaaagg atcttcacct agatcctttt aaattaaaaa tgaagtttta
     1561 aatcaatcta aagtatatat gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg
     1621 aggcacctat ctcagcgatc tgtctatttc gttcatccat agttgcctga ctccccgtcg
     1681 tgtagataac tacgatacgg gagggcttac catctggccc cagtgctgca atgataccgc
     1741 gagacccacg ctcaccggct ccagatttat cagcaataaa ccagccagcc ggaagggccg
     1801 agcgcagaag tggtcctgca actttatccg cctccatcca gtctattaat tgttgccggg
     1861 aagctagagt aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc attgctacag
     1921 gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt cagctccggt tcccaacgat
     1981 caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc
     2041 cgatcgttgt cagaagtaag ttggccgcag tgttatcact catggttatg gcagcactgc
     2101 ataattctct tactgtcatg ccatccgtaa gatgcttttc tgtgactggt gagtactcaa
     2161 ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac
     2221 gggataatac cgcgccacat agcagaactt taaaagtgct catcattgga aaacgttctt
     2281 cggggcgaaa actctcaagg atcttaccgc tgttgagatc cagttcgatg taacccactc
     2341 gtgcacccaa ctgatcttca gcatctttta ctttcaccag cgtttctggg tgagcaaaaa
     2401 caggaaggca aaatgccgca aaaaagggaa taagggcgac acggaaatgt tgaatactca
     2461 tactcttcct ttttcaatat tattgaagca tttatcaggg ttattgtctc atgagcggat
     2521 acatatttga atgtatttag aaaaataaac aaataggggt tccgcgcaca tttccccgaa
     2581 aagtgccacc tgacgtctaa gaaaccatta ttatcatgac attaacctat aaaaataggc
     2641 gtatcacgag gccctttcgt ctcgcgcgtt tcggtgatga cggtgaaaac ctctgacaca
     2701 tgcagctccc ggagacggtc acagcttgtc tgtaagcgga tgccgggagc agacaagccc
     2761 gtcagggcgc gtcagcgggt gttggcgggt gtcggggctg gcttaactat gcggcatcag
     2821 agcagattgt actgagagtg caccataaaa ttgtaaacgt taatattttg ttaaaattcg
     2881 cgttaaattt ttgttaaatc agctcatttt ttaaccaata gaccgaaatc ggcaaaatcc
     2941 cttataaatc aaaagaatag cccgagatag agttgagtgt tgttccagtt tggaacaaga
     3001 gtccactatt aaagaacgtg gactccaacg tcaaagggcg aaaaaccgtc tatcagggcg
     3061 atggcccact acgtgaacca tcacccaaat caagtttttt ggggtcgagg tgccgtaaag
     3121 cactaaatcg gaaccctaaa gggagccccc gatttagagc ttgacgggga aagccggcga
     3181 acgtggcgag aaaggaaggg aagaaagcga aaggagcggg cgctaaggcg ctggcaagtg
     3241 tagcggtcac gctgcgcgta accaccacac ccgccgcgct taatgcgccg ctacagggcg
     3301 cgtactatgg ttgctttgac gtatgcggtg tgaaataccg cacagatgcg taaggagaaa
     3361 ataccgcatc aggcgcccct gcaggcagct gcgcgctcgc tcgctcactg aggccgcccg
     3421 ggcaaagccc gggcgtcggg cgacctttgg tcgcccggcc tcagtgagcg agcgagcgcg
     3481 cagagaggga gtggccaact ccatcactag gggttcctgc ggccgctcgg tccgcacgtg
     3541 agatctcatg cctgctattg tcttcccaat cctccccctt gctgtcctgc cccaccccac
     3601 cccccagaat agaatgacac ctactcagac aatgcgatgc aatttcctca ttttattagg
     3661 aaaggacagt gggagtggca ccttccaggg tcaaggaagg cacgggggag gggcaaacaa
     3721 cagatggctg gcaactagaa ggcacagtcg aagatctctc gagccacgga attgtcagtg
     3781 cccaacagcc gagcccctgt ccagcagcgg gcaaggcagg cggcgatgag ttccgccgtg
     3841 gcaagaacta accaggattt atacaaggag gagaaaatga aagccatacg ggaagcaata
     3901 gcatgataca aaggcattaa agcagcgtat ccacatagcg taaaaggagc aacatagtta
     3961 agaataccag tcaatctttc acaaattttg taatccagag gttgattatg atatcgaatt
     4021 cgcggccgcg gcgcgccgtc gacttacttg tacagctcgt ccatgccgag agtgatcccg
     4081 gcggcggtca cgaactccag caggaccatg tgatcgcgct tctcgttggg gtctttgctc
     4141 agcttggact ggtagctcag gtagtggttg tcgggcagca gcacggggcc gtcgccgatg
     4201 ggggtgttct gctggtagtg gtcggcgagc tgcacgccgc cgtcctcgat gttgtggcgg
     4261 atcttgaagt tggccttgat gccgttcttc tgcttgtcgg cggtgatata gacgttgtgg
     4321 ctgttgtagt tgtactccag cttgtgcccc aggatgttgc cgtcctcctt gaagtcgatg
     4381 cccttcagct cgatgcggtt caccagggtg tcgccctcga acttcacctc ggcgcgggtc
     4441 ttgtagttgc cgtcgtcctt gaagaagatg gtgcgctcct ggacgtagcc ttcgggcatg
     4501 gcggacttga agaagtcgtg ctgcttcatg tggtcggggt agcgggcgaa gcactgcacg
     4561 ccgtagccca gggtggtcac gagggtgggc cagggcacgg gcagcttgcc ggtggtgcag
     4621 atcagcttca gggtcagctt gccgtaggtg gcatcgccct cgccctcgcc ggacacgctg
     4681 aacttgtggc cgtttacgtc gccgtccagc tcgaccagga tgggcaccac cccggtgaac
     4741 agctcctcgc ccttgctcac catggtggcg accggtagcg ctagcttcga aagatctgga
     4801 tcccagaagc aaatgtaagc aatagatggc tctgccctga cttttatgcc cagcccgagc
     4861 tccgccggcg cgtgaaagtc tttctcatga ccttatatag ctgtccccat tttacagatg
     4921 aggagactga ggtggggttc taacttggat tgtgtcaggt gcttacacga ggttggctgt
     4981 tgggtgtttg agtttcctag atggtctgac tccatcctgc ttgaccttga tttgggagat
     5041 tgctatagga ggttccttgc caattgtgct gtggaggtcc aagggagatg gcactaccaa
     5101 atgctaccaa cagccacctc ctcctatgtg ccaagcactg ttgtgtgctt ccttggctgc
     5161 tcttcaggaa agtggcatga gctggcatta tccccacttc acaaagaaag acagcacggt
     5221 ggcccatggc tacctagcca gtaataatgg aacaagtcac aggtcagggt ggctgtgtgg
     5281 tcatgggagg atgcctccgt ggggacagat gatgactcac actagcaatg ggagtgacat
     5341 tggcagatga agacgacaga ccccttctgc tgtgccgggg agcagttggg catctgtagg
     5401 ttggtcgagc agtttgaggt atagaagtgg cttgatgaca gcagcttcca ggagctgaga
     5461 gctttctcag caccagacct gtgacaggga gctgtggtgc tagcgggatg aacagcccca
     5521 agaagcagct aatctcctgg gttccgcttt gaaggtgagg aaacggagac aggaacat
//
