LOCUS       sequence-502291-        4993 bp    DNA     circular SYN 10-MAR-2026
DEFINITION  AiE2440m 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 4993)
  TITLE     Allen Institute AAV Enhancer Plasmids
REFERENCE   2  (bases 1 to 4993)
  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: AiP1957 - pAAV-AiE2440m-minBG-SYFP2-WPRE3-BGHpA
FEATURES             Location/Qualifiers
     source          1..4993
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     regulatory      519..528
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(525..527,528..530,531..1241)
                     /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(570..591)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(831..850)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1178..1199
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(1334..1354)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     polyA_signal    1541..1652
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   1770..1910
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   1770..1899
                     /label=AAV2 ITR
     rep_origin      1985..2440
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2072..2091)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2282..2303
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(2457..2476)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     2576..2598
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(2636..2654)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        2722..2826
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2827..2895,2896..3687)
                     /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(3045..3064)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3858..4446
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4347..4366
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     repeat_region   4508..4637
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
ORIGIN
        1 agatgcagag attggagttt gcccagctga attttggttt tgctttggtc cagtattcct
       61 cactatgatg ttctggaata gtaattctgt gatgttggag gcaggtgatt tggtttttgc
      121 ttttgatttt atatgcggtt acagttaagt gattgaatga atctcagaag agactttgaa
      181 cattggactt ttaacattgt tgagactgtt atagactacg ggactttctt ttttacattt
      241 aaaacttgta tatttctatg tttattttca aagaaaagtt caacttttga agttgaacta
      301 agtgtattct gtattattct atggcaaggt atggccccca tagatatgca tttgaacaag
      361 cttattggga ccagggagta gaatgtggtg gcttgaatat gcttgtccca gtgattggcc
      421 ttacggagct cgggctgggc ataaaagtca gggcagagcc atctattgct tacatttgct
      481 tctgggatcc agatctttcg aagctagcgc taccggtcgc caccatggtg agcaagggcg
      541 aggagctgtt caccggggtg gtgcccatcc tggtcgagct ggacggcgac gtaaacggcc
      601 acaagttcag cgtgtccggc gagggcgagg gcgatgccac ctacggcaag ctgaccctga
      661 agctgatctg caccaccggc aagctgcccg tgccctggcc caccctcgtg accaccctgg
      721 gctacggcgt gcagtgcttc gcccgctacc ccgaccacat gaagcagcac gacttcttca
      781 agtccgccat gcccgaaggc tacgtccagg agcgcaccat cttcttcaag gacgacggca
      841 actacaagac ccgcgccgag gtgaagttcg agggcgacac cctggtgaac cgcatcgagc
      901 tgaagggcat cgacttcaag gaggacggca acatcctggg gcacaagctg gagtacaact
      961 acaacagcca caacgtctat atcaccgccg acaagcagaa gaacggcatc aaggccaact
     1021 tcaagatccg ccacaacatc gaggacggcg gcgtgcagct cgccgaccac taccagcaga
     1081 acacccccat cggcgacggc cccgtgctgc tgcccgacaa ccactacctg agctaccagt
     1141 ccaagctgag caaagacccc aacgagaagc gcgatcacat ggtcctgctg gagttcgtga
     1201 ccgccgccgg gatcactctc ggcatggacg agctgtacaa gtaagtcgac ggcgcgccgc
     1261 ggccgcgaat tcgatatcat aatcaacctc tggattacaa aatttgtgaa agattgactg
     1321 gtattcttaa ctatgttgct ccttttacgc tatgtggata cgctgcttta atgcctttgt
     1381 atcatgctat tgcttcccgt atggctttca ttttctcctc cttgtataaa tcctggttag
     1441 ttcttgccac ggcggaactc atcgccgcct gccttgcccg ctgctggaca ggggctcggc
     1501 tgttgggcac tgacaattcc gtggctcgag agatcttcga ctgtgccttc tagttgccag
     1561 ccatctgttg tttgcccctc ccccgtgcct tccttgaccc tggaaggtgc cactcccact
     1621 gtcctttcct aataaaatga ggaaattgca tcgcattgtc tgagtaggtg tcattctatt
     1681 ctggggggtg gggtggggca ggacagcaag ggggaggatt gggaagacaa tagcaggcat
     1741 gagatctcac gtgcggaccg agcggccgca ggaaccccta gtgatggagt tggccactcc
     1801 ctctctgcgc gctcgctcgc tcactgaggc cgggcgacca aaggtcgccc gacgcccggg
     1861 ctttgcccgg gcggcctcag tgagcgagcg agcgcgcagc tgcctgcagg ggcgcctgat
     1921 gcggtatttt ctccttacgc atctgtgcgg tatttcacac cgcatacgtc aaagcaacca
     1981 tagtacgcgc cctgtagcgg cgcattaagc gcggcgggtg tggtggttac gcgcagcgtg
     2041 accgctacac ttgccagcgc cttagcgccc gctcctttcg ctttcttccc ttcctttctc
     2101 gccacgttcg ccggctttcc ccgtcaagct ctaaatcggg ggctcccttt agggttccga
     2161 tttagtgctt tacggcacct cgaccccaaa aaacttgatt tgggtgatgg ttcacgtagt
     2221 gggccatcgc cctgatagac ggtttttcgc cctttgacgt tggagtccac gttctttaat
     2281 agtggactct tgttccaaac tggaacaaca ctcaactcta tctcgggcta ttcttttgat
     2341 ttataaggga ttttgccgat ttcggtctat tggttaaaaa atgagctgat ttaacaaaaa
     2401 tttaacgcga attttaacaa aatattaacg tttacaattt tatggtgcac tctcagtaca
     2461 atctgctctg atgccgcata gttaagccag ccccgacacc cgccaacacc cgctgacgcg
     2521 ccctgacggg cttgtctgct cccggcatcc gcttacagac aagctgtgac cgtctccggg
     2581 agctgcatgt gtcagaggtt ttcaccgtca tcaccgaaac gcgcgagacg aaagggcctc
     2641 gtgatacgcc tatttttata ggttaatgtc atgataataa tggtttctta gacgtcaggt
     2701 ggcacttttc ggggaaatgt gcgcggaacc cctatttgtt tatttttcta aatacattca
     2761 aatatgtatc cgctcatgag acaataaccc tgataaatgc ttcaataata ttgaaaaagg
     2821 aagagtatga gtattcaaca tttccgtgtc gcccttattc ccttttttgc ggcattttgc
     2881 cttcctgttt ttgctcaccc agaaacgctg gtgaaagtaa aagatgctga agatcagttg
     2941 ggtgcacgag tgggttacat cgaactggat ctcaacagcg gtaagatcct tgagagtttt
     3001 cgccccgaag aacgttttcc aatgatgagc acttttaaag ttctgctatg tggcgcggta
     3061 ttatcccgta ttgacgccgg gcaagagcaa ctcggtcgcc gcatacacta ttctcagaat
     3121 gacttggttg agtactcacc agtcacagaa aagcatctta cggatggcat gacagtaaga
     3181 gaattatgca gtgctgccat aaccatgagt gataacactg cggccaactt acttctgaca
     3241 acgatcggag gaccgaagga gctaaccgct tttttgcaca acatggggga tcatgtaact
     3301 cgccttgatc gttgggaacc ggagctgaat gaagccatac caaacgacga gcgtgacacc
     3361 acgatgcctg tagcaatggc aacaacgttg cgcaaactat taactggcga actacttact
     3421 ctagcttccc ggcaacaatt aatagactgg atggaggcgg ataaagttgc aggaccactt
     3481 ctgcgctcgg cccttccggc tggctggttt attgctgata aatctggagc cggtgagcgt
     3541 gggtctcgcg gtatcattgc agcactgggg ccagatggta agccctcccg tatcgtagtt
     3601 atctacacga cggggagtca ggcaactatg gatgaacgaa atagacagat cgctgagata
     3661 ggtgcctcac tgattaagca ttggtaactg tcagaccaag tttactcata tatactttag
     3721 attgatttaa aacttcattt ttaatttaaa aggatctagg tgaagatcct ttttgataat
     3781 ctcatgacca aaatccctta acgtgagttt tcgttccact gagcgtcaga ccccgtagaa
     3841 aagatcaaag gatcttcttg agatcctttt tttctgcgcg taatctgctg cttgcaaaca
     3901 aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc aagagctacc aactcttttt
     3961 ccgaaggtaa ctggcttcag cagagcgcag ataccaaata ctgttcttct agtgtagccg
     4021 tagttaggcc accacttcaa gaactctgta gcaccgccta catacctcgc tctgctaatc
     4081 ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga
     4141 cgatagttac cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc
     4201 agcttggagc gaacgaccta caccgaactg agatacctac agcgtgagct atgagaaagc
     4261 gccacgcttc ccgaagggag aaaggcggac aggtatccgg taagcggcag ggtcggaaca
     4321 ggagagcgca cgagggagct tccaggggga aacgcctggt atctttatag tcctgtcggg
     4381 tttcgccacc tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta
     4441 tggaaaaacg ccagcaacgc ggccttttta cggttcctgg ccttttgctg gccttttgct
     4501 cacatgtcct gcaggcagct gcgcgctcgc tcgctcactg aggccgcccg ggcgtcgggc
     4561 gacctttggt cgcccggcct cagtgagcga gcgagcgcgc agagagggag tggccaactc
     4621 catcactagg ggttcctgcg gccgcaccag tactgtggtg ggatgattat caagaacagt
     4681 agcagcagtg gagtggtgtc ttccagagcc tagagggata cagagggcag agctggagga
     4741 gcccagaata tcaggtgtga atgccagaca ttagaacaaa aagatattag aacaaaaagt
     4801 tgaagttgcc ttggataccc caagacgtta gagatgtcag tcatgggata cctgccaagg
     4861 aaagttgcta acagttaatg gaaccagccc aaaagaacaa agtttgctac tgttgaatgc
     4921 taggtttgcc aaaatcagtc aataaggatg aaaggagttg gagatctgaa tagcactttg
     4981 atatcagaca tgg
//
