LOCUS       sequence-502279-        4778 bp    DNA     circular SYN 10-MAR-2026
DEFINITION  AiE2284m 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 4778)
  TITLE     Allen Institute AAV Enhancer Plasmids
REFERENCE   2  (bases 1 to 4778)
  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: AiP1617 - pAAV-AiE2284m-minBG-SYFP2-WPRE3-BGHpA
FEATURES             Location/Qualifiers
     source          1..4778
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     47..66
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(285..389)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     457..475
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(513..535)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     635..654
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     rep_origin      complement(671..1126)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(808..829)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     1020..1039
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     repeat_region   1201..1341
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   1212..1341
                     /label=AAV2 ITR
     polyA_signal    1459..1570
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     1757..1777
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(join(1870..2580,2581..2583,2584..2586))
                     /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(1912..1933)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     2261..2280
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2520..2541
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     regulatory      2583..2592
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     repeat_region   3252..3381
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     rep_origin      complement(3443..4031)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3523..3542)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4202..4778,1..215,216..284))
                     /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 tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca
       61 catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg aaaactctca
      121 aggatcttac cgctgttgag atccagttcg atgtaaccca ctcgtgcacc caactgatct
      181 tcagcatctt ttactttcac cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc
      241 gcaaaaaagg gaataagggc gacacggaaa tgttgaatac tcatactctt cctttttcaa
      301 tattattgaa gcatttatca gggttattgt ctcatgagcg gatacatatt tgaatgtatt
      361 tagaaaaata aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc acctgacgtc
      421 taagaaacca ttattatcat gacattaacc tataaaaata ggcgtatcac gaggcccttt
      481 cgtctcgcgc gtttcggtga tgacggtgaa aacctctgac acatgcagct cccggagacg
      541 gtcacagctt gtctgtaagc ggatgccggg agcagacaag cccgtcaggg cgcgtcagcg
      601 ggtgttggcg ggtgtcgggg ctggcttaac tatgcggcat cagagcagat tgtactgaga
      661 gtgcaccata aaattgtaaa cgttaatatt ttgttaaaat tcgcgttaaa tttttgttaa
      721 atcagctcat tttttaacca atagaccgaa atcggcaaaa tcccttataa atcaaaagaa
      781 tagcccgaga tagagttgag tgttgttcca gtttggaaca agagtccact attaaagaac
      841 gtggactcca acgtcaaagg gcgaaaaacc gtctatcagg gcgatggccc actacgtgaa
      901 ccatcaccca aatcaagttt tttggggtcg aggtgccgta aagcactaaa tcggaaccct
      961 aaagggagcc cccgatttag agcttgacgg ggaaagccgg cgaacgtggc gagaaaggaa
     1021 gggaagaaag cgaaaggagc gggcgctaag gcgctggcaa gtgtagcggt cacgctgcgc
     1081 gtaaccacca cacccgccgc gcttaatgcg ccgctacagg gcgcgtacta tggttgcttt
     1141 gacgtatgcg gtgtgaaata ccgcacagat gcgtaaggag aaaataccgc atcaggcgcc
     1201 cctgcaggca gctgcgcgct cgctcgctca ctgaggccgc ccgggcaaag cccgggcgtc
     1261 gggcgacctt tggtcgcccg gcctcagtga gcgagcgagc gcgcagagag ggagtggcca
     1321 actccatcac taggggttcc tgcggccgct cggtccgcac gtgagatctc atgcctgcta
     1381 ttgtcttccc aatcctcccc cttgctgtcc tgccccaccc caccccccag aatagaatga
     1441 cacctactca gacaatgcga tgcaatttcc tcattttatt aggaaaggac agtgggagtg
     1501 gcaccttcca gggtcaagga aggcacgggg gaggggcaaa caacagatgg ctggcaacta
     1561 gaaggcacag tcgaagatct ctcgagccac ggaattgtca gtgcccaaca gccgagcccc
     1621 tgtccagcag cgggcaaggc aggcggcgat gagttccgcc gtggcaagaa ctaaccagga
     1681 tttatacaag gaggagaaaa tgaaagccat acgggaagca atagcatgat acaaaggcat
     1741 taaagcagcg tatccacata gcgtaaaagg agcaacatag ttaagaatac cagtcaatct
     1801 ttcacaaatt ttgtaatcca gaggttgatt atgatatcga attcgcggcc gcggcgcgcc
     1861 gtcgacttac ttgtacagct cgtccatgcc gagagtgatc ccggcggcgg tcacgaactc
     1921 cagcaggacc atgtgatcgc gcttctcgtt ggggtctttg ctcagcttgg actggtagct
     1981 caggtagtgg ttgtcgggca gcagcacggg gccgtcgccg atgggggtgt tctgctggta
     2041 gtggtcggcg agctgcacgc cgccgtcctc gatgttgtgg cggatcttga agttggcctt
     2101 gatgccgttc ttctgcttgt cggcggtgat atagacgttg tggctgttgt agttgtactc
     2161 cagcttgtgc cccaggatgt tgccgtcctc cttgaagtcg atgcccttca gctcgatgcg
     2221 gttcaccagg gtgtcgccct cgaacttcac ctcggcgcgg gtcttgtagt tgccgtcgtc
     2281 cttgaagaag atggtgcgct cctggacgta gccttcgggc atggcggact tgaagaagtc
     2341 gtgctgcttc atgtggtcgg ggtagcgggc gaagcactgc acgccgtagc ccagggtggt
     2401 cacgagggtg ggccagggca cgggcagctt gccggtggtg cagatcagct tcagggtcag
     2461 cttgccgtag gtggcatcgc cctcgccctc gccggacacg ctgaacttgt ggccgtttac
     2521 gtcgccgtcc agctcgacca ggatgggcac caccccggtg aacagctcct cgcccttgct
     2581 caccatggtg gcgaccggta gcgctagctt cgaaagatct ggatcccaga agcaaatgta
     2641 agcaatagat ggctctgccc tgacttttat gcccagcccg agctccgccg gcggcatctg
     2701 caaggtggtt agagttttca aaattcactc tctgagcatg gctctggctc acagagtaaa
     2761 gtctaaacgg agctgtagaa atcagagcgg ccgacagggc acaggccttg catcctttcc
     2821 agccccctgc acacccgcac atgcgcgcgc acacatgctt attcttctat ttatttccct
     2881 ccactattca tattcattgg catcatcctt ccctcccagt gatcatccgc cccattgtct
     2941 tctcgccaca ccattcaatt caagacaatc gcgtcttatc cgctgtgtgc caggcactga
     3001 gcttagagtc atctgttctc ggagtcatct tggagccgag tcatcttgga atctgccctg
     3061 tcgtgcacag catctaactg atcacaaagc cctctccact ctgcctctgg aacagcacct
     3121 aaataccact ctgtctcttc cctgattgcc cttcttcttg tttctcatcc agagaactgt
     3181 ggcagcctta agtgtgccag cttcccgaac cgatcgccgg cgattattac tcttacaacg
     3241 cgtgcggccg caggaacccc tagtgatgga gttggccact ccctctctgc gcgctcgctc
     3301 gctcactgag gccgggcgac caaaggtcgc ccgacgcccg ggcggcctca gtgagcgagc
     3361 gagcgcgcag ctgcctgcag gacatgtgag caaaaggcca gcaaaaggcc aggaaccgta
     3421 aaaaggccgc gttgctggcg tttttccata ggctccgccc ccctgacgag catcacaaaa
     3481 atcgacgctc aagtcagagg tggcgaaacc cgacaggact ataaagatac caggcgtttc
     3541 cccctggaag ctccctcgtg cgctctcctg ttccgaccct gccgcttacc ggatacctgt
     3601 ccgcctttct cccttcggga agcgtggcgc tttctcatag ctcacgctgt aggtatctca
     3661 gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc gttcagcccg
     3721 accgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggtaaga cacgacttat
     3781 cgccactggc agcagccact ggtaacagga ttagcagagc gaggtatgta ggcggtgcta
     3841 cagagttctt gaagtggtgg cctaactacg gctacactag aagaacagta tttggtatct
     3901 gcgctctgct gaagccagtt accttcggaa aaagagttgg tagctcttga tccggcaaac
     3961 aaaccaccgc tggtagcggt ggtttttttg tttgcaagca gcagattacg cgcagaaaaa
     4021 aaggatctca agaagatcct ttgatctttt ctacggggtc tgacgctcag tggaacgaaa
     4081 actcacgtta agggattttg gtcatgagat tatcaaaaag gatcttcacc tagatccttt
     4141 taaattaaaa atgaagtttt aaatcaatct aaagtatata tgagtaaact tggtctgaca
     4201 gttaccaatg cttaatcagt gaggcaccta tctcagcgat ctgtctattt cgttcatcca
     4261 tagttgcctg actccccgtc gtgtagataa ctacgatacg ggagggctta ccatctggcc
     4321 ccagtgctgc aatgataccg cgagacccac gctcaccggc tccagattta tcagcaataa
     4381 accagccagc cggaagggcc gagcgcagaa gtggtcctgc aactttatcc gcctccatcc
     4441 agtctattaa ttgttgccgg gaagctagag taagtagttc gccagttaat agtttgcgca
     4501 acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat
     4561 tcagctccgg ttcccaacga tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag
     4621 cggttagctc cttcggtcct ccgatcgttg tcagaagtaa gttggccgca gtgttatcac
     4681 tcatggttat ggcagcactg cataattctc ttactgtcat gccatccgta agatgctttt
     4741 ctgtgactgg tgagtactca accaagtcat tctgagaa
//
