LOCUS       sequence-502264-        4758 bp    DNA     circular SYN 10-MAR-2026
DEFINITION  AiE2251m 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: AiP1584 - pAAV-AiE2251m-minBG-SYFP2-WPRE3-BGHpA
FEATURES             Location/Qualifiers
     source          1..4758
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(join(564..1355,1356..1424))
                     /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     1187..1206
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(1425..1529)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     1597..1615
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(1653..1675)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     1775..1794
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     rep_origin      complement(1811..2266)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1948..1969)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2160..2179
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     repeat_region   2341..2481
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   2352..2481
                     /label=AAV2 ITR
     polyA_signal    2599..2710
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     2897..2917
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(join(3010..3720,3721..3723,3724..3726))
                     /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(3052..3073)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     3401..3420
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     3660..3681
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     regulatory      3723..3732
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     repeat_region   4372..4501
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     rep_origin      complement(join(4563..4758,1..393))
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4643..4662)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 agctcacgct gtaggtatct cagttcggtg taggtcgttc gctccaagct gggctgtgtg
       61 cacgaacccc ccgttcagcc cgaccgctgc gccttatccg gtaactatcg tcttgagtcc
      121 aacccggtaa gacacgactt atcgccactg gcagcagcca ctggtaacag gattagcaga
      181 gcgaggtatg taggcggtgc tacagagttc ttgaagtggt ggcctaacta cggctacact
      241 agaagaacag tatttggtat ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt
      301 ggtagctctt gatccggcaa acaaaccacc gctggtagcg gtggtttttt tgtttgcaag
      361 cagcagatta cgcgcagaaa aaaaggatct caagaagatc ctttgatctt ttctacgggg
      421 tctgacgctc agtggaacga aaactcacgt taagggattt tggtcatgag attatcaaaa
      481 aggatcttca cctagatcct tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata
      541 tatgagtaaa cttggtctga cagttaccaa tgcttaatca gtgaggcacc tatctcagcg
      601 atctgtctat ttcgttcatc catagttgcc tgactccccg tcgtgtagat aactacgata
      661 cgggagggct taccatctgg ccccagtgct gcaatgatac cgcgagaccc acgctcaccg
      721 gctccagatt tatcagcaat aaaccagcca gccggaaggg ccgagcgcag aagtggtcct
      781 gcaactttat ccgcctccat ccagtctatt aattgttgcc gggaagctag agtaagtagt
      841 tcgccagtta atagtttgcg caacgttgtt gccattgcta caggcatcgt ggtgtcacgc
      901 tcgtcgtttg gtatggcttc attcagctcc ggttcccaac gatcaaggcg agttacatga
      961 tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt tgtcagaagt
     1021 aagttggccg cagtgttatc actcatggtt atggcagcac tgcataattc tcttactgtc
     1081 atgccatccg taagatgctt ttctgtgact ggtgagtact caaccaagtc attctgagaa
     1141 tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca
     1201 catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg aaaactctca
     1261 aggatcttac cgctgttgag atccagttcg atgtaaccca ctcgtgcacc caactgatct
     1321 tcagcatctt ttactttcac cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc
     1381 gcaaaaaagg gaataagggc gacacggaaa tgttgaatac tcatactctt cctttttcaa
     1441 tattattgaa gcatttatca gggttattgt ctcatgagcg gatacatatt tgaatgtatt
     1501 tagaaaaata aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc acctgacgtc
     1561 taagaaacca ttattatcat gacattaacc tataaaaata ggcgtatcac gaggcccttt
     1621 cgtctcgcgc gtttcggtga tgacggtgaa aacctctgac acatgcagct cccggagacg
     1681 gtcacagctt gtctgtaagc ggatgccggg agcagacaag cccgtcaggg cgcgtcagcg
     1741 ggtgttggcg ggtgtcgggg ctggcttaac tatgcggcat cagagcagat tgtactgaga
     1801 gtgcaccata aaattgtaaa cgttaatatt ttgttaaaat tcgcgttaaa tttttgttaa
     1861 atcagctcat tttttaacca atagaccgaa atcggcaaaa tcccttataa atcaaaagaa
     1921 tagcccgaga tagagttgag tgttgttcca gtttggaaca agagtccact attaaagaac
     1981 gtggactcca acgtcaaagg gcgaaaaacc gtctatcagg gcgatggccc actacgtgaa
     2041 ccatcaccca aatcaagttt tttggggtcg aggtgccgta aagcactaaa tcggaaccct
     2101 aaagggagcc cccgatttag agcttgacgg ggaaagccgg cgaacgtggc gagaaaggaa
     2161 gggaagaaag cgaaaggagc gggcgctaag gcgctggcaa gtgtagcggt cacgctgcgc
     2221 gtaaccacca cacccgccgc gcttaatgcg ccgctacagg gcgcgtacta tggttgcttt
     2281 gacgtatgcg gtgtgaaata ccgcacagat gcgtaaggag aaaataccgc atcaggcgcc
     2341 cctgcaggca gctgcgcgct cgctcgctca ctgaggccgc ccgggcaaag cccgggcgtc
     2401 gggcgacctt tggtcgcccg gcctcagtga gcgagcgagc gcgcagagag ggagtggcca
     2461 actccatcac taggggttcc tgcggccgct cggtccgcac gtgagatctc atgcctgcta
     2521 ttgtcttccc aatcctcccc cttgctgtcc tgccccaccc caccccccag aatagaatga
     2581 cacctactca gacaatgcga tgcaatttcc tcattttatt aggaaaggac agtgggagtg
     2641 gcaccttcca gggtcaagga aggcacgggg gaggggcaaa caacagatgg ctggcaacta
     2701 gaaggcacag tcgaagatct ctcgagccac ggaattgtca gtgcccaaca gccgagcccc
     2761 tgtccagcag cgggcaaggc aggcggcgat gagttccgcc gtggcaagaa ctaaccagga
     2821 tttatacaag gaggagaaaa tgaaagccat acgggaagca atagcatgat acaaaggcat
     2881 taaagcagcg tatccacata gcgtaaaagg agcaacatag ttaagaatac cagtcaatct
     2941 ttcacaaatt ttgtaatcca gaggttgatt atgatatcga attcgcggcc gcggcgcgcc
     3001 gtcgacttac ttgtacagct cgtccatgcc gagagtgatc ccggcggcgg tcacgaactc
     3061 cagcaggacc atgtgatcgc gcttctcgtt ggggtctttg ctcagcttgg actggtagct
     3121 caggtagtgg ttgtcgggca gcagcacggg gccgtcgccg atgggggtgt tctgctggta
     3181 gtggtcggcg agctgcacgc cgccgtcctc gatgttgtgg cggatcttga agttggcctt
     3241 gatgccgttc ttctgcttgt cggcggtgat atagacgttg tggctgttgt agttgtactc
     3301 cagcttgtgc cccaggatgt tgccgtcctc cttgaagtcg atgcccttca gctcgatgcg
     3361 gttcaccagg gtgtcgccct cgaacttcac ctcggcgcgg gtcttgtagt tgccgtcgtc
     3421 cttgaagaag atggtgcgct cctggacgta gccttcgggc atggcggact tgaagaagtc
     3481 gtgctgcttc atgtggtcgg ggtagcgggc gaagcactgc acgccgtagc ccagggtggt
     3541 cacgagggtg ggccagggca cgggcagctt gccggtggtg cagatcagct tcagggtcag
     3601 cttgccgtag gtggcatcgc cctcgccctc gccggacacg ctgaacttgt ggccgtttac
     3661 gtcgccgtcc agctcgacca ggatgggcac caccccggtg aacagctcct cgcccttgct
     3721 caccatggtg gcgaccggta gcgctagctt cgaaagatct ggatcccaga agcaaatgta
     3781 agcaatagat ggctctgccc tgacttttat gcccagcccg agctccgccg gcgacacagt
     3841 atgttatata ggcttaatta ttgccattga cagtgagaaa agacactagg aaaactcagg
     3901 gcagggtgac ctggatatca tcagagggac atgggaaaca tagttgcata gacatggaag
     3961 gtaagggtta tggaaatatc acaagagaga aagagcgtac aagttcaaag gaacagcatg
     4021 tgcaaagact gaaatgggag aaggcatgag gttgtcaaag gtactaggag agtgtggcga
     4081 cattcaagca cagagaatga gagagcatct gaggtcatca aaaatttgga aaagagtagg
     4141 actttgatgt accaggctca gaagcagatg aaggatggtg gggctttaat aaaaaggcat
     4201 gcagttttca gactccaatt ctaagcattc ttgctagaga ctccagcatg cttccagcct
     4261 aaatattctc aacatctttt cttgagcttc tattgctatt gcaaatgccc aaatgcagat
     4321 tgtgccaaga cgaccgccgg cgattattac tcttacaacg cgtgcggccg caggaacccc
     4381 tagtgatgga gttggccact ccctctctgc gcgctcgctc gctcactgag gccgggcgac
     4441 caaaggtcgc ccgacgcccg ggcggcctca gtgagcgagc gagcgcgcag ctgcctgcag
     4501 gacatgtgag caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg
     4561 tttttccata ggctccgccc ccctgacgag catcacaaaa atcgacgctc aagtcagagg
     4621 tggcgaaacc cgacaggact ataaagatac caggcgtttc cccctggaag ctccctcgtg
     4681 cgctctcctg ttccgaccct gccgcttacc ggatacctgt ccgcctttct cccttcggga
     4741 agcgtggcgc tttctcat
//
