LOCUS       sequence-502231-        5157 bp    DNA     circular SYN 10-MAR-2026
DEFINITION  AiE2197m 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 5157)
  TITLE     Allen Institute AAV Enhancer Plasmids
REFERENCE   2  (bases 1 to 5157)
  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: AiP1522 - pAAV-AiE2197m-minBG-SYFP2-WPRE3-BGHpA
FEATURES             Location/Qualifiers
     source          1..5157
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      739..1327
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1228..1247
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     repeat_region   1389..1518
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     regulatory      2557..2566
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(2563..2565,2566..2568,2569..3279)
                     /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(2608..2629)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2869..2888)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     3216..3237
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(3372..3392)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     polyA_signal    3579..3690
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   3808..3948
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   3808..3937
                     /label=AAV2 ITR
     rep_origin      4023..4478
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4110..4129)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4320..4341
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4495..4514)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4614..4636
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(4674..4692)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4760..4864
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4865..4933,4934..5157,1..568)
                     /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(5083..5102)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
ORIGIN
        1 tgacttggtt gagtactcac cagtcacaga aaagcatctt acggatggca tgacagtaag
       61 agaattatgc agtgctgcca taaccatgag tgataacact gcggccaact tacttctgac
      121 aacgatcgga ggaccgaagg agctaaccgc ttttttgcac aacatggggg atcatgtaac
      181 tcgccttgat cgttgggaac cggagctgaa tgaagccata ccaaacgacg agcgtgacac
      241 cacgatgcct gtagcaatgg caacaacgtt gcgcaaacta ttaactggcg aactacttac
      301 tctagcttcc cggcaacaat taatagactg gatggaggcg gataaagttg caggaccact
      361 tctgcgctcg gcccttccgg ctggctggtt tattgctgat aaatctggag ccggtgagcg
      421 tgggtctcgc ggtatcattg cagcactggg gccagatggt aagccctccc gtatcgtagt
      481 tatctacacg acggggagtc aggcaactat ggatgaacga aatagacaga tcgctgagat
      541 aggtgcctca ctgattaagc attggtaact gtcagaccaa gtttactcat atatacttta
      601 gattgattta aaacttcatt tttaatttaa aaggatctag gtgaagatcc tttttgataa
      661 tctcatgacc aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag accccgtaga
      721 aaagatcaaa ggatcttctt gagatccttt ttttctgcgc gtaatctgct gcttgcaaac
      781 aaaaaaacca ccgctaccag cggtggtttg tttgccggat caagagctac caactctttt
      841 tccgaaggta actggcttca gcagagcgca gataccaaat actgttcttc tagtgtagcc
      901 gtagttaggc caccacttca agaactctgt agcaccgcct acatacctcg ctctgctaat
      961 cctgttacca gtggctgctg ccagtggcga taagtcgtgt cttaccgggt tggactcaag
     1021 acgatagtta ccggataagg cgcagcggtc gggctgaacg gggggttcgt gcacacagcc
     1081 cagcttggag cgaacgacct acaccgaact gagataccta cagcgtgagc tatgagaaag
     1141 cgccacgctt cccgaaggga gaaaggcgga caggtatccg gtaagcggca gggtcggaac
     1201 aggagagcgc acgagggagc ttccaggggg aaacgcctgg tatctttata gtcctgtcgg
     1261 gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct
     1321 atggaaaaac gccagcaacg cggccttttt acggttcctg gccttttgct ggccttttgc
     1381 tcacatgtcc tgcaggcagc tgcgcgctcg ctcgctcact gaggccgccc gggcgtcggg
     1441 cgacctttgg tcgcccggcc tcagtgagcg agcgagcgcg cagagaggga gtggccaact
     1501 ccatcactag gggttcctgc ggccgcacgc gttgtaagag taataatcgc cggcgggagg
     1561 tgccgagggc tatagacagg tactccaggg acccaagcca cataagccct cagtggcaac
     1621 agcattccct tacctcttcc ccctccccca cctttcaccc ttccacatcc aacctctact
     1681 gtaccaccac aaccactctt gcagacagtc ccctagtgcc actgtcattg tgcactagtg
     1741 tggacagctt gcagccttgc ctgccctagg gaagaatttc ccagcaggga gtgggaggtg
     1801 ccaggcctcc aagtgattct gacctgacta taaaagaccc agctctattc ctgggtcatg
     1861 taaccgccta gaggctgcca agacactgac acctgtcctg gcattgcctt gctggtgagg
     1921 acccaaagct ggaggtggca gatgggtagg gagagcccta gaaacctaaa agacataaag
     1981 atctgttggc cgttttgagt cctttaaagt aaaggaaaca gccccactta cagaagaaag
     2041 cagaagtgtt ttgtaactgc acagatctcc acagtctgat ctgcccactc ctctcctcag
     2101 aaggaagtgg ctgcccttta ctcctccaca cagaatagag cagcctggag ctcgggacct
     2161 ctttaccctt gagactggga agttgagctg ggacttctga cagattttgc ctggcatccc
     2221 ctggcagggc tctgcgcagt ggcaatgggt agtgcccaag agaagtggtt gggccagttg
     2281 gaccagcaga gcccaaccca gcaagtcaag tcctccagag atctgagagg ccctgggtta
     2341 cttttaccaa gaccctgctc tcgctctcca ctccctagtc tcagctcagt ttgtgctttg
     2401 cccctggaat ggcttccatc cagctcttag tagtcaacaa accccatccc caccccgccg
     2461 gcggagctcg ggctgggcat aaaagtcagg gcagagccat ctattgctta catttgcttc
     2521 tgggatccag atctttcgaa gctagcgcta ccggtcgcca ccatggtgag caagggcgag
     2581 gagctgttca ccggggtggt gcccatcctg gtcgagctgg acggcgacgt aaacggccac
     2641 aagttcagcg tgtccggcga gggcgagggc gatgccacct acggcaagct gaccctgaag
     2701 ctgatctgca ccaccggcaa gctgcccgtg ccctggccca ccctcgtgac caccctgggc
     2761 tacggcgtgc agtgcttcgc ccgctacccc gaccacatga agcagcacga cttcttcaag
     2821 tccgccatgc ccgaaggcta cgtccaggag cgcaccatct tcttcaagga cgacggcaac
     2881 tacaagaccc gcgccgaggt gaagttcgag ggcgacaccc tggtgaaccg catcgagctg
     2941 aagggcatcg acttcaagga ggacggcaac atcctggggc acaagctgga gtacaactac
     3001 aacagccaca acgtctatat caccgccgac aagcagaaga acggcatcaa ggccaacttc
     3061 aagatccgcc acaacatcga ggacggcggc gtgcagctcg ccgaccacta ccagcagaac
     3121 acccccatcg gcgacggccc cgtgctgctg cccgacaacc actacctgag ctaccagtcc
     3181 aagctgagca aagaccccaa cgagaagcgc gatcacatgg tcctgctgga gttcgtgacc
     3241 gccgccggga tcactctcgg catggacgag ctgtacaagt aagtcgacgg cgcgccgcgg
     3301 ccgcgaattc gatatcataa tcaacctctg gattacaaaa tttgtgaaag attgactggt
     3361 attcttaact atgttgctcc ttttacgcta tgtggatacg ctgctttaat gcctttgtat
     3421 catgctattg cttcccgtat ggctttcatt ttctcctcct tgtataaatc ctggttagtt
     3481 cttgccacgg cggaactcat cgccgcctgc cttgcccgct gctggacagg ggctcggctg
     3541 ttgggcactg acaattccgt ggctcgagag atcttcgact gtgccttcta gttgccagcc
     3601 atctgttgtt tgcccctccc ccgtgccttc cttgaccctg gaaggtgcca ctcccactgt
     3661 cctttcctaa taaaatgagg aaattgcatc gcattgtctg agtaggtgtc attctattct
     3721 ggggggtggg gtggggcagg acagcaaggg ggaggattgg gaagacaata gcaggcatga
     3781 gatctcacgt gcggaccgag cggccgcagg aacccctagt gatggagttg gccactccct
     3841 ctctgcgcgc tcgctcgctc actgaggccg ggcgaccaaa ggtcgcccga cgcccgggct
     3901 ttgcccgggc ggcctcagtg agcgagcgag cgcgcagctg cctgcagggg cgcctgatgc
     3961 ggtattttct ccttacgcat ctgtgcggta tttcacaccg catacgtcaa agcaaccata
     4021 gtacgcgccc tgtagcggcg cattaagcgc ggcgggtgtg gtggttacgc gcagcgtgac
     4081 cgctacactt gccagcgcct tagcgcccgc tcctttcgct ttcttccctt cctttctcgc
     4141 cacgttcgcc ggctttcccc gtcaagctct aaatcggggg ctccctttag ggttccgatt
     4201 tagtgcttta cggcacctcg accccaaaaa acttgatttg ggtgatggtt cacgtagtgg
     4261 gccatcgccc tgatagacgg tttttcgccc tttgacgttg gagtccacgt tctttaatag
     4321 tggactcttg ttccaaactg gaacaacact caactctatc tcgggctatt cttttgattt
     4381 ataagggatt ttgccgattt cggtctattg gttaaaaaat gagctgattt aacaaaaatt
     4441 taacgcgaat tttaacaaaa tattaacgtt tacaatttta tggtgcactc tcagtacaat
     4501 ctgctctgat gccgcatagt taagccagcc ccgacacccg ccaacacccg ctgacgcgcc
     4561 ctgacgggct tgtctgctcc cggcatccgc ttacagacaa gctgtgaccg tctccgggag
     4621 ctgcatgtgt cagaggtttt caccgtcatc accgaaacgc gcgagacgaa agggcctcgt
     4681 gatacgccta tttttatagg ttaatgtcat gataataatg gtttcttaga cgtcaggtgg
     4741 cacttttcgg ggaaatgtgc gcggaacccc tatttgttta tttttctaaa tacattcaaa
     4801 tatgtatccg ctcatgagac aataaccctg ataaatgctt caataatatt gaaaaaggaa
     4861 gagtatgagt attcaacatt tccgtgtcgc ccttattccc ttttttgcgg cattttgcct
     4921 tcctgttttt gctcacccag aaacgctggt gaaagtaaaa gatgctgaag atcagttggg
     4981 tgcacgagtg ggttacatcg aactggatct caacagcggt aagatccttg agagttttcg
     5041 ccccgaagaa cgttttccaa tgatgagcac ttttaaagtt ctgctatgtg gcgcggtatt
     5101 atcccgtatt gacgccgggc aagagcaact cggtcgccgc atacactatt ctcagaa
//
