LOCUS       sequence-492122-        4680 bp    DNA     circular SYN 06-NOV-2025
DEFINITION  AiE2650m is an enhancer sequence designed to drive AAV-mediated 
            transgene expression in specific population sof brain cells..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4680)
  TITLE     Basal Ganglia/CERP Enhancer AAVs
REFERENCE   2  (bases 1 to 4680)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Nov 6, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"
COMMENT     Sequence Label: AiP2124 - pAAV-AiE2650m-minBG-SYFP2-WPRE3-BGHpA
FEATURES             Location/Qualifiers
     source          1..4680
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   131..271
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   142..271
                     /label=AAV2 ITR
     polyA_signal    389..500
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     687..707
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(join(800..1510,1511..1513,1514..1516))
                     /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(842..863)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     1191..1210
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1450..1471
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     regulatory      1513..1522
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     repeat_region   2084..2213
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     rep_origin      complement(2275..2863)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2355..2374)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3034..3825,3826..3894))
                     /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     3657..3676
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3895..3999)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     4067..4085
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(4123..4145)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     4245..4264
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     rep_origin      complement(join(4281..4680,1..56))
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4418..4439)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4630..4649
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
ORIGIN
        1 cacgctgcgc gtaaccacca cacccgccgc gcttaatgcg ccgctacagg gcgcgtacta
       61 tggttgcttt gacgtatgcg gtgtgaaata ccgcacagat gcgtaaggag aaaataccgc
      121 atcaggcgcc cctgcaggca gctgcgcgct cgctcgctca ctgaggccgc ccgggcaaag
      181 cccgggcgtc gggcgacctt tggtcgcccg gcctcagtga gcgagcgagc gcgcagagag
      241 ggagtggcca actccatcac taggggttcc tgcggccgct cggtccgcac gtgagatctc
      301 atgcctgcta ttgtcttccc aatcctcccc cttgctgtcc tgccccaccc caccccccag
      361 aatagaatga cacctactca gacaatgcga tgcaatttcc tcattttatt aggaaaggac
      421 agtgggagtg gcaccttcca gggtcaagga aggcacgggg gaggggcaaa caacagatgg
      481 ctggcaacta gaaggcacag tcgaagatct ctcgagccac ggaattgtca gtgcccaaca
      541 gccgagcccc tgtccagcag cgggcaaggc aggcggcgat gagttccgcc gtggcaagaa
      601 ctaaccagga tttatacaag gaggagaaaa tgaaagccat acgggaagca atagcatgat
      661 acaaaggcat taaagcagcg tatccacata gcgtaaaagg agcaacatag ttaagaatac
      721 cagtcaatct ttcacaaatt ttgtaatcca gaggttgatt atgatatcga attcgcggcc
      781 gcggcgcgcc gtcgacttac ttgtacagct cgtccatgcc gagagtgatc ccggcggcgg
      841 tcacgaactc cagcaggacc atgtgatcgc gcttctcgtt ggggtctttg ctcagcttgg
      901 actggtagct caggtagtgg ttgtcgggca gcagcacggg gccgtcgccg atgggggtgt
      961 tctgctggta gtggtcggcg agctgcacgc cgccgtcctc gatgttgtgg cggatcttga
     1021 agttggcctt gatgccgttc ttctgcttgt cggcggtgat atagacgttg tggctgttgt
     1081 agttgtactc cagcttgtgc cccaggatgt tgccgtcctc cttgaagtcg atgcccttca
     1141 gctcgatgcg gttcaccagg gtgtcgccct cgaacttcac ctcggcgcgg gtcttgtagt
     1201 tgccgtcgtc cttgaagaag atggtgcgct cctggacgta gccttcgggc atggcggact
     1261 tgaagaagtc gtgctgcttc atgtggtcgg ggtagcgggc gaagcactgc acgccgtagc
     1321 ccagggtggt cacgagggtg ggccagggca cgggcagctt gccggtggtg cagatcagct
     1381 tcagggtcag cttgccgtag gtggcatcgc cctcgccctc gccggacacg ctgaacttgt
     1441 ggccgtttac gtcgccgtcc agctcgacca ggatgggcac caccccggtg aacagctcct
     1501 cgcccttgct caccatggtg gcgaccggta gcgctagctt cgaaagatct ggatcccaga
     1561 agcaaatgta agcaatagat ggctctgccc tgacttttat gcccagcccg agctccgccg
     1621 gcgaaactgt taaatttgga tggcccgagt gttgagcaac catggaggtg ctcacctggc
     1681 tcaggtcgaa agctaataca cctttctgtt gacagctccc gtcgcccatc cggtctggaa
     1741 gcaaatatca ctgagctaag tagccggaat gagccaaaag cacgacctga gagagtgggg
     1801 atgaattaag cagctccacg gagaggaatg caaccttccc tgtgtctcca agctccataa
     1861 agccagggta aagttcaagc aaagactcta tgctaattga tccatttcca cccacttccc
     1921 gccccccccc ccaaatgtcc tggttttccc aggaatgtag acagcacagc acgtagaaaa
     1981 cacttcacac tgtctgcagt gctgccaagc agagctgaaa gagagaaggg ggccaggcag
     2041 gaatcagagc tgacagccag cttctgacta cgcgtgcggc cgcaggaacc cctagtgatg
     2101 gagttggcca ctccctctct gcgcgctcgc tcgctcactg aggccgggcg accaaaggtc
     2161 gcccgacgcc cgggcggcct cagtgagcga gcgagcgcgc agctgcctgc aggacatgtg
     2221 agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca
     2281 taggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa
     2341 cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg tgcgctctcc
     2401 tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg gaagcgtggc
     2461 gctttctcat agctcacgct gtaggtatct cagttcggtg taggtcgttc gctccaagct
     2521 gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg gtaactatcg
     2581 tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca ctggtaacag
     2641 gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt ggcctaacta
     2701 cggctacact agaagaacag tatttggtat ctgcgctctg ctgaagccag ttaccttcgg
     2761 aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg gtggtttttt
     2821 tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct caagaagatc ctttgatctt
     2881 ttctacgggg tctgacgctc agtggaacga aaactcacgt taagggattt tggtcatgag
     2941 attatcaaaa aggatcttca cctagatcct tttaaattaa aaatgaagtt ttaaatcaat
     3001 ctaaagtata tatgagtaaa cttggtctga cagttaccaa tgcttaatca gtgaggcacc
     3061 tatctcagcg atctgtctat ttcgttcatc catagttgcc tgactccccg tcgtgtagat
     3121 aactacgata cgggagggct taccatctgg ccccagtgct gcaatgatac cgcgagaccc
     3181 acgctcaccg gctccagatt tatcagcaat aaaccagcca gccggaaggg ccgagcgcag
     3241 aagtggtcct gcaactttat ccgcctccat ccagtctatt aattgttgcc gggaagctag
     3301 agtaagtagt tcgccagtta atagtttgcg caacgttgtt gccattgcta caggcatcgt
     3361 ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc ggttcccaac gatcaaggcg
     3421 agttacatga tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt
     3481 tgtcagaagt aagttggccg cagtgttatc actcatggtt atggcagcac tgcataattc
     3541 tcttactgtc atgccatccg taagatgctt ttctgtgact ggtgagtact caaccaagtc
     3601 attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa
     3661 taccgcgcca catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg
     3721 aaaactctca aggatcttac cgctgttgag atccagttcg atgtaaccca ctcgtgcacc
     3781 caactgatct tcagcatctt ttactttcac cagcgtttct gggtgagcaa aaacaggaag
     3841 gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa tgttgaatac tcatactctt
     3901 cctttttcaa tattattgaa gcatttatca gggttattgt ctcatgagcg gatacatatt
     3961 tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc
     4021 acctgacgtc taagaaacca ttattatcat gacattaacc tataaaaata ggcgtatcac
     4081 gaggcccttt cgtctcgcgc gtttcggtga tgacggtgaa aacctctgac acatgcagct
     4141 cccggagacg gtcacagctt gtctgtaagc ggatgccggg agcagacaag cccgtcaggg
     4201 cgcgtcagcg ggtgttggcg ggtgtcgggg ctggcttaac tatgcggcat cagagcagat
     4261 tgtactgaga gtgcaccata aaattgtaaa cgttaatatt ttgttaaaat tcgcgttaaa
     4321 tttttgttaa atcagctcat tttttaacca atagaccgaa atcggcaaaa tcccttataa
     4381 atcaaaagaa tagcccgaga tagagttgag tgttgttcca gtttggaaca agagtccact
     4441 attaaagaac gtggactcca acgtcaaagg gcgaaaaacc gtctatcagg gcgatggccc
     4501 actacgtgaa ccatcaccca aatcaagttt tttggggtcg aggtgccgta aagcactaaa
     4561 tcggaaccct aaagggagcc cccgatttag agcttgacgg ggaaagccgg cgaacgtggc
     4621 gagaaaggaa gggaagaaag cgaaaggagc gggcgctaag gcgctggcaa gtgtagcggt
//
