LOCUS       sequence-492133-        4746 bp    DNA     circular SYN 06-NOV-2025
DEFINITION  AiE2303m 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 4746)
  TITLE     Basal Ganglia/CERP Enhancer AAVs
REFERENCE   2  (bases 1 to 4746)
  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: AiP1638 - pAAV-AiE2303m-minBG-SYFP2-WPRE3-BGHpA
FEATURES             Location/Qualifiers
     source          1..4746
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      complement(166..754)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(246..265)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(925..1716,1717..1785))
                     /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     1548..1567
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(1786..1890)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     1958..1976
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(2014..2036)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     2136..2155
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     rep_origin      complement(2172..2627)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2309..2330)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2521..2540
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     repeat_region   2702..2842
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   2713..2842
                     /label=AAV2 ITR
     polyA_signal    2960..3071
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     3258..3278
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(join(3371..4081,4082..4084,4085..4087))
                     /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(3413..3434)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     3762..3781
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     4021..4042
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     regulatory      4084..4093
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     repeat_region   join(4721..4746,1..104)
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
ORIGIN
        1 actccctctc tgcgcgctcg ctcgctcact gaggccgggc gaccaaaggt cgcccgacgc
       61 ccgggcggcc tcagtgagcg agcgagcgcg cagctgcctg caggacatgt gagcaaaagg
      121 ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg
      181 cccccctgac gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg
      241 actataaaga taccaggcgt ttccccctgg aagctccctc gtgcgctctc ctgttccgac
      301 cctgccgctt accggatacc tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca
      361 tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt
      421 gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc
      481 caacccggta agacacgact tatcgccact ggcagcagcc actggtaaca ggattagcag
      541 agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg tggcctaact acggctacac
      601 tagaagaaca gtatttggta tctgcgctct gctgaagcca gttaccttcg gaaaaagagt
      661 tggtagctct tgatccggca aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa
      721 gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat cctttgatct tttctacggg
      781 gtctgacgct cagtggaacg aaaactcacg ttaagggatt ttggtcatga gattatcaaa
      841 aaggatcttc acctagatcc ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat
      901 atatgagtaa acttggtctg acagttacca atgcttaatc agtgaggcac ctatctcagc
      961 gatctgtcta tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga taactacgat
     1021 acgggagggc ttaccatctg gccccagtgc tgcaatgata ccgcgagacc cacgctcacc
     1081 ggctccagat ttatcagcaa taaaccagcc agccggaagg gccgagcgca gaagtggtcc
     1141 tgcaacttta tccgcctcca tccagtctat taattgttgc cgggaagcta gagtaagtag
     1201 ttcgccagtt aatagtttgc gcaacgttgt tgccattgct acaggcatcg tggtgtcacg
     1261 ctcgtcgttt ggtatggctt cattcagctc cggttcccaa cgatcaaggc gagttacatg
     1321 atcccccatg ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg ttgtcagaag
     1381 taagttggcc gcagtgttat cactcatggt tatggcagca ctgcataatt ctcttactgt
     1441 catgccatcc gtaagatgct tttctgtgac tggtgagtac tcaaccaagt cattctgaga
     1501 atagtgtatg cggcgaccga gttgctcttg cccggcgtca atacgggata ataccgcgcc
     1561 acatagcaga actttaaaag tgctcatcat tggaaaacgt tcttcggggc gaaaactctc
     1621 aaggatctta ccgctgttga gatccagttc gatgtaaccc actcgtgcac ccaactgatc
     1681 ttcagcatct tttactttca ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc
     1741 cgcaaaaaag ggaataaggg cgacacggaa atgttgaata ctcatactct tcctttttca
     1801 atattattga agcatttatc agggttattg tctcatgagc ggatacatat ttgaatgtat
     1861 ttagaaaaat aaacaaatag gggttccgcg cacatttccc cgaaaagtgc cacctgacgt
     1921 ctaagaaacc attattatca tgacattaac ctataaaaat aggcgtatca cgaggccctt
     1981 tcgtctcgcg cgtttcggtg atgacggtga aaacctctga cacatgcagc tcccggagac
     2041 ggtcacagct tgtctgtaag cggatgccgg gagcagacaa gcccgtcagg gcgcgtcagc
     2101 gggtgttggc gggtgtcggg gctggcttaa ctatgcggca tcagagcaga ttgtactgag
     2161 agtgcaccat aaaattgtaa acgttaatat tttgttaaaa ttcgcgttaa atttttgtta
     2221 aatcagctca ttttttaacc aatagaccga aatcggcaaa atcccttata aatcaaaaga
     2281 atagcccgag atagagttga gtgttgttcc agtttggaac aagagtccac tattaaagaa
     2341 cgtggactcc aacgtcaaag ggcgaaaaac cgtctatcag ggcgatggcc cactacgtga
     2401 accatcaccc aaatcaagtt ttttggggtc gaggtgccgt aaagcactaa atcggaaccc
     2461 taaagggagc ccccgattta gagcttgacg gggaaagccg gcgaacgtgg cgagaaagga
     2521 agggaagaaa gcgaaaggag cgggcgctaa ggcgctggca agtgtagcgg tcacgctgcg
     2581 cgtaaccacc acacccgccg cgcttaatgc gccgctacag ggcgcgtact atggttgctt
     2641 tgacgtatgc ggtgtgaaat accgcacaga tgcgtaagga gaaaataccg catcaggcgc
     2701 ccctgcaggc agctgcgcgc tcgctcgctc actgaggccg cccgggcaaa gcccgggcgt
     2761 cgggcgacct ttggtcgccc ggcctcagtg agcgagcgag cgcgcagaga gggagtggcc
     2821 aactccatca ctaggggttc ctgcggccgc tcggtccgca cgtgagatct catgcctgct
     2881 attgtcttcc caatcctccc ccttgctgtc ctgccccacc ccacccccca gaatagaatg
     2941 acacctactc agacaatgcg atgcaatttc ctcattttat taggaaagga cagtgggagt
     3001 ggcaccttcc agggtcaagg aaggcacggg ggaggggcaa acaacagatg gctggcaact
     3061 agaaggcaca gtcgaagatc tctcgagcca cggaattgtc agtgcccaac agccgagccc
     3121 ctgtccagca gcgggcaagg caggcggcga tgagttccgc cgtggcaaga actaaccagg
     3181 atttatacaa ggaggagaaa atgaaagcca tacgggaagc aatagcatga tacaaaggca
     3241 ttaaagcagc gtatccacat agcgtaaaag gagcaacata gttaagaata ccagtcaatc
     3301 tttcacaaat tttgtaatcc agaggttgat tatgatatcg aattcgcggc cgcggcgcgc
     3361 cgtcgactta cttgtacagc tcgtccatgc cgagagtgat cccggcggcg gtcacgaact
     3421 ccagcaggac catgtgatcg cgcttctcgt tggggtcttt gctcagcttg gactggtagc
     3481 tcaggtagtg gttgtcgggc agcagcacgg ggccgtcgcc gatgggggtg ttctgctggt
     3541 agtggtcggc gagctgcacg ccgccgtcct cgatgttgtg gcggatcttg aagttggcct
     3601 tgatgccgtt cttctgcttg tcggcggtga tatagacgtt gtggctgttg tagttgtact
     3661 ccagcttgtg ccccaggatg ttgccgtcct ccttgaagtc gatgcccttc agctcgatgc
     3721 ggttcaccag ggtgtcgccc tcgaacttca cctcggcgcg ggtcttgtag ttgccgtcgt
     3781 ccttgaagaa gatggtgcgc tcctggacgt agccttcggg catggcggac ttgaagaagt
     3841 cgtgctgctt catgtggtcg gggtagcggg cgaagcactg cacgccgtag cccagggtgg
     3901 tcacgagggt gggccagggc acgggcagct tgccggtggt gcagatcagc ttcagggtca
     3961 gcttgccgta ggtggcatcg ccctcgccct cgccggacac gctgaacttg tggccgttta
     4021 cgtcgccgtc cagctcgacc aggatgggca ccaccccggt gaacagctcc tcgcccttgc
     4081 tcaccatggt ggcgaccggt agcgctagct tcgaaagatc tggatcccag aagcaaatgt
     4141 aagcaataga tggctctgcc ctgactttta tgcccagccc gagctccgcc taatctatat
     4201 agtgagttac aggagtgagg gcaacgtagt gaaaccacac ttcaacaata acaataataa
     4261 caacagagat gccatgtgtt ctaagcatct ttatgagata ggatgattca aatgggttaa
     4321 tgttttaaga agaaaaaggc agaggaggta gccaaataac cacttgtact tatgactgcc
     4381 acttggacag ggaccttaga agccgcttct ccctccagac cagacacaga tgtctggctg
     4441 ggctgaggac acattcgtgg tcatgaacct tttgcagata acagaagaga tattcactga
     4501 gccggatatt actggtggcc tctctcagac acaaaggcta gagatggtac agtgcccctg
     4561 ctaaaggcac gggaaaatgg ggcatggaga aacagcacac acagtaggga agaagggcta
     4621 tttggatatc tctaaaagca tttacattac attgatttga gtgtgcaaac atgccatggg
     4681 gtagcagtcc gattattact cttacaacgc gtgcggccgc aggaacccct agtgatggag
     4741 ttggcc
//
