LOCUS       sequence-490240-        4745 bp    DNA     circular SYN 22-OCT-2025
DEFINITION  AiE2305m 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 4745)
  TITLE     Basal Ganglia/CERP Enhancer AAVs
REFERENCE   2  (bases 1 to 4745)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Oct 22, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"
COMMENT     Sequence Label: AiP1753 - pAAV-AiE2305m-minBG-SYFP2-WPRE3-BGHpA
FEATURES             Location/Qualifiers
     source          1..4745
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   1..130
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     regulatory      757..766
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(763..765,766..768,769..1479)
                     /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(808..829)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1069..1088)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1416..1437
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(1572..1592)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     polyA_signal    1779..1890
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   2008..2148
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   2008..2137
                     /label=AAV2 ITR
     rep_origin      2223..2678
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2310..2329)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2520..2541
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(2695..2714)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     2814..2836
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(2874..2892)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        2960..3064
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3065..3133,3134..3925)
                     /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(3283..3302)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4096..4684
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4585..4604
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 cctgcaggca gctgcgcgct cgctcgctca ctgaggccgc ccgggcgtcg ggcgaccttt
       61 ggtcgcccgg cctcagtgag cgagcgagcg cgcagagagg gagtggccaa ctccatcact
      121 aggggttcct gcggccgcac gcgttgtaag agtaataatc ggggaaaaga gcaaaaactg
      181 ccttttttgc attttgaagt aaatggatct tttttttttt tttcaagaca agactcactg
      241 ttcgtcatta gcaatcattt ccttaccacc tatcacagtg tggtgtgtgt gtagtgtgcc
      301 tctatgggct cacacatttc aacacttgct ttccagtggt ggcacagatt gggaggtgtg
      361 gagctttcag ggggcggagc ctagatggaa gaagtaaatc gctgcctcca gagttgcagc
      421 ccagcactgc tcttgcttct gggcctgctg ggctggcgag gtgtgagcaa ggcacatgat
      481 catctccctc cacctgagtc accagctact cccaccgcca tgccttcccc agatgatgta
      541 ccgttccccc cacccccacc cccaccaccg gaacctgaaa atcaaatgaa acctttcctc
      601 cctgacgttg ctctagcctg gtgacaaggg gatataacgc atacccctcc caaagggaac
      661 acggagctcg ggctgggcat aaaagtcagg gcagagccat ctattgctta catttgcttc
      721 tgggatccag atctttcgaa gctagcgcta ccggtcgcca ccatggtgag caagggcgag
      781 gagctgttca ccggggtggt gcccatcctg gtcgagctgg acggcgacgt aaacggccac
      841 aagttcagcg tgtccggcga gggcgagggc gatgccacct acggcaagct gaccctgaag
      901 ctgatctgca ccaccggcaa gctgcccgtg ccctggccca ccctcgtgac caccctgggc
      961 tacggcgtgc agtgcttcgc ccgctacccc gaccacatga agcagcacga cttcttcaag
     1021 tccgccatgc ccgaaggcta cgtccaggag cgcaccatct tcttcaagga cgacggcaac
     1081 tacaagaccc gcgccgaggt gaagttcgag ggcgacaccc tggtgaaccg catcgagctg
     1141 aagggcatcg acttcaagga ggacggcaac atcctggggc acaagctgga gtacaactac
     1201 aacagccaca acgtctatat caccgccgac aagcagaaga acggcatcaa ggccaacttc
     1261 aagatccgcc acaacatcga ggacggcggc gtgcagctcg ccgaccacta ccagcagaac
     1321 acccccatcg gcgacggccc cgtgctgctg cccgacaacc actacctgag ctaccagtcc
     1381 aagctgagca aagaccccaa cgagaagcgc gatcacatgg tcctgctgga gttcgtgacc
     1441 gccgccggga tcactctcgg catggacgag ctgtacaagt aagtcgacgg cgcgccgcgg
     1501 ccgcgaattc gatatcataa tcaacctctg gattacaaaa tttgtgaaag attgactggt
     1561 attcttaact atgttgctcc ttttacgcta tgtggatacg ctgctttaat gcctttgtat
     1621 catgctattg cttcccgtat ggctttcatt ttctcctcct tgtataaatc ctggttagtt
     1681 cttgccacgg cggaactcat cgccgcctgc cttgcccgct gctggacagg ggctcggctg
     1741 ttgggcactg acaattccgt ggctcgagag atcttcgact gtgccttcta gttgccagcc
     1801 atctgttgtt tgcccctccc ccgtgccttc cttgaccctg gaaggtgcca ctcccactgt
     1861 cctttcctaa taaaatgagg aaattgcatc gcattgtctg agtaggtgtc attctattct
     1921 ggggggtggg gtggggcagg acagcaaggg ggaggattgg gaagacaata gcaggcatga
     1981 gatctcacgt gcggaccgag cggccgcagg aacccctagt gatggagttg gccactccct
     2041 ctctgcgcgc tcgctcgctc actgaggccg ggcgaccaaa ggtcgcccga cgcccgggct
     2101 ttgcccgggc ggcctcagtg agcgagcgag cgcgcagctg cctgcagggg cgcctgatgc
     2161 ggtattttct ccttacgcat ctgtgcggta tttcacaccg catacgtcaa agcaaccata
     2221 gtacgcgccc tgtagcggcg cattaagcgc ggcgggtgtg gtggttacgc gcagcgtgac
     2281 cgctacactt gccagcgcct tagcgcccgc tcctttcgct ttcttccctt cctttctcgc
     2341 cacgttcgcc ggctttcccc gtcaagctct aaatcggggg ctccctttag ggttccgatt
     2401 tagtgcttta cggcacctcg accccaaaaa acttgatttg ggtgatggtt cacgtagtgg
     2461 gccatcgccc tgatagacgg tttttcgccc tttgacgttg gagtccacgt tctttaatag
     2521 tggactcttg ttccaaactg gaacaacact caactctatc tcgggctatt cttttgattt
     2581 ataagggatt ttgccgattt cggtctattg gttaaaaaat gagctgattt aacaaaaatt
     2641 taacgcgaat tttaacaaaa tattaacgtt tacaatttta tggtgcactc tcagtacaat
     2701 ctgctctgat gccgcatagt taagccagcc ccgacacccg ccaacacccg ctgacgcgcc
     2761 ctgacgggct tgtctgctcc cggcatccgc ttacagacaa gctgtgaccg tctccgggag
     2821 ctgcatgtgt cagaggtttt caccgtcatc accgaaacgc gcgagacgaa agggcctcgt
     2881 gatacgccta tttttatagg ttaatgtcat gataataatg gtttcttaga cgtcaggtgg
     2941 cacttttcgg ggaaatgtgc gcggaacccc tatttgttta tttttctaaa tacattcaaa
     3001 tatgtatccg ctcatgagac aataaccctg ataaatgctt caataatatt gaaaaaggaa
     3061 gagtatgagt attcaacatt tccgtgtcgc ccttattccc ttttttgcgg cattttgcct
     3121 tcctgttttt gctcacccag aaacgctggt gaaagtaaaa gatgctgaag atcagttggg
     3181 tgcacgagtg ggttacatcg aactggatct caacagcggt aagatccttg agagttttcg
     3241 ccccgaagaa cgttttccaa tgatgagcac ttttaaagtt ctgctatgtg gcgcggtatt
     3301 atcccgtatt gacgccgggc aagagcaact cggtcgccgc atacactatt ctcagaatga
     3361 cttggttgag tactcaccag tcacagaaaa gcatcttacg gatggcatga cagtaagaga
     3421 attatgcagt gctgccataa ccatgagtga taacactgcg gccaacttac ttctgacaac
     3481 gatcggagga ccgaaggagc taaccgcttt tttgcacaac atgggggatc atgtaactcg
     3541 ccttgatcgt tgggaaccgg agctgaatga agccatacca aacgacgagc gtgacaccac
     3601 gatgcctgta gcaatggcaa caacgttgcg caaactatta actggcgaac tacttactct
     3661 agcttcccgg caacaattaa tagactggat ggaggcggat aaagttgcag gaccacttct
     3721 gcgctcggcc cttccggctg gctggtttat tgctgataaa tctggagccg gtgagcgtgg
     3781 gtctcgcggt atcattgcag cactggggcc agatggtaag ccctcccgta tcgtagttat
     3841 ctacacgacg gggagtcagg caactatgga tgaacgaaat agacagatcg ctgagatagg
     3901 tgcctcactg attaagcatt ggtaactgtc agaccaagtt tactcatata tactttagat
     3961 tgatttaaaa cttcattttt aatttaaaag gatctaggtg aagatccttt ttgataatct
     4021 catgaccaaa atcccttaac gtgagttttc gttccactga gcgtcagacc ccgtagaaaa
     4081 gatcaaagga tcttcttgag atcctttttt tctgcgcgta atctgctgct tgcaaacaaa
     4141 aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa gagctaccaa ctctttttcc
     4201 gaaggtaact ggcttcagca gagcgcagat accaaatact gttcttctag tgtagccgta
     4261 gttaggccac cacttcaaga actctgtagc accgcctaca tacctcgctc tgctaatcct
     4321 gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt accgggttgg actcaagacg
     4381 atagttaccg gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca cacagcccag
     4441 cttggagcga acgacctaca ccgaactgag atacctacag cgtgagctat gagaaagcgc
     4501 cacgcttccc gaagggagaa aggcggacag gtatccggta agcggcaggg tcggaacagg
     4561 agagcgcacg agggagcttc cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt
     4621 tcgccacctc tgacttgagc gtcgattttt gtgatgctcg tcaggggggc ggagcctatg
     4681 gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc ttttgctggc cttttgctca
     4741 catgt
//
