LOCUS       sequence-492121-        4786 bp    DNA     circular SYN 06-NOV-2025
DEFINITION  AiE2653m 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 4786)
  TITLE     Basal Ganglia/CERP Enhancer AAVs
REFERENCE   2  (bases 1 to 4786)
  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: AiP2127 - pAAV-AiE2653m-minBG-SYFP2-WPRE3-BGHpA
FEATURES             Location/Qualifiers
     source          1..4786
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     regulatory      547..556
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(553..555,556..558,559..1269)
                     /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(598..619)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(859..878)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1206..1227
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(1362..1382)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     polyA_signal    1569..1680
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   1798..1938
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   1798..1927
                     /label=AAV2 ITR
     rep_origin      2013..2468
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2100..2119)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2310..2331
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(2485..2504)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     2604..2626
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(2664..2682)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        2750..2854
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2855..2923,2924..3715)
                     /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(3073..3092)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3886..4474
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4375..4394
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     repeat_region   4536..4665
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
ORIGIN
        1 gcctcttcac catgaacctg atagggtatc atgggattct ctgggagaaa gtaaagctag
       61 tcactgggtg atgaaaaatg agggccaata atgcaaatgg tataaaagaa acaaacaaag
      121 cttgtgttga ccaatgctta gagttgccgg cagagttcaa acagagtaga cagactcaca
      181 ctccagggaa taagtggaat ggaacattac ctttctgatc cctttaaaaa tgttggatgc
      241 taaaaacagt gcagggaaat gtctttagct tgacatgagt gcataagcaa cagtgatgaa
      301 gtccaaatgc agccggaatg agaggcttct cagggaagac cgtgctcgtg gaaatttgaa
      361 aaattcagaa gagataagag gagaacaacg ctttgttttc cttgttcact gtgaagatat
      421 tatacaagat gcatgagatg tgaaacgccg gcggagctcg ggctgggcat aaaagtcagg
      481 gcagagccat ctattgctta catttgcttc tgggatccag atctttcgaa gctagcgcta
      541 ccggtcgcca ccatggtgag caagggcgag gagctgttca ccggggtggt gcccatcctg
      601 gtcgagctgg acggcgacgt aaacggccac aagttcagcg tgtccggcga gggcgagggc
      661 gatgccacct acggcaagct gaccctgaag ctgatctgca ccaccggcaa gctgcccgtg
      721 ccctggccca ccctcgtgac caccctgggc tacggcgtgc agtgcttcgc ccgctacccc
      781 gaccacatga agcagcacga cttcttcaag tccgccatgc ccgaaggcta cgtccaggag
      841 cgcaccatct tcttcaagga cgacggcaac tacaagaccc gcgccgaggt gaagttcgag
      901 ggcgacaccc tggtgaaccg catcgagctg aagggcatcg acttcaagga ggacggcaac
      961 atcctggggc acaagctgga gtacaactac aacagccaca acgtctatat caccgccgac
     1021 aagcagaaga acggcatcaa ggccaacttc aagatccgcc acaacatcga ggacggcggc
     1081 gtgcagctcg ccgaccacta ccagcagaac acccccatcg gcgacggccc cgtgctgctg
     1141 cccgacaacc actacctgag ctaccagtcc aagctgagca aagaccccaa cgagaagcgc
     1201 gatcacatgg tcctgctgga gttcgtgacc gccgccggga tcactctcgg catggacgag
     1261 ctgtacaagt aagtcgacgg cgcgccgcgg ccgcgaattc gatatcataa tcaacctctg
     1321 gattacaaaa tttgtgaaag attgactggt attcttaact atgttgctcc ttttacgcta
     1381 tgtggatacg ctgctttaat gcctttgtat catgctattg cttcccgtat ggctttcatt
     1441 ttctcctcct tgtataaatc ctggttagtt cttgccacgg cggaactcat cgccgcctgc
     1501 cttgcccgct gctggacagg ggctcggctg ttgggcactg acaattccgt ggctcgagag
     1561 atcttcgact gtgccttcta gttgccagcc atctgttgtt tgcccctccc ccgtgccttc
     1621 cttgaccctg gaaggtgcca ctcccactgt cctttcctaa taaaatgagg aaattgcatc
     1681 gcattgtctg agtaggtgtc attctattct ggggggtggg gtggggcagg acagcaaggg
     1741 ggaggattgg gaagacaata gcaggcatga gatctcacgt gcggaccgag cggccgcagg
     1801 aacccctagt gatggagttg gccactccct ctctgcgcgc tcgctcgctc actgaggccg
     1861 ggcgaccaaa ggtcgcccga cgcccgggct ttgcccgggc ggcctcagtg agcgagcgag
     1921 cgcgcagctg cctgcagggg cgcctgatgc ggtattttct ccttacgcat ctgtgcggta
     1981 tttcacaccg catacgtcaa agcaaccata gtacgcgccc tgtagcggcg cattaagcgc
     2041 ggcgggtgtg gtggttacgc gcagcgtgac cgctacactt gccagcgcct tagcgcccgc
     2101 tcctttcgct ttcttccctt cctttctcgc cacgttcgcc ggctttcccc gtcaagctct
     2161 aaatcggggg ctccctttag ggttccgatt tagtgcttta cggcacctcg accccaaaaa
     2221 acttgatttg ggtgatggtt cacgtagtgg gccatcgccc tgatagacgg tttttcgccc
     2281 tttgacgttg gagtccacgt tctttaatag tggactcttg ttccaaactg gaacaacact
     2341 caactctatc tcgggctatt cttttgattt ataagggatt ttgccgattt cggtctattg
     2401 gttaaaaaat gagctgattt aacaaaaatt taacgcgaat tttaacaaaa tattaacgtt
     2461 tacaatttta tggtgcactc tcagtacaat ctgctctgat gccgcatagt taagccagcc
     2521 ccgacacccg ccaacacccg ctgacgcgcc ctgacgggct tgtctgctcc cggcatccgc
     2581 ttacagacaa gctgtgaccg tctccgggag ctgcatgtgt cagaggtttt caccgtcatc
     2641 accgaaacgc gcgagacgaa agggcctcgt gatacgccta tttttatagg ttaatgtcat
     2701 gataataatg gtttcttaga cgtcaggtgg cacttttcgg ggaaatgtgc gcggaacccc
     2761 tatttgttta tttttctaaa tacattcaaa tatgtatccg ctcatgagac aataaccctg
     2821 ataaatgctt caataatatt gaaaaaggaa gagtatgagt attcaacatt tccgtgtcgc
     2881 ccttattccc ttttttgcgg cattttgcct tcctgttttt gctcacccag aaacgctggt
     2941 gaaagtaaaa gatgctgaag atcagttggg tgcacgagtg ggttacatcg aactggatct
     3001 caacagcggt aagatccttg agagttttcg ccccgaagaa cgttttccaa tgatgagcac
     3061 ttttaaagtt ctgctatgtg gcgcggtatt atcccgtatt gacgccgggc aagagcaact
     3121 cggtcgccgc atacactatt ctcagaatga cttggttgag tactcaccag tcacagaaaa
     3181 gcatcttacg gatggcatga cagtaagaga attatgcagt gctgccataa ccatgagtga
     3241 taacactgcg gccaacttac ttctgacaac gatcggagga ccgaaggagc taaccgcttt
     3301 tttgcacaac atgggggatc atgtaactcg ccttgatcgt tgggaaccgg agctgaatga
     3361 agccatacca aacgacgagc gtgacaccac gatgcctgta gcaatggcaa caacgttgcg
     3421 caaactatta actggcgaac tacttactct agcttcccgg caacaattaa tagactggat
     3481 ggaggcggat aaagttgcag gaccacttct gcgctcggcc cttccggctg gctggtttat
     3541 tgctgataaa tctggagccg gtgagcgtgg gtctcgcggt atcattgcag cactggggcc
     3601 agatggtaag ccctcccgta tcgtagttat ctacacgacg gggagtcagg caactatgga
     3661 tgaacgaaat agacagatcg ctgagatagg tgcctcactg attaagcatt ggtaactgtc
     3721 agaccaagtt tactcatata tactttagat tgatttaaaa cttcattttt aatttaaaag
     3781 gatctaggtg aagatccttt ttgataatct catgaccaaa atcccttaac gtgagttttc
     3841 gttccactga gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag atcctttttt
     3901 tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt
     3961 gccggatcaa gagctaccaa ctctttttcc gaaggtaact ggcttcagca gagcgcagat
     4021 accaaatact gttcttctag tgtagccgta gttaggccac cacttcaaga actctgtagc
     4081 accgcctaca tacctcgctc tgctaatcct gttaccagtg gctgctgcca gtggcgataa
     4141 gtcgtgtctt accgggttgg actcaagacg atagttaccg gataaggcgc agcggtcggg
     4201 ctgaacgggg ggttcgtgca cacagcccag cttggagcga acgacctaca ccgaactgag
     4261 atacctacag cgtgagctat gagaaagcgc cacgcttccc gaagggagaa aggcggacag
     4321 gtatccggta agcggcaggg tcggaacagg agagcgcacg agggagcttc cagggggaaa
     4381 cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt
     4441 gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg
     4501 gttcctggcc ttttgctggc cttttgctca catgtcctgc aggcagctgc gcgctcgctc
     4561 gctcactgag gccgcccggg cgtcgggcga cctttggtcg cccggcctca gtgagcgagc
     4621 gagcgcgcag agagggagtg gccaactcca tcactagggg ttcctgcggc cgcacgcgta
     4681 tagaaatgct gatagtcttg tagctaggta gatttgaaga acagtggaga ggggacactc
     4741 tccccttccc catcccctgc ctttattgac agcagccttt gacgta
//
