LOCUS       sequence-489679-        5225 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AiE0071h_3xC2 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 5225)
  TITLE     Basal Ganglia/CERP Enhancer AAVs
REFERENCE   2  (bases 1 to 5225)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"
COMMENT     Sequence Label: AiP14494 -
            pAAV-AiE0071h_3xC2-minBG-SYFP2-WPRE...
FEATURES             Location/Qualifiers
     source          1..5225
                     /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      1237..1246
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(1243..1245,1246..1248,1249..1959)
                     /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(1288..1309)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1549..1568)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1896..1917
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(2052..2072)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     polyA_signal    2259..2370
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   2488..2628
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   2488..2617
                     /label=AAV2 ITR
     rep_origin      2703..3158
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2790..2809)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3000..3021
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3175..3194)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3294..3316
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(3354..3372)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3440..3544
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3545..3613,3614..4405)
                     /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(3763..3782)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4576..5164
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5065..5084
                     /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 gcgtctccca gtgtctacct caaggagaaa agaaaggtaa
      181 cagtaggata attgaatttt ttaagaagag aaagaagcag aatgcctaag tgtttaaaaa
      241 gctctattta aatttcttgt ctgtagcatg tactctaacc tataattatg gttttgaaaa
      301 aaaggataat tatgaaaatt tctgggctca ttaaagacat acaattgaca tccaaaatta
      361 tttgagttag aaatgatgct ggaatgtgtc tcttttaaga gactccgagg aaggaatttt
      421 gctttttaaa actctaagtt ccagattaac cctgagtgtc acgggtcacc ttaggtactc
      481 ccagtgtcta cctcaaggag aaaagaaagg taacagtagg ataattgaat tttttaagaa
      541 gagaaagaag cagaatgcct aagtgtttaa aaagctctat ttaaatttct tgtctgtagc
      601 atgtactcta acctataatt atggttttga aaaaaaggat aattatgaaa atttctgggc
      661 tcattaaaga catacaattg acatccaaaa ttatttgagt tagaaatgat gctggaatgt
      721 gtctctttta agagactccg aggaaggaat tttgcttttt aaaactctaa gttccagatt
      781 aaccctgagt gtcacgggtc accttaggta ctcccagtgt ctacctcaag gagaaaagaa
      841 aggtaacagt aggataattg aattttttaa gaagagaaag aagcagaatg cctaagtgtt
      901 taaaaagctc tatttaaatt tcttgtctgt agcatgtact ctaacctata attatggttt
      961 tgaaaaaaag gataattatg aaaatttctg ggctcattaa agacatacaa ttgacatcca
     1021 aaattatttg agttagaaat gatgctggaa tgtgtctctt ttaagagact ccgaggaagg
     1081 aattttgctt tttaaaactc taagttccag attaaccctg agtgtcacgg gtcaccttag
     1141 gtagagctcg ggctgggcat aaaagtcagg gcagagccat ctattgctta catttgcttc
     1201 tgggatccag atctttcgaa gctagcgcta ccggtcgcca ccatggtgag caagggcgag
     1261 gagctgttca ccggggtggt gcccatcctg gtcgagctgg acggcgacgt aaacggccac
     1321 aagttcagcg tgtccggcga gggcgagggc gatgccacct acggcaagct gaccctgaag
     1381 ctgatctgca ccaccggcaa gctgcccgtg ccctggccca ccctcgtgac caccctgggc
     1441 tacggcgtgc agtgcttcgc ccgctacccc gaccacatga agcagcacga cttcttcaag
     1501 tccgccatgc ccgaaggcta cgtccaggag cgcaccatct tcttcaagga cgacggcaac
     1561 tacaagaccc gcgccgaggt gaagttcgag ggcgacaccc tggtgaaccg catcgagctg
     1621 aagggcatcg acttcaagga ggacggcaac atcctggggc acaagctgga gtacaactac
     1681 aacagccaca acgtctatat caccgccgac aagcagaaga acggcatcaa ggccaacttc
     1741 aagatccgcc acaacatcga ggacggcggc gtgcagctcg ccgaccacta ccagcagaac
     1801 acccccatcg gcgacggccc cgtgctgctg cccgacaacc actacctgag ctaccagtcc
     1861 aagctgagca aagaccccaa cgagaagcgc gatcacatgg tcctgctgga gttcgtgacc
     1921 gccgccggga tcactctcgg catggacgag ctgtacaagt aagtcgacgg cgcgccgcgg
     1981 ccgcgaattc gatatcataa tcaacctctg gattacaaaa tttgtgaaag attgactggt
     2041 attcttaact atgttgctcc ttttacgcta tgtggatacg ctgctttaat gcctttgtat
     2101 catgctattg cttcccgtat ggctttcatt ttctcctcct tgtataaatc ctggttagtt
     2161 cttgccacgg cggaactcat cgccgcctgc cttgcccgct gctggacagg ggctcggctg
     2221 ttgggcactg acaattccgt ggctcgagag atcttcgact gtgccttcta gttgccagcc
     2281 atctgttgtt tgcccctccc ccgtgccttc cttgaccctg gaaggtgcca ctcccactgt
     2341 cctttcctaa taaaatgagg aaattgcatc gcattgtctg agtaggtgtc attctattct
     2401 ggggggtggg gtggggcagg acagcaaggg ggaggattgg gaagacaata gcaggcatga
     2461 gatctcacgt gcggaccgag cggccgcagg aacccctagt gatggagttg gccactccct
     2521 ctctgcgcgc tcgctcgctc actgaggccg ggcgaccaaa ggtcgcccga cgcccgggct
     2581 ttgcccgggc ggcctcagtg agcgagcgag cgcgcagctg cctgcagggg cgcctgatgc
     2641 ggtattttct ccttacgcat ctgtgcggta tttcacaccg catacgtcaa agcaaccata
     2701 gtacgcgccc tgtagcggcg cattaagcgc ggcgggtgtg gtggttacgc gcagcgtgac
     2761 cgctacactt gccagcgcct tagcgcccgc tcctttcgct ttcttccctt cctttctcgc
     2821 cacgttcgcc ggctttcccc gtcaagctct aaatcggggg ctccctttag ggttccgatt
     2881 tagtgcttta cggcacctcg accccaaaaa acttgatttg ggtgatggtt cacgtagtgg
     2941 gccatcgccc tgatagacgg tttttcgccc tttgacgttg gagtccacgt tctttaatag
     3001 tggactcttg ttccaaactg gaacaacact caactctatc tcgggctatt cttttgattt
     3061 ataagggatt ttgccgattt cggtctattg gttaaaaaat gagctgattt aacaaaaatt
     3121 taacgcgaat tttaacaaaa tattaacgtt tacaatttta tggtgcactc tcagtacaat
     3181 ctgctctgat gccgcatagt taagccagcc ccgacacccg ccaacacccg ctgacgcgcc
     3241 ctgacgggct tgtctgctcc cggcatccgc ttacagacaa gctgtgaccg tctccgggag
     3301 ctgcatgtgt cagaggtttt caccgtcatc accgaaacgc gcgagacgaa agggcctcgt
     3361 gatacgccta tttttatagg ttaatgtcat gataataatg gtttcttaga cgtcaggtgg
     3421 cacttttcgg ggaaatgtgc gcggaacccc tatttgttta tttttctaaa tacattcaaa
     3481 tatgtatccg ctcatgagac aataaccctg ataaatgctt caataatatt gaaaaaggaa
     3541 gagtatgagt attcaacatt tccgtgtcgc ccttattccc ttttttgcgg cattttgcct
     3601 tcctgttttt gctcacccag aaacgctggt gaaagtaaaa gatgctgaag atcagttggg
     3661 tgcacgagtg ggttacatcg aactggatct caacagcggt aagatccttg agagttttcg
     3721 ccccgaagaa cgttttccaa tgatgagcac ttttaaagtt ctgctatgtg gcgcggtatt
     3781 atcccgtatt gacgccgggc aagagcaact cggtcgccgc atacactatt ctcagaatga
     3841 cttggttgag tactcaccag tcacagaaaa gcatcttacg gatggcatga cagtaagaga
     3901 attatgcagt gctgccataa ccatgagtga taacactgcg gccaacttac ttctgacaac
     3961 gatcggagga ccgaaggagc taaccgcttt tttgcacaac atgggggatc atgtaactcg
     4021 ccttgatcgt tgggaaccgg agctgaatga agccatacca aacgacgagc gtgacaccac
     4081 gatgcctgta gcaatggcaa caacgttgcg caaactatta actggcgaac tacttactct
     4141 agcttcccgg caacaattaa tagactggat ggaggcggat aaagttgcag gaccacttct
     4201 gcgctcggcc cttccggctg gctggtttat tgctgataaa tctggagccg gtgagcgtgg
     4261 gtctcgcggt atcattgcag cactggggcc agatggtaag ccctcccgta tcgtagttat
     4321 ctacacgacg gggagtcagg caactatgga tgaacgaaat agacagatcg ctgagatagg
     4381 tgcctcactg attaagcatt ggtaactgtc agaccaagtt tactcatata tactttagat
     4441 tgatttaaaa cttcattttt aatttaaaag gatctaggtg aagatccttt ttgataatct
     4501 catgaccaaa atcccttaac gtgagttttc gttccactga gcgtcagacc ccgtagaaaa
     4561 gatcaaagga tcttcttgag atcctttttt tctgcgcgta atctgctgct tgcaaacaaa
     4621 aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa gagctaccaa ctctttttcc
     4681 gaaggtaact ggcttcagca gagcgcagat accaaatact gttcttctag tgtagccgta
     4741 gttaggccac cacttcaaga actctgtagc accgcctaca tacctcgctc tgctaatcct
     4801 gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt accgggttgg actcaagacg
     4861 atagttaccg gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca cacagcccag
     4921 cttggagcga acgacctaca ccgaactgag atacctacag cgtgagctat gagaaagcgc
     4981 cacgcttccc gaagggagaa aggcggacag gtatccggta agcggcaggg tcggaacagg
     5041 agagcgcacg agggagcttc cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt
     5101 tcgccacctc tgacttgagc gtcgattttt gtgatgctcg tcaggggggc ggagcctatg
     5161 gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc ttttgctggc cttttgctca
     5221 catgt
//
