LOCUS       sequence-492126-        4861 bp    DNA     circular SYN 06-NOV-2025
DEFINITION  AiE2658m 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 4861)
  TITLE     Basal Ganglia/CERP Enhancer AAVs
REFERENCE   2  (bases 1 to 4861)
  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: AiP2131 - pAAV-AiE2658m-minBG-SYFP2-WPRE3-BGHpA
FEATURES             Location/Qualifiers
     source          1..4861
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     regulatory      453..462
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(459..461,462..464,465..1175)
                     /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(504..525)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(765..784)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1112..1133
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(1268..1288)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     polyA_signal    1475..1586
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   1704..1844
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   1704..1833
                     /label=AAV2 ITR
     rep_origin      1919..2374
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2006..2025)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2216..2237
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(2391..2410)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     2510..2532
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(2570..2588)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        2656..2760
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2761..2829,2830..3621)
                     /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(2979..2998)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3792..4380
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4281..4300
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     repeat_region   4442..4571
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
ORIGIN
        1 gtgtactttc tcctgctttc atccctactt caagggttgc tcagtcattc tgacccaccc
       61 cagacgtgag ccagagaaat ccaaagagtc tgctggacta taacaaaaag catctccgaa
      121 taagtcttgg gatgatacta ggagtacaca gagctgctca ctatcttaca gagtaagttc
      181 tagtgagcag taagtagtat atacactcac gtgggtcaca aacaaccttg agtgtaaacg
      241 gttcccagtg gtgctcttga ggggccttgt gagctcactt agggtatatg tgttgttagg
      301 gaaggatcat aagactcagg gccaaagagg ctgtaggagg ctgtgagctc acgccggcgg
      361 agctcgggct gggcataaaa gtcagggcag agccatctat tgcttacatt tgcttctggg
      421 atccagatct ttcgaagcta gcgctaccgg tcgccaccat ggtgagcaag ggcgaggagc
      481 tgttcaccgg ggtggtgccc atcctggtcg agctggacgg cgacgtaaac ggccacaagt
      541 tcagcgtgtc cggcgagggc gagggcgatg ccacctacgg caagctgacc ctgaagctga
      601 tctgcaccac cggcaagctg cccgtgccct ggcccaccct cgtgaccacc ctgggctacg
      661 gcgtgcagtg cttcgcccgc taccccgacc acatgaagca gcacgacttc ttcaagtccg
      721 ccatgcccga aggctacgtc caggagcgca ccatcttctt caaggacgac ggcaactaca
      781 agacccgcgc cgaggtgaag ttcgagggcg acaccctggt gaaccgcatc gagctgaagg
      841 gcatcgactt caaggaggac ggcaacatcc tggggcacaa gctggagtac aactacaaca
      901 gccacaacgt ctatatcacc gccgacaagc agaagaacgg catcaaggcc aacttcaaga
      961 tccgccacaa catcgaggac ggcggcgtgc agctcgccga ccactaccag cagaacaccc
     1021 ccatcggcga cggccccgtg ctgctgcccg acaaccacta cctgagctac cagtccaagc
     1081 tgagcaaaga ccccaacgag aagcgcgatc acatggtcct gctggagttc gtgaccgccg
     1141 ccgggatcac tctcggcatg gacgagctgt acaagtaagt cgacggcgcg ccgcggccgc
     1201 gaattcgata tcataatcaa cctctggatt acaaaatttg tgaaagattg actggtattc
     1261 ttaactatgt tgctcctttt acgctatgtg gatacgctgc tttaatgcct ttgtatcatg
     1321 ctattgcttc ccgtatggct ttcattttct cctccttgta taaatcctgg ttagttcttg
     1381 ccacggcgga actcatcgcc gcctgccttg cccgctgctg gacaggggct cggctgttgg
     1441 gcactgacaa ttccgtggct cgagagatct tcgactgtgc cttctagttg ccagccatct
     1501 gttgtttgcc cctcccccgt gccttccttg accctggaag gtgccactcc cactgtcctt
     1561 tcctaataaa atgaggaaat tgcatcgcat tgtctgagta ggtgtcattc tattctgggg
     1621 ggtggggtgg ggcaggacag caagggggag gattgggaag acaatagcag gcatgagatc
     1681 tcacgtgcgg accgagcggc cgcaggaacc cctagtgatg gagttggcca ctccctctct
     1741 gcgcgctcgc tcgctcactg aggccgggcg accaaaggtc gcccgacgcc cgggctttgc
     1801 ccgggcggcc tcagtgagcg agcgagcgcg cagctgcctg caggggcgcc tgatgcggta
     1861 ttttctcctt acgcatctgt gcggtatttc acaccgcata cgtcaaagca accatagtac
     1921 gcgccctgta gcggcgcatt aagcgcggcg ggtgtggtgg ttacgcgcag cgtgaccgct
     1981 acacttgcca gcgccttagc gcccgctcct ttcgctttct tcccttcctt tctcgccacg
     2041 ttcgccggct ttccccgtca agctctaaat cgggggctcc ctttagggtt ccgatttagt
     2101 gctttacggc acctcgaccc caaaaaactt gatttgggtg atggttcacg tagtgggcca
     2161 tcgccctgat agacggtttt tcgccctttg acgttggagt ccacgttctt taatagtgga
     2221 ctcttgttcc aaactggaac aacactcaac tctatctcgg gctattcttt tgatttataa
     2281 gggattttgc cgatttcggt ctattggtta aaaaatgagc tgatttaaca aaaatttaac
     2341 gcgaatttta acaaaatatt aacgtttaca attttatggt gcactctcag tacaatctgc
     2401 tctgatgccg catagttaag ccagccccga cacccgccaa cacccgctga cgcgccctga
     2461 cgggcttgtc tgctcccggc atccgcttac agacaagctg tgaccgtctc cgggagctgc
     2521 atgtgtcaga ggttttcacc gtcatcaccg aaacgcgcga gacgaaaggg cctcgtgata
     2581 cgcctatttt tataggttaa tgtcatgata ataatggttt cttagacgtc aggtggcact
     2641 tttcggggaa atgtgcgcgg aacccctatt tgtttatttt tctaaataca ttcaaatatg
     2701 tatccgctca tgagacaata accctgataa atgcttcaat aatattgaaa aaggaagagt
     2761 atgagtattc aacatttccg tgtcgccctt attccctttt ttgcggcatt ttgccttcct
     2821 gtttttgctc acccagaaac gctggtgaaa gtaaaagatg ctgaagatca gttgggtgca
     2881 cgagtgggtt acatcgaact ggatctcaac agcggtaaga tccttgagag ttttcgcccc
     2941 gaagaacgtt ttccaatgat gagcactttt aaagttctgc tatgtggcgc ggtattatcc
     3001 cgtattgacg ccgggcaaga gcaactcggt cgccgcatac actattctca gaatgacttg
     3061 gttgagtact caccagtcac agaaaagcat cttacggatg gcatgacagt aagagaatta
     3121 tgcagtgctg ccataaccat gagtgataac actgcggcca acttacttct gacaacgatc
     3181 ggaggaccga aggagctaac cgcttttttg cacaacatgg gggatcatgt aactcgcctt
     3241 gatcgttggg aaccggagct gaatgaagcc ataccaaacg acgagcgtga caccacgatg
     3301 cctgtagcaa tggcaacaac gttgcgcaaa ctattaactg gcgaactact tactctagct
     3361 tcccggcaac aattaataga ctggatggag gcggataaag ttgcaggacc acttctgcgc
     3421 tcggcccttc cggctggctg gtttattgct gataaatctg gagccggtga gcgtgggtct
     3481 cgcggtatca ttgcagcact ggggccagat ggtaagccct cccgtatcgt agttatctac
     3541 acgacgggga gtcaggcaac tatggatgaa cgaaatagac agatcgctga gataggtgcc
     3601 tcactgatta agcattggta actgtcagac caagtttact catatatact ttagattgat
     3661 ttaaaacttc atttttaatt taaaaggatc taggtgaaga tcctttttga taatctcatg
     3721 accaaaatcc cttaacgtga gttttcgttc cactgagcgt cagaccccgt agaaaagatc
     3781 aaaggatctt cttgagatcc tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa
     3841 ccaccgctac cagcggtggt ttgtttgccg gatcaagagc taccaactct ttttccgaag
     3901 gtaactggct tcagcagagc gcagatacca aatactgttc ttctagtgta gccgtagtta
     3961 ggccaccact tcaagaactc tgtagcaccg cctacatacc tcgctctgct aatcctgtta
     4021 ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg ggttggactc aagacgatag
     4081 ttaccggata aggcgcagcg gtcgggctga acggggggtt cgtgcacaca gcccagcttg
     4141 gagcgaacga cctacaccga actgagatac ctacagcgtg agctatgaga aagcgccacg
     4201 cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg aacaggagag
     4261 cgcacgaggg agcttccagg gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc
     4321 cacctctgac ttgagcgtcg atttttgtga tgctcgtcag gggggcggag cctatggaaa
     4381 aacgccagca acgcggcctt tttacggttc ctggcctttt gctggccttt tgctcacatg
     4441 tcctgcaggc agctgcgcgc tcgctcgctc actgaggccg cccgggcgtc gggcgacctt
     4501 tggtcgcccg gcctcagtga gcgagcgagc gcgcagagag ggagtggcca actccatcac
     4561 taggggttcc tgcggccgca cgcgttccgt ttccagtctg atgtctggcg tctgtcctct
     4621 ctggtttttg gaataggcca gttttcctgg aatatgcttt tattgaactc taaaaaggct
     4681 ggatctttta tgcagattta cttactattt caaagactct ttgaagttgg caaaacattc
     4741 tgttatttcc gagagttatt accgctgaaa tatgggcata gcctgggagg tcagtctgca
     4801 atcttgttta gatcctttta aatttagaaa gtttacagtg gcttaaaaat agatcacatt
     4861 t
//
