LOCUS       sequence-492132-        4746 bp    DNA     circular SYN 06-NOV-2025
DEFINITION  AiE2313m 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: AiP1646 - pAAV-AiE2313m-minBG-SYFP2-WPRE3-BGHpA
FEATURES             Location/Qualifiers
     source          1..4746
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     588..607
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(826..930)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     998..1016
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(1054..1076)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     1176..1195
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     rep_origin      complement(1212..1667)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1349..1370)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     1561..1580
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     repeat_region   1742..1882
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   1753..1882
                     /label=AAV2 ITR
     polyA_signal    2000..2111
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     2298..2318
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(join(2411..3121,3122..3124,3125..3127))
                     /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(2453..2474)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     2802..2821
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     3061..3082
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     regulatory      3124..3133
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     repeat_region   3761..3890
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     rep_origin      complement(3952..4540)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4032..4051)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4711..4746,1..756,757..825))
                     /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"
ORIGIN
        1 gatctgtcta tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga taactacgat
       61 acgggagggc ttaccatctg gccccagtgc tgcaatgata ccgcgagacc cacgctcacc
      121 ggctccagat ttatcagcaa taaaccagcc agccggaagg gccgagcgca gaagtggtcc
      181 tgcaacttta tccgcctcca tccagtctat taattgttgc cgggaagcta gagtaagtag
      241 ttcgccagtt aatagtttgc gcaacgttgt tgccattgct acaggcatcg tggtgtcacg
      301 ctcgtcgttt ggtatggctt cattcagctc cggttcccaa cgatcaaggc gagttacatg
      361 atcccccatg ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg ttgtcagaag
      421 taagttggcc gcagtgttat cactcatggt tatggcagca ctgcataatt ctcttactgt
      481 catgccatcc gtaagatgct tttctgtgac tggtgagtac tcaaccaagt cattctgaga
      541 atagtgtatg cggcgaccga gttgctcttg cccggcgtca atacgggata ataccgcgcc
      601 acatagcaga actttaaaag tgctcatcat tggaaaacgt tcttcggggc gaaaactctc
      661 aaggatctta ccgctgttga gatccagttc gatgtaaccc actcgtgcac ccaactgatc
      721 ttcagcatct tttactttca ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc
      781 cgcaaaaaag ggaataaggg cgacacggaa atgttgaata ctcatactct tcctttttca
      841 atattattga agcatttatc agggttattg tctcatgagc ggatacatat ttgaatgtat
      901 ttagaaaaat aaacaaatag gggttccgcg cacatttccc cgaaaagtgc cacctgacgt
      961 ctaagaaacc attattatca tgacattaac ctataaaaat aggcgtatca cgaggccctt
     1021 tcgtctcgcg cgtttcggtg atgacggtga aaacctctga cacatgcagc tcccggagac
     1081 ggtcacagct tgtctgtaag cggatgccgg gagcagacaa gcccgtcagg gcgcgtcagc
     1141 gggtgttggc gggtgtcggg gctggcttaa ctatgcggca tcagagcaga ttgtactgag
     1201 agtgcaccat aaaattgtaa acgttaatat tttgttaaaa ttcgcgttaa atttttgtta
     1261 aatcagctca ttttttaacc aatagaccga aatcggcaaa atcccttata aatcaaaaga
     1321 atagcccgag atagagttga gtgttgttcc agtttggaac aagagtccac tattaaagaa
     1381 cgtggactcc aacgtcaaag ggcgaaaaac cgtctatcag ggcgatggcc cactacgtga
     1441 accatcaccc aaatcaagtt ttttggggtc gaggtgccgt aaagcactaa atcggaaccc
     1501 taaagggagc ccccgattta gagcttgacg gggaaagccg gcgaacgtgg cgagaaagga
     1561 agggaagaaa gcgaaaggag cgggcgctaa ggcgctggca agtgtagcgg tcacgctgcg
     1621 cgtaaccacc acacccgccg cgcttaatgc gccgctacag ggcgcgtact atggttgctt
     1681 tgacgtatgc ggtgtgaaat accgcacaga tgcgtaagga gaaaataccg catcaggcgc
     1741 ccctgcaggc agctgcgcgc tcgctcgctc actgaggccg cccgggcaaa gcccgggcgt
     1801 cgggcgacct ttggtcgccc ggcctcagtg agcgagcgag cgcgcagaga gggagtggcc
     1861 aactccatca ctaggggttc ctgcggccgc tcggtccgca cgtgagatct catgcctgct
     1921 attgtcttcc caatcctccc ccttgctgtc ctgccccacc ccacccccca gaatagaatg
     1981 acacctactc agacaatgcg atgcaatttc ctcattttat taggaaagga cagtgggagt
     2041 ggcaccttcc agggtcaagg aaggcacggg ggaggggcaa acaacagatg gctggcaact
     2101 agaaggcaca gtcgaagatc tctcgagcca cggaattgtc agtgcccaac agccgagccc
     2161 ctgtccagca gcgggcaagg caggcggcga tgagttccgc cgtggcaaga actaaccagg
     2221 atttatacaa ggaggagaaa atgaaagcca tacgggaagc aatagcatga tacaaaggca
     2281 ttaaagcagc gtatccacat agcgtaaaag gagcaacata gttaagaata ccagtcaatc
     2341 tttcacaaat tttgtaatcc agaggttgat tatgatatcg aattcgcggc cgcggcgcgc
     2401 cgtcgactta cttgtacagc tcgtccatgc cgagagtgat cccggcggcg gtcacgaact
     2461 ccagcaggac catgtgatcg cgcttctcgt tggggtcttt gctcagcttg gactggtagc
     2521 tcaggtagtg gttgtcgggc agcagcacgg ggccgtcgcc gatgggggtg ttctgctggt
     2581 agtggtcggc gagctgcacg ccgccgtcct cgatgttgtg gcggatcttg aagttggcct
     2641 tgatgccgtt cttctgcttg tcggcggtga tatagacgtt gtggctgttg tagttgtact
     2701 ccagcttgtg ccccaggatg ttgccgtcct ccttgaagtc gatgcccttc agctcgatgc
     2761 ggttcaccag ggtgtcgccc tcgaacttca cctcggcgcg ggtcttgtag ttgccgtcgt
     2821 ccttgaagaa gatggtgcgc tcctggacgt agccttcggg catggcggac ttgaagaagt
     2881 cgtgctgctt catgtggtcg gggtagcggg cgaagcactg cacgccgtag cccagggtgg
     2941 tcacgagggt gggccagggc acgggcagct tgccggtggt gcagatcagc ttcagggtca
     3001 gcttgccgta ggtggcatcg ccctcgccct cgccggacac gctgaacttg tggccgttta
     3061 cgtcgccgtc cagctcgacc aggatgggca ccaccccggt gaacagctcc tcgcccttgc
     3121 tcaccatggt ggcgaccggt agcgctagct tcgaaagatc tggatcccag aagcaaatgt
     3181 aagcaataga tggctctgcc ctgactttta tgcccagccc gagctccgtg aattatctca
     3241 gaccgcattt ttcattatgt ctaagggaag tgagtacatc atacagtgta agcttcgccc
     3301 ttgcttatta caacagaatc ctcccagggc tggcagaaga tgtggtctgt ttccttttct
     3361 tggggctcag acataaattc ttgtttgtta ggtgtggata gcggtaggcg aggatgcagg
     3421 ctttcacatt tacaaaggtt gacctgttct ctcactaggt ctgtaacaag aacacaaata
     3481 cgatcttacc tttcctatcc ttctctcctg gggactctct gggtcttcac actgatccag
     3541 cctgaacctt ggctaccatg agtgttttcc gggtaaactc ttcctaaagt ccagtctggg
     3601 cctggtttca atgtaagagt tgcacctagc ccggaaagga ctggaggccc cagggagttt
     3661 agaggtttgg tggggtgaga gttggggtgg agccattctc atggagacag gggtcaggaa
     3721 tgaggtatgc gattattact cttacaacgc gtgcggccgc aggaacccct agtgatggag
     3781 ttggccactc cctctctgcg cgctcgctcg ctcactgagg ccgggcgacc aaaggtcgcc
     3841 cgacgcccgg gcggcctcag tgagcgagcg agcgcgcagc tgcctgcagg acatgtgagc
     3901 aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag
     3961 gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc
     4021 gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt
     4081 tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct
     4141 ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg
     4201 ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct
     4261 tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat
     4321 tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg
     4381 ctacactaga agaacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa
     4441 aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt
     4501 ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc
     4561 tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt
     4621 atcaaaaagg atcttcacct agatcctttt aaattaaaaa tgaagtttta aatcaatcta
     4681 aagtatatat gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg aggcacctat
     4741 ctcagc
//
