LOCUS       sequence-492128-        4746 bp    DNA     circular SYN 06-NOV-2025
DEFINITION  AiE2318m 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: AiP1649 - pAAV-AiE2318m-minBG-SYFP2-WPRE3-BGHpA
FEATURES             Location/Qualifiers
     source          1..4746
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     regulatory      395..404
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(401..403,404..406,407..1117)
                     /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(446..467)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(707..726)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1054..1075
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(1210..1230)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     polyA_signal    1417..1528
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   1646..1786
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   1646..1775
                     /label=AAV2 ITR
     rep_origin      1861..2316
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1948..1967)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2158..2179
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(2333..2352)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     2452..2474
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(2512..2530)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        2598..2702
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2703..2771,2772..3563)
                     /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(2921..2940)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3734..4322
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4223..4242
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     repeat_region   4384..4513
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
ORIGIN
        1 tgtggaatcc gggtggggtg tgcaagctga gtcagctggc aggctttggc cagctctggt
       61 ctgcataggc agcggagtca ccgatctggt cccagcccgc atgcatgaca acggaaagtg
      121 ctgaggcgca catcgtgtct gccggagccc aggctgactc tgcgctttct ttcactgtgg
      181 tctaagcctg gtagagactc ctagcctagg ggcagaacaa gcaatccctg gcacagacac
      241 tcaggcgtga ccgtaagcgt acggtttagc cttttctaag actgcgtaag cctggccagc
      301 ggagctcggg ctgggcataa aagtcagggc agagccatct attgcttaca tttgcttctg
      361 ggatccagat ctttcgaagc tagcgctacc ggtcgccacc atggtgagca agggcgagga
      421 gctgttcacc ggggtggtgc ccatcctggt cgagctggac ggcgacgtaa acggccacaa
      481 gttcagcgtg tccggcgagg gcgagggcga tgccacctac ggcaagctga ccctgaagct
      541 gatctgcacc accggcaagc tgcccgtgcc ctggcccacc ctcgtgacca ccctgggcta
      601 cggcgtgcag tgcttcgccc gctaccccga ccacatgaag cagcacgact tcttcaagtc
      661 cgccatgccc gaaggctacg tccaggagcg caccatcttc ttcaaggacg acggcaacta
      721 caagacccgc gccgaggtga agttcgaggg cgacaccctg gtgaaccgca tcgagctgaa
      781 gggcatcgac ttcaaggagg acggcaacat cctggggcac aagctggagt acaactacaa
      841 cagccacaac gtctatatca ccgccgacaa gcagaagaac ggcatcaagg ccaacttcaa
      901 gatccgccac aacatcgagg acggcggcgt gcagctcgcc gaccactacc agcagaacac
      961 ccccatcggc gacggccccg tgctgctgcc cgacaaccac tacctgagct accagtccaa
     1021 gctgagcaaa gaccccaacg agaagcgcga tcacatggtc ctgctggagt tcgtgaccgc
     1081 cgccgggatc actctcggca tggacgagct gtacaagtaa gtcgacggcg cgccgcggcc
     1141 gcgaattcga tatcataatc aacctctgga ttacaaaatt tgtgaaagat tgactggtat
     1201 tcttaactat gttgctcctt ttacgctatg tggatacgct gctttaatgc ctttgtatca
     1261 tgctattgct tcccgtatgg ctttcatttt ctcctccttg tataaatcct ggttagttct
     1321 tgccacggcg gaactcatcg ccgcctgcct tgcccgctgc tggacagggg ctcggctgtt
     1381 gggcactgac aattccgtgg ctcgagagat cttcgactgt gccttctagt tgccagccat
     1441 ctgttgtttg cccctccccc gtgccttcct tgaccctgga aggtgccact cccactgtcc
     1501 tttcctaata aaatgaggaa attgcatcgc attgtctgag taggtgtcat tctattctgg
     1561 ggggtggggt ggggcaggac agcaaggggg aggattggga agacaatagc aggcatgaga
     1621 tctcacgtgc ggaccgagcg gccgcaggaa cccctagtga tggagttggc cactccctct
     1681 ctgcgcgctc gctcgctcac tgaggccggg cgaccaaagg tcgcccgacg cccgggcttt
     1741 gcccgggcgg cctcagtgag cgagcgagcg cgcagctgcc tgcaggggcg cctgatgcgg
     1801 tattttctcc ttacgcatct gtgcggtatt tcacaccgca tacgtcaaag caaccatagt
     1861 acgcgccctg tagcggcgca ttaagcgcgg cgggtgtggt ggttacgcgc agcgtgaccg
     1921 ctacacttgc cagcgcctta gcgcccgctc ctttcgcttt cttcccttcc tttctcgcca
     1981 cgttcgccgg ctttccccgt caagctctaa atcgggggct ccctttaggg ttccgattta
     2041 gtgctttacg gcacctcgac cccaaaaaac ttgatttggg tgatggttca cgtagtgggc
     2101 catcgccctg atagacggtt tttcgccctt tgacgttgga gtccacgttc tttaatagtg
     2161 gactcttgtt ccaaactgga acaacactca actctatctc gggctattct tttgatttat
     2221 aagggatttt gccgatttcg gtctattggt taaaaaatga gctgatttaa caaaaattta
     2281 acgcgaattt taacaaaata ttaacgttta caattttatg gtgcactctc agtacaatct
     2341 gctctgatgc cgcatagtta agccagcccc gacacccgcc aacacccgct gacgcgccct
     2401 gacgggcttg tctgctcccg gcatccgctt acagacaagc tgtgaccgtc tccgggagct
     2461 gcatgtgtca gaggttttca ccgtcatcac cgaaacgcgc gagacgaaag ggcctcgtga
     2521 tacgcctatt tttataggtt aatgtcatga taataatggt ttcttagacg tcaggtggca
     2581 cttttcgggg aaatgtgcgc ggaaccccta tttgtttatt tttctaaata cattcaaata
     2641 tgtatccgct catgagacaa taaccctgat aaatgcttca ataatattga aaaaggaaga
     2701 gtatgagtat tcaacatttc cgtgtcgccc ttattccctt ttttgcggca ttttgccttc
     2761 ctgtttttgc tcacccagaa acgctggtga aagtaaaaga tgctgaagat cagttgggtg
     2821 cacgagtggg ttacatcgaa ctggatctca acagcggtaa gatccttgag agttttcgcc
     2881 ccgaagaacg ttttccaatg atgagcactt ttaaagttct gctatgtggc gcggtattat
     2941 cccgtattga cgccgggcaa gagcaactcg gtcgccgcat acactattct cagaatgact
     3001 tggttgagta ctcaccagtc acagaaaagc atcttacgga tggcatgaca gtaagagaat
     3061 tatgcagtgc tgccataacc atgagtgata acactgcggc caacttactt ctgacaacga
     3121 tcggaggacc gaaggagcta accgcttttt tgcacaacat gggggatcat gtaactcgcc
     3181 ttgatcgttg ggaaccggag ctgaatgaag ccataccaaa cgacgagcgt gacaccacga
     3241 tgcctgtagc aatggcaaca acgttgcgca aactattaac tggcgaacta cttactctag
     3301 cttcccggca acaattaata gactggatgg aggcggataa agttgcagga ccacttctgc
     3361 gctcggccct tccggctggc tggtttattg ctgataaatc tggagccggt gagcgtgggt
     3421 ctcgcggtat cattgcagca ctggggccag atggtaagcc ctcccgtatc gtagttatct
     3481 acacgacggg gagtcaggca actatggatg aacgaaatag acagatcgct gagataggtg
     3541 cctcactgat taagcattgg taactgtcag accaagttta ctcatatata ctttagattg
     3601 atttaaaact tcatttttaa tttaaaagga tctaggtgaa gatccttttt gataatctca
     3661 tgaccaaaat cccttaacgt gagttttcgt tccactgagc gtcagacccc gtagaaaaga
     3721 tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa
     3781 aaccaccgct accagcggtg gtttgtttgc cggatcaaga gctaccaact ctttttccga
     3841 aggtaactgg cttcagcaga gcgcagatac caaatactgt tcttctagtg tagccgtagt
     3901 taggccacca cttcaagaac tctgtagcac cgcctacata cctcgctctg ctaatcctgt
     3961 taccagtggc tgctgccagt ggcgataagt cgtgtcttac cgggttggac tcaagacgat
     4021 agttaccgga taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca cagcccagct
     4081 tggagcgaac gacctacacc gaactgagat acctacagcg tgagctatga gaaagcgcca
     4141 cgcttcccga agggagaaag gcggacaggt atccggtaag cggcagggtc ggaacaggag
     4201 agcgcacgag ggagcttcca gggggaaacg cctggtatct ttatagtcct gtcgggtttc
     4261 gccacctctg acttgagcgt cgatttttgt gatgctcgtc aggggggcgg agcctatgga
     4321 aaaacgccag caacgcggcc tttttacggt tcctggcctt ttgctggcct tttgctcaca
     4381 tgtcctgcag gcagctgcgc gctcgctcgc tcactgaggc cgcccgggcg tcgggcgacc
     4441 tttggtcgcc cggcctcagt gagcgagcga gcgcgcagag agggagtggc caactccatc
     4501 actaggggtt cctgcggccg cacgcgttgt aagagtaata atcggtaggg tgtgaggaat
     4561 ctagggtctg gatccggaca caggctgccg ctgtgcggag cgggtgggcg tgaggggagg
     4621 ttcccttgtt actccctctc cccccagtac tcaccgagtg agtgagcacc gcaccgacgg
     4681 aaaatggaga ggctgcgcgc gagcctggtg ttcgctgctt tatagtcatg tctggctgcc
     4741 tctccc
//
