LOCUS       sequence-492113-        4681 bp    DNA     circular SYN 06-NOV-2025
DEFINITION  AiE2647m 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 4681)
  TITLE     Basal Ganglia/CERP Enhancer AAVs
REFERENCE   2  (bases 1 to 4681)
  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: AiP2118 - pAAV-AiE2647m-minBG-SYFP2-WPRE3-BGHpA
FEATURES             Location/Qualifiers
     source          1..4681
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     regulatory      235..244
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(241..243,244..246,247..957)
                     /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(286..307)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(547..566)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     894..915
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(1050..1070)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     polyA_signal    1257..1368
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   1486..1626
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   1486..1615
                     /label=AAV2 ITR
     rep_origin      1701..2156
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1788..1807)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1998..2019
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(2173..2192)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     2292..2314
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(2352..2370)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        2438..2542
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2543..2611,2612..3403)
                     /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(2761..2780)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3574..4162
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4063..4082
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     repeat_region   4224..4353
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
ORIGIN
        1 caacattgcg tattttgagg aattgaggaa attagtgcca ttttcaagga ctttgttttc
       61 acaaggatga aagatataag gatgttggtt ttcaccacat ctctcttaaa cttttctaac
      121 tggtctgtca caacgccggc ggagctcggg ctgggcataa aagtcagggc agagccatct
      181 attgcttaca tttgcttctg ggatccagat ctttcgaagc tagcgctacc ggtcgccacc
      241 atggtgagca agggcgagga gctgttcacc ggggtggtgc ccatcctggt cgagctggac
      301 ggcgacgtaa acggccacaa gttcagcgtg tccggcgagg gcgagggcga tgccacctac
      361 ggcaagctga ccctgaagct gatctgcacc accggcaagc tgcccgtgcc ctggcccacc
      421 ctcgtgacca ccctgggcta cggcgtgcag tgcttcgccc gctaccccga ccacatgaag
      481 cagcacgact tcttcaagtc cgccatgccc gaaggctacg tccaggagcg caccatcttc
      541 ttcaaggacg acggcaacta caagacccgc gccgaggtga agttcgaggg cgacaccctg
      601 gtgaaccgca tcgagctgaa gggcatcgac ttcaaggagg acggcaacat cctggggcac
      661 aagctggagt acaactacaa cagccacaac gtctatatca ccgccgacaa gcagaagaac
      721 ggcatcaagg ccaacttcaa gatccgccac aacatcgagg acggcggcgt gcagctcgcc
      781 gaccactacc agcagaacac ccccatcggc gacggccccg tgctgctgcc cgacaaccac
      841 tacctgagct accagtccaa gctgagcaaa gaccccaacg agaagcgcga tcacatggtc
      901 ctgctggagt tcgtgaccgc cgccgggatc actctcggca tggacgagct gtacaagtaa
      961 gtcgacggcg cgccgcggcc gcgaattcga tatcataatc aacctctgga ttacaaaatt
     1021 tgtgaaagat tgactggtat tcttaactat gttgctcctt ttacgctatg tggatacgct
     1081 gctttaatgc ctttgtatca tgctattgct tcccgtatgg ctttcatttt ctcctccttg
     1141 tataaatcct ggttagttct tgccacggcg gaactcatcg ccgcctgcct tgcccgctgc
     1201 tggacagggg ctcggctgtt gggcactgac aattccgtgg ctcgagagat cttcgactgt
     1261 gccttctagt tgccagccat ctgttgtttg cccctccccc gtgccttcct tgaccctgga
     1321 aggtgccact cccactgtcc tttcctaata aaatgaggaa attgcatcgc attgtctgag
     1381 taggtgtcat tctattctgg ggggtggggt ggggcaggac agcaaggggg aggattggga
     1441 agacaatagc aggcatgaga tctcacgtgc ggaccgagcg gccgcaggaa cccctagtga
     1501 tggagttggc cactccctct ctgcgcgctc gctcgctcac tgaggccggg cgaccaaagg
     1561 tcgcccgacg cccgggcttt gcccgggcgg cctcagtgag cgagcgagcg cgcagctgcc
     1621 tgcaggggcg cctgatgcgg tattttctcc ttacgcatct gtgcggtatt tcacaccgca
     1681 tacgtcaaag caaccatagt acgcgccctg tagcggcgca ttaagcgcgg cgggtgtggt
     1741 ggttacgcgc agcgtgaccg ctacacttgc cagcgcctta gcgcccgctc ctttcgcttt
     1801 cttcccttcc tttctcgcca cgttcgccgg ctttccccgt caagctctaa atcgggggct
     1861 ccctttaggg ttccgattta gtgctttacg gcacctcgac cccaaaaaac ttgatttggg
     1921 tgatggttca cgtagtgggc catcgccctg atagacggtt tttcgccctt tgacgttgga
     1981 gtccacgttc tttaatagtg gactcttgtt ccaaactgga acaacactca actctatctc
     2041 gggctattct tttgatttat aagggatttt gccgatttcg gtctattggt taaaaaatga
     2101 gctgatttaa caaaaattta acgcgaattt taacaaaata ttaacgttta caattttatg
     2161 gtgcactctc agtacaatct gctctgatgc cgcatagtta agccagcccc gacacccgcc
     2221 aacacccgct gacgcgccct gacgggcttg tctgctcccg gcatccgctt acagacaagc
     2281 tgtgaccgtc tccgggagct gcatgtgtca gaggttttca ccgtcatcac cgaaacgcgc
     2341 gagacgaaag ggcctcgtga tacgcctatt tttataggtt aatgtcatga taataatggt
     2401 ttcttagacg tcaggtggca cttttcgggg aaatgtgcgc ggaaccccta tttgtttatt
     2461 tttctaaata cattcaaata tgtatccgct catgagacaa taaccctgat aaatgcttca
     2521 ataatattga aaaaggaaga gtatgagtat tcaacatttc cgtgtcgccc ttattccctt
     2581 ttttgcggca ttttgccttc ctgtttttgc tcacccagaa acgctggtga aagtaaaaga
     2641 tgctgaagat cagttgggtg cacgagtggg ttacatcgaa ctggatctca acagcggtaa
     2701 gatccttgag agttttcgcc ccgaagaacg ttttccaatg atgagcactt ttaaagttct
     2761 gctatgtggc gcggtattat cccgtattga cgccgggcaa gagcaactcg gtcgccgcat
     2821 acactattct cagaatgact tggttgagta ctcaccagtc acagaaaagc atcttacgga
     2881 tggcatgaca gtaagagaat tatgcagtgc tgccataacc atgagtgata acactgcggc
     2941 caacttactt ctgacaacga tcggaggacc gaaggagcta accgcttttt tgcacaacat
     3001 gggggatcat gtaactcgcc ttgatcgttg ggaaccggag ctgaatgaag ccataccaaa
     3061 cgacgagcgt gacaccacga tgcctgtagc aatggcaaca acgttgcgca aactattaac
     3121 tggcgaacta cttactctag cttcccggca acaattaata gactggatgg aggcggataa
     3181 agttgcagga ccacttctgc gctcggccct tccggctggc tggtttattg ctgataaatc
     3241 tggagccggt gagcgtgggt ctcgcggtat cattgcagca ctggggccag atggtaagcc
     3301 ctcccgtatc gtagttatct acacgacggg gagtcaggca actatggatg aacgaaatag
     3361 acagatcgct gagataggtg cctcactgat taagcattgg taactgtcag accaagttta
     3421 ctcatatata ctttagattg atttaaaact tcatttttaa tttaaaagga tctaggtgaa
     3481 gatccttttt gataatctca tgaccaaaat cccttaacgt gagttttcgt tccactgagc
     3541 gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat
     3601 ctgctgcttg caaacaaaaa aaccaccgct accagcggtg gtttgtttgc cggatcaaga
     3661 gctaccaact ctttttccga aggtaactgg cttcagcaga gcgcagatac caaatactgt
     3721 tcttctagtg tagccgtagt taggccacca cttcaagaac tctgtagcac cgcctacata
     3781 cctcgctctg ctaatcctgt taccagtggc tgctgccagt ggcgataagt cgtgtcttac
     3841 cgggttggac tcaagacgat agttaccgga taaggcgcag cggtcgggct gaacgggggg
     3901 ttcgtgcaca cagcccagct tggagcgaac gacctacacc gaactgagat acctacagcg
     3961 tgagctatga gaaagcgcca cgcttcccga agggagaaag gcggacaggt atccggtaag
     4021 cggcagggtc ggaacaggag agcgcacgag ggagcttcca gggggaaacg cctggtatct
     4081 ttatagtcct gtcgggtttc gccacctctg acttgagcgt cgatttttgt gatgctcgtc
     4141 aggggggcgg agcctatgga aaaacgccag caacgcggcc tttttacggt tcctggcctt
     4201 ttgctggcct tttgctcaca tgtcctgcag gcagctgcgc gctcgctcgc tcactgaggc
     4261 cgcccgggcg tcgggcgacc tttggtcgcc cggcctcagt gagcgagcga gcgcgcagag
     4321 agggagtggc caactccatc actaggggtt cctgcggccg cacgcgtgag atatttgata
     4381 ttgattctga attgacactg actacagtag agcccaaaag cattttggcc ctgcagaaaa
     4441 agtatgtatt atgggagtca ggtgattaat gaagtttgga ctgaagtcaa atttacataa
     4501 catccgtgga tgcacaattt atcctgcttt ttccagttac agaatgcata tttgtgttag
     4561 atattcttag acctcatcaa aattcataaa tttgttccct gacatttgga acacaagcta
     4621 ctttattgga aaggccaatt ggaaggcatt ggtaatgcca gggtaaaaaa tgaactaaaa
     4681 t
//
