LOCUS       sequence-492131-        4746 bp    DNA     circular SYN 06-NOV-2025
DEFINITION  AiE2306m 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: AiP1641 - pAAV-AiE2306m-minBG-SYFP2-WPRE3-BGHpA
FEATURES             Location/Qualifiers
     source          1..4746
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     148..169
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(323..342)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     442..464
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(502..520)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        588..692
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(693..761,762..1553)
                     /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(911..930)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      1724..2312
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2213..2232
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     repeat_region   2374..2503
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     regulatory      3131..3140
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(3137..3139,3140..3142,3143..3853)
                     /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(3182..3203)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(3443..3462)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     3790..3811
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(3946..3966)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     polyA_signal    4153..4264
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   4382..4522
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   4382..4511
                     /label=AAV2 ITR
     rep_origin      join(4597..4746,1..306)
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4684..4703)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
ORIGIN
        1 atcgggggct ccctttaggg ttccgattta gtgctttacg gcacctcgac cccaaaaaac
       61 ttgatttggg tgatggttca cgtagtgggc catcgccctg atagacggtt tttcgccctt
      121 tgacgttgga gtccacgttc tttaatagtg gactcttgtt ccaaactgga acaacactca
      181 actctatctc gggctattct tttgatttat aagggatttt gccgatttcg gtctattggt
      241 taaaaaatga gctgatttaa caaaaattta acgcgaattt taacaaaata ttaacgttta
      301 caattttatg gtgcactctc agtacaatct gctctgatgc cgcatagtta agccagcccc
      361 gacacccgcc aacacccgct gacgcgccct gacgggcttg tctgctcccg gcatccgctt
      421 acagacaagc tgtgaccgtc tccgggagct gcatgtgtca gaggttttca ccgtcatcac
      481 cgaaacgcgc gagacgaaag ggcctcgtga tacgcctatt tttataggtt aatgtcatga
      541 taataatggt ttcttagacg tcaggtggca cttttcgggg aaatgtgcgc ggaaccccta
      601 tttgtttatt tttctaaata cattcaaata tgtatccgct catgagacaa taaccctgat
      661 aaatgcttca ataatattga aaaaggaaga gtatgagtat tcaacatttc cgtgtcgccc
      721 ttattccctt ttttgcggca ttttgccttc ctgtttttgc tcacccagaa acgctggtga
      781 aagtaaaaga tgctgaagat cagttgggtg cacgagtggg ttacatcgaa ctggatctca
      841 acagcggtaa gatccttgag agttttcgcc ccgaagaacg ttttccaatg atgagcactt
      901 ttaaagttct gctatgtggc gcggtattat cccgtattga cgccgggcaa gagcaactcg
      961 gtcgccgcat acactattct cagaatgact tggttgagta ctcaccagtc acagaaaagc
     1021 atcttacgga tggcatgaca gtaagagaat tatgcagtgc tgccataacc atgagtgata
     1081 acactgcggc caacttactt ctgacaacga tcggaggacc gaaggagcta accgcttttt
     1141 tgcacaacat gggggatcat gtaactcgcc ttgatcgttg ggaaccggag ctgaatgaag
     1201 ccataccaaa cgacgagcgt gacaccacga tgcctgtagc aatggcaaca acgttgcgca
     1261 aactattaac tggcgaacta cttactctag cttcccggca acaattaata gactggatgg
     1321 aggcggataa agttgcagga ccacttctgc gctcggccct tccggctggc tggtttattg
     1381 ctgataaatc tggagccggt gagcgtgggt ctcgcggtat cattgcagca ctggggccag
     1441 atggtaagcc ctcccgtatc gtagttatct acacgacggg gagtcaggca actatggatg
     1501 aacgaaatag acagatcgct gagataggtg cctcactgat taagcattgg taactgtcag
     1561 accaagttta ctcatatata ctttagattg atttaaaact tcatttttaa tttaaaagga
     1621 tctaggtgaa gatccttttt gataatctca tgaccaaaat cccttaacgt gagttttcgt
     1681 tccactgagc gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat cctttttttc
     1741 tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct accagcggtg gtttgtttgc
     1801 cggatcaaga gctaccaact ctttttccga aggtaactgg cttcagcaga gcgcagatac
     1861 caaatactgt tcttctagtg tagccgtagt taggccacca cttcaagaac tctgtagcac
     1921 cgcctacata cctcgctctg ctaatcctgt taccagtggc tgctgccagt ggcgataagt
     1981 cgtgtcttac cgggttggac tcaagacgat agttaccgga taaggcgcag cggtcgggct
     2041 gaacgggggg ttcgtgcaca cagcccagct tggagcgaac gacctacacc gaactgagat
     2101 acctacagcg tgagctatga gaaagcgcca cgcttcccga agggagaaag gcggacaggt
     2161 atccggtaag cggcagggtc ggaacaggag agcgcacgag ggagcttcca gggggaaacg
     2221 cctggtatct ttatagtcct gtcgggtttc gccacctctg acttgagcgt cgatttttgt
     2281 gatgctcgtc aggggggcgg agcctatgga aaaacgccag caacgcggcc tttttacggt
     2341 tcctggcctt ttgctggcct tttgctcaca tgtcctgcag gcagctgcgc gctcgctcgc
     2401 tcactgaggc cgcccgggcg tcgggcgacc tttggtcgcc cggcctcagt gagcgagcga
     2461 gcgcgcagag agggagtggc caactccatc actaggggtt cctgcggccg cacgcgttgt
     2521 aagagtaata atcgagcttc ttccatttca ggtaacttat tccctggtcc agcccaccaa
     2581 tcagagccta tacctttggg gatattctcc ctgcttattc tttttcatgt agggcttgga
     2641 caataggcag aagatcagct gaatatctgt gctaatctct ggggggtttg catttcagaa
     2701 acaaaccggg gctcttacca gggctgtcat ttatcatggc ccattgtgtt tggagaatat
     2761 aagaaaaggt ggcgctctgg ggagagctcc agctgcagga tgatgaatca ctgggctgcc
     2821 actgaaggta tacccagtcc tttatttcct gttcctcggc aaaggctgtg agtgtatctg
     2881 tatcaggctt ggcgatcaga taacatagat aaagttaaag cgccaaatga cttgttattt
     2941 tatcaaagct cgttttgaaa ttgttatcca tccttgttat tgtatttaac aaaggcattc
     3001 agattatcat tacaacaaag cccaacactg ttttacggag ctcgggctgg gcataaaagt
     3061 cagggcagag ccatctattg cttacatttg cttctgggat ccagatcttt cgaagctagc
     3121 gctaccggtc gccaccatgg tgagcaaggg cgaggagctg ttcaccgggg tggtgcccat
     3181 cctggtcgag ctggacggcg acgtaaacgg ccacaagttc agcgtgtccg gcgagggcga
     3241 gggcgatgcc acctacggca agctgaccct gaagctgatc tgcaccaccg gcaagctgcc
     3301 cgtgccctgg cccaccctcg tgaccaccct gggctacggc gtgcagtgct tcgcccgcta
     3361 ccccgaccac atgaagcagc acgacttctt caagtccgcc atgcccgaag gctacgtcca
     3421 ggagcgcacc atcttcttca aggacgacgg caactacaag acccgcgccg aggtgaagtt
     3481 cgagggcgac accctggtga accgcatcga gctgaagggc atcgacttca aggaggacgg
     3541 caacatcctg gggcacaagc tggagtacaa ctacaacagc cacaacgtct atatcaccgc
     3601 cgacaagcag aagaacggca tcaaggccaa cttcaagatc cgccacaaca tcgaggacgg
     3661 cggcgtgcag ctcgccgacc actaccagca gaacaccccc atcggcgacg gccccgtgct
     3721 gctgcccgac aaccactacc tgagctacca gtccaagctg agcaaagacc ccaacgagaa
     3781 gcgcgatcac atggtcctgc tggagttcgt gaccgccgcc gggatcactc tcggcatgga
     3841 cgagctgtac aagtaagtcg acggcgcgcc gcggccgcga attcgatatc ataatcaacc
     3901 tctggattac aaaatttgtg aaagattgac tggtattctt aactatgttg ctccttttac
     3961 gctatgtgga tacgctgctt taatgccttt gtatcatgct attgcttccc gtatggcttt
     4021 cattttctcc tccttgtata aatcctggtt agttcttgcc acggcggaac tcatcgccgc
     4081 ctgccttgcc cgctgctgga caggggctcg gctgttgggc actgacaatt ccgtggctcg
     4141 agagatcttc gactgtgcct tctagttgcc agccatctgt tgtttgcccc tcccccgtgc
     4201 cttccttgac cctggaaggt gccactccca ctgtcctttc ctaataaaat gaggaaattg
     4261 catcgcattg tctgagtagg tgtcattcta ttctgggggg tggggtgggg caggacagca
     4321 agggggagga ttgggaagac aatagcaggc atgagatctc acgtgcggac cgagcggccg
     4381 caggaacccc tagtgatgga gttggccact ccctctctgc gcgctcgctc gctcactgag
     4441 gccgggcgac caaaggtcgc ccgacgcccg ggctttgccc gggcggcctc agtgagcgag
     4501 cgagcgcgca gctgcctgca ggggcgcctg atgcggtatt ttctccttac gcatctgtgc
     4561 ggtatttcac accgcatacg tcaaagcaac catagtacgc gccctgtagc ggcgcattaa
     4621 gcgcggcggg tgtggtggtt acgcgcagcg tgaccgctac acttgccagc gccttagcgc
     4681 ccgctccttt cgctttcttc ccttcctttc tcgccacgtt cgccggcttt ccccgtcaag
     4741 ctctaa
//
