LOCUS       sequence-492114-        4746 bp    DNA     circular SYN 06-NOV-2025
DEFINITION  AiE2324m 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: AiP1685 - pAAV-AiE2324m-minBG-SYFP2-WPRE3-BGHpA
FEATURES             Location/Qualifiers
     source          1..4746
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     2..21
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     repeat_region   163..292
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     regulatory      920..929
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(926..928,929..931,932..1642)
                     /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(971..992)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1232..1251)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1579..1600
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(1735..1755)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     polyA_signal    1942..2053
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   2171..2311
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   2171..2300
                     /label=AAV2 ITR
     rep_origin      2386..2841
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2473..2492)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2683..2704
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(2858..2877)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     2977..2999
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(3037..3055)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3123..3227
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3228..3296,3297..4088)
                     /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(3446..3465)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      join(4259..4746,1..101)
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
ORIGIN
        1 ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg ccacctctga cttgagcgtc
       61 gatttttgtg atgctcgtca ggggggcgga gcctatggaa aaacgccagc aacgcggcct
      121 ttttacggtt cctggccttt tgctggcctt ttgctcacat gtcctgcagg cagctgcgcg
      181 ctcgctcgct cactgaggcc gcccgggcgt cgggcgacct ttggtcgccc ggcctcagtg
      241 agcgagcgag cgcgcagaga gggagtggcc aactccatca ctaggggttc ctgcggccgc
      301 acgcgttgta agagtaataa tcgtcatttc atccagaaat agtcagactg attcaaaacc
      361 catctcctag caggagtgat gcaggttgga aattcctgaa aaaggatcaa agggtctcaa
      421 aggttgattg ttaggtagct atttagatgt tagcagagtt taaggtcatg gggctgaacc
      481 cctccttcaa tatcagcagc ttattaaacc aggaggtcag actgcaggac tctgattccc
      541 tgcaccggga cacggtgttc tgaactgctc aggaagctat gataagatta ctgtctagcc
      601 agaggcttcc taataaagaa caaagacgta ccattgttgt tccctcttcc cccaagagtc
      661 agtccacgtg cacctgtcaa tcacagggac aggacaatgc gcttctgctt tagcagcttc
      721 agaaaggata caatgctctc tcccgccatt tagtccttct cacctggaca ttctctccag
      781 ttcagtttgt aaaaactacc ttcttctcta ctttcctgtg tgcccggagc tcgggctggg
      841 cataaaagtc agggcagagc catctattgc ttacatttgc ttctgggatc cagatctttc
      901 gaagctagcg ctaccggtcg ccaccatggt gagcaagggc gaggagctgt tcaccggggt
      961 ggtgcccatc ctggtcgagc tggacggcga cgtaaacggc cacaagttca gcgtgtccgg
     1021 cgagggcgag ggcgatgcca cctacggcaa gctgaccctg aagctgatct gcaccaccgg
     1081 caagctgccc gtgccctggc ccaccctcgt gaccaccctg ggctacggcg tgcagtgctt
     1141 cgcccgctac cccgaccaca tgaagcagca cgacttcttc aagtccgcca tgcccgaagg
     1201 ctacgtccag gagcgcacca tcttcttcaa ggacgacggc aactacaaga cccgcgccga
     1261 ggtgaagttc gagggcgaca ccctggtgaa ccgcatcgag ctgaagggca tcgacttcaa
     1321 ggaggacggc aacatcctgg ggcacaagct ggagtacaac tacaacagcc acaacgtcta
     1381 tatcaccgcc gacaagcaga agaacggcat caaggccaac ttcaagatcc gccacaacat
     1441 cgaggacggc ggcgtgcagc tcgccgacca ctaccagcag aacaccccca tcggcgacgg
     1501 ccccgtgctg ctgcccgaca accactacct gagctaccag tccaagctga gcaaagaccc
     1561 caacgagaag cgcgatcaca tggtcctgct ggagttcgtg accgccgccg ggatcactct
     1621 cggcatggac gagctgtaca agtaagtcga cggcgcgccg cggccgcgaa ttcgatatca
     1681 taatcaacct ctggattaca aaatttgtga aagattgact ggtattctta actatgttgc
     1741 tccttttacg ctatgtggat acgctgcttt aatgcctttg tatcatgcta ttgcttcccg
     1801 tatggctttc attttctcct ccttgtataa atcctggtta gttcttgcca cggcggaact
     1861 catcgccgcc tgccttgccc gctgctggac aggggctcgg ctgttgggca ctgacaattc
     1921 cgtggctcga gagatcttcg actgtgcctt ctagttgcca gccatctgtt gtttgcccct
     1981 cccccgtgcc ttccttgacc ctggaaggtg ccactcccac tgtcctttcc taataaaatg
     2041 aggaaattgc atcgcattgt ctgagtaggt gtcattctat tctggggggt ggggtggggc
     2101 aggacagcaa gggggaggat tgggaagaca atagcaggca tgagatctca cgtgcggacc
     2161 gagcggccgc aggaacccct agtgatggag ttggccactc cctctctgcg cgctcgctcg
     2221 ctcactgagg ccgggcgacc aaaggtcgcc cgacgcccgg gctttgcccg ggcggcctca
     2281 gtgagcgagc gagcgcgcag ctgcctgcag gggcgcctga tgcggtattt tctccttacg
     2341 catctgtgcg gtatttcaca ccgcatacgt caaagcaacc atagtacgcg ccctgtagcg
     2401 gcgcattaag cgcggcgggt gtggtggtta cgcgcagcgt gaccgctaca cttgccagcg
     2461 ccttagcgcc cgctcctttc gctttcttcc cttcctttct cgccacgttc gccggctttc
     2521 cccgtcaagc tctaaatcgg gggctccctt tagggttccg atttagtgct ttacggcacc
     2581 tcgaccccaa aaaacttgat ttgggtgatg gttcacgtag tgggccatcg ccctgataga
     2641 cggtttttcg ccctttgacg ttggagtcca cgttctttaa tagtggactc ttgttccaaa
     2701 ctggaacaac actcaactct atctcgggct attcttttga tttataaggg attttgccga
     2761 tttcggtcta ttggttaaaa aatgagctga tttaacaaaa atttaacgcg aattttaaca
     2821 aaatattaac gtttacaatt ttatggtgca ctctcagtac aatctgctct gatgccgcat
     2881 agttaagcca gccccgacac ccgccaacac ccgctgacgc gccctgacgg gcttgtctgc
     2941 tcccggcatc cgcttacaga caagctgtga ccgtctccgg gagctgcatg tgtcagaggt
     3001 tttcaccgtc atcaccgaaa cgcgcgagac gaaagggcct cgtgatacgc ctatttttat
     3061 aggttaatgt catgataata atggtttctt agacgtcagg tggcactttt cggggaaatg
     3121 tgcgcggaac ccctatttgt ttatttttct aaatacattc aaatatgtat ccgctcatga
     3181 gacaataacc ctgataaatg cttcaataat attgaaaaag gaagagtatg agtattcaac
     3241 atttccgtgt cgcccttatt cccttttttg cggcattttg ccttcctgtt tttgctcacc
     3301 cagaaacgct ggtgaaagta aaagatgctg aagatcagtt gggtgcacga gtgggttaca
     3361 tcgaactgga tctcaacagc ggtaagatcc ttgagagttt tcgccccgaa gaacgttttc
     3421 caatgatgag cacttttaaa gttctgctat gtggcgcggt attatcccgt attgacgccg
     3481 ggcaagagca actcggtcgc cgcatacact attctcagaa tgacttggtt gagtactcac
     3541 cagtcacaga aaagcatctt acggatggca tgacagtaag agaattatgc agtgctgcca
     3601 taaccatgag tgataacact gcggccaact tacttctgac aacgatcgga ggaccgaagg
     3661 agctaaccgc ttttttgcac aacatggggg atcatgtaac tcgccttgat cgttgggaac
     3721 cggagctgaa tgaagccata ccaaacgacg agcgtgacac cacgatgcct gtagcaatgg
     3781 caacaacgtt gcgcaaacta ttaactggcg aactacttac tctagcttcc cggcaacaat
     3841 taatagactg gatggaggcg gataaagttg caggaccact tctgcgctcg gcccttccgg
     3901 ctggctggtt tattgctgat aaatctggag ccggtgagcg tgggtctcgc ggtatcattg
     3961 cagcactggg gccagatggt aagccctccc gtatcgtagt tatctacacg acggggagtc
     4021 aggcaactat ggatgaacga aatagacaga tcgctgagat aggtgcctca ctgattaagc
     4081 attggtaact gtcagaccaa gtttactcat atatacttta gattgattta aaacttcatt
     4141 tttaatttaa aaggatctag gtgaagatcc tttttgataa tctcatgacc aaaatccctt
     4201 aacgtgagtt ttcgttccac tgagcgtcag accccgtaga aaagatcaaa ggatcttctt
     4261 gagatccttt ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag
     4321 cggtggtttg tttgccggat caagagctac caactctttt tccgaaggta actggcttca
     4381 gcagagcgca gataccaaat actgttcttc tagtgtagcc gtagttaggc caccacttca
     4441 agaactctgt agcaccgcct acatacctcg ctctgctaat cctgttacca gtggctgctg
     4501 ccagtggcga taagtcgtgt cttaccgggt tggactcaag acgatagtta ccggataagg
     4561 cgcagcggtc gggctgaacg gggggttcgt gcacacagcc cagcttggag cgaacgacct
     4621 acaccgaact gagataccta cagcgtgagc tatgagaaag cgccacgctt cccgaaggga
     4681 gaaaggcgga caggtatccg gtaagcggca gggtcggaac aggagagcgc acgagggagc
     4741 ttccag
//
