LOCUS       sequence-492119-        4874 bp    DNA     circular SYN 06-NOV-2025
DEFINITION  AiE2203m 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 4874)
  TITLE     Basal Ganglia/CERP Enhancer AAVs
REFERENCE   2  (bases 1 to 4874)
  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: AiP1528 - pAAV-AiE2203m-minBG-SYFP2-WPRE3-BGHpA
FEATURES             Location/Qualifiers
     source          1..4874
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     regulatory      499..508
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(505..507,508..510,511..1221)
                     /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(550..571)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(811..830)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1158..1179
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(1314..1334)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     polyA_signal    1521..1632
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   1750..1890
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   1750..1879
                     /label=AAV2 ITR
     rep_origin      1965..2420
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2052..2071)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2262..2283
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(2437..2456)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     2556..2578
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(2616..2634)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        2702..2806
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2807..2875,2876..3667)
                     /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(3025..3044)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3838..4426
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4327..4346
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     repeat_region   4488..4617
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
ORIGIN
        1 aatatcctgt caaaatcctc atgataatga acgcagcctg ggccggctgg cacggcagct
       61 ccacactgca gccggcaaga tagcgcagct aatttatgag cgagggattg ggctgtgctg
      121 gcaaagcagg ctgggtccag caacggaggg cggtccttgc ttgatggaga gccatcagct
      181 agggatatca ttgggttcca gtgacccaca gcatgagtga cccttgtaca gctcgttcct
      241 tggggccctc cctggcttgc aggaaaggat gaggggcaac gcctttcaaa aactgaccca
      301 actccatctg cctaagatcc cggggtggtg tttgccatct gaggtagaag gtgggcacag
      361 agtgctcatg aagaggtcag aggctcactg tatgttgcgc cggcggagct cgggctgggc
      421 ataaaagtca gggcagagcc atctattgct tacatttgct tctgggatcc agatctttcg
      481 aagctagcgc taccggtcgc caccatggtg agcaagggcg aggagctgtt caccggggtg
      541 gtgcccatcc tggtcgagct ggacggcgac gtaaacggcc acaagttcag cgtgtccggc
      601 gagggcgagg gcgatgccac ctacggcaag ctgaccctga agctgatctg caccaccggc
      661 aagctgcccg tgccctggcc caccctcgtg accaccctgg gctacggcgt gcagtgcttc
      721 gcccgctacc ccgaccacat gaagcagcac gacttcttca agtccgccat gcccgaaggc
      781 tacgtccagg agcgcaccat cttcttcaag gacgacggca actacaagac ccgcgccgag
      841 gtgaagttcg agggcgacac cctggtgaac cgcatcgagc tgaagggcat cgacttcaag
      901 gaggacggca acatcctggg gcacaagctg gagtacaact acaacagcca caacgtctat
      961 atcaccgccg acaagcagaa gaacggcatc aaggccaact tcaagatccg ccacaacatc
     1021 gaggacggcg gcgtgcagct cgccgaccac taccagcaga acacccccat cggcgacggc
     1081 cccgtgctgc tgcccgacaa ccactacctg agctaccagt ccaagctgag caaagacccc
     1141 aacgagaagc gcgatcacat ggtcctgctg gagttcgtga ccgccgccgg gatcactctc
     1201 ggcatggacg agctgtacaa gtaagtcgac ggcgcgccgc ggccgcgaat tcgatatcat
     1261 aatcaacctc tggattacaa aatttgtgaa agattgactg gtattcttaa ctatgttgct
     1321 ccttttacgc tatgtggata cgctgcttta atgcctttgt atcatgctat tgcttcccgt
     1381 atggctttca ttttctcctc cttgtataaa tcctggttag ttcttgccac ggcggaactc
     1441 atcgccgcct gccttgcccg ctgctggaca ggggctcggc tgttgggcac tgacaattcc
     1501 gtggctcgag agatcttcga ctgtgccttc tagttgccag ccatctgttg tttgcccctc
     1561 ccccgtgcct tccttgaccc tggaaggtgc cactcccact gtcctttcct aataaaatga
     1621 ggaaattgca tcgcattgtc tgagtaggtg tcattctatt ctggggggtg gggtggggca
     1681 ggacagcaag ggggaggatt gggaagacaa tagcaggcat gagatctcac gtgcggaccg
     1741 agcggccgca ggaaccccta gtgatggagt tggccactcc ctctctgcgc gctcgctcgc
     1801 tcactgaggc cgggcgacca aaggtcgccc gacgcccggg ctttgcccgg gcggcctcag
     1861 tgagcgagcg agcgcgcagc tgcctgcagg ggcgcctgat gcggtatttt ctccttacgc
     1921 atctgtgcgg tatttcacac cgcatacgtc aaagcaacca tagtacgcgc cctgtagcgg
     1981 cgcattaagc gcggcgggtg tggtggttac gcgcagcgtg accgctacac ttgccagcgc
     2041 cttagcgccc gctcctttcg ctttcttccc ttcctttctc gccacgttcg ccggctttcc
     2101 ccgtcaagct ctaaatcggg ggctcccttt agggttccga tttagtgctt tacggcacct
     2161 cgaccccaaa aaacttgatt tgggtgatgg ttcacgtagt gggccatcgc cctgatagac
     2221 ggtttttcgc cctttgacgt tggagtccac gttctttaat agtggactct tgttccaaac
     2281 tggaacaaca ctcaactcta tctcgggcta ttcttttgat ttataaggga ttttgccgat
     2341 ttcggtctat tggttaaaaa atgagctgat ttaacaaaaa tttaacgcga attttaacaa
     2401 aatattaacg tttacaattt tatggtgcac tctcagtaca atctgctctg atgccgcata
     2461 gttaagccag ccccgacacc cgccaacacc cgctgacgcg ccctgacggg cttgtctgct
     2521 cccggcatcc gcttacagac aagctgtgac cgtctccggg agctgcatgt gtcagaggtt
     2581 ttcaccgtca tcaccgaaac gcgcgagacg aaagggcctc gtgatacgcc tatttttata
     2641 ggttaatgtc atgataataa tggtttctta gacgtcaggt ggcacttttc ggggaaatgt
     2701 gcgcggaacc cctatttgtt tatttttcta aatacattca aatatgtatc cgctcatgag
     2761 acaataaccc tgataaatgc ttcaataata ttgaaaaagg aagagtatga gtattcaaca
     2821 tttccgtgtc gcccttattc ccttttttgc ggcattttgc cttcctgttt ttgctcaccc
     2881 agaaacgctg gtgaaagtaa aagatgctga agatcagttg ggtgcacgag tgggttacat
     2941 cgaactggat ctcaacagcg gtaagatcct tgagagtttt cgccccgaag aacgttttcc
     3001 aatgatgagc acttttaaag ttctgctatg tggcgcggta ttatcccgta ttgacgccgg
     3061 gcaagagcaa ctcggtcgcc gcatacacta ttctcagaat gacttggttg agtactcacc
     3121 agtcacagaa aagcatctta cggatggcat gacagtaaga gaattatgca gtgctgccat
     3181 aaccatgagt gataacactg cggccaactt acttctgaca acgatcggag gaccgaagga
     3241 gctaaccgct tttttgcaca acatggggga tcatgtaact cgccttgatc gttgggaacc
     3301 ggagctgaat gaagccatac caaacgacga gcgtgacacc acgatgcctg tagcaatggc
     3361 aacaacgttg cgcaaactat taactggcga actacttact ctagcttccc ggcaacaatt
     3421 aatagactgg atggaggcgg ataaagttgc aggaccactt ctgcgctcgg cccttccggc
     3481 tggctggttt attgctgata aatctggagc cggtgagcgt gggtctcgcg gtatcattgc
     3541 agcactgggg ccagatggta agccctcccg tatcgtagtt atctacacga cggggagtca
     3601 ggcaactatg gatgaacgaa atagacagat cgctgagata ggtgcctcac tgattaagca
     3661 ttggtaactg tcagaccaag tttactcata tatactttag attgatttaa aacttcattt
     3721 ttaatttaaa aggatctagg tgaagatcct ttttgataat ctcatgacca aaatccctta
     3781 acgtgagttt tcgttccact gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg
     3841 agatcctttt tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac cgctaccagc
     3901 ggtggtttgt ttgccggatc aagagctacc aactcttttt ccgaaggtaa ctggcttcag
     3961 cagagcgcag ataccaaata ctgttcttct agtgtagccg tagttaggcc accacttcaa
     4021 gaactctgta gcaccgccta catacctcgc tctgctaatc ctgttaccag tggctgctgc
     4081 cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac cggataaggc
     4141 gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc agcttggagc gaacgaccta
     4201 caccgaactg agatacctac agcgtgagct atgagaaagc gccacgcttc ccgaagggag
     4261 aaaggcggac aggtatccgg taagcggcag ggtcggaaca ggagagcgca cgagggagct
     4321 tccaggggga aacgcctggt atctttatag tcctgtcggg tttcgccacc tctgacttga
     4381 gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc
     4441 ggccttttta cggttcctgg ccttttgctg gccttttgct cacatgtcct gcaggcagct
     4501 gcgcgctcgc tcgctcactg aggccgcccg ggcgtcgggc gacctttggt cgcccggcct
     4561 cagtgagcga gcgagcgcgc agagagggag tggccaactc catcactagg ggttcctgcg
     4621 gccgcacgcg ttgtaagagt aataatcgcc ggcgatctgc acagcaggga aatgggctgg
     4681 cagctgatac tggtagcgtg tgtggccctg tgcacctgag ggaggcttga gttgcaagga
     4741 caggagcacg tgtggtatcc ttggagaaac gaggcagaag ggagagagaa aacagaaagc
     4801 atcgcacagg cagacaccag cattgtctgt tcccagtgag gggtcagttc ggcaagttca
     4861 ggattagtaa attt
//
