LOCUS       sequence-492136-        4664 bp    DNA     circular SYN 06-NOV-2025
DEFINITION  AiE2208m 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 4664)
  TITLE     Basal Ganglia/CERP Enhancer AAVs
REFERENCE   2  (bases 1 to 4664)
  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: AiP1541 - pAAV-AiE2208m-minBG-SYFP2-WPRE3-BGHpA
FEATURES             Location/Qualifiers
     source          1..4664
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(117..136)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     236..258
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(296..314)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        382..486
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(487..555,556..1347)
                     /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(705..724)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      1518..2106
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2007..2026
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     repeat_region   2168..2297
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     regulatory      2843..2852
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(2849..2851,2852..2854,2855..3565)
                     /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(2894..2915)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(3155..3174)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     3502..3523
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(3658..3678)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     polyA_signal    3865..3976
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   4094..4234
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   4094..4223
                     /label=AAV2 ITR
     rep_origin      join(4309..4664,1..100)
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4396..4415)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4606..4627
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
ORIGIN
        1 ttataaggga ttttgccgat ttcggtctat tggttaaaaa atgagctgat ttaacaaaaa
       61 tttaacgcga attttaacaa aatattaacg tttacaattt tatggtgcac tctcagtaca
      121 atctgctctg atgccgcata gttaagccag ccccgacacc cgccaacacc cgctgacgcg
      181 ccctgacggg cttgtctgct cccggcatcc gcttacagac aagctgtgac cgtctccggg
      241 agctgcatgt gtcagaggtt ttcaccgtca tcaccgaaac gcgcgagacg aaagggcctc
      301 gtgatacgcc tatttttata ggttaatgtc atgataataa tggtttctta gacgtcaggt
      361 ggcacttttc ggggaaatgt gcgcggaacc cctatttgtt tatttttcta aatacattca
      421 aatatgtatc cgctcatgag acaataaccc tgataaatgc ttcaataata ttgaaaaagg
      481 aagagtatga gtattcaaca tttccgtgtc gcccttattc ccttttttgc ggcattttgc
      541 cttcctgttt ttgctcaccc agaaacgctg gtgaaagtaa aagatgctga agatcagttg
      601 ggtgcacgag tgggttacat cgaactggat ctcaacagcg gtaagatcct tgagagtttt
      661 cgccccgaag aacgttttcc aatgatgagc acttttaaag ttctgctatg tggcgcggta
      721 ttatcccgta ttgacgccgg gcaagagcaa ctcggtcgcc gcatacacta ttctcagaat
      781 gacttggttg agtactcacc agtcacagaa aagcatctta cggatggcat gacagtaaga
      841 gaattatgca gtgctgccat aaccatgagt gataacactg cggccaactt acttctgaca
      901 acgatcggag gaccgaagga gctaaccgct tttttgcaca acatggggga tcatgtaact
      961 cgccttgatc gttgggaacc ggagctgaat gaagccatac caaacgacga gcgtgacacc
     1021 acgatgcctg tagcaatggc aacaacgttg cgcaaactat taactggcga actacttact
     1081 ctagcttccc ggcaacaatt aatagactgg atggaggcgg ataaagttgc aggaccactt
     1141 ctgcgctcgg cccttccggc tggctggttt attgctgata aatctggagc cggtgagcgt
     1201 gggtctcgcg gtatcattgc agcactgggg ccagatggta agccctcccg tatcgtagtt
     1261 atctacacga cggggagtca ggcaactatg gatgaacgaa atagacagat cgctgagata
     1321 ggtgcctcac tgattaagca ttggtaactg tcagaccaag tttactcata tatactttag
     1381 attgatttaa aacttcattt ttaatttaaa aggatctagg tgaagatcct ttttgataat
     1441 ctcatgacca aaatccctta acgtgagttt tcgttccact gagcgtcaga ccccgtagaa
     1501 aagatcaaag gatcttcttg agatcctttt tttctgcgcg taatctgctg cttgcaaaca
     1561 aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc aagagctacc aactcttttt
     1621 ccgaaggtaa ctggcttcag cagagcgcag ataccaaata ctgttcttct agtgtagccg
     1681 tagttaggcc accacttcaa gaactctgta gcaccgccta catacctcgc tctgctaatc
     1741 ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga
     1801 cgatagttac cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc
     1861 agcttggagc gaacgaccta caccgaactg agatacctac agcgtgagct atgagaaagc
     1921 gccacgcttc ccgaagggag aaaggcggac aggtatccgg taagcggcag ggtcggaaca
     1981 ggagagcgca cgagggagct tccaggggga aacgcctggt atctttatag tcctgtcggg
     2041 tttcgccacc tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta
     2101 tggaaaaacg ccagcaacgc ggccttttta cggttcctgg ccttttgctg gccttttgct
     2161 cacatgtcct gcaggcagct gcgcgctcgc tcgctcactg aggccgcccg ggcgtcgggc
     2221 gacctttggt cgcccggcct cagtgagcga gcgagcgcgc agagagggag tggccaactc
     2281 catcactagg ggttcctgcg gccgcacgcg ttgtaagagt aataatcgcc ggcggtttcc
     2341 agtgcccctt agaggagttc ccggctgtca ctagaaaaga aaaaaaaaag aggagggatg
     2401 tggaaggggg gagtgagcct tcataaagag agtgagataa ggagaagttt gctttcttgc
     2461 ttatgaattt tcgtatcttt ccctttgccc ggatcttatg ggttacaaga tcggtcacag
     2521 cagcagtgtc gatgactaat gtggttattg agaaggattt tttaatttcc tggaaggcaa
     2581 acgccattac ttcccatccc cgcacgtctc agaccggcga tgcaggaacg actggggagt
     2641 cggtgatggg agttattata aaactgatga attattggaa aaaggacgaa ctgctgggca
     2701 aggagaactg cacttccctt ccatttccgg ttccatggta gcgccggcgg agctcgggct
     2761 gggcataaaa gtcagggcag agccatctat tgcttacatt tgcttctggg atccagatct
     2821 ttcgaagcta gcgctaccgg tcgccaccat ggtgagcaag ggcgaggagc tgttcaccgg
     2881 ggtggtgccc atcctggtcg agctggacgg cgacgtaaac ggccacaagt tcagcgtgtc
     2941 cggcgagggc gagggcgatg ccacctacgg caagctgacc ctgaagctga tctgcaccac
     3001 cggcaagctg cccgtgccct ggcccaccct cgtgaccacc ctgggctacg gcgtgcagtg
     3061 cttcgcccgc taccccgacc acatgaagca gcacgacttc ttcaagtccg ccatgcccga
     3121 aggctacgtc caggagcgca ccatcttctt caaggacgac ggcaactaca agacccgcgc
     3181 cgaggtgaag ttcgagggcg acaccctggt gaaccgcatc gagctgaagg gcatcgactt
     3241 caaggaggac ggcaacatcc tggggcacaa gctggagtac aactacaaca gccacaacgt
     3301 ctatatcacc gccgacaagc agaagaacgg catcaaggcc aacttcaaga tccgccacaa
     3361 catcgaggac ggcggcgtgc agctcgccga ccactaccag cagaacaccc ccatcggcga
     3421 cggccccgtg ctgctgcccg acaaccacta cctgagctac cagtccaagc tgagcaaaga
     3481 ccccaacgag aagcgcgatc acatggtcct gctggagttc gtgaccgccg ccgggatcac
     3541 tctcggcatg gacgagctgt acaagtaagt cgacggcgcg ccgcggccgc gaattcgata
     3601 tcataatcaa cctctggatt acaaaatttg tgaaagattg actggtattc ttaactatgt
     3661 tgctcctttt acgctatgtg gatacgctgc tttaatgcct ttgtatcatg ctattgcttc
     3721 ccgtatggct ttcattttct cctccttgta taaatcctgg ttagttcttg ccacggcgga
     3781 actcatcgcc gcctgccttg cccgctgctg gacaggggct cggctgttgg gcactgacaa
     3841 ttccgtggct cgagagatct tcgactgtgc cttctagttg ccagccatct gttgtttgcc
     3901 cctcccccgt gccttccttg accctggaag gtgccactcc cactgtcctt tcctaataaa
     3961 atgaggaaat tgcatcgcat tgtctgagta ggtgtcattc tattctgggg ggtggggtgg
     4021 ggcaggacag caagggggag gattgggaag acaatagcag gcatgagatc tcacgtgcgg
     4081 accgagcggc cgcaggaacc cctagtgatg gagttggcca ctccctctct gcgcgctcgc
     4141 tcgctcactg aggccgggcg accaaaggtc gcccgacgcc cgggctttgc ccgggcggcc
     4201 tcagtgagcg agcgagcgcg cagctgcctg caggggcgcc tgatgcggta ttttctcctt
     4261 acgcatctgt gcggtatttc acaccgcata cgtcaaagca accatagtac gcgccctgta
     4321 gcggcgcatt aagcgcggcg ggtgtggtgg ttacgcgcag cgtgaccgct acacttgcca
     4381 gcgccttagc gcccgctcct ttcgctttct tcccttcctt tctcgccacg ttcgccggct
     4441 ttccccgtca agctctaaat cgggggctcc ctttagggtt ccgatttagt gctttacggc
     4501 acctcgaccc caaaaaactt gatttgggtg atggttcacg tagtgggcca tcgccctgat
     4561 agacggtttt tcgccctttg acgttggagt ccacgttctt taatagtgga ctcttgttcc
     4621 aaactggaac aacactcaac tctatctcgg gctattcttt tgat
//
