LOCUS       sequence-490123-        4911 bp    DNA     circular SYN 21-OCT-2025
DEFINITION  AiE2645m is an enhancer sequence designed to drive AAV-mediated 
            transgene expression in specific populations of brain cells..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4911)
  TITLE     Basal Ganglia/CERP Enhancer AAVs
REFERENCE   2  (bases 1 to 4911)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Oct 21, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"
COMMENT     Sequence Label: AiP2116 - pAAV-AiE2645m-minBG-SYFP2-WPRE3-BGHpA
FEATURES             Location/Qualifiers
     source          1..4911
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   1..130
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     regulatory      923..932
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(929..931,932..934,935..1645)
                     /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(974..995)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1235..1254)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1582..1603
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(1738..1758)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     polyA_signal    1945..2056
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   2174..2314
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   2174..2303
                     /label=AAV2 ITR
     rep_origin      2389..2844
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2476..2495)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2686..2707
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(2861..2880)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     2980..3002
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(3040..3058)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3126..3230
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3231..3299,3300..4091)
                     /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(3449..3468)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4262..4850
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4751..4770
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 cctgcaggca gctgcgcgct cgctcgctca ctgaggccgc ccgggcgtcg ggcgaccttt
       61 ggtcgcccgg cctcagtgag cgagcgagcg cgcagagagg gagtggccaa ctccatcact
      121 aggggttcct gcggccgcac gcgtcacaaa tgtgtgcatg tatgaaagct ctggctaatg
      181 gtctgagaat tgtaaggaag gtggctgtct ggtcctgtat gtggcccgtc ctttgttgtt
      241 tggagagaca gcgagtgtgt gttcacactg tcctttgctt tcatctaggc tgtgtgtttt
      301 cttgtcttta ctgtcatctt gagccttgtt ttggagggca gaagaatgct taggtcatgt
      361 ttagttttta tagcctcatt tagtgaaaat ggatcctttg ggccccgtct atgcccattg
      421 cctaaattta aagttccttt tcttgcaggt ggcttaattt atatttgttg atggcacaac
      481 gaatgcaaat aactacacgg gtgaccttta ggaggctgcc agagccaaac cagttattcg
      541 aatgacttga attcagaaaa gcttctggac atatttttag catttctaca aatactcttt
      601 tcttgaggaa agggtgtttt gggaagtgta tagaaagctc atccatagtg aattgtcgga
      661 tatttcaacc ctataccact ctgggaggag gaaagctgag cagaattaga gaccccagtc
      721 gatcacacag ggatttatgg gttatccatt ttatcagatg ccttcatatt cctggcaggt
      781 gcatacctat acctcaactg tgaacattac tgaagcgaac acgccggcgg agctcgggct
      841 gggcataaaa gtcagggcag agccatctat tgcttacatt tgcttctggg atccagatct
      901 ttcgaagcta gcgctaccgg tcgccaccat ggtgagcaag ggcgaggagc tgttcaccgg
      961 ggtggtgccc atcctggtcg agctggacgg cgacgtaaac ggccacaagt tcagcgtgtc
     1021 cggcgagggc gagggcgatg ccacctacgg caagctgacc ctgaagctga tctgcaccac
     1081 cggcaagctg cccgtgccct ggcccaccct cgtgaccacc ctgggctacg gcgtgcagtg
     1141 cttcgcccgc taccccgacc acatgaagca gcacgacttc ttcaagtccg ccatgcccga
     1201 aggctacgtc caggagcgca ccatcttctt caaggacgac ggcaactaca agacccgcgc
     1261 cgaggtgaag ttcgagggcg acaccctggt gaaccgcatc gagctgaagg gcatcgactt
     1321 caaggaggac ggcaacatcc tggggcacaa gctggagtac aactacaaca gccacaacgt
     1381 ctatatcacc gccgacaagc agaagaacgg catcaaggcc aacttcaaga tccgccacaa
     1441 catcgaggac ggcggcgtgc agctcgccga ccactaccag cagaacaccc ccatcggcga
     1501 cggccccgtg ctgctgcccg acaaccacta cctgagctac cagtccaagc tgagcaaaga
     1561 ccccaacgag aagcgcgatc acatggtcct gctggagttc gtgaccgccg ccgggatcac
     1621 tctcggcatg gacgagctgt acaagtaagt cgacggcgcg ccgcggccgc gaattcgata
     1681 tcataatcaa cctctggatt acaaaatttg tgaaagattg actggtattc ttaactatgt
     1741 tgctcctttt acgctatgtg gatacgctgc tttaatgcct ttgtatcatg ctattgcttc
     1801 ccgtatggct ttcattttct cctccttgta taaatcctgg ttagttcttg ccacggcgga
     1861 actcatcgcc gcctgccttg cccgctgctg gacaggggct cggctgttgg gcactgacaa
     1921 ttccgtggct cgagagatct tcgactgtgc cttctagttg ccagccatct gttgtttgcc
     1981 cctcccccgt gccttccttg accctggaag gtgccactcc cactgtcctt tcctaataaa
     2041 atgaggaaat tgcatcgcat tgtctgagta ggtgtcattc tattctgggg ggtggggtgg
     2101 ggcaggacag caagggggag gattgggaag acaatagcag gcatgagatc tcacgtgcgg
     2161 accgagcggc cgcaggaacc cctagtgatg gagttggcca ctccctctct gcgcgctcgc
     2221 tcgctcactg aggccgggcg accaaaggtc gcccgacgcc cgggctttgc ccgggcggcc
     2281 tcagtgagcg agcgagcgcg cagctgcctg caggggcgcc tgatgcggta ttttctcctt
     2341 acgcatctgt gcggtatttc acaccgcata cgtcaaagca accatagtac gcgccctgta
     2401 gcggcgcatt aagcgcggcg ggtgtggtgg ttacgcgcag cgtgaccgct acacttgcca
     2461 gcgccttagc gcccgctcct ttcgctttct tcccttcctt tctcgccacg ttcgccggct
     2521 ttccccgtca agctctaaat cgggggctcc ctttagggtt ccgatttagt gctttacggc
     2581 acctcgaccc caaaaaactt gatttgggtg atggttcacg tagtgggcca tcgccctgat
     2641 agacggtttt tcgccctttg acgttggagt ccacgttctt taatagtgga ctcttgttcc
     2701 aaactggaac aacactcaac tctatctcgg gctattcttt tgatttataa gggattttgc
     2761 cgatttcggt ctattggtta aaaaatgagc tgatttaaca aaaatttaac gcgaatttta
     2821 acaaaatatt aacgtttaca attttatggt gcactctcag tacaatctgc tctgatgccg
     2881 catagttaag ccagccccga cacccgccaa cacccgctga cgcgccctga cgggcttgtc
     2941 tgctcccggc atccgcttac agacaagctg tgaccgtctc cgggagctgc atgtgtcaga
     3001 ggttttcacc gtcatcaccg aaacgcgcga gacgaaaggg cctcgtgata cgcctatttt
     3061 tataggttaa tgtcatgata ataatggttt cttagacgtc aggtggcact tttcggggaa
     3121 atgtgcgcgg aacccctatt tgtttatttt tctaaataca ttcaaatatg tatccgctca
     3181 tgagacaata accctgataa atgcttcaat aatattgaaa aaggaagagt atgagtattc
     3241 aacatttccg tgtcgccctt attccctttt ttgcggcatt ttgccttcct gtttttgctc
     3301 acccagaaac gctggtgaaa gtaaaagatg ctgaagatca gttgggtgca cgagtgggtt
     3361 acatcgaact ggatctcaac agcggtaaga tccttgagag ttttcgcccc gaagaacgtt
     3421 ttccaatgat gagcactttt aaagttctgc tatgtggcgc ggtattatcc cgtattgacg
     3481 ccgggcaaga gcaactcggt cgccgcatac actattctca gaatgacttg gttgagtact
     3541 caccagtcac agaaaagcat cttacggatg gcatgacagt aagagaatta tgcagtgctg
     3601 ccataaccat gagtgataac actgcggcca acttacttct gacaacgatc ggaggaccga
     3661 aggagctaac cgcttttttg cacaacatgg gggatcatgt aactcgcctt gatcgttggg
     3721 aaccggagct gaatgaagcc ataccaaacg acgagcgtga caccacgatg cctgtagcaa
     3781 tggcaacaac gttgcgcaaa ctattaactg gcgaactact tactctagct tcccggcaac
     3841 aattaataga ctggatggag gcggataaag ttgcaggacc acttctgcgc tcggcccttc
     3901 cggctggctg gtttattgct gataaatctg gagccggtga gcgtgggtct cgcggtatca
     3961 ttgcagcact ggggccagat ggtaagccct cccgtatcgt agttatctac acgacgggga
     4021 gtcaggcaac tatggatgaa cgaaatagac agatcgctga gataggtgcc tcactgatta
     4081 agcattggta actgtcagac caagtttact catatatact ttagattgat ttaaaacttc
     4141 atttttaatt taaaaggatc taggtgaaga tcctttttga taatctcatg accaaaatcc
     4201 cttaacgtga gttttcgttc cactgagcgt cagaccccgt agaaaagatc aaaggatctt
     4261 cttgagatcc tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac
     4321 cagcggtggt ttgtttgccg gatcaagagc taccaactct ttttccgaag gtaactggct
     4381 tcagcagagc gcagatacca aatactgttc ttctagtgta gccgtagtta ggccaccact
     4441 tcaagaactc tgtagcaccg cctacatacc tcgctctgct aatcctgtta ccagtggctg
     4501 ctgccagtgg cgataagtcg tgtcttaccg ggttggactc aagacgatag ttaccggata
     4561 aggcgcagcg gtcgggctga acggggggtt cgtgcacaca gcccagcttg gagcgaacga
     4621 cctacaccga actgagatac ctacagcgtg agctatgaga aagcgccacg cttcccgaag
     4681 ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg
     4741 agcttccagg gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc cacctctgac
     4801 ttgagcgtcg atttttgtga tgctcgtcag gggggcggag cctatggaaa aacgccagca
     4861 acgcggcctt tttacggttc ctggcctttt gctggccttt tgctcacatg t
//
