LOCUS       sequence-492116-        4888 bp    DNA     circular SYN 06-NOV-2025
DEFINITION  AiE2640m 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 4888)
  TITLE     Basal Ganglia/CERP Enhancer AAVs
REFERENCE   2  (bases 1 to 4888)
  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: AiP20005 - pAAV-AiE2640m-minBG-SYFP2-WPRE3-BGHpA
FEATURES             Location/Qualifiers
     source          1..4888
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(join(649..1440,1441..1509))
                     /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     1272..1291
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(1510..1614)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     1682..1700
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(1738..1760)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     1860..1879
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     rep_origin      complement(1896..2351)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2033..2054)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2245..2264
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     repeat_region   2426..2566
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   2437..2566
                     /label=AAV2 ITR
     polyA_signal    2684..2795
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     2982..3002
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(join(3095..3805,3806..3808,3809..3811))
                     /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(3137..3158)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     3486..3505
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     3745..3766
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     regulatory      3808..3817
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     repeat_region   4587..4716
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     rep_origin      complement(join(4778..4888,1..478))
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4858..4877)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga tacctgtccg cctttctccc
       61 ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg tatctcagtt cggtgtaggt
      121 cgttcgctcc aagctgggct gtgtgcacga accccccgtt cagcccgacc gctgcgcctt
      181 atccggtaac tatcgtcttg agtccaaccc ggtaagacac gacttatcgc cactggcagc
      241 agccactggt aacaggatta gcagagcgag gtatgtaggc ggtgctacag agttcttgaa
      301 gtggtggcct aactacggct acactagaag aacagtattt ggtatctgcg ctctgctgaa
      361 gccagttacc ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg
      421 tagcggtggt ttttttgttt gcaagcagca gattacgcgc agaaaaaaag gatctcaaga
      481 agatcctttg atcttttcta cggggtctga cgctcagtgg aacgaaaact cacgttaagg
      541 gattttggtc atgagattat caaaaaggat cttcacctag atccttttaa attaaaaatg
      601 aagttttaaa tcaatctaaa gtatatatga gtaaacttgg tctgacagtt accaatgctt
      661 aatcagtgag gcacctatct cagcgatctg tctatttcgt tcatccatag ttgcctgact
      721 ccccgtcgtg tagataacta cgatacggga gggcttacca tctggcccca gtgctgcaat
      781 gataccgcga gacccacgct caccggctcc agatttatca gcaataaacc agccagccgg
      841 aagggccgag cgcagaagtg gtcctgcaac tttatccgcc tccatccagt ctattaattg
      901 ttgccgggaa gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat
      961 tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg gcttcattca gctccggttc
     1021 ccaacgatca aggcgagtta catgatcccc catgttgtgc aaaaaagcgg ttagctcctt
     1081 cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg ttatcactca tggttatggc
     1141 agcactgcat aattctctta ctgtcatgcc atccgtaaga tgcttttctg tgactggtga
     1201 gtactcaacc aagtcattct gagaatagtg tatgcggcga ccgagttgct cttgcccggc
     1261 gtcaatacgg gataataccg cgccacatag cagaacttta aaagtgctca tcattggaaa
     1321 acgttcttcg gggcgaaaac tctcaaggat cttaccgctg ttgagatcca gttcgatgta
     1381 acccactcgt gcacccaact gatcttcagc atcttttact ttcaccagcg tttctgggtg
     1441 agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata agggcgacac ggaaatgttg
     1501 aatactcata ctcttccttt ttcaatatta ttgaagcatt tatcagggtt attgtctcat
     1561 gagcggatac atatttgaat gtatttagaa aaataaacaa ataggggttc cgcgcacatt
     1621 tccccgaaaa gtgccacctg acgtctaaga aaccattatt atcatgacat taacctataa
     1681 aaataggcgt atcacgaggc cctttcgtct cgcgcgtttc ggtgatgacg gtgaaaacct
     1741 ctgacacatg cagctcccgg agacggtcac agcttgtctg taagcggatg ccgggagcag
     1801 acaagcccgt cagggcgcgt cagcgggtgt tggcgggtgt cggggctggc ttaactatgc
     1861 ggcatcagag cagattgtac tgagagtgca ccataaaatt gtaaacgtta atattttgtt
     1921 aaaattcgcg ttaaattttt gttaaatcag ctcatttttt aaccaataga ccgaaatcgg
     1981 caaaatccct tataaatcaa aagaatagcc cgagatagag ttgagtgttg ttccagtttg
     2041 gaacaagagt ccactattaa agaacgtgga ctccaacgtc aaagggcgaa aaaccgtcta
     2101 tcagggcgat ggcccactac gtgaaccatc acccaaatca agttttttgg ggtcgaggtg
     2161 ccgtaaagca ctaaatcgga accctaaagg gagcccccga tttagagctt gacggggaaa
     2221 gccggcgaac gtggcgagaa aggaagggaa gaaagcgaaa ggagcgggcg ctaaggcgct
     2281 ggcaagtgta gcggtcacgc tgcgcgtaac caccacaccc gccgcgctta atgcgccgct
     2341 acagggcgcg tactatggtt gctttgacgt atgcggtgtg aaataccgca cagatgcgta
     2401 aggagaaaat accgcatcag gcgcccctgc aggcagctgc gcgctcgctc gctcactgag
     2461 gccgcccggg caaagcccgg gcgtcgggcg acctttggtc gcccggcctc agtgagcgag
     2521 cgagcgcgca gagagggagt ggccaactcc atcactaggg gttcctgcgg ccgctcggtc
     2581 cgcacgtgag atctcatgcc tgctattgtc ttcccaatcc tcccccttgc tgtcctgccc
     2641 caccccaccc cccagaatag aatgacacct actcagacaa tgcgatgcaa tttcctcatt
     2701 ttattaggaa aggacagtgg gagtggcacc ttccagggtc aaggaaggca cgggggaggg
     2761 gcaaacaaca gatggctggc aactagaagg cacagtcgaa gatctctcga gccacggaat
     2821 tgtcagtgcc caacagccga gcccctgtcc agcagcgggc aaggcaggcg gcgatgagtt
     2881 ccgccgtggc aagaactaac caggatttat acaaggagga gaaaatgaaa gccatacggg
     2941 aagcaatagc atgatacaaa ggcattaaag cagcgtatcc acatagcgta aaaggagcaa
     3001 catagttaag aataccagtc aatctttcac aaattttgta atccagaggt tgattatgat
     3061 atcgaattcg cggccgcggc gcgccgtcga cttacttgta cagctcgtcc atgccgagag
     3121 tgatcccggc ggcggtcacg aactccagca ggaccatgtg atcgcgcttc tcgttggggt
     3181 ctttgctcag cttggactgg tagctcaggt agtggttgtc gggcagcagc acggggccgt
     3241 cgccgatggg ggtgttctgc tggtagtggt cggcgagctg cacgccgccg tcctcgatgt
     3301 tgtggcggat cttgaagttg gccttgatgc cgttcttctg cttgtcggcg gtgatataga
     3361 cgttgtggct gttgtagttg tactccagct tgtgccccag gatgttgccg tcctccttga
     3421 agtcgatgcc cttcagctcg atgcggttca ccagggtgtc gccctcgaac ttcacctcgg
     3481 cgcgggtctt gtagttgccg tcgtccttga agaagatggt gcgctcctgg acgtagcctt
     3541 cgggcatggc ggacttgaag aagtcgtgct gcttcatgtg gtcggggtag cgggcgaagc
     3601 actgcacgcc gtagcccagg gtggtcacga gggtgggcca gggcacgggc agcttgccgg
     3661 tggtgcagat cagcttcagg gtcagcttgc cgtaggtggc atcgccctcg ccctcgccgg
     3721 acacgctgaa cttgtggccg tttacgtcgc cgtccagctc gaccaggatg ggcaccaccc
     3781 cggtgaacag ctcctcgccc ttgctcacca tggtggcgac cggtagcgct agcttcgaaa
     3841 gatctggatc ccagaagcaa atgtaagcaa tagatggctc tgccctgact tttatgccca
     3901 gcccgagctc cgccggcgca cacctggcaa ttttcaggcc aacagtatgg tcaaagggcc
     3961 aaagttcttt gtcatcctag aagttagtct gtgtctaaaa tattcctgtt ttttttccca
     4021 gatgaggagt cctaagctaa gatgactgtg tcctgcagct tagacatgac tttcagaagt
     4081 cttctttcta ttgcactgaa aatattgcca ttttgaacag attggccttg acctttgaac
     4141 agattggcct tgactttgaa aaatgtcaga ggggattttg attatggcca aagcatctca
     4201 cctgtacaag acattgtttg ttaaagagct ttggagttca tggcattgcc aggtaccttt
     4261 tttgcctgtt ttcttttagc aaacatgttt tagcaccaag gcaattcccg tcttacaaag
     4321 caattatcat taggaaatgt cacagtataa actgttggca gtaggaaaac agatctgatt
     4381 agtggctgac ttaggggatg ctcctcccaa gtttagaaga ttgtcatcga gcaggtgatt
     4441 agacacacct actgagcaat ggagtggtta ctgggattct ctaaaattct aaacacccaa
     4501 tttgaagacc accccccaat ctttgttgaa aatagttaat acttagtata ttttgttttc
     4561 atatataaaa ttacgcgtgc ggccgcagga acccctagtg atggagttgg ccactccctc
     4621 tctgcgcgct cgctcgctca ctgaggccgg gcgaccaaag gtcgcccgac gcccgggcgg
     4681 cctcagtgag cgagcgagcg cgcagctgcc tgcaggacat gtgagcaaaa ggccagcaaa
     4741 aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc cgcccccctg
     4801 acgagcatca caaaaatcga cgctcaagtc agaggtggcg aaacccgaca ggactataaa
     4861 gataccaggc gtttccccct ggaagctc
//
