LOCUS       sequence-492125-        4909 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AiE2657m 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 4909)
  TITLE     Basal Ganglia/CERP Enhancer AAVs
REFERENCE   2  (bases 1 to 4909)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"
COMMENT     Sequence Label: AiP2130 - pAAV-AiE2657m-minBG-SYFP2-WPRE3-BGHpA
FEATURES             Location/Qualifiers
     source          1..4909
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(join(309..1100,1101..1169))
                     /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     932..951
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(1170..1274)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     1342..1360
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(1398..1420)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     1520..1539
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     rep_origin      complement(1556..2011)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1693..1714)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     1905..1924
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     repeat_region   2086..2226
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   2097..2226
                     /label=AAV2 ITR
     polyA_signal    2344..2455
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     2642..2662
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(join(2755..3465,3466..3468,3469..3471))
                     /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(2797..2818)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     3146..3165
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     3405..3426
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     regulatory      3468..3477
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     repeat_region   4268..4397
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     rep_origin      complement(join(4459..4909,1..138))
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4539..4558)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa gagttggtag ctcttgatcc
       61 ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt gcaagcagca gattacgcgc
      121 agaaaaaaag gatctcaaga agatcctttg atcttttcta cggggtctga cgctcagtgg
      181 aacgaaaact cacgttaagg gattttggtc atgagattat caaaaaggat cttcacctag
      241 atccttttaa attaaaaatg aagttttaaa tcaatctaaa gtatatatga gtaaacttgg
      301 tctgacagtt accaatgctt aatcagtgag gcacctatct cagcgatctg tctatttcgt
      361 tcatccatag ttgcctgact ccccgtcgtg tagataacta cgatacggga gggcttacca
      421 tctggcccca gtgctgcaat gataccgcga gacccacgct caccggctcc agatttatca
      481 gcaataaacc agccagccgg aagggccgag cgcagaagtg gtcctgcaac tttatccgcc
      541 tccatccagt ctattaattg ttgccgggaa gctagagtaa gtagttcgcc agttaatagt
      601 ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg
      661 gcttcattca gctccggttc ccaacgatca aggcgagtta catgatcccc catgttgtgc
      721 aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg
      781 ttatcactca tggttatggc agcactgcat aattctctta ctgtcatgcc atccgtaaga
      841 tgcttttctg tgactggtga gtactcaacc aagtcattct gagaatagtg tatgcggcga
      901 ccgagttgct cttgcccggc gtcaatacgg gataataccg cgccacatag cagaacttta
      961 aaagtgctca tcattggaaa acgttcttcg gggcgaaaac tctcaaggat cttaccgctg
     1021 ttgagatcca gttcgatgta acccactcgt gcacccaact gatcttcagc atcttttact
     1081 ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata
     1141 agggcgacac ggaaatgttg aatactcata ctcttccttt ttcaatatta ttgaagcatt
     1201 tatcagggtt attgtctcat gagcggatac atatttgaat gtatttagaa aaataaacaa
     1261 ataggggttc cgcgcacatt tccccgaaaa gtgccacctg acgtctaaga aaccattatt
     1321 atcatgacat taacctataa aaataggcgt atcacgaggc cctttcgtct cgcgcgtttc
     1381 ggtgatgacg gtgaaaacct ctgacacatg cagctcccgg agacggtcac agcttgtctg
     1441 taagcggatg ccgggagcag acaagcccgt cagggcgcgt cagcgggtgt tggcgggtgt
     1501 cggggctggc ttaactatgc ggcatcagag cagattgtac tgagagtgca ccataaaatt
     1561 gtaaacgtta atattttgtt aaaattcgcg ttaaattttt gttaaatcag ctcatttttt
     1621 aaccaataga ccgaaatcgg caaaatccct tataaatcaa aagaatagcc cgagatagag
     1681 ttgagtgttg ttccagtttg gaacaagagt ccactattaa agaacgtgga ctccaacgtc
     1741 aaagggcgaa aaaccgtcta tcagggcgat ggcccactac gtgaaccatc acccaaatca
     1801 agttttttgg ggtcgaggtg ccgtaaagca ctaaatcgga accctaaagg gagcccccga
     1861 tttagagctt gacggggaaa gccggcgaac gtggcgagaa aggaagggaa gaaagcgaaa
     1921 ggagcgggcg ctaaggcgct ggcaagtgta gcggtcacgc tgcgcgtaac caccacaccc
     1981 gccgcgctta atgcgccgct acagggcgcg tactatggtt gctttgacgt atgcggtgtg
     2041 aaataccgca cagatgcgta aggagaaaat accgcatcag gcgcccctgc aggcagctgc
     2101 gcgctcgctc gctcactgag gccgcccggg caaagcccgg gcgtcgggcg acctttggtc
     2161 gcccggcctc agtgagcgag cgagcgcgca gagagggagt ggccaactcc atcactaggg
     2221 gttcctgcgg ccgctcggtc cgcacgtgag atctcatgcc tgctattgtc ttcccaatcc
     2281 tcccccttgc tgtcctgccc caccccaccc cccagaatag aatgacacct actcagacaa
     2341 tgcgatgcaa tttcctcatt ttattaggaa aggacagtgg gagtggcacc ttccagggtc
     2401 aaggaaggca cgggggaggg gcaaacaaca gatggctggc aactagaagg cacagtcgaa
     2461 gatctctcga gccacggaat tgtcagtgcc caacagccga gcccctgtcc agcagcgggc
     2521 aaggcaggcg gcgatgagtt ccgccgtggc aagaactaac caggatttat acaaggagga
     2581 gaaaatgaaa gccatacggg aagcaatagc atgatacaaa ggcattaaag cagcgtatcc
     2641 acatagcgta aaaggagcaa catagttaag aataccagtc aatctttcac aaattttgta
     2701 atccagaggt tgattatgat atcgaattcg cggccgcggc gcgccgtcga cttacttgta
     2761 cagctcgtcc atgccgagag tgatcccggc ggcggtcacg aactccagca ggaccatgtg
     2821 atcgcgcttc tcgttggggt ctttgctcag cttggactgg tagctcaggt agtggttgtc
     2881 gggcagcagc acggggccgt cgccgatggg ggtgttctgc tggtagtggt cggcgagctg
     2941 cacgccgccg tcctcgatgt tgtggcggat cttgaagttg gccttgatgc cgttcttctg
     3001 cttgtcggcg gtgatataga cgttgtggct gttgtagttg tactccagct tgtgccccag
     3061 gatgttgccg tcctccttga agtcgatgcc cttcagctcg atgcggttca ccagggtgtc
     3121 gccctcgaac ttcacctcgg cgcgggtctt gtagttgccg tcgtccttga agaagatggt
     3181 gcgctcctgg acgtagcctt cgggcatggc ggacttgaag aagtcgtgct gcttcatgtg
     3241 gtcggggtag cgggcgaagc actgcacgcc gtagcccagg gtggtcacga gggtgggcca
     3301 gggcacgggc agcttgccgg tggtgcagat cagcttcagg gtcagcttgc cgtaggtggc
     3361 atcgccctcg ccctcgccgg acacgctgaa cttgtggccg tttacgtcgc cgtccagctc
     3421 gaccaggatg ggcaccaccc cggtgaacag ctcctcgccc ttgctcacca tggtggcgac
     3481 cggtagcgct agcttcgaaa gatctggatc ccagaagcaa atgtaagcaa tagatggctc
     3541 tgccctgact tttatgccca gcccgagctc cgccggcgct tcacagatac aaatagatga
     3601 ggaagtttag ttcaccttgt atgtgatggt ccaagcctgg gaacaactcc aaccacacca
     3661 agtctaggag atagatatga agaacacaga acagctaacg atgcagccat ctagagggct
     3721 cgttaagaca ccgcttcctg gactccatcg tcagaatctc agatgcaaga aggagcaggg
     3781 gcgggggtgg agggctgagg atttgcattg caacaagttc ccagatgagg ctgagagaag
     3841 tttaccagcc acttgtctag actgcagaat cctatgcaga cattcaaaat agtgccctag
     3901 aagaatgtat gttgatttga gccaacgcac acaggagacg atttaaagtg cagcagagcc
     3961 tggccgctca gcagcctggt gatgccgtca gcataagtat aaatataagt gcacacactc
     4021 gcaaagagaa gcaggaggca cgcatgggac ctgggcagga agcaggatgg ggcttcaagg
     4081 agctcagagc caggctggct taggcgtgtt cctgcctctg cctctgtttg cctagtgccc
     4141 ctgaatagac tggagagcgg cctgaactat taaaactctt taagaaagtc ctgtcagcaa
     4201 tcccatttca cagatgacag accttcatcc caagactgct cagtgcctgt tgaacgcgtg
     4261 cggccgcagg aacccctagt gatggagttg gccactccct ctctgcgcgc tcgctcgctc
     4321 actgaggccg ggcgaccaaa ggtcgcccga cgcccgggcg gcctcagtga gcgagcgagc
     4381 gcgcagctgc ctgcaggaca tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa
     4441 ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc acaaaaatcg
     4501 acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg cgtttccccc
     4561 tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat acctgtccgc
     4621 ctttctccct tcgggaagcg tggcgctttc tcatagctca cgctgtaggt atctcagttc
     4681 ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc agcccgaccg
     4741 ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg acttatcgcc
     4801 actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg gtgctacaga
     4861 gttcttgaag tggtggccta actacggcta cactagaaga acagtattt
//
