LOCUS       sequence-489677-        4806 bp    DNA     circular SYN 17-OCT-2025
DEFINITION  AiE1494m 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 4806)
  TITLE     Basal Ganglia/CERP Enhancer AAVs
REFERENCE   2  (bases 1 to 4806)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Oct 17, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"
COMMENT     Sequence Label: AiP15214 - pAAV-AiE1494m-minBG-SYFP2-WPRE3-BGHpA
FEATURES             Location/Qualifiers
     source          1..4806
                     /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      818..827
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(824..826,827..829,830..1540)
                     /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(869..890)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1130..1149)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1477..1498
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(1633..1653)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     polyA_signal    1840..1951
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   2069..2209
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   2069..2198
                     /label=AAV2 ITR
     rep_origin      2284..2739
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2371..2390)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2581..2602
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(2756..2775)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     2875..2897
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(2935..2953)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3021..3125
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3126..3194,3195..3986)
                     /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(3344..3363)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4157..4745
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4646..4665
                     /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 gcgtcagagc cccaccatct agtatgaaag gccgatgctc
      181 aagaagcaag gctgggcgtc aaacctcttc agtctaagcc aacggtgtat ccagtctagg
      241 actaatggtt gtaggcagca tagctgttac tgctgtggat gagcacaggg taagaagagc
      301 tagaccctca tctctagtgg ggtaatggac agctggattt cttagggaat tcagacatcc
      361 agaggagagg gaaaggggga aagagggtgg gagcctgaga gccagcttga aggactctag
      421 gaggtaagag tatcaacacc acggtcaatc aagtggacct aggagcgatc tcacttcccg
      481 gctgtgtgac ctcaaaccga gggtctcacc taagcctcag aaatctcaac aaaacttaac
      541 aggaaatgcc cagttcatat tcagcacttg acaggctgag gcaggaggat ttcatatgtg
      601 tgtgtgtgtg tgtgtgtgtg tgtgtgtgtc ctgggacaca aagcaagcaa gcaaacagat
      661 aaagcagtag aacttagtcc agccttgctt cttattaggg ggttcagttg agcacagctg
      721 ttgcgagctc gggctgggca taaaagtcag ggcagagcca tctattgctt acatttgctt
      781 ctgggatcca gatctttcga agctagcgct accggtcgcc accatggtga gcaagggcga
      841 ggagctgttc accggggtgg tgcccatcct ggtcgagctg gacggcgacg taaacggcca
      901 caagttcagc gtgtccggcg agggcgaggg cgatgccacc tacggcaagc tgaccctgaa
      961 gctgatctgc accaccggca agctgcccgt gccctggccc accctcgtga ccaccctggg
     1021 ctacggcgtg cagtgcttcg cccgctaccc cgaccacatg aagcagcacg acttcttcaa
     1081 gtccgccatg cccgaaggct acgtccagga gcgcaccatc ttcttcaagg acgacggcaa
     1141 ctacaagacc cgcgccgagg tgaagttcga gggcgacacc ctggtgaacc gcatcgagct
     1201 gaagggcatc gacttcaagg aggacggcaa catcctgggg cacaagctgg agtacaacta
     1261 caacagccac aacgtctata tcaccgccga caagcagaag aacggcatca aggccaactt
     1321 caagatccgc cacaacatcg aggacggcgg cgtgcagctc gccgaccact accagcagaa
     1381 cacccccatc ggcgacggcc ccgtgctgct gcccgacaac cactacctga gctaccagtc
     1441 caagctgagc aaagacccca acgagaagcg cgatcacatg gtcctgctgg agttcgtgac
     1501 cgccgccggg atcactctcg gcatggacga gctgtacaag taagtcgacg gcgcgccgcg
     1561 gccgcgaatt cgatatcata atcaacctct ggattacaaa atttgtgaaa gattgactgg
     1621 tattcttaac tatgttgctc cttttacgct atgtggatac gctgctttaa tgcctttgta
     1681 tcatgctatt gcttcccgta tggctttcat tttctcctcc ttgtataaat cctggttagt
     1741 tcttgccacg gcggaactca tcgccgcctg ccttgcccgc tgctggacag gggctcggct
     1801 gttgggcact gacaattccg tggctcgaga gatcttcgac tgtgccttct agttgccagc
     1861 catctgttgt ttgcccctcc cccgtgcctt ccttgaccct ggaaggtgcc actcccactg
     1921 tcctttccta ataaaatgag gaaattgcat cgcattgtct gagtaggtgt cattctattc
     1981 tggggggtgg ggtggggcag gacagcaagg gggaggattg ggaagacaat agcaggcatg
     2041 agatctcacg tgcggaccga gcggccgcag gaacccctag tgatggagtt ggccactccc
     2101 tctctgcgcg ctcgctcgct cactgaggcc gggcgaccaa aggtcgcccg acgcccgggc
     2161 tttgcccggg cggcctcagt gagcgagcga gcgcgcagct gcctgcaggg gcgcctgatg
     2221 cggtattttc tccttacgca tctgtgcggt atttcacacc gcatacgtca aagcaaccat
     2281 agtacgcgcc ctgtagcggc gcattaagcg cggcgggtgt ggtggttacg cgcagcgtga
     2341 ccgctacact tgccagcgcc ttagcgcccg ctcctttcgc tttcttccct tcctttctcg
     2401 ccacgttcgc cggctttccc cgtcaagctc taaatcgggg gctcccttta gggttccgat
     2461 ttagtgcttt acggcacctc gaccccaaaa aacttgattt gggtgatggt tcacgtagtg
     2521 ggccatcgcc ctgatagacg gtttttcgcc ctttgacgtt ggagtccacg ttctttaata
     2581 gtggactctt gttccaaact ggaacaacac tcaactctat ctcgggctat tcttttgatt
     2641 tataagggat tttgccgatt tcggtctatt ggttaaaaaa tgagctgatt taacaaaaat
     2701 ttaacgcgaa ttttaacaaa atattaacgt ttacaatttt atggtgcact ctcagtacaa
     2761 tctgctctga tgccgcatag ttaagccagc cccgacaccc gccaacaccc gctgacgcgc
     2821 cctgacgggc ttgtctgctc ccggcatccg cttacagaca agctgtgacc gtctccggga
     2881 gctgcatgtg tcagaggttt tcaccgtcat caccgaaacg cgcgagacga aagggcctcg
     2941 tgatacgcct atttttatag gttaatgtca tgataataat ggtttcttag acgtcaggtg
     3001 gcacttttcg gggaaatgtg cgcggaaccc ctatttgttt atttttctaa atacattcaa
     3061 atatgtatcc gctcatgaga caataaccct gataaatgct tcaataatat tgaaaaagga
     3121 agagtatgag tattcaacat ttccgtgtcg cccttattcc cttttttgcg gcattttgcc
     3181 ttcctgtttt tgctcaccca gaaacgctgg tgaaagtaaa agatgctgaa gatcagttgg
     3241 gtgcacgagt gggttacatc gaactggatc tcaacagcgg taagatcctt gagagttttc
     3301 gccccgaaga acgttttcca atgatgagca cttttaaagt tctgctatgt ggcgcggtat
     3361 tatcccgtat tgacgccggg caagagcaac tcggtcgccg catacactat tctcagaatg
     3421 acttggttga gtactcacca gtcacagaaa agcatcttac ggatggcatg acagtaagag
     3481 aattatgcag tgctgccata accatgagtg ataacactgc ggccaactta cttctgacaa
     3541 cgatcggagg accgaaggag ctaaccgctt ttttgcacaa catgggggat catgtaactc
     3601 gccttgatcg ttgggaaccg gagctgaatg aagccatacc aaacgacgag cgtgacacca
     3661 cgatgcctgt agcaatggca acaacgttgc gcaaactatt aactggcgaa ctacttactc
     3721 tagcttcccg gcaacaatta atagactgga tggaggcgga taaagttgca ggaccacttc
     3781 tgcgctcggc ccttccggct ggctggttta ttgctgataa atctggagcc ggtgagcgtg
     3841 ggtctcgcgg tatcattgca gcactggggc cagatggtaa gccctcccgt atcgtagtta
     3901 tctacacgac ggggagtcag gcaactatgg atgaacgaaa tagacagatc gctgagatag
     3961 gtgcctcact gattaagcat tggtaactgt cagaccaagt ttactcatat atactttaga
     4021 ttgatttaaa acttcatttt taatttaaaa ggatctaggt gaagatcctt tttgataatc
     4081 tcatgaccaa aatcccttaa cgtgagtttt cgttccactg agcgtcagac cccgtagaaa
     4141 agatcaaagg atcttcttga gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa
     4201 aaaaaccacc gctaccagcg gtggtttgtt tgccggatca agagctacca actctttttc
     4261 cgaaggtaac tggcttcagc agagcgcaga taccaaatac tgttcttcta gtgtagccgt
     4321 agttaggcca ccacttcaag aactctgtag caccgcctac atacctcgct ctgctaatcc
     4381 tgttaccagt ggctgctgcc agtggcgata agtcgtgtct taccgggttg gactcaagac
     4441 gatagttacc ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc acacagccca
     4501 gcttggagcg aacgacctac accgaactga gatacctaca gcgtgagcta tgagaaagcg
     4561 ccacgcttcc cgaagggaga aaggcggaca ggtatccggt aagcggcagg gtcggaacag
     4621 gagagcgcac gagggagctt ccagggggaa acgcctggta tctttatagt cctgtcgggt
     4681 ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg cggagcctat
     4741 ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc cttttgctgg ccttttgctc
     4801 acatgt
//
