LOCUS       sequence-471582-        5965 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Beta-Secretase to be expressed (1:1) with either ATLAS protein..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5965)
  AUTHORS   Rivera JF, Huang H, Weng W, Sohn H, Girasole AE, Li S, Albanese MA, 
            Qin M, Tao C, Klug ME, Rao S, Paletzki R, Herring BE, Kanoski SE, 
            Zhang LI, Gerfen CR, Sabatini BL, Arnold DB
  TITLE     ATLAS: a rationally designed anterograde transsynaptic tracer.
  JOURNAL   Nat Methods. 2025 May;22(5):1101-1111. doi: 
            10.1038/s41592-025-02670-x. Epub 2025 May 1.
  PUBMED    40312509
REFERENCE   2  (bases 1 to 5965)
  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"; doi: 
            "10.1038/s41592-025-02670-x"; journalName: "Nat Methods"; date: 
            "2025-05"; volume: "22"; issue: "5"; pages: "1101-1111"
COMMENT     Sequence Label: pAAV-DIO-BACE-HA
FEATURES             Location/Qualifiers
     source          1..5965
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   1..130
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     promoter        175..622
                     /label=hSyn promoter
                     /note="human synapsin I promoter; confers neuron-specific 
                     expression (Kugler et al., 2003)"
     protein_bind    complement(660..693)
                     /label=lox2272
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (AAGTATCC) (Shaw et al., 2021). lox2272 sites are 
                     compatible with each other, but incompatible with loxP or 
                     loxN sites (Lee and Saito, 1988)."
     protein_bind    complement(744..777)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     CDS             complement(2157..2183)
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     protein_bind    2325..2358
                     /label=lox2272
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (AAGTATCC) (Shaw et al., 2021). lox2272 sites are 
                     compatible with each other, but incompatible with loxP or 
                     loxN sites (Lee and Saito, 1988)."
     protein_bind    2409..2442
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     misc_feature    2467..3055
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(2520..2540)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(2938..2949)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(3121..3140)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3175..3194
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     repeat_region   3228..3368
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   3228..3357
                     /label=AAV2 ITR
     rep_origin      3443..3898
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3530..3549)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3740..3761
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3915..3934)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4034..4056
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(4094..4112)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4180..4284
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4285..4353,4354..5145)
                     /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(4503..4522)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5316..5904
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5805..5824
                     /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 gcgtgtgtct agactgcaga gggccctgcg tatgagtgca
      181 agtgggtttt aggaccagga tgaggcgggg tgggggtgcc tacctgacga ccgaccccga
      241 cccactggac aagcacccaa cccccattcc ccaaattgcg catcccctat cagagagggg
      301 gaggggaaac aggatgcggc gaggcgcgtg cgcactgcca gcttcagcac cgcggacagt
      361 gccttcgccc ccgcctggcg gcgcgcgcca ccgccgcctc agcactgaag gcgcgctgac
      421 gtcactcgcc ggtcccccgc aaactcccct tcccggccac cttggtcgcg tccgcgccgc
      481 cgccggccca gccggaccgc accacgcgag gcgcgagata ggggggcacg ggcgcgacca
      541 tctgcgctgc ggcgccggcg actcagcgct gcctcagtct gcggtgggca gcggaggagt
      601 cgtgtcgtgc ctgagagcgc agtcgagaag gtaccggatc ctctagagtc gactccggaa
      661 taacttcgta taggatactt tatacgaagt tatgcagaat ggtagctgga ttgtagctgc
      721 tattagcaat atgaaacctc ttaataactt cgtatagcat acattatacg aagttatggc
      781 gcgccctact tcagcaggga gatgtcatca gcaaagtcat catgctgctg gcgcaggcag
      841 cggaggcagc gccactgaca caccatgagg cagagtggca gcatgaagag ggcgcagatg
      901 gcagccatga cataggctat ggtcatgagg gttgactcat ctgtctgtgg aatgttgtag
      961 ccacagtctt ccatgtccaa ggtgacaaaa gggccttcca ccgctgccgt cctgaactca
     1021 tcgtgcacat ggcaagcgct gacagcaaag ccaattcgtt ttcgggcccg atcaaagaca
     1081 acgtagaagc cctccatgat aacagctccc ataacagtgc ccgtggatga ctgtgagatg
     1141 gcaaacttgt aacagtcgtc ttgggacgtg gccacatctt ccactggccg caggtattgc
     1201 tgcggaagga tggtgatgcg gaaggactgg ttggtaacct cacccattag gtagagtgag
     1261 atgactggga aaatgttcca aggggtggtg cctgcttgcc agcacaccag ctgctctcct
     1321 agccagaaac catcagggaa cttctccgtg gaggaggctg ccttgatgga tttgactgca
     1381 gcttcaaaca ctttcttggg caaacgaagg ttggtggtgc cactgtccac aatgctcttg
     1441 tcatagttgt actccttgca gtccattttc agatcctgtc cattgatctc cacccgcaca
     1501 atgatcacct cataatacca ctcccgccgg atgggtgtat accagagact gcctgtgtac
     1561 agcgagtggt cgatacctcc aatgatcatg ctccctccga cagaggccag cacttcagac
     1621 tggttgaggg ggaagccagc accacaaagc tgcagggaga agaggttggg aacgtgggtc
     1681 tgctttacca gagagtcaaa gaaaggctcc agggagtcgt caggcctggc aatctcagca
     1741 taggccagcc ccaggatgcc ttcccagttg gagccgttga tgaagaactt gtctgattca
     1801 gtgatggcag caatgttggc acgcacagtg acgttggggc catgggggat gcttaccagg
     1861 tcggtgccca gctccccttc ccacttgccc tgggtgtagg gcacatacac acccttccgg
     1921 aggtcccggt atgtgctgga cagctgcctc tggtagtagc gatgcaggaa ggggtggggg
     1981 gcagcaccca ctgcaaagtt actgctgcct gtatccacca ggatgttgag cgtctgcggg
     2041 gggctgccca cggtcatctc cacgtagtag ccctgccccg acttgcccct caggttgtcc
     2101 accatctcca caaagctgcc cctccggccg ggctcctcgg gctcttcgtc ggtctcagcg
     2161 taatctggta catcgtaagg gtaccggggc agccgcagcc ccaggggggc gccccccagg
     2221 ccgctgcgca ggggcagccg gatgccgtgc tgggtgccgt gggcaggcag cactcccgcg
     2281 cccatccaca gcaggagcca gggcagggct tgggccatgc tagcataact tcgtataaag
     2341 tatcctatac gaagttattt gccttaaccc agaaattatc actgttattc tttagaatgg
     2401 tgcaaagaat aacttcgtat aatgtatgct atacgaagtt atgaattcga tatcaagctt
     2461 atcgataatc aacctctgga ttacaaaatt tgtgaaagat tgactggtat tcttaactat
     2521 gttgctcctt ttacgctatg tggatacgct gctttaatgc ctttgtatca tgctattgct
     2581 tcccgtatgg ctttcatttt ctcctccttg tataaatcct ggttgctgtc tctttatgag
     2641 gagttgtggc ccgttgtcag gcaacgtggc gtggtgtgca ctgtgtttgc tgacgcaacc
     2701 cccactggtt ggggcattgc caccacctgt cagctccttt ccgggacttt cgctttcccc
     2761 ctccctattg ccacggcgga actcatcgcc gcctgccttg cccgctgctg gacaggggct
     2821 cggctgttgg gcactgacaa ttccgtggtg ttgtcgggga aatcatcgtc ctttccttgg
     2881 ctgctcgcct atgttgccac ctggattctg cgcgggacgt ccttctgcta cgtcccttcg
     2941 gccctcaatc cagcggacct tccttcccgc ggcctgctgc cggctctgcg gcctcttccg
     3001 cgtcttcgcc ttcgccctca gacgagtcgg atctcccttt gggccgcctc cccgcatcga
     3061 taccgagcgc tgctcgagag atctacttgt ttattgcagc ttataatggt tacaaataaa
     3121 gcaatagcat cacaaatttc acaaataaag catttttttc actgcattct agttgtggtt
     3181 tgtccaaact catcaatgta tcttacacgt gcggaccgag cggccgcagg aacccctagt
     3241 gatggagttg gccactccct ctctgcgcgc tcgctcgctc actgaggccg ggcgaccaaa
     3301 ggtcgcccga cgcccgggct ttgcccgggc ggcctcagtg agcgagcgag cgcgcagctg
     3361 cctgcagggg cgcctgatgc ggtattttct ccttacgcat ctgtgcggta tttcacaccg
     3421 catacgtcaa agcaaccata gtacgcgccc tgtagcggcg cattaagcgc ggcgggtgtg
     3481 gtggttacgc gcagcgtgac cgctacactt gccagcgcct tagcgcccgc tcctttcgct
     3541 ttcttccctt cctttctcgc cacgttcgcc ggctttcccc gtcaagctct aaatcggggg
     3601 ctccctttag ggttccgatt tagtgcttta cggcacctcg accccaaaaa acttgatttg
     3661 ggtgatggtt cacgtagtgg gccatcgccc tgatagacgg tttttcgccc tttgacgttg
     3721 gagtccacgt tctttaatag tggactcttg ttccaaactg gaacaacact caactctatc
     3781 tcgggctatt cttttgattt ataagggatt ttgccgattt cggtctattg gttaaaaaat
     3841 gagctgattt aacaaaaatt taacgcgaat tttaacaaaa tattaacgtt tacaatttta
     3901 tggtgcactc tcagtacaat ctgctctgat gccgcatagt taagccagcc ccgacacccg
     3961 ccaacacccg ctgacgcgcc ctgacgggct tgtctgctcc cggcatccgc ttacagacaa
     4021 gctgtgaccg tctccgggag ctgcatgtgt cagaggtttt caccgtcatc accgaaacgc
     4081 gcgagacgaa agggcctcgt gatacgccta tttttatagg ttaatgtcat gataataatg
     4141 gtttcttaga cgtcaggtgg cacttttcgg ggaaatgtgc gcggaacccc tatttgttta
     4201 tttttctaaa tacattcaaa tatgtatccg ctcatgagac aataaccctg ataaatgctt
     4261 caataatatt gaaaaaggaa gagtatgagt attcaacatt tccgtgtcgc ccttattccc
     4321 ttttttgcgg cattttgcct tcctgttttt gctcacccag aaacgctggt gaaagtaaaa
     4381 gatgctgaag atcagttggg tgcacgagtg ggttacatcg aactggatct caacagcggt
     4441 aagatccttg agagttttcg ccccgaagaa cgttttccaa tgatgagcac ttttaaagtt
     4501 ctgctatgtg gcgcggtatt atcccgtatt gacgccgggc aagagcaact cggtcgccgc
     4561 atacactatt ctcagaatga cttggttgag tactcaccag tcacagaaaa gcatcttacg
     4621 gatggcatga cagtaagaga attatgcagt gctgccataa ccatgagtga taacactgcg
     4681 gccaacttac ttctgacaac gatcggagga ccgaaggagc taaccgcttt tttgcacaac
     4741 atgggggatc atgtaactcg ccttgatcgt tgggaaccgg agctgaatga agccatacca
     4801 aacgacgagc gtgacaccac gatgcctgta gcaatggcaa caacgttgcg caaactatta
     4861 actggcgaac tacttactct agcttcccgg caacaattaa tagactggat ggaggcggat
     4921 aaagttgcag gaccacttct gcgctcggcc cttccggctg gctggtttat tgctgataaa
     4981 tctggagccg gtgagcgtgg gtctcgcggt atcattgcag cactggggcc agatggtaag
     5041 ccctcccgta tcgtagttat ctacacgacg gggagtcagg caactatgga tgaacgaaat
     5101 agacagatcg ctgagatagg tgcctcactg attaagcatt ggtaactgtc agaccaagtt
     5161 tactcatata tactttagat tgatttaaaa cttcattttt aatttaaaag gatctaggtg
     5221 aagatccttt ttgataatct catgaccaaa atcccttaac gtgagttttc gttccactga
     5281 gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag atcctttttt tctgcgcgta
     5341 atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa
     5401 gagctaccaa ctctttttcc gaaggtaact ggcttcagca gagcgcagat accaaatact
     5461 gttcttctag tgtagccgta gttaggccac cacttcaaga actctgtagc accgcctaca
     5521 tacctcgctc tgctaatcct gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt
     5581 accgggttgg actcaagacg atagttaccg gataaggcgc agcggtcggg ctgaacgggg
     5641 ggttcgtgca cacagcccag cttggagcga acgacctaca ccgaactgag atacctacag
     5701 cgtgagctat gagaaagcgc cacgcttccc gaagggagaa aggcggacag gtatccggta
     5761 agcggcaggg tcggaacagg agagcgcacg agggagcttc cagggggaaa cgcctggtat
     5821 ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt gtgatgctcg
     5881 tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc
     5941 ttttgctggc cttttgctca catgt
//
