LOCUS       sequence-471577-        6981 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Anterograde transsynaptic tracer protein to express in presynaptic 
            glutamatergic neurons, Cre payload. Must use with AAV8-DIO-Reporter 
            viruses infected in postsynaptic cells..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6981)
  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 6981)
  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-ATLASsnCre
FEATURES             Location/Qualifiers
     source          1..6981
                     /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)"
     regulatory      636..645
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1098..1136
                     /codon_start=1
                     /product="rationally designed epitope tag that forms a 
                     stable alpha-helix (Gotzke et al., 2019)"
                     /label=ALFA-tag
                     /note="It may be beneficial to flank the ALFA-tag sequence 
                     with proline residues to preserve the helical structure."
                     /translation="SRLEEELRRRLTE"
     CDS             1464..1487
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             1656..1676
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             1677..2702
                     /codon_start=1
                     /product="site-specific recombinase"
                     /label=Cre
                     /note="Cre is a site-specific recombinase from 
                     bacteriophage P1. Recombination occurs at loxP sequences."
                     /translation="SNLLTVHQNLPALPVDATSDEVRKNLMDMFRDRQAFSEHTWKMLQ
                     SVCRSWAAWCKLNNRKWFPAEPEDVRDYLLYLQARGLAVKTIQQHLGQLNMLHRRSGLP
                     RPSDSNAVSLVMRRIRKENVDAGERAKQALAFERTDFDQVRSLMENSDRCQDIRNLAFL
                     GIAYNTLLRIAEIARIRVKDISRTDGGRMLIHIGRTKTLVSTAGVEKALSLGVTKLVER
                     WISVSGVADDPNNYLFCRVRKNGVAAPSATSQLSTRALEGIFEATHRLIYGAKDDSGQR
                     YLAWSGHSARVGAARDMARAGVSIPEIMQAGGWTNVNIVMNYIRNLDSETGAMVRLLED
                     GD"
     CDS             2715..3056
                     /codon_start=1
                     /product="zinc-finger domain from the CCR5-224 (+) 
                     zinc-finger nuclease that recognizes the left half-site 
                     target sequence in the human CCR5 gene (Perez et al., 2008;
                     Gross et al., 2013)"
                     /label=CCR5TC
                     /translation="FQCRICMRNFSDRSNLSRHIRTHTGEKPFACDICGRKFAISSNLN
                     SHTKIHTGSQKPFQCRICMRNFSRSDNLARHIRTHTGEKPFACDICGRKFATSGNLTRH
                     TKIHLRGSQL"
     misc_feature    3108..3696
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(3161..3181)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3579..3590)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    3728..4204
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(3927..3946)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     repeat_region   4244..4384
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   4244..4373
                     /label=AAV2 ITR
     rep_origin      4459..4914
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4546..4565)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4756..4777
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4931..4950)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5050..5072
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(5110..5128)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5196..5300
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5301..5369,5370..6161)
                     /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(5519..5538)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6332..6920
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6821..6840
                     /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 gtaccgccac catggaggtg cagctggagg
      661 agtctggggg cggcttggtg cggcctgggg ggtctctaag actctcctgt gcagcctctg
      721 gattcccctt cagtaaatat tttatgagct gggtccgcca ggctccagga aaggggctcg
      781 aatgggtctc aactattagt gctagtggca attatgaaac ctatacagag tccgtgaagg
      841 gccgattcac catcgccaga gacaacgcca agaacacgct gtatctgcag atgaacagcc
      901 tgaaaccgga ggacacggcc gtctattact gtgcaaaagg gagctgggcc cgggatatga
      961 cgaggggtca ggggacccag gtcactgtct cctcagaacc caagggcgga gggtcccaat
     1021 tgatgatgat catcttggga gtgatttgcg ccatcatcct catcatcatc atcgtttact
     1081 tcagcactac tagtccgagc cgcctggaag aagaactgcg ccgccgcctg accgaaccgg
     1141 agctcgctag tgaaattagt gaagtaaacc tagacgctga atttagagct agcatgctcg
     1201 aggtcaagga agcatcacca accagcatcc agatcagctg gagcaggagg gaccaggtga
     1261 ttcgctacta ccgcatcacc tacggcgaaa ctggtggcaa tagccctgtc caaaaattca
     1321 ccgtgcctgg cagcaaatcc actgctacca tcagcggcct gaaacctggt gttgactata
     1381 ccatcacggt gtacgccgtc acgcccttcg ccaacttctg ggactggcac ggcccgatct
     1441 ccatcaacta ccgcaccgga tccgactaca aggatgacga cgataagggt agcggctcct
     1501 ctagaggaag cggaagcgga tatgtaagca ggcatcccgg aggcgggtgc ggttacgttt
     1561 cacggcaccc cggcggtggt tgcggttatg taagtcgaca tccgggagga ggctgcggtt
     1621 atgtgagccg tcatcccgga gggggatgca agcttcccaa gaagaagagg aaggtgtcca
     1681 atttactgac cgtacaccaa aatttgcctg cattaccggt cgatgcaacg agtgatgagg
     1741 ttcgcaagaa cctgatggac atgttcaggg atcgccaggc gttttctgag catacctgga
     1801 aaatgcttca gtccgtttgc cggtcgtggg cggcatggtg caagttgaat aaccggaaat
     1861 ggtttcccgc agaacctgaa gatgttcgcg attatcttct atatcttcag gcgcgcggtc
     1921 tggcagtaaa aactatccag caacatttgg gccagctaaa catgcttcat cgtcggtccg
     1981 ggctgccacg accaagtgac agcaatgctg tttcactggt tatgcggcgg atccgaaaag
     2041 aaaacgttga tgccggtgaa cgtgcaaaac aggctctagc gttcgaacgc actgatttcg
     2101 accaggttcg ttcactcatg gaaaatagcg atcgctgcca ggatatacgt aatctggcat
     2161 ttctggggat tgcttataac accctgttac gtatagccga aattgccagg atcagggtta
     2221 aagatatctc acgtactgac ggtgggagaa tgttaatcca tattggcaga acgaaaacgc
     2281 tggttagcac cgcaggtgta gagaaggcac ttagcctggg ggtaactaaa ctggtcgagc
     2341 gatggatttc cgtctctggt gtagctgatg atccgaataa ctacctgttt tgccgggtca
     2401 gaaaaaatgg tgttgccgcg ccatctgcca ccagccagct atcaactcgc gccctggaag
     2461 ggatttttga agcaactcat cgattgattt acggcgctaa ggatgactct ggtcagagat
     2521 acctggcctg gtctggacac agtgcccgtg tcggagccgc gcgagatatg gcccgcgctg
     2581 gagtttcaat accggagatc atgcaagctg gtggctggac caatgtaaat attgtcatga
     2641 actatatccg taacctggat agtgaaacag gggcaatggt gcgcctgctg gaagatggcg
     2701 atacgcgtgg tagcttccag tgccggatct gcatgcggaa cttcagcgac cggtccaacc
     2761 tgagcaggca catcagaacc cacaccggag aaaagccctt cgcctgcgac atttgcggcc
     2821 ggaagttcgc catcagcagc aacctgaaca gccacaccaa gatccacact ggcagccaga
     2881 aacctttcca gtgcagaatt tgtatgagaa actttagcag aagcgacaac ctggccagac
     2941 acatccggac acatactggt gaaaaacctt ttgcctgtga tatctgtggc agaaagtttg
     3001 ccacctccgg caatctgacc cggcacacaa agattcacct gcggggcagc cagctatagt
     3061 gtacaagatc tctcgagacg cgtgctagcg gcgccaagct tatcgataat caacctctgg
     3121 attacaaaat ttgtgaaaga ttgactggta ttcttaacta tgttgctcct tttacgctat
     3181 gtggatacgc tgctttaatg cctttgtatc atgctattgc ttcccgtatg gctttcattt
     3241 tctcctcctt gtataaatcc tggttgctgt ctctttatga ggagttgtgg cccgttgtca
     3301 ggcaacgtgg cgtggtgtgc actgtgtttg ctgacgcaac ccccactggt tggggcattg
     3361 ccaccacctg tcagctcctt tccgggactt tcgctttccc cctccctatt gccacggcgg
     3421 aactcatcgc cgcctgcctt gcccgctgct ggacaggggc tcggctgttg ggcactgaca
     3481 attccgtggt gttgtcgggg aaatcatcgt cctttccttg gctgctcgcc tatgttgcca
     3541 cctggattct gcgcgggacg tccttctgct acgtcccttc ggccctcaat ccagcggacc
     3601 ttccttcccg cggcctgctg ccggctctgc ggcctcttcc gcgtcttcgc cttcgccctc
     3661 agacgagtcg gatctccctt tgggccgcct ccccgcatcg ataccgagcg ctgctcgaga
     3721 gatctacggg tggcatccct gtgacccctc cccagtgcct ctcctggccc tggaagttgc
     3781 cactccagtg cccaccagcc ttgtcctaat aaaattaagt tgcatcattt tgtctgacta
     3841 ggtgtccttc tataatatta tggggtggag gggggtggta tggagcaagg ggcaagttgg
     3901 gaagacaacc tgtagggcct gcggggtcta ttgggaacca agctggagtg cagtggcaca
     3961 atcttggctc actgcaatct ccgcctcctg ggttcaagcg attctcctgc ctcagcctcc
     4021 cgagttgttg ggattccagg catgcatgac caggctcagc taatttttgt ttttttggta
     4081 gagacggggt ttcaccatat tggccaggct ggtctccaac tcctaatctc aggtgatcta
     4141 cccaccttgg cctcccaaat tgctgggatt acaggcgtga accactgctc ccttccctgt
     4201 ccttctgatt ttgtaggtaa ccacgtgcgg accgagcggc cgcaggaacc cctagtgatg
     4261 gagttggcca ctccctctct gcgcgctcgc tcgctcactg aggccgggcg accaaaggtc
     4321 gcccgacgcc cgggctttgc ccgggcggcc tcagtgagcg agcgagcgcg cagctgcctg
     4381 caggggcgcc tgatgcggta ttttctcctt acgcatctgt gcggtatttc acaccgcata
     4441 cgtcaaagca accatagtac gcgccctgta gcggcgcatt aagcgcggcg ggtgtggtgg
     4501 ttacgcgcag cgtgaccgct acacttgcca gcgccttagc gcccgctcct ttcgctttct
     4561 tcccttcctt tctcgccacg ttcgccggct ttccccgtca agctctaaat cgggggctcc
     4621 ctttagggtt ccgatttagt gctttacggc acctcgaccc caaaaaactt gatttgggtg
     4681 atggttcacg tagtgggcca tcgccctgat agacggtttt tcgccctttg acgttggagt
     4741 ccacgttctt taatagtgga ctcttgttcc aaactggaac aacactcaac tctatctcgg
     4801 gctattcttt tgatttataa gggattttgc cgatttcggt ctattggtta aaaaatgagc
     4861 tgatttaaca aaaatttaac gcgaatttta acaaaatatt aacgtttaca attttatggt
     4921 gcactctcag tacaatctgc tctgatgccg catagttaag ccagccccga cacccgccaa
     4981 cacccgctga cgcgccctga cgggcttgtc tgctcccggc atccgcttac agacaagctg
     5041 tgaccgtctc cgggagctgc atgtgtcaga ggttttcacc gtcatcaccg aaacgcgcga
     5101 gacgaaaggg cctcgtgata cgcctatttt tataggttaa tgtcatgata ataatggttt
     5161 cttagacgtc aggtggcact tttcggggaa atgtgcgcgg aacccctatt tgtttatttt
     5221 tctaaataca ttcaaatatg tatccgctca tgagacaata accctgataa atgcttcaat
     5281 aatattgaaa aaggaagagt atgagtattc aacatttccg tgtcgccctt attccctttt
     5341 ttgcggcatt ttgccttcct gtttttgctc acccagaaac gctggtgaaa gtaaaagatg
     5401 ctgaagatca gttgggtgca cgagtgggtt acatcgaact ggatctcaac agcggtaaga
     5461 tccttgagag ttttcgcccc gaagaacgtt ttccaatgat gagcactttt aaagttctgc
     5521 tatgtggcgc ggtattatcc cgtattgacg ccgggcaaga gcaactcggt cgccgcatac
     5581 actattctca gaatgacttg gttgagtact caccagtcac agaaaagcat cttacggatg
     5641 gcatgacagt aagagaatta tgcagtgctg ccataaccat gagtgataac actgcggcca
     5701 acttacttct gacaacgatc ggaggaccga aggagctaac cgcttttttg cacaacatgg
     5761 gggatcatgt aactcgcctt gatcgttggg aaccggagct gaatgaagcc ataccaaacg
     5821 acgagcgtga caccacgatg cctgtagcaa tggcaacaac gttgcgcaaa ctattaactg
     5881 gcgaactact tactctagct tcccggcaac aattaataga ctggatggag gcggataaag
     5941 ttgcaggacc acttctgcgc tcggcccttc cggctggctg gtttattgct gataaatctg
     6001 gagccggtga gcgtgggtct cgcggtatca ttgcagcact ggggccagat ggtaagccct
     6061 cccgtatcgt agttatctac acgacgggga gtcaggcaac tatggatgaa cgaaatagac
     6121 agatcgctga gataggtgcc tcactgatta agcattggta actgtcagac caagtttact
     6181 catatatact ttagattgat ttaaaacttc atttttaatt taaaaggatc taggtgaaga
     6241 tcctttttga taatctcatg accaaaatcc cttaacgtga gttttcgttc cactgagcgt
     6301 cagaccccgt agaaaagatc aaaggatctt cttgagatcc tttttttctg cgcgtaatct
     6361 gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg gatcaagagc
     6421 taccaactct ttttccgaag gtaactggct tcagcagagc gcagatacca aatactgttc
     6481 ttctagtgta gccgtagtta ggccaccact tcaagaactc tgtagcaccg cctacatacc
     6541 tcgctctgct aatcctgtta ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg
     6601 ggttggactc aagacgatag ttaccggata aggcgcagcg gtcgggctga acggggggtt
     6661 cgtgcacaca gcccagcttg gagcgaacga cctacaccga actgagatac ctacagcgtg
     6721 agctatgaga aagcgccacg cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg
     6781 gcagggtcgg aacaggagag cgcacgaggg agcttccagg gggaaacgcc tggtatcttt
     6841 atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg atttttgtga tgctcgtcag
     6901 gggggcggag cctatggaaa aacgccagca acgcggcctt tttacggttc ctggcctttt
     6961 gctggccttt tgctcacatg t
//
