LOCUS       sequence-337525-        7279 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AAV floxed Soma-targeted Cal-Light with KA2.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7279)
  AUTHORS   Hyun JH, Nagahama K, Namkung H, Mignocchi N, Roh SE, Hannan P, 
            Krussel S, Kwak C, McElroy A, Liu B, Cui M, Lee S, Lee D, Huganir 
            RL, Worley PF, Sawa A, Kwon HB
  TITLE     Tagging active neurons by soma-targeted Cal-Light.
  JOURNAL   Nat Commun. 2022 Dec 12;13(1):7692. doi: 10.1038/s41467-022-35406-y.
  PUBMED    36509775
REFERENCE   2  (bases 1 to 7279)
  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"; journalName: "Nat 
            Commun."; date: "2022-12-12"; volume: "13(1):7692. doi"; pages: " 
            10.1038/s41467-022-35406-y"
COMMENT     Sequence Label: 
            pAAV-CMV-IE94-Flex-TM-KA2-Cam-NES-TeVN-AsLOV2-TEVseq-bGH-PA
FEATURES             Location/Qualifiers
     source          1..7279
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   62..202
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   73..202
                     /label=AAV2 ITR
     promoter        209..1186
                     /label=CMV IE94 promoter
                     /note="enhancer/promoter region of simian cytomegalovirus 
                     major immediate early transcription unit IE94"
     protein_bind    complement(1199..1232)
                     /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(1283..1316)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     CDS             complement(join(1319..1705,1706..2323))
                     /codon_start=1
                     /product="tetracycline-controlled transactivator, 
                     comprising a fusion of the tetracycline repressor TetR with
                     the C-terminal activation domain of herpes simplex virus 
                     VP16"
                     /label=tTA
                     /note="In the Tet-Off(R) system, tTA binds to the 
                     Tet-responsive element and stimulates transcription only in
                     the absence of tetracycline or doxycycline."
                     /translation="SRLDKSKVINSALELLNEVGIEGLTTRKLAQKLGVEQPTLYWHVK
                     NKRALLDALAIEMLDRHHTHFCPLEGESWQDFLRNNAKSFRCALLSHRDGAKVHLGTRP
                     TEKQYETLENQLAFLCQQGFSLENALYALSAVGHFTLGCVLEDQEHQVAKEERETPTTD
                     SMPPLLRQAIELFDHQGAEPAFLFGLELIICGLEKQLKCESGSAYSRARTKNNYGSTIE
                     GLLDLPDDDAPEEAGLAAPRLSFLPAGHTRRLSTAPPTDVSLGDELHLDGEDVAMAHAD
                     ALDDFDLDMLGDGDSPGPGFTPHDSAPYGALDMADFEFEQMFTDALGIDEYGG"
     primer_bind     2234..2253
                     /label=Tet-R
                     /note="Tetracycline resistance gene, reverse primer"
     CDS             complement(2324..2344)
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     CDS             complement(3137..3163)
                     /codon_start=1
                     /product="nuclear export signal from the HIV Rev protein 
                     (Fischer et al., 1995)"
                     /label=NES
                     /translation="LPPLERLTL"
     CDS             complement(3698..4147)
                     /codon_start=1
                     /product="first 150 residues of the mouse kainate receptor 
                     subunit 2"
                     /label=KA2
                     /note="mediates targeting to the neuronal soma (Shemesh et 
                     al., 2017)"
                     /translation="MPAELLLLLIVAFANPSCQVLSSLRMAAILDDQTVCGRGERLALA
                     LAREQINGIIEVPAKARVEVDIFELQRDSQYETTDTMCQILPKGVVSVLGPSSSPASAS
                     TVSHICGEKEIPHIKVGPEETPRLQYLRFASVSLYPSNEDVSLAVS"
     CDS             complement(4178..4324)
                     /codon_start=1
                     /product="transmembrane domain from platelet derived growth
                     factor receptor beta"
                     /label=PDGFR-beta TM domain
                     /translation="AVGQDTQEVIVVPHSLPFKVVVISAILALVVLTIISLIILIMLWQ
                     KKPR"
     CDS             complement(4328..4357)
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     sig_peptide     complement(4361..4420)
                     /codon_start=1
                     /product="leader sequence from mouse immunoglobulin kappa 
                     light chain"
                     /label=Ig-kappa leader
                     /translation="METDTLLLWVLLLWVPGSTG"
     protein_bind    4432..4465
                     /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    4516..4549
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     repeat_region   4603..4743
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   4603..4732
                     /label=AAV2 ITR
     rep_origin      4818..5273
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4905..4924)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5115..5136
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(5290..5309)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5409..5431
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(5469..5487)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5555..5659
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5660..5728,5729..6520)
                     /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(5878..5897)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6691..7279
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7180..7199
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 aacgccagca acgcggcctt tttacggttc ctggcctttt gctggccttt tgctcacatg
       61 tcctgcaggc agctgcgcgc tcgctcgctc actgaggccg cccgggcaaa gcccgggcgt
      121 cgggcgacct ttggtcgccc ggcctcagtg agcgagcgag cgcgcagaga gggagtggcc
      181 aactccatca ctaggggttc ctacgcgtgc caattcaata tggcgtatat ggactcatgc
      241 caattcaata tggtggatct ggacctgtgc caattcaata tggcgtatat ggactcgtgc
      301 caattcaata tggtggatct ggaccccagc caattcaata tggcggactt ggcaccatgc
      361 caattcaata tggcggacct ggcactgtgc caactgggga ggggtctact tggcacggtg
      421 ccaagtttga ggaggggtct tggccctgtg ccaagtccgc catattgaat tggcatggtg
      481 ccaataatgg cggccatatt ggctatatgc caggatcaat atataggcaa tatccaatat
      541 ggccctatgc caatatggct attggccagg ttcaatacta tgtattggcc ctatgccata
      601 tagtattcca tatatgggtt ttcctattga cgtagatagc ccctcccaat gggcggtccc
      661 atataccata tatggggctt cctaataccg cccatagcca ctcccccatt gacgtcaatg
      721 gtctctatat atggtctttc ctattgacgt catatgggcg gtcctattga cgtatatggc
      781 gcctccccca ttgacgtcaa ttacggtaaa tggcccgcct ggctcaatgc ccattgacgt
      841 caataggacc acccaccatt gacgtcaatg ggatggctca ttgcccattc atatccgttc
      901 tcacgccccc tattgacgtc aatgacggta aatggcccac ttggcagtac atcaatatct
      961 attaatagta acttggcaag tacattacta ttggaagtac gccagggtac attggcagta
     1021 ctcccattga cgtcaatggc ggtaaatggc ccgcgatggc tgccaagtac atccccattg
     1081 acgtcaatgg ggaggggcaa tgacgcaaat gggcgttcca ttgacgtaaa tgggcggtag
     1141 gcgtgcctaa tgggaggtct atataagcaa tgctcgttta gggaacgtcg actccggaat
     1201 aacttcgtat aggatacttt atacgaagtt atgcagaatg gtagctggat tgtagctgct
     1261 attagcaata tgaaacctct taataacttc gtatagcata cattatacga agttatggct
     1321 acccaccgta ctcgtcaatt ccaagggcat cggtaaacat ctgctcaaac tcgaagtcgg
     1381 ccatatccag agcgccgtag ggggcggagt cgtggggggt aaatcccgga cccggggaat
     1441 ccccgtcccc caacatgtcc agatcgaaat cgtctagcgc gtcggcatgc gccatcgcca
     1501 cgtcctcgcc gtctaagtgg agctcgtccc ccaggctgac atcggtcggg ggggccgtcg
     1561 acagtctgcg cgtgtgtccc gcggggagaa aggacaggcg cggagccgcc agccccgcct
     1621 cttcgggggc gtcgtcgtcc gggagatcga gcaggccctc gatggtagac ccgtaattgt
     1681 ttttcgtacg cgcgcggctg tacgcggacc cactttcaca tttaagttgt ttttctaatc
     1741 cgcatatgat caattcaagg ccgaataaga aggctggctc tgcaccttgg tgatcaaata
     1801 attcgatagc ttgtcgtaat aatggcggca tactatcagt agtaggtgtt tccctttctt
     1861 ctttagcgac ttgatgctct tgatcttcca atacgcaacc taaagtaaaa tgccccacag
     1921 cgctgagtgc atataatgca ttctctagtg aaaaaccttg ttggcataaa aaggctaatt
     1981 gattttcgag agtttcatac tgtttttctg taggccgtgt acctaaatgt acttttgctc
     2041 catcgcgatg acttagtaaa gcacatctaa aacttttagc gttattacgt aaaaaatctt
     2101 gccagctttc cccttctaaa gggcaaaagt gagtatggtg cctatctaac atctcaatgg
     2161 ctaaggcgtc gagcaaagcc cgcttatttt ttacatgcca atacaatgta ggctgctcta
     2221 cacctagctt ctgggcgagt ttacgggttg ttaaaccttc gattccgacc tcattaagca
     2281 gctctaatgc gctgttaatc actttacttt tatctaatct agagccctgg aagtacaggt
     2341 tctcaatctg aaaggctgtc tttttgatca ggcagaccgc ctcacgctca gcggcgccat
     2401 gaagacgttc ggtgccatcc agctgcacgc caatgaagta ctggacgtcg cccttataat
     2461 cgcgcatcgg ctgcaggtgg aacacgttcc agaacttctt gccgctcttg gtataattga
     2521 tcagctgaac ggtcacttca gtctggttgt caatagcatc tctgattttc cgaacagtgg
     2581 cacggtctgt ctcgggtccc tgcagaaaac ggcaattgcg ccccaggatc tcttcccgag
     2641 aatattctgt cagctgcagg aaggaatcgc tcgcaaaaat gattgggttg tcaggcagtc
     2701 tcggatcagt aatcacgaaa tttttctcga tccgttccag tgtggttgcc agaaactccc
     2761 cactaccact accgagctcc tgaaagtttg tggtcaccag acagattctc tcctctctct
     2821 ggggctctcg gaacttcagt ttctgaggaa aaggggggaa gtccttgggc atcctaatga
     2881 tgatcatatc ccgcccatcg atcaggtgct gctgcagagt ggttgtgttc ttcactttga
     2941 acaccccgtg caggctctgc accagcagag tgccattatt ccgcctgaac aggtgcttgt
     3001 tggtgatgat aaagggtccg aacccgatgc cgtacaggct tgttgtgtgg ccatcactct
     3061 cgttggtcag gtgacaaatt gtgctactga tagggttgta gtccctaggt cccttgaaca
     3121 ggctctcgcc catttcgaga gtaagacgtt caagaggagg aagttgaagc atggatccac
     3181 cggagcggcc gcctcctccg gacccaccac ctccagagcc accgccaccg catgccttcg
     3241 ctgtcatcat ttgtacaaac tcttcgtagt ttacctgacc atccccatcg atgtctgctt
     3301 ccctgatcat ttcatcaacc tcttcatctg ttaacttctc tccaaggttt gtcatcacgt
     3361 ggcgaagctc tgctgcactg atgtagccat tgccatcctt atcaaacaca ccgaacgctt
     3421 ctctaatttc ttcttccgtg tccctgtatt tcatttttct tgccatcatt gtcaggaact
     3481 cagggaagtc gattgtgccg tcaccgtcgg catctacttc attgatcatg tcttgcagct
     3541 ctgcttctgt ggggttctgc cccagagacc gcatcaccgt ccccagctcc ttggttgtta
     3601 ttgtcccatc cccgtccttg tcaaataggg agaaagcctc tttaaattct gcgatctgct
     3661 cttcagtcag ttggtccggc atctcgaggc tacctccgct gactgccagt gacacatcct
     3721 cgtttgatgg atacaggctc actgaggcga agcgcaggta ctgcagcctg ggtgtttcct
     3781 ctggcccgac cttgatatgg gggatttcct tctcgccaca aatgtgagac acggtacttg
     3841 cagaagcagg agaggagctt ggtcccagga cggagaccac tccttttggc aggatctggc
     3901 acatggtgtc tgtggtctcg tactggctat ctctctgcag ttcgaaaatg tccacctcga
     3961 cccgggcctt tgcaggcact tcaatgatgc cattgatctg ctccctagcc agtgccagtg
     4021 ccagcctctc tcctcttccg cacacggtct gatcgtccag aatagcggcc atcctcagag
     4081 aagacagcac ctggcaagaa gggttagcaa aagcgacaat cagcagcagc agcagttcgg
     4141 cgggcatggt acctccctcg agacctccag agccaccacg tggcttcttc tgccaaagca
     4201 tgatgaggat gataagggag atgatggtga gcaccaccag ggccaggatg gctgagatca
     4261 ccaccacctt aaagggcaag gagtgtggca ccacgatgac ctcctgcgtg tcctggccca
     4321 cagcattcag atcctcttct gagatgagtt tttgttcgtc accagtggaa cctggaaccc
     4381 agagcagcag tacccatagc aggagtgtgt ctgtctccat gtacctctag cataacttcg
     4441 tataaagtat cctatacgaa gttatttgcc ttaacccaga aattatcact gttattcttt
     4501 agaatggtgc aaagaataac ttcgtataat gtatgctata cgaagttatg aattcaaata
     4561 aaatacgaaa tgcacgtaac cacgtgcgga ccgagcggcc gcaggaaccc ctagtgatgg
     4621 agttggccac tccctctctg cgcgctcgct cgctcactga ggccgggcga ccaaaggtcg
     4681 cccgacgccc gggctttgcc cgggcggcct cagtgagcga gcgagcgcgc agctgcctgc
     4741 aggggcgcct gatgcggtat tttctcctta cgcatctgtg cggtatttca caccgcatac
     4801 gtcaaagcaa ccatagtacg cgccctgtag cggcgcatta agcgcggcgg gtgtggtggt
     4861 tacgcgcagc gtgaccgcta cacttgccag cgccttagcg cccgctcctt tcgctttctt
     4921 cccttccttt ctcgccacgt tcgccggctt tccccgtcaa gctctaaatc gggggctccc
     4981 tttagggttc cgatttagtg ctttacggca cctcgacccc aaaaaacttg atttgggtga
     5041 tggttcacgt agtgggccat cgccctgata gacggttttt cgccctttga cgttggagtc
     5101 cacgttcttt aatagtggac tcttgttcca aactggaaca acactcaact ctatctcggg
     5161 ctattctttt gatttataag ggattttgcc gatttcggtc tattggttaa aaaatgagct
     5221 gatttaacaa aaatttaacg cgaattttaa caaaatatta acgtttacaa ttttatggtg
     5281 cactctcagt acaatctgct ctgatgccgc atagttaagc cagccccgac acccgccaac
     5341 acccgctgac gcgccctgac gggcttgtct gctcccggca tccgcttaca gacaagctgt
     5401 gaccgtctcc gggagctgca tgtgtcagag gttttcaccg tcatcaccga aacgcgcgag
     5461 acgaaagggc ctcgtgatac gcctattttt ataggttaat gtcatgataa taatggtttc
     5521 ttagacgtca ggtggcactt ttcggggaaa tgtgcgcgga acccctattt gtttattttt
     5581 ctaaatacat tcaaatatgt atccgctcat gagacaataa ccctgataaa tgcttcaata
     5641 atattgaaaa aggaagagta tgagtattca acatttccgt gtcgccctta ttcccttttt
     5701 tgcggcattt tgccttcctg tttttgctca cccagaaacg ctggtgaaag taaaagatgc
     5761 tgaagatcag ttgggtgcac gagtgggtta catcgaactg gatctcaaca gcggtaagat
     5821 ccttgagagt tttcgccccg aagaacgttt tccaatgatg agcactttta aagttctgct
     5881 atgtggcgcg gtattatccc gtattgacgc cgggcaagag caactcggtc gccgcataca
     5941 ctattctcag aatgacttgg ttgagtactc accagtcaca gaaaagcatc ttacggatgg
     6001 catgacagta agagaattat gcagtgctgc cataaccatg agtgataaca ctgcggccaa
     6061 cttacttctg acaacgatcg gaggaccgaa ggagctaacc gcttttttgc acaacatggg
     6121 ggatcatgta actcgccttg atcgttggga accggagctg aatgaagcca taccaaacga
     6181 cgagcgtgac accacgatgc ctgtagcaat ggcaacaacg ttgcgcaaac tattaactgg
     6241 cgaactactt actctagctt cccggcaaca attaatagac tggatggagg cggataaagt
     6301 tgcaggacca cttctgcgct cggcccttcc ggctggctgg tttattgctg ataaatctgg
     6361 agccggtgag cgtgggtctc gcggtatcat tgcagcactg gggccagatg gtaagccctc
     6421 ccgtatcgta gttatctaca cgacggggag tcaggcaact atggatgaac gaaatagaca
     6481 gatcgctgag ataggtgcct cactgattaa gcattggtaa ctgtcagacc aagtttactc
     6541 atatatactt tagattgatt taaaacttca tttttaattt aaaaggatct aggtgaagat
     6601 cctttttgat aatctcatga ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc
     6661 agaccccgta gaaaagatca aaggatcttc ttgagatcct ttttttctgc gcgtaatctg
     6721 ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt tgtttgccgg atcaagagct
     6781 accaactctt tttccgaagg taactggctt cagcagagcg cagataccaa atactgttct
     6841 tctagtgtag ccgtagttag gccaccactt caagaactct gtagcaccgc ctacatacct
     6901 cgctctgcta atcctgttac cagtggctgc tgccagtggc gataagtcgt gtcttaccgg
     6961 gttggactca agacgatagt taccggataa ggcgcagcgg tcgggctgaa cggggggttc
     7021 gtgcacacag cccagcttgg agcgaacgac ctacaccgaa ctgagatacc tacagcgtga
     7081 gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg
     7141 cagggtcgga acaggagagc gcacgaggga gcttccaggg ggaaacgcct ggtatcttta
     7201 tagtcctgtc gggtttcgcc acctctgact tgagcgtcga tttttgtgat gctcgtcagg
     7261 ggggcggagc ctatggaaa
//
