LOCUS       sequence-337520-        7250 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AAV Soma-targeted Cal-Light with KA2.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7250)
  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 7250)
  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-pCMV-TM-KA2-CaM-NES-TevN-AsLOV2-TEVseq-tTA
FEATURES             Location/Qualifiers
     source          1..7250
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   1..126
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     promoter        159..1143
                     /label=CMV IE94 promoter
                     /note="enhancer/promoter region of simian cytomegalovirus 
                     major immediate early transcription unit IE94"
     promoter        1179..1197
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     1179..1196
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     sig_peptide     1216..1275
                     /codon_start=1
                     /product="leader sequence from mouse immunoglobulin kappa 
                     light chain"
                     /label=Ig-kappa leader
                     /translation="METDTLLLWVLLLWVPGSTG"
     CDS             1279..1308
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             1312..1458
                     /codon_start=1
                     /product="transmembrane domain from platelet derived growth
                     factor receptor beta"
                     /label=PDGFR-beta TM domain
                     /translation="AVGQDTQEVIVVPHSLPFKVVVISAILALVVLTIISLIILIMLWQ
                     KKPR"
     CDS             1489..1938
                     /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             2473..2499
                     /codon_start=1
                     /product="nuclear export signal from the HIV Rev protein 
                     (Fischer et al., 1995)"
                     /label=NES
                     /translation="LPPLERLTL"
     CDS             3292..3312
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     CDS             join(3313..3930,3931..4317)
                     /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     complement(3383..3402)
                     /label=Tet-R
                     /note="Tetracycline resistance gene, reverse primer"
     primer_bind     complement(4326..4345)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(4327..4345)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     polyA_signal    4350..4484
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4374..4393)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     4428..4447
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     repeat_region   4513..4653
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   4513..4642
                     /label=AAV2 ITR
     rep_origin      4728..5183
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4815..4834)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5025..5046
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(5200..5219)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5319..5341
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(5379..5397)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5465..5569
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5570..5638,5639..6430)
                     /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(5788..5807)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6601..7189
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7090..7109
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 cctgcaggca gctgcgcgct cgctcgctca ctgaggccgc ccgggcgtcg ggcgaccttt
       61 ggtcgcccgg cctcagtgag cgagcgcgca gagagggagt ggccaactcc atcactaggg
      121 gttcctgcgg ccgcacgcgt ctcttcgcta ttacgccagt cgaccgccaa ttcaatatgg
      181 cgtatatgga ctcatgccaa ttcaatatgg tggatctgga cctgtgccaa ttcaatatgg
      241 cgtatatgga ctcgtgccaa ttcaatatgg tggatctgga ccccagccaa ttcaatatgg
      301 cggacttggc accatgccaa ttcaatatgg cggacctggc actgtgccaa ctggggaggg
      361 gtctacttgg cacggtgcca agtttgagga ggggtcttgg ccctgtgcca agtccgccat
      421 attgaattgg catggtgcca ataatggcgg ccatattggc tatatgccag gatcaatata
      481 taggcaatat ccaatatggc cctatgccaa tatggctatt ggccaggttc aatactatgt
      541 attggcccta tgccatatag tattccatat atgggttttc ctattgacgt agatagcccc
      601 tcccaatggg cggtcccata taccatatat ggggcttcct aataccgccc atagccactc
      661 ccccattgac gtcaatggtc tctatatatg gtctttccta ttgacgtcat atgggcggtc
      721 ctattgacgt atatggcgcc tcccccattg acgtcaatta cggtaaatgg cccgcctggc
      781 tcaatgccca ttgacgtcaa taggaccacc caccattgac gtcaatggga tggctcattg
      841 cccattcata tccgttctca cgccccctat tgacgtcaat gacggtaaat ggcccacttg
      901 gcagtacatc aatatctatt aatagtaact tggcaagtac attactattg gaagtacgcc
      961 agggtacatt ggcagtactc ccattgacgt caatggcggt aaatggcccg cgatggctgc
     1021 caagtacatc cccattgacg tcaatgggga ggggcaatga cgcaaatggg cgttccattg
     1081 acgtaaatgg gcggtaggcg tgcctaatgg gaggtctata taagcaatgc tcgtttaggg
     1141 aaccgccatt ctgcctgggg acgtcggagc aagctttcat ttaggtgaca ctatagaata
     1201 caagctactt gttctatgga gacagacaca ctcctgctat gggtactgct gctctgggtt
     1261 ccaggttcca ctggtgacga acaaaaactc atctcagaag aggatctgaa tgctgtgggc
     1321 caggacacgc aggaggtcat cgtggtgcca cactccttgc cctttaaggt ggtggtgatc
     1381 tcagccatcc tggccctggt ggtgctcacc atcatctccc ttatcatcct catcatgctt
     1441 tggcagaaga agccacgtgg tggctctgga ggtctcgagg gaggtaccat gcccgccgaa
     1501 ctgctgctgc tgctgattgt cgcttttgct aacccttctt gccaggtgct gtcttctctg
     1561 aggatggccg ctattctgga cgatcagacc gtgtgcggaa gaggagagag gctggcactg
     1621 gcactggcta gggagcagat caatggcatc attgaagtgc ctgcaaaggc ccgggtcgag
     1681 gtggacattt tcgaactgca gagagatagc cagtacgaga ccacagacac catgtgccag
     1741 atcctgccaa aaggagtggt ctccgtcctg ggaccaagct cctctcctgc ttctgcaagt
     1801 accgtgtctc acatttgtgg cgagaaggaa atcccccata tcaaggtcgg gccagaggaa
     1861 acacccaggc tgcagtacct gcgcttcgcc tcagtgagcc tgtatccatc aaacgaggat
     1921 gtgtcactgg cagtcagcgg aggtagcctc gagatgccgg accaactgac tgaagagcag
     1981 atcgcagaat ttaaagaggc tttctcccta tttgacaagg acggggatgg gacaataaca
     2041 accaaggagc tggggacggt gatgcggtct ctggggcaga accccacaga agcagagctg
     2101 caagacatga tcaatgaagt agatgccgac ggtgacggca caatcgactt ccctgagttc
     2161 ctgacaatga tggcaagaaa aatgaaatac agggacacgg aagaagaaat tagagaagcg
     2221 ttcggtgtgt ttgataagga tggcaatggc tacatcagtg cagcagagct tcgccacgtg
     2281 atgacaaacc ttggagagaa gttaacagat gaagaggttg atgaaatgat cagggaagca
     2341 gacatcgatg gggatggtca ggtaaactac gaagagtttg tacaaatgat gacagcgaag
     2401 gcatgcggtg gcggtggctc tggaggtggt gggtccggag gaggcggccg ctccggtgga
     2461 tccatgcttc aacttcctcc tcttgaacgt cttactctcg aaatgggcga gagcctgttc
     2521 aagggaccta gggactacaa ccctatcagt agcacaattt gtcacctgac caacgagagt
     2581 gatggccaca caacaagcct gtacggcatc gggttcggac cctttatcat caccaacaag
     2641 cacctgttca ggcggaataa tggcactctg ctggtgcaga gcctgcacgg ggtgttcaaa
     2701 gtgaagaaca caaccactct gcagcagcac ctgatcgatg ggcgggatat gatcatcatt
     2761 aggatgccca aggacttccc cccttttcct cagaaactga agttccgaga gccccagaga
     2821 gaggagagaa tctgtctggt gaccacaaac tttcaggagc tcggtagtgg tagtggggag
     2881 tttctggcaa ccacactgga acggatcgag aaaaatttcg tgattactga tccgagactg
     2941 cctgacaacc caatcatttt tgcgagcgat tccttcctgc agctgacaga atattctcgg
     3001 gaagagatcc tggggcgcaa ttgccgtttt ctgcagggac ccgagacaga ccgtgccact
     3061 gttcggaaaa tcagagatgc tattgacaac cagactgaag tgaccgttca gctgatcaat
     3121 tataccaaga gcggcaagaa gttctggaac gtgttccacc tgcagccgat gcgcgattat
     3181 aagggcgacg tccagtactt cattggcgtg cagctggatg gcaccgaacg tcttcatggc
     3241 gccgctgagc gtgaggcggt ctgcctgatc aaaaagacag cctttcagat tgagaacctg
     3301 tacttccagg gctctagatt agataaaagt aaagtgatta acagcgcatt agagctgctt
     3361 aatgaggtcg gaatcgaagg tttaacaacc cgtaaactcg cccagaagct aggtgtagag
     3421 cagcctacat tgtattggca tgtaaaaaat aagcgggctt tgctcgacgc cttagccatt
     3481 gagatgttag ataggcacca tactcacttt tgccctttag aaggggaaag ctggcaagat
     3541 tttttacgta ataacgctaa aagttttaga tgtgctttac taagtcatcg cgatggagca
     3601 aaagtacatt taggtacacg gcctacagaa aaacagtatg aaactctcga aaatcaatta
     3661 gcctttttat gccaacaagg tttttcacta gagaatgcat tatatgcact cagcgctgtg
     3721 gggcatttta ctttaggttg cgtattggaa gatcaagagc atcaagtcgc taaagaagaa
     3781 agggaaacac ctactactga tagtatgccg ccattattac gacaagctat cgaattattt
     3841 gatcaccaag gtgcagagcc agccttctta ttcggccttg aattgatcat atgcggatta
     3901 gaaaaacaac ttaaatgtga aagtgggtcc gcgtacagcc gcgcgcgtac gaaaaacaat
     3961 tacgggtcta ccatcgaggg cctgctcgat ctcccggacg acgacgcccc cgaagaggcg
     4021 gggctggcgg ctccgcgcct gtcctttctc cccgcgggac acacgcgcag actgtcgacg
     4081 gcccccccga ccgatgtcag cctgggggac gagctccact tagacggcga ggacgtggcg
     4141 atggcgcatg ccgacgcgct agacgatttc gatctggaca tgttggggga cggggattcc
     4201 ccgggtccgg gatttacccc ccacgactcc gccccctacg gcgctctgga tatggccgac
     4261 ttcgagtttg agcagatgtt taccgatgcc cttggaattg acgagtacgg tgggtaggct
     4321 agcctcccta tagtgagtcg tattacgtag atccagacat gataagatac attgatgagt
     4381 ttggacaaac cacaactaga atgcagtgaa aaaaatgctt tatttgtgaa atttgtgatg
     4441 ctattgcttt atttgtaacc attataagct gcaataaaca agttggtaac cacgtgcgga
     4501 ccgagcggcc gcaggaaccc ctagtgatgg agttggccac tccctctctg cgcgctcgct
     4561 cgctcactga ggccgggcga ccaaaggtcg cccgacgccc gggctttgcc cgggcggcct
     4621 cagtgagcga gcgagcgcgc agctgcctgc aggggcgcct gatgcggtat tttctcctta
     4681 cgcatctgtg cggtatttca caccgcatac gtcaaagcaa ccatagtacg cgccctgtag
     4741 cggcgcatta agcgcggcgg gtgtggtggt tacgcgcagc gtgaccgcta cacttgccag
     4801 cgccttagcg cccgctcctt tcgctttctt cccttccttt ctcgccacgt tcgccggctt
     4861 tccccgtcaa gctctaaatc gggggctccc tttagggttc cgatttagtg ctttacggca
     4921 cctcgacccc aaaaaacttg atttgggtga tggttcacgt agtgggccat cgccctgata
     4981 gacggttttt cgccctttga cgttggagtc cacgttcttt aatagtggac tcttgttcca
     5041 aactggaaca acactcaact ctatctcggg ctattctttt gatttataag ggattttgcc
     5101 gatttcggtc tattggttaa aaaatgagct gatttaacaa aaatttaacg cgaattttaa
     5161 caaaatatta acgtttacaa ttttatggtg cactctcagt acaatctgct ctgatgccgc
     5221 atagttaagc cagccccgac acccgccaac acccgctgac gcgccctgac gggcttgtct
     5281 gctcccggca tccgcttaca gacaagctgt gaccgtctcc gggagctgca tgtgtcagag
     5341 gttttcaccg tcatcaccga aacgcgcgag acgaaagggc ctcgtgatac gcctattttt
     5401 ataggttaat gtcatgataa taatggtttc ttagacgtca ggtggcactt ttcggggaaa
     5461 tgtgcgcgga acccctattt gtttattttt ctaaatacat tcaaatatgt atccgctcat
     5521 gagacaataa ccctgataaa tgcttcaata atattgaaaa aggaagagta tgagtattca
     5581 acatttccgt gtcgccctta ttcccttttt tgcggcattt tgccttcctg tttttgctca
     5641 cccagaaacg ctggtgaaag taaaagatgc tgaagatcag ttgggtgcac gagtgggtta
     5701 catcgaactg gatctcaaca gcggtaagat ccttgagagt tttcgccccg aagaacgttt
     5761 tccaatgatg agcactttta aagttctgct atgtggcgcg gtattatccc gtattgacgc
     5821 cgggcaagag caactcggtc gccgcataca ctattctcag aatgacttgg ttgagtactc
     5881 accagtcaca gaaaagcatc ttacggatgg catgacagta agagaattat gcagtgctgc
     5941 cataaccatg agtgataaca ctgcggccaa cttacttctg acaacgatcg gaggaccgaa
     6001 ggagctaacc gcttttttgc acaacatggg ggatcatgta actcgccttg atcgttggga
     6061 accggagctg aatgaagcca taccaaacga cgagcgtgac accacgatgc ctgtagcaat
     6121 ggcaacaacg ttgcgcaaac tattaactgg cgaactactt actctagctt cccggcaaca
     6181 attaatagac tggatggagg cggataaagt tgcaggacca cttctgcgct cggcccttcc
     6241 ggctggctgg tttattgctg ataaatctgg agccggtgag cgtgggtctc gcggtatcat
     6301 tgcagcactg gggccagatg gtaagccctc ccgtatcgta gttatctaca cgacggggag
     6361 tcaggcaact atggatgaac gaaatagaca gatcgctgag ataggtgcct cactgattaa
     6421 gcattggtaa ctgtcagacc aagtttactc atatatactt tagattgatt taaaacttca
     6481 tttttaattt aaaaggatct aggtgaagat cctttttgat aatctcatga ccaaaatccc
     6541 ttaacgtgag ttttcgttcc actgagcgtc agaccccgta gaaaagatca aaggatcttc
     6601 ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc
     6661 agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt
     6721 cagcagagcg cagataccaa atactgttct tctagtgtag ccgtagttag gccaccactt
     6781 caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc
     6841 tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa
     6901 ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac
     6961 ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg
     7021 gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga
     7081 gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact
     7141 tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa
     7201 cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgt
//
