LOCUS       sequence-337498-        7154 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Soma-targeted Cal-Light with KA2.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7154)
  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 7154)
  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: pCMV-TM-KA2-CaM-NES-TevN-AsLOV2-TEVseq-tTA
FEATURES             Location/Qualifiers
     source          1..7154
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        1..978
                     /label=CMV IE94 promoter
                     /note="enhancer/promoter region of simian cytomegalovirus 
                     major immediate early transcription unit IE94"
     promoter        1014..1032
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     1014..1031
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     sig_peptide     1051..1110
                     /codon_start=1
                     /product="leader sequence from mouse immunoglobulin kappa 
                     light chain"
                     /label=Ig-kappa leader
                     /translation="METDTLLLWVLLLWVPGSTG"
     CDS             1114..1143
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             1147..1293
                     /codon_start=1
                     /product="transmembrane domain from platelet derived growth
                     factor receptor beta"
                     /label=PDGFR-beta TM domain
                     /translation="AVGQDTQEVIVVPHSLPFKVVVISAILALVVLTIISLIILIMLWQ
                     KKPR"
     CDS             1324..1773
                     /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             2308..2334
                     /codon_start=1
                     /product="nuclear export signal from the HIV Rev protein 
                     (Fischer et al., 1995)"
                     /label=NES
                     /translation="LPPLERLTL"
     CDS             3127..3147
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     CDS             join(3148..3765,3766..4152)
                     /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(3218..3237)
                     /label=Tet-R
                     /note="Tetracycline resistance gene, reverse primer"
     primer_bind     complement(4161..4180)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(4162..4180)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     polyA_signal    4185..4319
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4209..4228)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     4263..4282
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(4432..4452)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(4432..4450)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(4471..4487)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4471..4487)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4484..4506)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4495..4511
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(join(4519..4525,4526..4543,4544..4549))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4564..4585
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4702..4719)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4872..5460)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4952..4971)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5631..6422,6423..6491))
                     /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     6254..6273
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6492..6596)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(6622..7077)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6759..6780)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     6971..6990
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
ORIGIN
        1 gccaattcaa tatggcgtat atggactcat gccaattcaa tatggtggat ctggacctgt
       61 gccaattcaa tatggcgtat atggactcgt gccaattcaa tatggtggat ctggacccca
      121 gccaattcaa tatggcggac ttggcaccat gccaattcaa tatggcggac ctggcactgt
      181 gccaactggg gaggggtcta cttggcacgg tgccaagttt gaggaggggt cttggccctg
      241 tgccaagtcc gccatattga attggcatgg tgccaataat ggcggccata ttggctatat
      301 gccaggatca atatataggc aatatccaat atggccctat gccaatatgg ctattggcca
      361 ggttcaatac tatgtattgg ccctatgcca tatagtattc catatatggg ttttcctatt
      421 gacgtagata gcccctccca atgggcggtc ccatatacca tatatggggc ttcctaatac
      481 cgcccatagc cactccccca ttgacgtcaa tggtctctat atatggtctt tcctattgac
      541 gtcatatggg cggtcctatt gacgtatatg gcgcctcccc cattgacgtc aattacggta
      601 aatggcccgc ctggctcaat gcccattgac gtcaatagga ccacccacca ttgacgtcaa
      661 tgggatggct cattgcccat tcatatccgt tctcacgccc cctattgacg tcaatgacgg
      721 taaatggccc acttggcagt acatcaatat ctattaatag taacttggca agtacattac
      781 tattggaagt acgccagggt acattggcag tactcccatt gacgtcaatg gcggtaaatg
      841 gcccgcgatg gctgccaagt acatccccat tgacgtcaat ggggaggggc aatgacgcaa
      901 atgggcgttc cattgacgta aatgggcggt aggcgtgcct aatgggaggt ctatataagc
      961 aatgctcgtt tagggaaccg ccattctgcc tggggacgtc ggagcaagct ttcatttagg
     1021 tgacactata gaatacaagc tacttgttct atggagacag acacactcct gctatgggta
     1081 ctgctgctct gggttccagg ttccactggt gacgaacaaa aactcatctc agaagaggat
     1141 ctgaatgctg tgggccagga cacgcaggag gtcatcgtgg tgccacactc cttgcccttt
     1201 aaggtggtgg tgatctcagc catcctggcc ctggtggtgc tcaccatcat ctcccttatc
     1261 atcctcatca tgctttggca gaagaagcca cgtggtggct ctggaggtct cgagggaggt
     1321 accatgcccg ccgaactgct gctgctgctg attgtcgctt ttgctaaccc ttcttgccag
     1381 gtgctgtctt ctctgaggat ggccgctatt ctggacgatc agaccgtgtg cggaagagga
     1441 gagaggctgg cactggcact ggctagggag cagatcaatg gcatcattga agtgcctgca
     1501 aaggcccggg tcgaggtgga cattttcgaa ctgcagagag atagccagta cgagaccaca
     1561 gacaccatgt gccagatcct gccaaaagga gtggtctccg tcctgggacc aagctcctct
     1621 cctgcttctg caagtaccgt gtctcacatt tgtggcgaga aggaaatccc ccatatcaag
     1681 gtcgggccag aggaaacacc caggctgcag tacctgcgct tcgcctcagt gagcctgtat
     1741 ccatcaaacg aggatgtgtc actggcagtc agcggaggta gcctcgagat gccggaccaa
     1801 ctgactgaag agcagatcgc agaatttaaa gaggctttct ccctatttga caaggacggg
     1861 gatgggacaa taacaaccaa ggagctgggg acggtgatgc ggtctctggg gcagaacccc
     1921 acagaagcag agctgcaaga catgatcaat gaagtagatg ccgacggtga cggcacaatc
     1981 gacttccctg agttcctgac aatgatggca agaaaaatga aatacaggga cacggaagaa
     2041 gaaattagag aagcgttcgg tgtgtttgat aaggatggca atggctacat cagtgcagca
     2101 gagcttcgcc acgtgatgac aaaccttgga gagaagttaa cagatgaaga ggttgatgaa
     2161 atgatcaggg aagcagacat cgatggggat ggtcaggtaa actacgaaga gtttgtacaa
     2221 atgatgacag cgaaggcatg cggtggcggt ggctctggag gtggtgggtc cggaggaggc
     2281 ggccgctccg gtggatccat gcttcaactt cctcctcttg aacgtcttac tctcgaaatg
     2341 ggcgagagcc tgttcaaggg acctagggac tacaacccta tcagtagcac aatttgtcac
     2401 ctgaccaacg agagtgatgg ccacacaaca agcctgtacg gcatcgggtt cggacccttt
     2461 atcatcacca acaagcacct gttcaggcgg aataatggca ctctgctggt gcagagcctg
     2521 cacggggtgt tcaaagtgaa gaacacaacc actctgcagc agcacctgat cgatgggcgg
     2581 gatatgatca tcattaggat gcccaaggac ttcccccctt ttcctcagaa actgaagttc
     2641 cgagagcccc agagagagga gagaatctgt ctggtgacca caaactttca ggagctcggt
     2701 agtggtagtg gggagtttct ggcaaccaca ctggaacgga tcgagaaaaa tttcgtgatt
     2761 actgatccga gactgcctga caacccaatc atttttgcga gcgattcctt cctgcagctg
     2821 acagaatatt ctcgggaaga gatcctgggg cgcaattgcc gttttctgca gggacccgag
     2881 acagaccgtg ccactgttcg gaaaatcaga gatgctattg acaaccagac tgaagtgacc
     2941 gttcagctga tcaattatac caagagcggc aagaagttct ggaacgtgtt ccacctgcag
     3001 ccgatgcgcg attataaggg cgacgtccag tacttcattg gcgtgcagct ggatggcacc
     3061 gaacgtcttc atggcgccgc tgagcgtgag gcggtctgcc tgatcaaaaa gacagccttt
     3121 cagattgaga acctgtactt ccagggctct agattagata aaagtaaagt gattaacagc
     3181 gcattagagc tgcttaatga ggtcggaatc gaaggtttaa caacccgtaa actcgcccag
     3241 aagctaggtg tagagcagcc tacattgtat tggcatgtaa aaaataagcg ggctttgctc
     3301 gacgccttag ccattgagat gttagatagg caccatactc acttttgccc tttagaaggg
     3361 gaaagctggc aagatttttt acgtaataac gctaaaagtt ttagatgtgc tttactaagt
     3421 catcgcgatg gagcaaaagt acatttaggt acacggccta cagaaaaaca gtatgaaact
     3481 ctcgaaaatc aattagcctt tttatgccaa caaggttttt cactagagaa tgcattatat
     3541 gcactcagcg ctgtggggca ttttacttta ggttgcgtat tggaagatca agagcatcaa
     3601 gtcgctaaag aagaaaggga aacacctact actgatagta tgccgccatt attacgacaa
     3661 gctatcgaat tatttgatca ccaaggtgca gagccagcct tcttattcgg ccttgaattg
     3721 atcatatgcg gattagaaaa acaacttaaa tgtgaaagtg ggtccgcgta cagccgcgcg
     3781 cgtacgaaaa acaattacgg gtctaccatc gagggcctgc tcgatctccc ggacgacgac
     3841 gcccccgaag aggcggggct ggcggctccg cgcctgtcct ttctccccgc gggacacacg
     3901 cgcagactgt cgacggcccc cccgaccgat gtcagcctgg gggacgagct ccacttagac
     3961 ggcgaggacg tggcgatggc gcatgccgac gcgctagacg atttcgatct ggacatgttg
     4021 ggggacgggg attccccggg tccgggattt accccccacg actccgcccc ctacggcgct
     4081 ctggatatgg ccgacttcga gtttgagcag atgtttaccg atgcccttgg aattgacgag
     4141 tacggtgggt aggctagcct ccctatagtg agtcgtatta cgtagatcca gacatgataa
     4201 gatacattga tgagtttgga caaaccacaa ctagaatgca gtgaaaaaaa tgctttattt
     4261 gtgaaatttg tgatgctatt gctttatttg taaccattat aagctgcaat aaacaagtta
     4321 acaacaacaa ttgcattcat tttatgtttc aggttcaggg ggaggtgtgg gaggtttttt
     4381 aattcgcgtc cgcggcgcca atgcattggg cccggtaccc agcttttgtt ccctttagtg
     4441 agggttaatt gcgcgcttgg cgtaatcatg gtcatagctg tttcctgtgt gaaattgtta
     4501 tccgctcaca attccacaca acatacgagc cgggagcata aagtgtaaag cctggggtgc
     4561 ctaatgagtg agctaactca cattaattgc gttgcgctca ctgcccgctt tccagtcggg
     4621 aaacctgtcg tgccagctgc attaatgaat cggccaacgc gcggggagag gcggtttgcg
     4681 tattgggcgc tcttccgctt cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg
     4741 ggagcggtat cagctcactc aaaggcggta atacggttat ccacagaatc aggggataac
     4801 gcaggaaaga acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg
     4861 ttgctggcgt ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca
     4921 agtcagaggt ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc
     4981 tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc
     5041 ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta ggtatctcag ttcggtgtag
     5101 gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc
     5161 ttatccggta actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca
     5221 gcagccactg gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg
     5281 aagtggtggc ctaactacgg ctacactaga agaacagtat ttggtatctg cgctctgctg
     5341 aagccagtta ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct
     5401 ggtagcggtg gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa
     5461 gaagatcctt tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa
     5521 gggattttgg tcatgagatt atcaaaaagg atcttcacct agatcctttt aaattaaaaa
     5581 tgaagtttta aatcaatcta aagtatatat gagtaaactt ggtctgacag ttaccaatgc
     5641 ttaatcagtg aggcacctat ctcagcgatc tgtctatttc gttcatccat agttgcctga
     5701 ctccccgtcg tgtagataac tacgatacgg gagggcttac catctggccc cagtgctgca
     5761 atgataccgc gagacccacg ctcaccggct ccagatttat cagcaataaa ccagccagcc
     5821 ggaagggccg agcgcagaag tggtcctgca actttatccg cctccatcca gtctattaat
     5881 tgttgccggg aagctagagt aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc
     5941 attgctacag gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt cagctccggt
     6001 tcccaacgat caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc
     6061 ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag tgttatcact catggttatg
     6121 gcagcactgc ataattctct tactgtcatg ccatccgtaa gatgcttttc tgtgactggt
     6181 gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg
     6241 gcgtcaatac gggataatac cgcgccacat agcagaactt taaaagtgct catcattgga
     6301 aaacgttctt cggggcgaaa actctcaagg atcttaccgc tgttgagatc cagttcgatg
     6361 taacccactc gtgcacccaa ctgatcttca gcatctttta ctttcaccag cgtttctggg
     6421 tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa taagggcgac acggaaatgt
     6481 tgaatactca tactcttcct ttttcaatat tattgaagca tttatcaggg ttattgtctc
     6541 atgagcggat acatatttga atgtatttag aaaaataaac aaataggggt tccgcgcaca
     6601 tttccccgaa aagtgccacc taaattgtaa gcgttaatat tttgttaaaa ttcgcgttaa
     6661 atttttgtta aatcagctca ttttttaacc aataggccga aatcggcaaa atcccttata
     6721 aatcaaaaga atagaccgag atagggttga gtgttgttcc agtttggaac aagagtccac
     6781 tattaaagaa cgtggactcc aacgtcaaag ggcgaaaaac cgtctatcag ggcgatggcc
     6841 cactacgtga accatcaccc taatcaagtt ttttggggtc gaggtgccgt aaagcactaa
     6901 atcggaaccc taaagggagc ccccgattta gagcttgacg gggaaagccg gcgaacgtgg
     6961 cgagaaagga agggaagaaa gcgaaaggag cgggcgctag ggcgctggca agtgtagcgg
     7021 tcacgctgcg cgtaaccacc acacccgccg cgcttaatgc gccgctacag ggcgcgtccc
     7081 attcgccatt caggctgcgc aactgttggg aagggcgatc ggtgcgggcc tcttcgctat
     7141 tacgccagtc gacc
//
