LOCUS       sequence-337501-        6859 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Soma-targeted Cal-Light with Kv2.1.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6859)
  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 6859)
  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-Kv2.1-CaM-NES-TevN-AsLOV2-TEVseq-tTA
FEATURES             Location/Qualifiers
     source          1..6859
                     /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        1013..1031
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     1013..1030
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     sig_peptide     1050..1109
                     /codon_start=1
                     /product="leader sequence from mouse immunoglobulin kappa 
                     light chain"
                     /label=Ig-kappa leader
                     /translation="METDTLLLWVLLLWVPGSTG"
     CDS             1113..1142
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             1146..1292
                     /codon_start=1
                     /product="transmembrane domain from platelet derived growth
                     factor receptor beta"
                     /label=PDGFR-beta TM domain
                     /translation="AVGQDTQEVIVVPHSLPFKVVVISAILALVVLTIISLIILIMLWQ
                     KKPR"
     CDS             2013..2039
                     /codon_start=1
                     /product="nuclear export signal from the HIV Rev protein 
                     (Fischer et al., 1995)"
                     /label=NES
                     /translation="LPPLERLTL"
     CDS             2832..2852
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     CDS             join(2853..3470,3471..3857)
                     /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(2923..2942)
                     /label=Tet-R
                     /note="Tetracycline resistance gene, reverse primer"
     primer_bind     complement(3866..3885)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(3867..3885)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     polyA_signal    3890..4024
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3914..3933)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     3968..3987
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(4137..4157)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(4137..4155)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(4176..4192)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4176..4192)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4189..4211)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4200..4216
                     /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(4224..4230,4231..4248,4249..4254))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4269..4290
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4407..4424)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4577..5165)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4657..4676)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5336..6127,6128..6196))
                     /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     5959..5978
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6197..6301)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(6327..6782)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6464..6485)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     6676..6695
                     /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 tgatttaggt
     1021 gacactatag aatacaagct acttgttcta tggagacaga cacactcctg ctatgggtac
     1081 tgctgctctg ggttccaggt tccactggtg acgaacaaaa actcatctca gaagaggatc
     1141 tgaatgctgt gggccaggac acgcaggagg tcatcgtggt gccacactcc ttgcccttta
     1201 aggtggtggt gatctcagcc atcctggccc tggtggtgct caccatcatc tcccttatca
     1261 tcctcatcat gctttggcag aagaagccac gtcagtccca gcctattctg aacaccaaag
     1321 agatggcccc ccagagtaaa ccccctgagg agctggaaat gtcctcaatg ccctcccctg
     1381 tggcacctct tcctgcaaga acagagggcg tgatcgacat gaggtccatg agttctattg
     1441 actcttttat ctcatgcgcc accgattttc cagaggctac gcggtttctc gagatgccgg
     1501 accaactgac tgaagagcag atcgcagaat ttaaagaggc tttctcccta tttgacaagg
     1561 acggggatgg gacaataaca accaaggagc tggggacggt gatgcggtct ctggggcaga
     1621 accccacaga agcagagctg caagacatga tcaatgaagt agatgccgac ggtgacggca
     1681 caatcgactt ccctgagttc ctgacaatga tggcaagaaa aatgaaatac agggacacgg
     1741 aagaagaaat tagagaagcg ttcggtgtgt ttgataagga tggcaatggc tacatcagtg
     1801 cagcagagct tcgccacgtg atgacaaacc ttggagagaa gttaacagat gaagaggttg
     1861 atgaaatgat cagggaagca gacatcgatg gggatggtca ggtaaactac gaagagtttg
     1921 tacaaatgat gacagcgaag gcatgcggtg gcggtggctc tggaggtggt gggtccggag
     1981 gaggcggccg ctccggtgga tccatgcttc aacttcctcc tcttgaacgt cttactctcg
     2041 aaatgggcga gagcctgttc aagggaccta gggactacaa ccctatcagt agcacaattt
     2101 gtcacctgac caacgagagt gatggccaca caacaagcct gtacggcatc gggttcggac
     2161 cctttatcat caccaacaag cacctgttca ggcggaataa tggcactctg ctggtgcaga
     2221 gcctgcacgg ggtgttcaaa gtgaagaaca caaccactct gcagcagcac ctgatcgatg
     2281 ggcgggatat gatcatcatt aggatgccca aggacttccc cccttttcct cagaaactga
     2341 agttccgaga gccccagaga gaggagagaa tctgtctggt gaccacaaac tttcaggagc
     2401 tcggtagtgg tagtggggag tttctggcaa ccacactgga acggatcgag aaaaatttcg
     2461 tgattactga tccgagactg cctgacaacc caatcatttt tgcgagcgat tccttcctgc
     2521 agctgacaga atattctcgg gaagagatcc tggggcgcaa ttgccgtttt ctgcagggac
     2581 ccgagacaga ccgtgccact gttcggaaaa tcagagatgc tattgacaac cagactgaag
     2641 tgaccgttca gctgatcaat tataccaaga gcggcaagaa gttctggaac gtgttccacc
     2701 tgcagccgat gcgcgattat aagggcgacg tccagtactt cattggcgtg cagctggatg
     2761 gcaccgaacg tcttcatggc gccgctgagc gtgaggcggt ctgcctgatc aaaaagacag
     2821 cctttcagat tgagaacctg tacttccagg gctctagatt agataaaagt aaagtgatta
     2881 acagcgcatt agagctgctt aatgaggtcg gaatcgaagg tttaacaacc cgtaaactcg
     2941 cccagaagct aggtgtagag cagcctacat tgtattggca tgtaaaaaat aagcgggctt
     3001 tgctcgacgc cttagccatt gagatgttag ataggcacca tactcacttt tgccctttag
     3061 aaggggaaag ctggcaagat tttttacgta ataacgctaa aagttttaga tgtgctttac
     3121 taagtcatcg cgatggagca aaagtacatt taggtacacg gcctacagaa aaacagtatg
     3181 aaactctcga aaatcaatta gcctttttat gccaacaagg tttttcacta gagaatgcat
     3241 tatatgcact cagcgctgtg gggcatttta ctttaggttg cgtattggaa gatcaagagc
     3301 atcaagtcgc taaagaagaa agggaaacac ctactactga tagtatgccg ccattattac
     3361 gacaagctat cgaattattt gatcaccaag gtgcagagcc agccttctta ttcggccttg
     3421 aattgatcat atgcggatta gaaaaacaac ttaaatgtga aagtgggtcc gcgtacagcc
     3481 gcgcgcgtac gaaaaacaat tacgggtcta ccatcgaggg cctgctcgat ctcccggacg
     3541 acgacgcccc cgaagaggcg gggctggcgg ctccgcgcct gtcctttctc cccgcgggac
     3601 acacgcgcag actgtcgacg gcccccccga ccgatgtcag cctgggggac gagctccact
     3661 tagacggcga ggacgtggcg atggcgcatg ccgacgcgct agacgatttc gatctggaca
     3721 tgttggggga cggggattcc ccgggtccgg gatttacccc ccacgactcc gccccctacg
     3781 gcgctctgga tatggccgac ttcgagtttg agcagatgtt taccgatgcc cttggaattg
     3841 acgagtacgg tgggtaggct agcctcccta tagtgagtcg tattacgtag atccagacat
     3901 gataagatac attgatgagt ttggacaaac cacaactaga atgcagtgaa aaaaatgctt
     3961 tatttgtgaa atttgtgatg ctattgcttt atttgtaacc attataagct gcaataaaca
     4021 agttaacaac aacaattgca ttcattttat gtttcaggtt cagggggagg tgtgggaggt
     4081 tttttaattc gcgtccgcgg cgccaatgca ttgggcccgg tacccagctt ttgttccctt
     4141 tagtgagggt taattgcgcg cttggcgtaa tcatggtcat agctgtttcc tgtgtgaaat
     4201 tgttatccgc tcacaattcc acacaacata cgagccggga gcataaagtg taaagcctgg
     4261 ggtgcctaat gagtgagcta actcacatta attgcgttgc gctcactgcc cgctttccag
     4321 tcgggaaacc tgtcgtgcca gctgcattaa tgaatcggcc aacgcgcggg gagaggcggt
     4381 ttgcgtattg ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg
     4441 ctgcgggagc ggtatcagct cactcaaagg cggtaatacg gttatccaca gaatcagggg
     4501 ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg
     4561 ccgcgttgct ggcgtttttc cataggctcc gcccccctga cgagcatcac aaaaatcgac
     4621 gctcaagtca gaggtggcga aacccgacag gactataaag ataccaggcg tttccccctg
     4681 gaagctccct cgtgcgctct cctgttccga ccctgccgct taccggatac ctgtccgcct
     4741 ttctcccttc gggaagcgtg gcgctttctc atagctcacg ctgtaggtat ctcagttcgg
     4801 tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct
     4861 gcgccttatc cggtaactat cgtcttgagt ccaacccggt aagacacgac ttatcgccac
     4921 tggcagcagc cactggtaac aggattagca gagcgaggta tgtaggcggt gctacagagt
     4981 tcttgaagtg gtggcctaac tacggctaca ctagaagaac agtatttggt atctgcgctc
     5041 tgctgaagcc agttaccttc ggaaaaagag ttggtagctc ttgatccggc aaacaaacca
     5101 ccgctggtag cggtggtttt tttgtttgca agcagcagat tacgcgcaga aaaaaaggat
     5161 ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc tcagtggaac gaaaactcac
     5221 gttaagggat tttggtcatg agattatcaa aaaggatctt cacctagatc cttttaaatt
     5281 aaaaatgaag ttttaaatca atctaaagta tatatgagta aacttggtct gacagttacc
     5341 aatgcttaat cagtgaggca cctatctcag cgatctgtct atttcgttca tccatagttg
     5401 cctgactccc cgtcgtgtag ataactacga tacgggaggg cttaccatct ggccccagtg
     5461 ctgcaatgat accgcgagac ccacgctcac cggctccaga tttatcagca ataaaccagc
     5521 cagccggaag ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc atccagtcta
     5581 ttaattgttg ccgggaagct agagtaagta gttcgccagt taatagtttg cgcaacgttg
     5641 ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct tcattcagct
     5701 ccggttccca acgatcaagg cgagttacat gatcccccat gttgtgcaaa aaagcggtta
     5761 gctccttcgg tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg
     5821 ttatggcagc actgcataat tctcttactg tcatgccatc cgtaagatgc ttttctgtga
     5881 ctggtgagta ctcaaccaag tcattctgag aatagtgtat gcggcgaccg agttgctctt
     5941 gcccggcgtc aatacgggat aataccgcgc cacatagcag aactttaaaa gtgctcatca
     6001 ttggaaaacg ttcttcgggg cgaaaactct caaggatctt accgctgttg agatccagtt
     6061 cgatgtaacc cactcgtgca cccaactgat cttcagcatc ttttactttc accagcgttt
     6121 ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga
     6181 aatgttgaat actcatactc ttcctttttc aatattattg aagcatttat cagggttatt
     6241 gtctcatgag cggatacata tttgaatgta tttagaaaaa taaacaaata ggggttccgc
     6301 gcacatttcc ccgaaaagtg ccacctaaat tgtaagcgtt aatattttgt taaaattcgc
     6361 gttaaatttt tgttaaatca gctcattttt taaccaatag gccgaaatcg gcaaaatccc
     6421 ttataaatca aaagaataga ccgagatagg gttgagtgtt gttccagttt ggaacaagag
     6481 tccactatta aagaacgtgg actccaacgt caaagggcga aaaaccgtct atcagggcga
     6541 tggcccacta cgtgaaccat caccctaatc aagttttttg gggtcgaggt gccgtaaagc
     6601 actaaatcgg aaccctaaag ggagcccccg atttagagct tgacggggaa agccggcgaa
     6661 cgtggcgaga aaggaaggga agaaagcgaa aggagcgggc gctagggcgc tggcaagtgt
     6721 agcggtcacg ctgcgcgtaa ccaccacacc cgccgcgctt aatgcgccgc tacagggcgc
     6781 gtcccattcg ccattcaggc tgcgcaactg ttgggaaggg cgatcggtgc gggcctcttc
     6841 gctattacgc cagtcgacc
//
