LOCUS       sequence-181393-        6680 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  expresses Cal-light TM-CaM-NES-TEV-N-AsLOV2-TEVseq-tTA component in 
            mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6680)
  AUTHORS   Lee D, Hyun JH, Jung K, Hannan P, Kwon HB
  TITLE     A calcium- and light-gated switch to induce gene expression in 
            activated neurons.
  JOURNAL   Nat Biotechnol. 2017 Jun 26. doi: 10.1038/nbt.3902.
  PUBMED    28650460
REFERENCE   2  (bases 1 to 6680)
  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 
            Biotechnol. 2017 Jun 26. doi: 10.1038/nbt.3902."
COMMENT     Sequence Label: pCMV-TM-CaM-NES-TEV-N-AsLOV2-TEVseq-tTA
FEATURES             Location/Qualifiers
     source          1..6680
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        1005..1023
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     1005..1022
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     sig_peptide     1042..1101
                     /codon_start=1
                     /product="leader sequence from mouse immunoglobulin kappa 
                     light chain"
                     /label=Ig-kappa leader
                     /translation="METDTLLLWVLLLWVPGSTG"
     CDS             1105..1134
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             1138..1284
                     /codon_start=1
                     /product="transmembrane domain from platelet derived growth
                     factor receptor beta"
                     /label=PDGFR-beta TM domain
                     /translation="AVGQDTQEVIVVPHSLPFKVVVISAILALVVLTIISLIILIMLWQ
                     KKPR"
     CDS             1825..1851
                     /codon_start=1
                     /product="nuclear export signal from the HIV Rev protein 
                     (Fischer et al., 1995)"
                     /label=NES
                     /translation="LPPLERLTL"
     CDS             2644..2664
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     CDS             join(2665..3282,3283..3669)
                     /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(2735..2754)
                     /label=Tet-R
                     /note="Tetracycline resistance gene, reverse primer"
     primer_bind     complement(3678..3697)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(3679..3697)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     polyA_signal    3702..3836
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3726..3745)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     3780..3799
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(3949..3969)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(3949..3967)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(3988..4004)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3988..4004)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4001..4023)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4012..4028
                     /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(4036..4042,4043..4060,4061..4066))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4081..4102
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4219..4236)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4389..4977)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4469..4488)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5148..5939,5940..6008))
                     /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     5771..5790
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6009..6113)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(6139..6594)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6276..6297)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     6488..6507
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        join(6672..6680,1..969)
                     /label=CMV IE94 promoter
                     /note="enhancer/promoter region of simian cytomegalovirus 
                     major immediate early transcription unit IE94"
ORIGIN
        1 atatggcgta tatggactca tgccaattca atatggtgga tctggacctg tgccaattca
       61 atatggcgta tatggactcg tgccaattca atatggtgga tctggacccc agccaattca
      121 atatggcgga cttggcacca tgccaattca atatggcgga cctggcactg tgccaactgg
      181 ggaggggtct acttggcacg gtgccaagtt tgaggagggg tcttggccct gtgccaagtc
      241 cgccatattg aattggcatg gtgccaataa tggcggccat attggctata tgccaggatc
      301 aatatatagg caatatccaa tatggcccta tgccaatatg gctattggcc aggttcaata
      361 ctatgtattg gccctatgcc atatagtatt ccatatatgg gttttcctat tgacgtagat
      421 agcccctccc aatgggcggt cccatatacc atatatgggg cttcctaata ccgcccatag
      481 ccactccccc attgacgtca atggtctcta tatatggtct ttcctattga cgtcatatgg
      541 gcggtcctat tgacgtatat ggcgcctccc ccattgacgt caattacggt aaatggcccg
      601 cctggctcaa tgcccattga cgtcaatagg accacccacc attgacgtca atgggatggc
      661 tcattgccca ttcatatccg ttctcacgcc ccctattgac gtcaatgacg gtaaatggcc
      721 cacttggcag tacatcaata tctattaata gtaacttggc aagtacatta ctattggaag
      781 tacgccaggg tacattggca gtactcccat tgacgtcaat ggcggtaaat ggcccgcgat
      841 ggctgccaag tacatcccca ttgacgtcaa tggggagggg caatgacgca aatgggcgtt
      901 ccattgacgt aaatgggcgg taggcgtgcc taatgggagg tctatataag caatgctcgt
      961 ttagggaacc gccattctgc ctggggacgt cggagcaagc tttcatttag gtgacactat
     1021 agaatacaag ctacttgttc tatggagaca gacacactcc tgctatgggt actgctgctc
     1081 tgggttccag gttccactgg tgacgaacaa aaactcatct cagaagagga tctgaatgct
     1141 gtgggccagg acacgcagga ggtcatcgtg gtgccacact ccttgccctt taaggtggtg
     1201 gtgatctcag ccatcctggc cctggtggtg ctcaccatca tctcccttat catcctcatc
     1261 atgctttggc agaagaagcc acgtggtggc tctggaggtc tcgagatgcc ggaccaactg
     1321 actgaagagc agatcgcaga atttaaagag gctttctccc tatttgacaa ggacggggat
     1381 gggacaataa caaccaagga gctggggacg gtgatgcggt ctctggggca gaaccccaca
     1441 gaagcagagc tgcaagacat gatcaatgaa gtagatgccg acggtgacgg cacaatcgac
     1501 ttccctgagt tcctgacaat gatggcaaga aaaatgaaat acagggacac ggaagaagaa
     1561 attagagaag cgttcggtgt gtttgataag gatggcaatg gctacatcag tgcagcagag
     1621 cttcgccacg tgatgacaaa ccttggagag aagttaacag atgaagaggt tgatgaaatg
     1681 atcagggaag cagacatcga tggggatggt caggtaaact acgaagagtt tgtacaaatg
     1741 atgacagcga aggcatgcgg tggcggtggc tctggaggtg gtgggtccgg aggaggcggc
     1801 cgctccggtg gatccatgct tcaacttcct cctcttgaac gtcttactct cgagatgggc
     1861 gagagcctgt tcaagggacc tagggactac aaccctatca gtagcacaat ttgtcacctg
     1921 accaacgaga gtgatggcca cacaacaagc ctgtacggca tcgggttcgg accctttatc
     1981 atcaccaaca agcacctgtt caggcggaat aatggcactc tgctggtgca gagcctgcac
     2041 ggggtgttca aagtgaagaa cacaaccact ctgcagcagc acctgatcga tgggcgggat
     2101 atgatcatca ttaggatgcc caaggacttc cccccttttc ctcagaaact gaagttccga
     2161 gagccccaga gagaggagag aatctgtctg gtgaccacaa actttcagga gctcggtagt
     2221 ggtagtgggg agtttctggc aaccacactg gaacggatcg agaaaaattt cgtgattact
     2281 gatccgagac tgcctgacaa cccaatcatt tttgcgagcg attccttcct gcagctgaca
     2341 gaatattctc gggaagagat cctggggcgc aattgccgtt ttctgcaggg acccgagaca
     2401 gaccgtgcca ctgttcggaa aatcagagat gctattgaca accagactga agtgaccgtt
     2461 cagctgatca attataccaa gagcggcaag aagttctgga acgtgttcca cctgcagccg
     2521 atgcgcgatt ataagggcga cgtccagtac ttcattggcg tgcagctgga tggcaccgaa
     2581 cgtcttcatg gcgccgctga gcgtgaggcg gtctgcctga tcaaaaagac agcctttcag
     2641 attgagaacc tgtacttcca gggctctaga ttagataaaa gtaaagtgat taacagcgca
     2701 ttagagctgc ttaatgaggt cggaatcgaa ggtttaacaa cccgtaaact cgcccagaag
     2761 ctaggtgtag agcagcctac attgtattgg catgtaaaaa ataagcgggc tttgctcgac
     2821 gccttagcca ttgagatgtt agataggcac catactcact tttgcccttt agaaggggaa
     2881 agctggcaag attttttacg taataacgct aaaagtttta gatgtgcttt actaagtcat
     2941 cgcgatggag caaaagtaca tttaggtaca cggcctacag aaaaacagta tgaaactctc
     3001 gaaaatcaat tagccttttt atgccaacaa ggtttttcac tagagaatgc attatatgca
     3061 ctcagcgctg tggggcattt tactttaggt tgcgtattgg aagatcaaga gcatcaagtc
     3121 gctaaagaag aaagggaaac acctactact gatagtatgc cgccattatt acgacaagct
     3181 atcgaattat ttgatcacca aggtgcagag ccagccttct tattcggcct tgaattgatc
     3241 atatgcggat tagaaaaaca acttaaatgt gaaagtgggt ccgcgtacag ccgcgcgcgt
     3301 acgaaaaaca attacgggtc taccatcgag ggcctgctcg atctcccgga cgacgacgcc
     3361 cccgaagagg cggggctggc ggctccgcgc ctgtcctttc tccccgcggg acacacgcgc
     3421 agactgtcga cggccccccc gaccgatgtc agcctggggg acgagctcca cttagacggc
     3481 gaggacgtgg cgatggcgca tgccgacgcg ctagacgatt tcgatctgga catgttgggg
     3541 gacggggatt ccccgggtcc gggatttacc ccccacgact ccgcccccta cggcgctctg
     3601 gatatggccg acttcgagtt tgagcagatg tttaccgatg cccttggaat tgacgagtac
     3661 ggtgggtagg ctagcctccc tatagtgagt cgtattacgt agatccagac atgataagat
     3721 acattgatga gtttggacaa accacaacta gaatgcagtg aaaaaaatgc tttatttgtg
     3781 aaatttgtga tgctattgct ttatttgtaa ccattataag ctgcaataaa caagttaaca
     3841 acaacaattg cattcatttt atgtttcagg ttcaggggga ggtgtgggag gttttttaat
     3901 tcgcgtccgc ggcgccaatg cattgggccc ggtacccagc ttttgttccc tttagtgagg
     3961 gttaattgcg cgcttggcgt aatcatggtc atagctgttt cctgtgtgaa attgttatcc
     4021 gctcacaatt ccacacaaca tacgagccgg gagcataaag tgtaaagcct ggggtgccta
     4081 atgagtgagc taactcacat taattgcgtt gcgctcactg cccgctttcc agtcgggaaa
     4141 cctgtcgtgc cagctgcatt aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat
     4201 tgggcgctct tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc ggctgcggga
     4261 gcggtatcag ctcactcaaa ggcggtaata cggttatcca cagaatcagg ggataacgca
     4321 ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa ggccgcgttg
     4381 ctggcgtttt tccataggct ccgcccccct gacgagcatc acaaaaatcg acgctcaagt
     4441 cagaggtggc gaaacccgac aggactataa agataccagg cgtttccccc tggaagctcc
     4501 ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat acctgtccgc ctttctccct
     4561 tcgggaagcg tggcgctttc tcatagctca cgctgtaggt atctcagttc ggtgtaggtc
     4621 gttcgctcca agctgggctg tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta
     4681 tccggtaact atcgtcttga gtccaacccg gtaagacacg acttatcgcc actggcagca
     4741 gccactggta acaggattag cagagcgagg tatgtaggcg gtgctacaga gttcttgaag
     4801 tggtggccta actacggcta cactagaaga acagtatttg gtatctgcgc tctgctgaag
     4861 ccagttacct tcggaaaaag agttggtagc tcttgatccg gcaaacaaac caccgctggt
     4921 agcggtggtt tttttgtttg caagcagcag attacgcgca gaaaaaaagg atctcaagaa
     4981 gatcctttga tcttttctac ggggtctgac gctcagtgga acgaaaactc acgttaaggg
     5041 attttggtca tgagattatc aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga
     5101 agttttaaat caatctaaag tatatatgag taaacttggt ctgacagtta ccaatgctta
     5161 atcagtgagg cacctatctc agcgatctgt ctatttcgtt catccatagt tgcctgactc
     5221 cccgtcgtgt agataactac gatacgggag ggcttaccat ctggccccag tgctgcaatg
     5281 ataccgcgag acccacgctc accggctcca gatttatcag caataaacca gccagccgga
     5341 agggccgagc gcagaagtgg tcctgcaact ttatccgcct ccatccagtc tattaattgt
     5401 tgccgggaag ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt
     5461 gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag ctccggttcc
     5521 caacgatcaa ggcgagttac atgatccccc atgttgtgca aaaaagcggt tagctccttc
     5581 ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt tatcactcat ggttatggca
     5641 gcactgcata attctcttac tgtcatgcca tccgtaagat gcttttctgt gactggtgag
     5701 tactcaacca agtcattctg agaatagtgt atgcggcgac cgagttgctc ttgcccggcg
     5761 tcaatacggg ataataccgc gccacatagc agaactttaa aagtgctcat cattggaaaa
     5821 cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa
     5881 cccactcgtg cacccaactg atcttcagca tcttttactt tcaccagcgt ttctgggtga
     5941 gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg gaaatgttga
     6001 atactcatac tcttcctttt tcaatattat tgaagcattt atcagggtta ttgtctcatg
     6061 agcggataca tatttgaatg tatttagaaa aataaacaaa taggggttcc gcgcacattt
     6121 ccccgaaaag tgccacctaa attgtaagcg ttaatatttt gttaaaattc gcgttaaatt
     6181 tttgttaaat cagctcattt tttaaccaat aggccgaaat cggcaaaatc ccttataaat
     6241 caaaagaata gaccgagata gggttgagtg ttgttccagt ttggaacaag agtccactat
     6301 taaagaacgt ggactccaac gtcaaagggc gaaaaaccgt ctatcagggc gatggcccac
     6361 tacgtgaacc atcaccctaa tcaagttttt tggggtcgag gtgccgtaaa gcactaaatc
     6421 ggaaccctaa agggagcccc cgatttagag cttgacgggg aaagccggcg aacgtggcga
     6481 gaaaggaagg gaagaaagcg aaaggagcgg gcgctagggc gctggcaagt gtagcggtca
     6541 cgctgcgcgt aaccaccaca cccgccgcgc ttaatgcgcc gctacagggc gcgtcccatt
     6601 cgccattcag gctgcgcaac tgttgggaag ggcgatcggt gcgggcctct tcgctattac
     6661 gccagtcgac cgccaattca
//
