LOCUS       sequence-325983-        5466 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses a nuclear localized genetically encoded calcium reporter..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5466)
  AUTHORS   Enyedi B, Jelcic M, Niethammer P
  TITLE     The Cell Nucleus Serves as a Mechanotransducer of Tissue 
            Damage-Induced Inflammation.
  JOURNAL   Cell. 2016 May 19;165(5):1160-1170. doi: 10.1016/j.cell.2016.04.016.
  PUBMED    27203112
REFERENCE   2  (bases 1 to 5466)
  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"; doi: 
            "10.1016/j.cell.2016.04"; journalName: "Cell"; date: "2016-05-19- 
            19"; volume: "165"; issue: "5"; pages: "1160-1170"
COMMENT     Sequence Label: pCS2+ GCamp6-3NLS
FEATURES             Location/Qualifiers
     source          1..5466
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        859..877
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     859..876
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     regulatory      916..925
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             937..993
                     /codon_start=1
                     /product="calmodulin-binding peptide from avian smooth 
                     muscle myosin light chain kinase"
                     /label=calmodulin-binding peptide
                     /translation="RRKWNKTGHAVRAIGRLSS"
     primer_bind     1206..1227
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(1333..1354)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1594..1613)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     CDS             2215..2235
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             2239..2259
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             2263..2283
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     polyA_signal    2342..2476
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2366..2385)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2420..2439
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(2589..2609)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(2589..2607)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(2628..2644)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2628..2644)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2641..2663)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2652..2668
                     /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(2676..2682,2683..2700,2701..2706))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2721..2742
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2859..2876)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3030..3618)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3110..3129)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3789..4580,4581..4649))
                     /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     4412..4431
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4650..4754)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(4780..5235)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4917..4938)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     5129..5148
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        join(5306..5466,1..824)
                     /label=CMV IE94 promoter
                     /note="enhancer/promoter region of simian cytomegalovirus 
                     major immediate early transcription unit IE94"
ORIGIN
        1 attcaatatg gcggacctgg cactgtgcca actggggagg ggtctacttg gcacggtgcc
       61 aagtttgagg aggggtcttg gccctgtgcc aagtccgcca tattgaattg gcatggtgcc
      121 aataatggcg gccatattgg ctatatgcca ggatcaatat ataggcaata tccaatatgg
      181 ccctatgcca atatggctat tggccaggtt caatactatg tattggccct atgccatata
      241 gtattccata tatgggtttt cctattgacg tagatagccc ctcccaatgg gcggtcccat
      301 ataccatata tggggcttcc taataccgcc catagccact cccccattga cgtcaatggt
      361 ctctatatat ggtctttcct attgacgtca tatgggcggt cctattgacg tatatggcgc
      421 ctcccccatt gacgtcaatt acggtaaatg gcccgcctgg ctcaatgccc attgacgtca
      481 ataggaccac ccaccattga cgtcaatggg atggctcatt gcccattcat atccgttctc
      541 acgcccccta ttgacgtcaa tgacggtaaa tggcccactt ggcagtacat caatatctat
      601 taatagtaac ttggcaagta cattactatt ggaagtacgc cagggtacat tggcagtact
      661 cccattgacg tcaatggcgg taaatggccc gcgatggctg ccaagtacat ccccattgac
      721 gtcaatgggg aggggcaatg acgcaaatgg gcgttccatt gacgtaaatg ggcggtaggc
      781 gtgcctaatg ggaggtctat ataagcaatg ctcgtttagg gaaccgccat tctgcctggg
      841 gacgtcggag caagcttgat ttaggtgaca ctatagaata caagctactt gttctttttg
      901 caggatccca tcgatgccac catggtggac tcatcacgtc gtaagtggaa taagacaggt
      961 cacgcagtca gagctatagg tcggctgagc tcactcgaga acgtctatat caaggccgac
     1021 aagcagaaga acggcatcaa ggcgaacttc cacatccgcc acaacatcga ggacggcggc
     1081 gtgcagctcg cctaccacta ccagcagaac acccccatcg gcgacggccc cgtgctgctg
     1141 cccgacaacc actacctgag cgtgcagtcc aaactttcga aagaccccaa cgagaagcgc
     1201 gatcacatgg tcctgctgga gttcgtgacc gccgccggga tcactctcgg catggacgag
     1261 ctgtacaagg gcggtaccgg agggagcatg gtgagcaagg gcgaggagct gttcaccggg
     1321 gtggtgccca tcctggtcga gctggacggc gacgtaaacg gccacaagtt cagcgtgtcc
     1381 ggcgagggtg agggcgatgc cacctacggc aagctgaccc tgaagttcat ctgcaccacc
     1441 ggcaagctgc ccgtgccctg gcccaccctc gtgaccaccc tgacctacgg cgtgcagtgc
     1501 ttcagccgct accccgacca catgaagcag cacgacttct tcaagtccgc catgcccgaa
     1561 ggctacatcc aggagcgcac catcttcttc aaggacgacg gcaactacaa gacccgcgcc
     1621 gaggtgaagt tcgagggcga caccctggtg aaccgcatcg agctgaaggg catcgacttc
     1681 aaggaggacg gcaacatcct ggggcacaag ctggagtaca acctgccgga ccaactgact
     1741 gaagagcaga tcgcagaatt taaagaggct ttctccctat ttgacaagga cggggatggg
     1801 acaataacaa ccaaggagct ggggacggtg atgcggtctc tggggcagaa ccccacagaa
     1861 gcagagctgc aggacatgat caatgaagta gatgccgacg gtgacggcac aatcgacttc
     1921 cctgagttcc tgacaatgat ggcaagaaaa atgaaataca gggacacgga agaagaaatt
     1981 agagaagcgt tcggtgtgtt tgataaggat ggcaatggct acatcagtgc agcagagctt
     2041 cgccacgtga tgacaaacct tggagagaag ttaacagatg aagaggttga tgaaatgatc
     2101 agggaagcag acatcgatgg ggatggtcag gtaaactacg aagagtttgt acaaatgatg
     2161 acagcgaagg gcggtggaag atctcgagct caagcttcga attcccgggt cgatccaaaa
     2221 aagaagagaa aggtagatcc aaaaaagaag agaaaggtag atccaaaaaa gaagagaaag
     2281 gtataggtac cgcgggcccg ggatccaccg gatctagaac tatagtgagt cgtattacgt
     2341 agatccagac atgataagat acattgatga gtttggacaa accacaacta gaatgcagtg
     2401 aaaaaaatgc tttatttgtg aaatttgtga tgctattgct ttatttgtaa ccattataag
     2461 ctgcaataaa caagttaaca acaacaattg cattcatttt atgtttcagg ttcaggggga
     2521 ggtgtgggag gttttttaat tcgcggccgc ggcgccaatg cattgggccc ggtacccagc
     2581 ttttgttccc tttagtgagg gttaattgcg cgcttggcgt aatcatggtc atagctgttt
     2641 cctgtgtgaa attgttatcc gctcacaatt ccacacaaca tacgagccgg gagcataaag
     2701 tgtaaagcct ggggtgccta atgagtgagc taactcacat taattgcgtt gcgctcactg
     2761 cccgctttcc agtcgggaaa cctgtcgtgc cagctgcatt aatgaatcgg ccaacgcgcg
     2821 gggagaggcg gtttgcgtat tgggcgctct tccgcttcct cgctcactga ctcgctgcgc
     2881 tcggtcgttc ggctgcggcg agcggtatca gctcactcaa aggcggtaat acggttatcc
     2941 acagaatcag gggataacgc aggaaagaac atgtgagcaa aaggccagca aaaggccagg
     3001 aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc tccgcccccc tgacgagcat
     3061 cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga caggactata aagataccag
     3121 gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga
     3181 tacctgtccg cctttctccc ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg
     3241 tatctcagtt cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga accccccgtt
     3301 cagcccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc ggtaagacac
     3361 gacttatcgc cactggcagc agccactggt aacaggatta gcagagcgag gtatgtaggc
     3421 ggtgctacag agttcttgaa gtggtggcct aactacggct acactagaag aacagtattt
     3481 ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa gagttggtag ctcttgatcc
     3541 ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt gcaagcagca gattacgcgc
     3601 agaaaaaaag gatctcaaga agatcctttg atcttttcta cggggtctga cgctcagtgg
     3661 aacgaaaact cacgttaagg gattttggtc atgagattat caaaaaggat cttcacctag
     3721 atccttttaa attaaaaatg aagttttaaa tcaatctaaa gtatatatga gtaaacttgg
     3781 tctgacagtt accaatgctt aatcagtgag gcacctatct cagcgatctg tctatttcgt
     3841 tcatccatag ttgcctgact ccccgtcgtg tagataacta cgatacggga gggcttacca
     3901 tctggcccca gtgctgcaat gataccgcga gacccacgct caccggctcc agatttatca
     3961 gcaataaacc agccagccgg aagggccgag cgcagaagtg gtcctgcaac tttatccgcc
     4021 tccatccagt ctattaattg ttgccgggaa gctagagtaa gtagttcgcc agttaatagt
     4081 ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg
     4141 gcttcattca gctccggttc ccaacgatca aggcgagtta catgatcccc catgttgtgc
     4201 aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg
     4261 ttatcactca tggttatggc agcactgcat aattctctta ctgtcatgcc atccgtaaga
     4321 tgcttttctg tgactggtga gtactcaacc aagtcattct gagaatagtg tatgcggcga
     4381 ccgagttgct cttgcccggc gtcaatacgg gataataccg cgccacatag cagaacttta
     4441 aaagtgctca tcattggaaa acgttcttcg gggcgaaaac tctcaaggat cttaccgctg
     4501 ttgagatcca gttcgatgta acccactcgt gcacccaact gatcttcagc atcttttact
     4561 ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata
     4621 agggcgacac ggaaatgttg aatactcata ctcttccttt ttcaatatta ttgaagcatt
     4681 tatcagggtt attgtctcat gagcggatac atatttgaat gtatttagaa aaataaacaa
     4741 ataggggttc cgcgcacatt tccccgaaaa gtgccaccta aattgtaagc gttaatattt
     4801 tgttaaaatt cgcgttaaat ttttgttaaa tcagctcatt ttttaaccaa taggccgaaa
     4861 tcggcaaaat cccttataaa tcaaaagaat agaccgagat agggttgagt gttgttccag
     4921 tttggaacaa gagtccacta ttaaagaacg tggactccaa cgtcaaaggg cgaaaaaccg
     4981 tctatcaggg cgatggccca ctacgtgaac catcacccta atcaagtttt ttggggtcga
     5041 ggtgccgtaa agcactaaat cggaacccta aagggagccc ccgatttaga gcttgacggg
     5101 gaaagccggc gaacgtggcg agaaaggaag ggaagaaagc gaaaggagcg ggcgctaggg
     5161 cgctggcaag tgtagcggtc acgctgcgcg taaccaccac acccgccgcg cttaatgcgc
     5221 cgctacaggg cgcgtcccat tcgccattca ggctgcgcaa ctgttgggaa gggcgatcgg
     5281 tgcgggcctc ttcgctatta cgccagtcga ccgccaattc aatatggcgt atatggactc
     5341 atgccaattc aatatggtgg atctggacct gtgccaattc aatatggcgt atatggactc
     5401 gtgccaattc aatatggtgg atctggaccc cagccaattc aatatggcgg acttggcacc
     5461 atgcca
//
