LOCUS       sequence-114685-        1083 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  Plasmid for tet-inducible expression of dCas9-EGFP in mammalian 
            cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 1083)
  AUTHORS   Ochiai H, Sugawara T, Yamamoto T
  TITLE     Simultaneous live imaging of the transcription and nuclear position 
            of specific genes.
  JOURNAL   Nucleic Acids Res. 2015 Jun 19. pii: gkv624.
  PUBMED    26092696
REFERENCE   2  (bases 1 to 1083)
  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: "Nucleic 
            Acids Res. 2015 Jun 19. pii: gkv624."
COMMENT     Sequence Label: pBSK-Delta-B-TRE-dCas9-EGFP
FEATURES             Location/Qualifiers
     source          1..1083
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     CDS             complement(315..335)
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             complement(339..365)
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     promoter        complement(405..719)
                     /label=tight TRE promoter
                     /note="Tet-responsive promoter PTight, consisting of seven 
                     tet operator sequences followed by the minimal CMV 
                     promoter"
     primer_bind     complement(408..432)
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     protein_bind    479..497
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    515..533
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    550..568
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    586..604
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    622..640
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    657..675
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    693..711
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     primer_bind     complement(779..799)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(779..797)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(818..834)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(818..834)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(831..853)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    842..858
                     /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(866..872,873..890,891..896))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    911..932
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(1049..1066)
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 tcttccagtc tgtggaagaa gctgtcgtcc accttggcca tctcgttgct gaagatctct
       61 tgcagatagc agatccggtt cttccgtctg gtgtatcttc ttctggcggt tctcttcagc
      121 cgggtggcct cggctgtttc gccgctgtcg aacagcaggg ctccgatcag gttcttcttg
      181 atgctgtgcc ggtcggtgtt gcccagcacc ttgaatttct tgctgggcac cttgtactcg
      241 tcggtgatca cggcccagcc cacagagttg gtgccgatgg ccaggccgat gctgtacttc
      301 ttgtcggacg cttcgacctt gcgctttttc ttcggcgaag cgtaatctgg aacatcgtat
      361 gggtacatgg tggcaccggt cgggtaccga gctcgaattc tccaggcgat ctgacggttc
      421 actaaacgag ctctgcttat ataggcctcc caccgtacac gcctacctcg acatacgttc
      481 tctatcactg atagggagta aactcgacat acgttctcta tcactgatag ggataaactc
      541 gacatacgtt ctctatcact gatagggagt aaactcgaca tacgttctct atcactgata
      601 gggagtaaac tcgacatacg ttctctatca ctgataggga gtaaactcga catcgttctc
      661 tatcactgat agggagtaaa ctcgacatac gttctctatc actgataggg agtaaactcg
      721 agtgaagacg actagttcta gagcggccgc caccgcggtg gagctccagc ttttgttccc
      781 tttagtgagg gttaattgcg cgcttggcgt aatcatggtc atagctgttt cctgtgtgaa
      841 attgttatcc gctcacaatt ccacacaaca tacgagccgg aagcataaag tgtaaagcct
      901 ggggtgccta atgagtgagc taactcacat taattgcgtt gcgctcactg cccgctttcc
      961 agtcgggaaa cctgtcgtgc cagctgcatt aatgaatcgg ccaacgcgcg gggagaggcg
     1021 gtttgcgtat tgggcgctct tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc
     1081 ggc
//
