LOCUS       sequence-96699-e         593 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  Co-expresses human optimized S. pyogenes dCas9 fused to two copies 
            of VP64 and GFP.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 593)
  AUTHORS   Kabadi AM, Ousterout DG, Hilton IB, Gersbach CA
  TITLE     Multiplex CRISPR/Cas9-based genome engineering from a single 
            lentiviral vector.
  JOURNAL   Nucleic Acids Res. 2014 Aug 13. pii: gku749.
  PUBMED    25122746
REFERENCE   2  (bases 1 to 593)
  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. 2014 Aug 13. pii: gku749."
COMMENT     Sequence Label: pLV hUbC-VP64 dCas9 VP64-T2A-GFP
FEATURES             Location/Qualifiers
     source          1..593
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     protein_bind    complement(49..82)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     CDS             complement(69..395)
                     /codon_start=1
                     /gene="lacZ fragment"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /label=lacZ-alpha
                     /translation="MTMITPSAQLTLTKGNKSWRTSRGGPVPNSPYSESYYARSLAVVL
                     QRRDWENPGVTQLNRLAAHPPFASWRNSEEARTDRPSQQLRSLNGEWDAPCSGATTVSY
                     HNFV"
     primer_bind     245..267
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     259..276
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     260..276
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     286..305
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        286..304
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(352..372)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(352..370)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(391..407)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(391..407)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(404..426)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    415..431
                     /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(439..445,446..463,464..469))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    484..505
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
ORIGIN
        1 ttttcgggtt tattacaggg acagcagaga tccagtttgg ttaattaaat aacttcgtat
       61 agcatacatt atacgaagtt atgataagag acggtggttg cgccgctaca gggcgcgtcc
      121 cattcgccat tcaggctgcg caactgttgg gaagggcgat cggtgcgggc ctcttcgcta
      181 ttacgccagc tggcgaaagg gggatgtgct gcaaggcgat taagttgggt aacgccaggg
      241 ttttcccagt cacgacgttg taaaacgacg gccagtgagc gcgcgtaata cgactcacta
      301 tagggcgaat tgggtaccgg gccccccctc gaggtcctcc agcttttgtt ccctttagtg
      361 agggttaatt gcgcgcttgg cgtaatcatg gtcatagctg tttcctgtgt gaaattgtta
      421 tccgctcaca attccacaca acatacgagc cggaagcata aagtgtaaag cctggggtgc
      481 ctaatgagtg agctaactca cattaattgc gttgcgctca ctgcccgctt tccactgcat
      541 gacgtctcca caattaatta acccgtgtcg gctccagatc tggcctccgc gcc
//
