pCas-Guide-EF1a-CD4 vector (V012291)

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LOCUS       pCas-Guide-EF1a-       11006 bp DNA     circular SYN 01-JAN-1980
DEFINITION  Mammalian vector for co-expressing a guide RNA (gRNA) together with 
            Cas9 and CD4, for genome editing using the S. pyogenes CRISPR/Cas9 
            system.
ACCESSION   .
VERSION     .
KEYWORDS    pCas-Guide-EF1a-CD4
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 11006)
  AUTHORS   OriGene
  TITLE     Direct Submission
REFERENCE   2  (bases 1 to 11006)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..11006
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        90..330
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA"
     misc_RNA        384..459
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     terminator      460..465
                     /note="polIII terminator"
                     /note="RNA polymerase III transcription terminator"
     promoter        480..1658
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation
                     factor EF-1-alpha"
     CDS             1679..2950
                     /label=CD4 (truncated)
                     /note="truncated human CD4 cell surface glycoprotein
                     lacking the cytoplasmic tail"
     polyA_signal    2977..3430
                     /label=PGK poly(A) signal
                     /note="mouse phosphoglycerate kinase 1 polyadenylation
                     signal"
     protein_bind    3457..3490
                     /label=loxP
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     enhancer        3560..3863
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        3864..4067
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
     promoter        4093..4111
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             4170..8273
                     /label=Cas9
                     /note="Cas9 (Csn1) endonuclease from the Streptococcus
                     pyogenes Type II CRISPR/Cas system"
     CDS             8274..8294
                     /codon_start=1
                     /product="nuclear localization signal of SV40 large T
                     antigen"
                     /note="SV40 NLS"
                     /translation="PKKKRKV"
     CDS             8301..8321
                     /label=SV40 NLS
                     /note="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
     CDS             8343..8372
                     /label=Myc
                     /note="Myc (human c-Myc proto-oncogene) epitope tag"
     CDS             8391..8414
                     /label=FLAG
                     /note="FLAG(R) epitope tag, followed by an enterokinase
                     cleavage site"
     polyA_signal    8461..9083
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     rep_origin      complement(9204..9792)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(9970..10827)
                     /label=AmpR
                     /note="beta-lactamase"
     primer_bind     10942..10958
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        10963..10981
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"