LOCUS       Exported                6624 bp ds-DNA     circular SYN 13-MAY-2021
DEFINITION  Expression of sgRNA from a imyc promoter (Pimyc).
ACCESSION   .
VERSION     .
KEYWORDS    pRH2521
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6624)
  AUTHORS   Singh AK, Carette X, Potluri LP, Sharp JD, Xu R, Prisic S, Husson RN
  TITLE     Investigating essential gene function in Mycobacterium tuberculosis 
            using an efficient CRISPR interference system.
  JOURNAL   Nucleic Acids Res. 2016 Jul 12. pii: gkw625.
  PUBMED    27407107
REFERENCE   2  (bases 1 to 6624)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Nucleic 
            Acids Res. 2016 Jul 12. pii: gkw625."
FEATURES             Location/Qualifiers
     source          1..6624
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     primer_bind     37..53
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     primer_bind     38..54
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     terminator      197..224
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     protein_bind    424..441
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     misc_RNA        503..578
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     terminator      1588..1615
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     primer_bind     complement(1705..1722)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1876..2464)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1956..1975)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"