pBM019 vector (V015680)

LOCUS       62056_4130        9826 bp DNA     circular SYN 28-JUN-2019
DEFINITION  Cloning vector pBM019, complete sequence.
ACCESSION   MN019123
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9826)
  AUTHORS   Markus BM, Bell GW, Lorenzi HA, Lourido S.
  TITLE     Optimizing Systems for Cas9 Expression in Toxoplasma gondii
  JOURNAL   mSphere 4 (3), e00386-19 (2019)
  PUBMED    31243081
REFERENCE   2  (bases 1 to 9826)
  AUTHORS   Markus BM.
  TITLE     Direct Submission
  JOURNAL   Submitted (04-JUN-2019) Whitehead Institute, 455 Main Street, 
            Cambridge, MA 02142, United States
REFERENCE   3  (bases 1 to 9826)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "mSphere"; 
            date: "2019"; volume: "4"; issue: "3"; pages: "e00386-19"
COMMENT     SGRef: number: 2; type: "Journal Article"; journalName: "Submitted 
            (04-JUN-2019) Whitehead Institute, 455 Main Street, Cambridge, MA 
            02142, United States"
COMMENT     ##Assembly-Data-START##
            Sequencing Technology :: Sanger dideoxy sequencing 
            ##Assembly-Data-END##
FEATURES             Location/Qualifiers
     source          1..9826
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(4..861)
                     /label=AmpR
                     /note="beta-lactamase"
     promoter        complement(862..966)
                     /label=AmpR promoter
     rep_origin      complement(992..1447)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     primer_bind     1589..1605
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        1612..1630
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     1656..1672
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     misc_feature    1684..2183
                     /label=U6 upstream region
                     /note="U6 upstream region"
     misc_feature    2184..2204
                     /label=sgRNA-1 protospacer
                     /note="sgRNA-1 protospacer"
     misc_RNA        2205..2280
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     misc_feature    2281..2295
                     /label=PolIII terminator
                     /note="PolIII terminator"
     5'UTR           2309..2806
                     /label=TgTUB1
                     /note="TgTUB1"
     misc_feature    2816..2881
                     /note="3xFLAG; three tandem FLAG(R) epitope tags, followed
                     by an enterokinase cleavage site"
     misc_feature    2888..2908
                     /note="SV40 NLS; nuclear localization signal of SV40 large
                     T antigen"
     CDS             2933..7033
                     /label=Cas9
                     /note="Cas9 (Csn1) endonuclease from the Streptococcus
                     pyogenes Type II CRISPR/Cas system"
     misc_feature    7034..7081
                     /note="nucleoplasmin NLS; bipartite nuclear localization
                     signal from nucleoplasmin"
     CDS             7097..7153
                     /label=P2A
                     /note="2A peptide from porcine teschovirus-1 polyprotein"
     CDS             7169..7822
                     /label=CmR
                     /note="chloramphenicol acetyltransferase"
     CDS             7823..7849
                     /label=HA
                     /note="HA (human influenza hemagglutinin) epitope tag"
     3'UTR           7859..8601
                     /label=DHFR
                     /note="DHFR"
     promoter        complement(8627..8645)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(8666..8682)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    complement(8690..8706)
                     /label=lac operator
                     /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(8714..8744)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    complement(8759..8780)
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     rep_origin      complement(9068..9656)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"