pBM064 vector (V016179)

LOCUS       62056_4140        5279 bp DNA     circular SYN 28-JUN-2019
DEFINITION  Cloning vector pBM064, complete sequence.
ACCESSION   MN019113
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5279)
  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 5279)
  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 5279)
  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..5279
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      complement(1..589)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(763..1554)
                     /label=NeoR/KanR
                     /note="aminoglycoside phosphotransferase"
     promoter        complement(1555..1659)
                     /label=AmpR promoter
     primer_bind     1779..1795
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     regulatory      1921..2420
                     /label=U6 promoter
                     /note="U6 promoter"
                     /regulatory_class="promoter"
     misc_feature    2421..2441
                     /label=sgRNA-1 protospacer
                     /note="sgRNA-1 protospacer"
     misc_RNA        2442..2517
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     misc_feature    2518..2532
                     /label=PolIII terminator
                     /note="PolIII terminator"
     misc_feature    2533..2540
                     /label=AscI site
                     /note="AscI site"
     misc_feature    2566..2573
                     /label=SbfI site
                     /note="SbfI site"
     regulatory      2574..3072
                     /label=TUB1 promoter
                     /note="TUB1 promoter"
                     /regulatory_class="promoter"
     misc_feature    3067..3075
                     /label=Kozak
                     /note="Kozak"
     misc_feature    3079..3084
                     /label=BamHI site
                     /note="BamHI site"
     CDS             3085..3792
                     /label=mRuby3
                     /note="monomeric red fluorescent protein derived from
                     mRuby2, with improved brightness and photostability (Bajar 
                     et al., 2016)"
     misc_feature    3808..3864
                     /note="eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides; P2A; 2A peptide from porcine teschovirus-1 
                     polyprotein"
     CDS             3880..4533
                     /label=CmR
                     /note="chloramphenicol acetyltransferase"
     3'UTR           4537..4852
                     /label=SAG1
                     /note="SAG1"
     primer_bind     complement(4976..4992)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"