pSAG1-mNeonGreen_TUB1-dTomato vector (V016188)

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LOCUS       62056_19180        6710 bp DNA     circular SYN 15-OCT-2021
DEFINITION  Cloning vector pSAG1-mNeonGreen_TUB1-dTomato, complete sequence.
ACCESSION   MZ090944
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6710)
  AUTHORS   Markus BM, Boydston EA, Lourido S.
  TITLE     CRISPR-Mediated Transcriptional Repression in Toxoplasma gondii
  JOURNAL   mSphere, e0047421 (2021) In press
  PUBMED    34643425
REFERENCE   2  (bases 1 to 6710)
  AUTHORS   Markus BM.
  TITLE     Direct Submission
  JOURNAL   Submitted (02-MAY-2021) Whitehead Institute, 455 Main Street, 
            Cambridge, MA 02142, United States
REFERENCE   3  (bases 1 to 6710)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "mSphere, 
            e0047421 (2021) In press"
COMMENT     SGRef: number: 2; type: "Journal Article"; journalName: "Submitted 
            (02-MAY-2021) 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..6710
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      complement(3..458)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     primer_bind     600..616
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        623..641
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     misc_feature    672..1428
                     /label=TgSAG1 upstream region
                     /note="TgSAG1 upstream region"
     CDS             1435..2142
                     /label=mNeonGreen
                     /note="bright monomeric yellow-green fluorescent protein
                     derived from LanYFP (Shaner et al., 2013)"
     misc_feature    2143..2172
                     /label=ty1 tag
                     /note="ty1 tag"
     gene            2184..2496
                     /gene="TgSAG1"
                     /label=TgSAG1
     3'UTR           2184..2496
                     /gene="TgSAG1"
     misc_feature    2540..3037
                     /label=TgTUB1 upstream region
                     /note="TgTUB1 upstream region"
     CDS             3044..3745
                     /label=dTomato
                     /note="dimeric variant of DsRed fluorescent protein (Shaner
                     et al., 2004)"
     gene            3755..4496
                     /gene="TgDHFR-TS"
                     /label=TgDHFR-TS
     3'UTR           3755..4496
                     /gene="TgDHFR-TS"
     promoter        complement(4522..4540)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(4561..4577)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    complement(4585..4601)
                     /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(4609..4639)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    complement(4654..4675)
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     rep_origin      complement(4963..5551)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(5725..6582)
                     /label=AmpR
                     /note="beta-lactamase"
     promoter        complement(6583..6687)
                     /label=AmpR promoter