pSG252 vector (V003274)

LOCUS       40924_39862       11625 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Expression vector pSG252, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 11625)
  AUTHORS   Goldfless SJ, Wagner JC, Niles JC.
  TITLE     Versatile control of Plasmodium falciparum gene expression with an 
            inducible protein-RNA interaction
  JOURNAL   Nat Commun 5, 5329 (2014)
  PUBMED    25370483
REFERENCE   2  (bases 1 to 11625)
  AUTHORS   Goldfless SJ, Niles JC.
  TITLE     Direct Submission
  JOURNAL   Submitted (10-AUG-2014) Biological Engineering, MIT, 77 
            Massachusetts Avenue, Cambridge, MA 02139, USA
REFERENCE   3  (bases 1 to 11625)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 11625)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Nat Commun 
            5, 5329 (2014)"
COMMENT     SGRef: number: 2; type: "Journal Article"; journalName: "Submitted 
            (10-AUG-2014) Biological Engineering, MIT, 77 Massachusetts Avenue, 
            Cambridge, MA 02139, USA"
COMMENT     SGRef: number: 3; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..11625
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     regulatory      complement(24..859)
                     /label=derived from P. falciparum hsp86 3' UTR
                     /note="derived from P. falciparum hsp86 3' UTR"
                     /regulatory_class="terminator"
     CDS             complement(869..2518)
                     /label=luciferase
                     /note="firefly luciferase"
     misc_feature    complement(2541..2587)
                     /label=mutant TetR aptamer
                     /note="mutant TetR aptamer"
     regulatory      complement(2590..3219)
                     /note="contains promoter and 5' UTR from P. falciparum 
                     calmodulin gene"
                     /regulatory_class="promoter"
     regulatory      3220..4089
                     /note="contains promoter and 5'UTR from P. falciparum Hsp86
                     (PF07_0029)"
                     /regulatory_class="promoter"
     CDS             4100..4720
                     /label=TetR
                     /note="tetracycline repressor TetR"
     CDS             4730..4783
                     /label=T2A
                     /note="2A peptide from Thosea asigna virus capsid protein"
     CDS             4784..5719
                     /label=RLuc
                     /note="Renilla luciferase"
     misc_feature    5717..5779
                     /note="Thosea asigna virus 2A-like peptide, P. falciparum 
                     optimized codon usage"
     CDS             5726..5779
                     /codon_start=1
                     /product="2A peptide from Thosea asigna virus capsid
                     protein"
                     /label=T2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="EGRGSLLTCGDVEENPGP"
     CDS             5780..6568
                     /label=NeoR/KanR
                     /note="aminoglycoside phosphotransferase"
     regulatory      6607..7170
                     /label=derived from P. falciparum HRPII 3' UTR
                     /note="derived from P. falciparum HRPII 3' UTR"
                     /regulatory_class="terminator"
     gene            complement(7218..8574)
                     /label=kanMX
                     /note="yeast selectable marker conferring kanamycin
                     resistance (Wach et al., 1994)"
     repeat_region   8582..9120
     repeat_region   9121..9675
     misc_feature    complement(9829..10332)
                     /label=CEN/ARS
                     /note="S. cerevisiae CEN6 centromere fused to an
                     autonomously replicating sequence"
     rep_origin      10583..11171
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     protein_bind    11459..11480
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     promoter        11495..11525
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    11533..11549
                     /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)."
     primer_bind     11557..11573
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        11591..11609
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"