LOCUS Exported 6458 bp ds-DNA circular SYN 17-DEC-2018
DEFINITION Cloning vector pDC-KHT, complete sequence.
ACCESSION JF430587
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 6458)
AUTHORS Crivat G, Tokumasu F, Sa JM, Hwang J, Wellems TE.
TITLE Tetracysteine-Based Fluorescent Tags to Study Protein Localization
and Trafficking in Plasmodium falciparum-Infected Erythrocytes
JOURNAL PLoS ONE 6 (8), E22975 (2011)
PUBMED 21860664
REFERENCE 2 (bases 1 to 6458)
AUTHORS Crivat G, Tokumasu F, Sa JM, Hwang J, Wellems TE.
TITLE Direct Submission
JOURNAL Submitted (23-FEB-2011) LMVR, NIAID, 12735 Twinbrook Pkwy, 3E10C,
Rockville, MD 20852, USA
REFERENCE 3 (bases 1 to 6458)
AUTHORS .
TITLE Direct Submission
FEATURES Location/Qualifiers
source 1..6458
/organism="Cloning vector pDC-KHT"
/mol_type="other DNA"
/clone="pDC-KHT"
/note="derived from Plasmodium falciparum; transfection
plasmid expressing KAHRP followed by TC tag"
/db_xref="taxon:1035313"
CDS complement(878..895)
/codon_start=1
/product="6xHis affinity tag"
/label=6xHis
/translation="HHHHHH"
CDS complement(896..925)
/codon_start=1
/product="Myc (human c-Myc proto-oncogene) epitope tag"
/label=Myc
/translation="EQKLISEEDL"
CDS complement(941..958)
/codon_start=1
/product="tetracysteine peptide that binds biarsenical
labeling reagents"
/label=tetracysteine tag
/translation="CCPGCC"
CDS complement(995..1021)
/codon_start=1
/product="9xHis affinity tag"
/label=9xHis
/translation="HHHHHHHHH"
CDS complement(1082..1099)
/codon_start=1
/product="6xHis affinity tag"
/label=6xHis
/translation="HHHHHH"
CDS complement(1115..1132)
/codon_start=1
/product="HQHQHQ metal affinity tag"
/label=HQ tag
/note="The HQ tag binds to nickel and other metal ions much
like the 6xHis tag, but may allow elution at lower
concentrations of imidazole (Godat et al., 2008)."
/translation="HQHQHQ"
CDS complement(1121..1138)
/codon_start=1
/product="HQHQHQ metal affinity tag"
/label=HQ tag
/note="The HQ tag binds to nickel and other metal ions much
like the 6xHis tag, but may allow elution at lower
concentrations of imidazole (Godat et al., 2008)."
/translation="HQHQHQ"
CDS complement(1136..1156)
/codon_start=1
/product="6xHis affinity tag"
/label=7xHis
/translation="HHHHHHH"
promoter complement(3776..3793)
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
primer_bind complement(3800..3816)
/label=M13 fwd
/note="common sequencing primer, one of multiple similar
variants"
promoter 4290..4394
/gene="bla"
/label=AmpR promoter
CDS 4395..5255
/codon_start=1
/gene="bla"
/product="beta-lactamase"
/label=AmpR
/note="confers resistance to ampicillin, carbenicillin, and
related antibiotics"
/translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
EPELNEAIPNDERDTTMPVAMPTTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
LIKHW"
rep_origin 5426..6014
/direction=RIGHT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
protein_bind 6302..6323
/label=CAP binding site
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding activates transcription in the presence
of cAMP."
promoter 6338..6368
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 6376..6392
/label=lac operator
/bound_moiety="lac repressor encoded by lacI"
/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 6400..6416
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
promoter 6434..6452
/label=SP6 promoter
/note="promoter for bacteriophage SP6 RNA polymerase"
LOCUS Exported 6458 bp ds-DNA circular SYN 17-DEC-2018
DEFINITION Cloning vector pDC-KHT, complete sequence.
ACCESSION JF430587
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 6458)
AUTHORS Crivat G, Tokumasu F, Sa JM, Hwang J, Wellems TE.
TITLE Tetracysteine-Based Fluorescent Tags to Study Protein Localization
and Trafficking in Plasmodium falciparum-Infected Erythrocytes
JOURNAL PLoS ONE 6 (8), E22975 (2011)
PUBMED 21860664
REFERENCE 2 (bases 1 to 6458)
AUTHORS Crivat G, Tokumasu F, Sa JM, Hwang J, Wellems TE.
TITLE Direct Submission
JOURNAL Submitted (23-FEB-2011) LMVR, NIAID, 12735 Twinbrook Pkwy, 3E10C,
Rockville, MD 20852, USA
REFERENCE 3 (bases 1 to 6458)
AUTHORS .
TITLE Direct Submission
FEATURES Location/Qualifiers
source 1..6458
/organism="Cloning vector pDC-KHT"
/mol_type="other DNA"
/clone="pDC-KHT"
/note="derived from Plasmodium falciparum; transfection
plasmid expressing KAHRP followed by TC tag"
/db_xref="taxon:1035313"
CDS complement(878..895)
/codon_start=1
/product="6xHis affinity tag"
/label=6xHis
/translation="HHHHHH"
CDS complement(896..925)
/codon_start=1
/product="Myc (human c-Myc proto-oncogene) epitope tag"
/label=Myc
/translation="EQKLISEEDL"
CDS complement(941..958)
/codon_start=1
/product="tetracysteine peptide that binds biarsenical
labeling reagents"
/label=tetracysteine tag
/translation="CCPGCC"
CDS complement(995..1021)
/codon_start=1
/product="9xHis affinity tag"
/label=9xHis
/translation="HHHHHHHHH"
CDS complement(1082..1099)
/codon_start=1
/product="6xHis affinity tag"
/label=6xHis
/translation="HHHHHH"
CDS complement(1115..1132)
/codon_start=1
/product="HQHQHQ metal affinity tag"
/label=HQ tag
/note="The HQ tag binds to nickel and other metal ions much
like the 6xHis tag, but may allow elution at lower
concentrations of imidazole (Godat et al., 2008)."
/translation="HQHQHQ"
CDS complement(1121..1138)
/codon_start=1
/product="HQHQHQ metal affinity tag"
/label=HQ tag
/note="The HQ tag binds to nickel and other metal ions much
like the 6xHis tag, but may allow elution at lower
concentrations of imidazole (Godat et al., 2008)."
/translation="HQHQHQ"
CDS complement(1136..1156)
/codon_start=1
/product="6xHis affinity tag"
/label=7xHis
/translation="HHHHHHH"
promoter complement(3776..3793)
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
primer_bind complement(3800..3816)
/label=M13 fwd
/note="common sequencing primer, one of multiple similar
variants"
promoter 4290..4394
/gene="bla"
/label=AmpR promoter
CDS 4395..5255
/codon_start=1
/gene="bla"
/product="beta-lactamase"
/label=AmpR
/note="confers resistance to ampicillin, carbenicillin, and
related antibiotics"
/translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
EPELNEAIPNDERDTTMPVAMPTTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
LIKHW"
rep_origin 5426..6014
/direction=RIGHT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
protein_bind 6302..6323
/label=CAP binding site
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding activates transcription in the presence
of cAMP."
promoter 6338..6368
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 6376..6392
/label=lac operator
/bound_moiety="lac repressor encoded by lacI"
/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 6400..6416
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
promoter 6434..6452
/label=SP6 promoter
/note="promoter for bacteriophage SP6 RNA polymerase"