Basic Vector Information
- Vector Name:
- pIR3HYG-GW (a)-GFP+ (b)
- Antibiotic Resistance:
- Ampicillin
- Length:
- 13417 bp
- Type:
- Leishmania vector
- Replication origin:
- ori
- Source/Author:
- Brettmann EA, Lye LF, Beverley SM.
- Promoter:
- lac UV5
pIR3HYG-GW (a)-GFP+ (b) vector Vector Map
pIR3HYG-GW (a)-GFP+ (b) vector Sequence
LOCUS 40924_25646 13417 bp DNA circular SYN 18-DEC-2018
DEFINITION Leishmania vector pIR3HYG-GW (a)-GFP+ (b), complete sequence.
ACCESSION .
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 13417)
AUTHORS Brettmann EA, Lye LF, Beverley SM.
TITLE Spontaneous excision and facilitated recovery as a control for
phenotypes arising from RNA interference and other dominant
transgenes
JOURNAL Mol. Biochem. Parasitol. (2018) In press
PUBMED 29357296
REFERENCE 2 (bases 1 to 13417)
AUTHORS Brettmann EA, Beverley SM, Lye L-F.
TITLE Direct Submission
JOURNAL Submitted (28-DEC-2017) Department of Molecular Microbiology,
Washington University School of Medicine, 660 S. Euclid Ave, Campus
Box 8230, St. Louis, MO 63110, USA
REFERENCE 3 (bases 1 to 13417)
TITLE Direct Submission
REFERENCE 4 (bases 1 to 13417)
AUTHORS .
TITLE Direct Submission
COMMENT SGRef: number: 1; type: "Journal Article"; journalName: "Mol.
Biochem. Parasitol. (2018) In press"
COMMENT SGRef: number: 2; type: "Journal Article"; journalName: "Submitted
(28-DEC-2017) Department of Molecular Microbiology, Washington
University School of Medicine, 660 S. Euclid Ave, Campus Box 8230,
St. Louis, MO 63110, USA"
COMMENT SGRef: number: 3; type: "Journal Article"
COMMENT ##Assembly-Data-START##
Assembly Method :: Lasergene v. 12
Sequencing Technology :: Sanger dideoxy sequencing; 454
##Assembly-Data-END##
FEATURES Location/Qualifiers
source 1..13417
/mol_type="other DNA"
/organism="synthetic DNA construct"
protein_bind 25..149
/label=attR2
/note="recombination site for the Gateway(R) LR reaction"
CDS complement(193..495)
/label=ccdB
/note="CcdB, a bacterial toxin that poisons DNA gyrase"
CDS complement(840..1496)
/label=CmR
/note="chloramphenicol acetyltransferase"
promoter complement(1550..1580)
/label=lac UV5 promoter
/note="E. coli lac promoter with an 'up' mutation"
protein_bind complement(1610..1729)
/label=attR1
/note="recombination site for the Gateway(R) LR reaction"
misc_feature 1741..2189
/label=MusPEX11- MusPEX11 loop sequences
/note="MusPEX11- MusPEX11 loop sequences"
CDS 2204..2233
/label=Myc
/note="Myc (human c-Myc proto-oncogene) epitope tag"
CDS 2240..2269
/codon_start=1
/product="Myc (human c-Myc proto-oncogene) epitope tag"
/label=Myc
/translation="EQKLISEEDL"
CDS 2276..2305
/codon_start=1
/product="Myc (human c-Myc proto-oncogene) epitope tag"
/label=Myc
/translation="EQKLISEEDL"
protein_bind 2321..2440
/label=attR1
/note="recombination site for the Gateway(R) LR reaction"
promoter 2470..2500
/label=lac UV5 promoter
/note="E. coli lac promoter with an 'up' mutation"
CDS 2554..3210
/label=CmR
/note="chloramphenicol acetyltransferase"
CDS 3555..3857
/label=ccdB
/note="CcdB, a bacterial toxin that poisons DNA gyrase"
protein_bind complement(3901..4025)
/label=attR2
/note="recombination site for the Gateway(R) LR reaction"
promoter complement(4227..4245)
/label=T3 promoter
/note="promoter for bacteriophage T3 RNA polymerase"
misc_feature 4322..5043
/note="alpha tub IR- Leishmania braziliensis strain M2903
alpha-tubulin intergenic region"
misc_feature 4973..4974
/label=SA- spice acceptor AG dinucleotide
/note="SA- spice acceptor AG dinucleotide"
CDS 5062..5778
/label=EGFP
/note="enhanced GFP"
promoter complement(5974..5992)
/label=T3 promoter
/note="promoter for bacteriophage T3 RNA polymerase"
misc_feature 6059..6607
/note="LPG1 IR- Leishmania donovani strain 1S2D LPG1
intergenic region"
misc_feature 6561..6562
/label=SA- spice acceptor AG dinucleotide
/note="SA- spice acceptor AG dinucleotide"
RBS 6619..6641
/label=RBS
/note="efficient ribosome binding site from bacteriophage
T7 gene 10 (Olins and Rangwala, 1989)"
CDS 6667..7689
/label=HygR
/note="aminoglycoside phosphotransferase from E. coli"
misc_feature 7728..8299
/note="1.7K IR- Leishmania major strain LT252 1.7K
intergenic region"
misc_feature 8296..9381
/note="3' SSU- segment containing 3' region of Leishmania
major strain Friedlin small subunit ribosomal RNA"
promoter complement(9398..9416)
/label=T3 promoter
/note="promoter for bacteriophage T3 RNA polymerase"
primer_bind complement(9437..9453)
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
protein_bind 9461..9477
/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)."
promoter complement(9485..9515)
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind complement(9530..9551)
/label=CAP binding site
/note="CAP binding activates transcription in the presence
of cAMP."
rep_origin complement(9839..10427)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
CDS complement(10601..11458)
/label=AmpR
/note="beta-lactamase"
promoter complement(11459..11563)
/label=AmpR promoter
rep_origin 11590..12045
/direction=RIGHT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
primer_bind 12186..12202
/label=M13 fwd
/note="common sequencing primer, one of multiple similar
variants"
promoter 12212..12230
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
misc_feature 12244..13156
/note="5' SSU- segment containing 5' region of Leishmania
major strain Friedlin small subunit ribosomal RNA"
misc_feature 13185..13417
/note="DST IR- Leishmania major strain DST intergenic
region"
misc_feature 13340..13341
/label=SA- spice acceptor AG dinucleotide
/note="SA- spice acceptor AG dinucleotide"
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