Basic Vector Information
- Vector Name:
- pEFBOS-creIRESPuro
- Antibiotic Resistance:
- Ampicillin
- Length:
- 8102 bp
- Type:
- Expression vector
- Replication origin:
- ori
- Source/Author:
- Davis RP, Ng ES, Costa M, Mossman AK, Sourris K, Elefanty AG, Stanley EG.
- Promoter:
- EF-1α
pEFBOS-creIRESPuro vector Vector Map
Plasmid Resuspension Protocol:
1. Centrifuge at 5,000×g for 5 min.
2. Carefully open the tube and add 20 μl of sterile water to dissolve the DNA.
3. Close the tube and incubate for 10 minutes at room temperature.
4. Briefly vortex the tube and then do a quick spin to concentrate the liquid at the bottom. Speed is less than 5000×g.
5.Store the plasmid at -20 ℃.
pEFBOS-creIRESPuro vector Sequence
LOCUS 40924_17089 8102 bp DNA circular SYN 17-DEC-2018
DEFINITION Expression vector pEFBOS-creIRESPuro, complete sequence.
ACCESSION .
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 8102)
AUTHORS Davis RP, Ng ES, Costa M, Mossman AK, Sourris K, Elefanty AG,
Stanley EG.
TITLE Targeting a GFP reporter gene to the MIXL1 locus of human embryonic
stem cells identifies human primitive streak-like cells and enables
isolation of primitive hematopoietic precursors
JOURNAL Blood 111 (4), 1876-1884 (2008)
PUBMED 18032708
REFERENCE 2 (bases 1 to 8102)
AUTHORS Davis RP, Costa M, Grandela C, Holland AM, Hatzistavrou T, Micallef
SJ, Li X, Goulburn AL, Azzola L, Elefanty AG, Stanley EG.
TITLE A protocol for removal of antibiotic resistance cassettes from human
embryonic stem cells genetically modified by homologous
recombination or transgenesis
JOURNAL Unpublished
REFERENCE 3 (bases 1 to 8102)
AUTHORS Davis RP, Stanley EG, Elefanty AG.
TITLE Direct Submission
JOURNAL Submitted (02-MAY-2008) MISCL, Monash University, Wellington,
Clayton, Victoria 3800, Australia
REFERENCE 4 (bases 1 to 8102)
TITLE Direct Submission
REFERENCE 5 (bases 1 to 8102)
AUTHORS .
TITLE Direct Submission
COMMENT SGRef: number: 1; type: "Journal Article"; journalName: "Blood";
date: "2008"; volume: "111"; issue: "4"; pages: "1876-1884"
COMMENT SGRef: number: 2; type: "Journal Article"; journalName:
"Unpublished"
COMMENT SGRef: number: 3; type: "Journal Article"; journalName: "Submitted
(02-MAY-2008) MISCL, Monash University, Wellington, Clayton,
Victoria 3800, Australia"
COMMENT SGRef: number: 4; type: "Journal Article"
FEATURES Location/Qualifiers
source 1..8102
/mol_type="other DNA"
/organism="synthetic DNA construct"
protein_bind 107..128
/label=CAP binding site
/note="CAP binding activates transcription in the presence
of cAMP."
promoter 143..173
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 181..197
/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 205..221
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
promoter complement(333..543)
/label=SV40 promoter
/note="SV40 early promoter"
promoter 562..1743
/label=EF-1-alpha promoter
/note="strong constitutive promoter for human elongation
factor EF-1-alpha"
CDS 1766..1786
/codon_start=1
/product="nuclear localization signal of SV40 (simian virus
40) large T antigen"
/label=SV40 NLS
/translation="PKKKRKV"
CDS 1784..2812
/codon_start=1
/label=Cre
/note="site-specific recombinase"
/translation="VSNLLTVHQNLPALPVDATSDEVRKNLMDMFRDRQAFSEHTWKML
LSVCRSWAAWCKLNNRKWFPAEPEDVRDYLLYLQARGLAVKTIQQHLGQLNMLHRRSGL
PRPSDSNAVSLVMRRIRKENVDAGERAKQALAFERTDFDQVRSLMENSDRCQDIRNLAF
LGIAYNTLLRIAEIARIRVKDISRTDGGRMLIHIGRTKTLVSTAGVEKALSLGVTKLVE
RWISVSGVADDPNNYLFCRVRKNGVAAPSATSQLSTRALEGIFEATHRLIYGAKDDSGQ
RYLAWSGHSARVGAARDMARAGVSIPEIMQAGGWTNVNIVMNYIRNLDSETGAMVRLLE
DGD"
misc_feature 2849..3435
/label=IRES2
/note="internal ribosome entry site (IRES) of the
encephalomyocarditis virus (EMCV)"
CDS 3445..4041
/codon_start=1
/label=PuroR
/note="puromycin N-acetyltransferase"
/translation="LTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
polyA_signal 4125..4349
/label=bGH poly(A) signal
/note="bovine growth hormone polyadenylation signal"
3'UTR 4522..5222
/label=bovine growth hormone 3' UTR
/note="bovine growth hormone 3' UTR"
primer_bind complement(5231..5247)
/label=M13 fwd
/note="common sequencing primer, one of multiple similar
variants"
rep_origin 5460..5915
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
promoter 6197..6301
/label=AmpR promoter
CDS 6302..7159
/codon_start=1
/label=AmpR
/note="beta-lactamase"
/translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
LIKHW"
rep_origin 7333..7921
/direction=RIGHT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
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