Basic Vector Information
GAPTrapT2AIresMuro 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 ℃.
GAPTrapT2AIresMuro vector Sequence
LOCUS 40924_1029 11810 bp DNA circular SYN 17-DEC-2018
DEFINITION Cloning vector GAPTrapT2AIresMuro, complete sequence.
ACCESSION .
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 11810)
AUTHORS Kao T, Labonne T, Niclis JC, Chaurasia R, Lokmic Z, Qian E, Bruveris
FF, Howden SE, Motazedian A, Schiesser JV, Costa M, Sourris K, Ng E,
Anderson D, Giudice A, Farlie P, Cheung M, Lamande SR, Penington AJ,
Parish CL, Thomson LH, Rafii A, Elliott DA, Elefanty AG, Stanley EG.
TITLE GAPTrap: A Simple Expression System for Pluripotent Stem Cells and
Their Derivatives
JOURNAL Stem Cell Reports (2016) In press
PUBMED 27594589
REFERENCE 2 (bases 1 to 11810)
AUTHORS Stanley E.
TITLE Direct Submission
JOURNAL Submitted (25-AUG-2016) Cell Biology, Murdoch Childrens Research
Institute, Royal Children's Hospital, Flemington Road, Melbourne,
Victoria 3052, Australia
REFERENCE 3 (bases 1 to 11810)
TITLE Direct Submission
REFERENCE 4 (bases 1 to 11810)
AUTHORS .
TITLE Direct Submission
COMMENT SGRef: number: 1; type: "Journal Article"; journalName: "Stem Cell
Reports (2016) In press"
COMMENT SGRef: number: 2; type: "Journal Article"; journalName: "Submitted
(25-AUG-2016) Cell Biology, Murdoch Childrens Research Institute,
Royal Children's Hospital, Flemington Road, Melbourne, Victoria
3052, Australia"
COMMENT SGRef: number: 3; type: "Journal Article"
COMMENT ##Assembly-Data-START##
Sequencing Technology :: Sanger dideoxy sequencing
##Assembly-Data-END##
FEATURES Location/Qualifiers
source 1..11810
/mol_type="other DNA"
/organism="synthetic DNA construct"
promoter 24..128
/label=AmpR promoter
CDS 129..986
/codon_start=1
/label=AmpR
/note="beta-lactamase"
/translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
LIKHW"
rep_origin 1160..1748
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
protein_bind 2036..2057
/label=CAP binding site
/note="CAP binding activates transcription in the presence
of cAMP."
promoter 2072..2102
/label=lac promoter
/note="promoter for the E. coli lac operon"
misc_feature 2110..2126
/label=lac operator
/note="lac operator"
protein_bind 2110..2126
/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 2134..2150
/label=M13 rev
/note="M13 rev"
promoter 2171..2189
/label=T3 promoter
/note="promoter for bacteriophage T3 RNA polymerase"
gene 2216..5559
/gene="GAPDH"
/label=GAPDH
/note="glyceraldehyde-3-phosphate dehydrogenase"
misc_feature 2219..5556
/gene="GAPDH"
/label=left arm
/note="left arm"
CDS 5581..5634
/codon_start=1
/label=T2A
/note="2A peptide from Thosea asigna virus capsid protein"
/translation="EGRGSLLTCGDVEENPGP"
misc_feature 5657..6243
/label=IRES2
/note="internal ribosome entry site (IRES) of the
encephalomyocarditis virus (EMCV)"
CDS 6262..6858
/codon_start=1
/label=PuroR
/note="puromycin N-acetyltransferase"
/translation="LTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
misc_feature 6897..11132
/label=right arm
/note="right arm"
promoter complement(11167..11185)
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
primer_bind complement(11195..11211)
/label=M13 fwd
/note="common sequencing primer, one of multiple similar
variants"
rep_origin complement(11352..11807)
/direction=LEFT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
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