Basic Vector Information
GAPTrapT2AIresMeo 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 ℃.
GAPTrapT2AIresMeo vector Sequence
LOCUS Exported 12005 bp ds-DNA circular SYN 17-DEC-2018 DEFINITION Cloning vector GAPTrapT2AIresMeo, complete sequence. ACCESSION KX783039 VERSION . KEYWORDS . SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 12005) 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 12005) AUTHORS Stanley EG, Elefanty AG, Kao T, Labonne T. 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 12005) AUTHORS . TITLE Direct Submission COMMENT ##Assembly-Data-START## Sequencing Technology :: Sanger dideoxy sequencing ##Assembly-Data-END## FEATURES Location/Qualifiers source 1..12005 /organism="Cloning vector GAPTrapT2AIresMeo" /mol_type="other DNA" /db_xref="taxon:1899385" gene 24..989 /gene="ampR" /label=ampR /note="codes ampicillin resistance" regulatory 24..128 /regulatory_class="promoter" /gene="ampR" promoter 24..128 /gene="bla" /label=AmpR promoter CDS 129..989 /codon_start=1 /gene="ampR" /product="AmpR" /label=ampR /protein_id="AOO95911.1" /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS LIKHW" rep_origin 1160..11748 rep_origin 1160..1748 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" protein_bind 2036..2057 /label=CAP binding site /bound_moiety="CAP" regulatory 2072..2102 /regulatory_class="promoter" /note="lac promoter" 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" misc_feature 2146..2200 /label=lacZ-alpha /note="lacZ-alpha" regulatory 2171..2189 /regulatory_class="promoter" /note="T3 promoter" 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" misc_feature 5560..5658 /label=T2A /note="T2A" CDS 5584..5637 /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" misc_feature 5660..6246 /label=IRES2 /note="internal ribosome entry site (IRES) of the encephalomyocarditis virus (EMCV)" regulatory 5684..6246 /regulatory_class="attenuator" /note="IRES" CDS 6247..7059 /codon_start=1 /product="Meo" /label=Meo /note="codes for G418 resistance" /protein_id="AOO95910.1" /translation="MARMARGIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRL SAQGRPVLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVP GQDLLSSHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDD LDEEHQGLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVAD RYQDIALATRDIAEELCGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF" misc_feature 7089..11324 /label=right arm /note="right arm" misc_feature 11348..12005 /label=pBluescript II KS(-) /note="pBluescript II KS(-)" promoter complement(11362..11380) /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" primer_bind complement(11390..11406) /label=M13 fwd /note="common sequencing primer, one of multiple similar variants" rep_origin complement(11547..12002) /direction=LEFT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis"
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