Basic Vector Information
pw+SNattB 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 ℃.
pw+SNattB vector Sequence
LOCUS 40924_46153 7517 bp DNA circular SYN 18-DEC-2018 DEFINITION Transformation vector pw+SNattB, complete sequence. ACCESSION . VERSION . KEYWORDS . SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 7517) AUTHORS Koch R, Ledermann R, Urwyler O, Heller M, Suter B. TITLE Systematic functional analysis of Bicaudal-D serine phosphorylation and intragenic suppression of a female sterile allele of BicD JOURNAL PLoS ONE 4 (2), E4552 (2009) PUBMED 19234596 REFERENCE 2 (bases 1 to 7517) AUTHORS Koch R, Ledermann R, Urwyler O, Heller M, Suter B. TITLE Direct Submission JOURNAL Submitted (19-MAY-2008) Institute for Cell Biology, University of Berne, Baltzerstrasse 4, Bern 3012, Switzerland REFERENCE 3 (bases 1 to 7517) TITLE Direct Submission REFERENCE 4 (bases 1 to 7517) AUTHORS . TITLE Direct Submission COMMENT SGRef: number: 1; type: "Journal Article"; journalName: "PLoS ONE"; date: "2009"; volume: "4"; issue: "2"; pages: "E4552" COMMENT SGRef: number: 2; type: "Journal Article"; journalName: "Submitted (19-MAY-2008) Institute for Cell Biology, University of Berne, Baltzerstrasse 4, Bern 3012, Switzerland" COMMENT SGRef: number: 3; type: "Journal Article" FEATURES Location/Qualifiers source 1..7517 /mol_type="other DNA" /organism="synthetic DNA construct" rep_origin 4..459 /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind 600..616 /label=M13 fwd /note="common sequencing primer, one of multiple similar variants" promoter 623..641 /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" gene 666..4802 /label=mini-white /note="This modified version of the white gene lacks part of the first intron." protein_bind 4809..4842 /label=loxP /note="Cre-mediated recombination occurs in the 8-bp core sequence (ATGTATGC) (Shaw et al., 2021)." misc_feature 4857..5005 /label=multiple cloning site /note="multiple cloning site" protein_bind 5041..5110 /label=attB /note="attB site for the phi-C31 integrase (Groth et al., 2000)" promoter complement(5329..5347) /label=T3 promoter /note="promoter for bacteriophage T3 RNA polymerase" primer_bind complement(5368..5384) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" protein_bind complement(5392..5408) /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)." promoter complement(5416..5446) /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind complement(5461..5482) /label=CAP binding site /note="CAP binding activates transcription in the presence of cAMP." rep_origin complement(5770..6358) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" CDS complement(6532..7389) /codon_start=1 /label=AmpR /note="beta-lactamase" /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS LIKHW" promoter complement(7390..7494) /label=AmpR promoter
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