Basic Vector Information
p34S-Km3 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 ℃.
p34S-Km3 vector Sequence
LOCUS 40924_2592 3714 bp DNA circular SYN 18-DEC-2018 DEFINITION Cloning vector p34S-Km3, complete sequence. ACCESSION . VERSION . KEYWORDS . SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 3714) AUTHORS Dennis JJ, Zylstra GJ. TITLE Plasposons: modular self-cloning minitransposon derivatives for rapid genetic analysis of gram-negative bacterial genomes JOURNAL Appl. Environ. Microbiol. 64 (7), 2710-2715 (1998) PUBMED 9647854 REFERENCE 2 (bases 1 to 3714) AUTHORS Dennis JJ, Zylstra GJ. TITLE Improved antibiotic-resistance cassettes through restriction site elimination using Pfu DNA polymerase PCR JOURNAL BioTechniques 25 (5), 772-774 (1998) PUBMED 9821576 REFERENCE 3 (bases 1 to 3714) AUTHORS Dennis JJ, Zylstra GJ. TITLE Direct Submission JOURNAL Submitted (12-AUG-1998) Biotechnology Center for Agriculture and the Environment, Cook College, Rutgers University, 59 Dudley Rd., New Brunswick, NJ 08901, USA REFERENCE 4 (bases 1 to 3714) TITLE Direct Submission REFERENCE 5 (bases 1 to 3714) AUTHORS . TITLE Direct Submission COMMENT SGRef: number: 1; type: "Journal Article"; journalName: "Appl. Environ. Microbiol."; date: "1998"; volume: "64"; issue: "7"; pages: "2710-2715" COMMENT SGRef: number: 2; type: "Journal Article"; journalName: "BioTechniques"; date: "1998"; volume: "25"; issue: "5"; pages: "772-774" COMMENT SGRef: number: 3; type: "Journal Article"; journalName: "Submitted (12-AUG-1998) Biotechnology Center for Agriculture and the Environment, Cook College, Rutgers University, 59 Dudley Rd., New Brunswick, NJ 08901, USA" COMMENT SGRef: number: 4; type: "Journal Article" FEATURES Location/Qualifiers source 1..3714 /mol_type="other DNA" /organism="synthetic DNA construct" misc_feature 1..51 /label=multiple cloning site cassette /note="multiple cloning site cassette" CDS 162..974 /codon_start=1 /label=KanR /note="aminoglycoside phosphotransferase" /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDEAWLLTTAIPGK TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF" misc_feature 978..1028 /label=multiple cloning site cassette /note="multiple cloning site cassette" promoter complement(1034..1052) /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" primer_bind complement(1071..1087) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" protein_bind complement(1095..1111) /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(1119..1149) /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind complement(1164..1185) /label=CAP binding site /note="CAP binding activates transcription in the presence of cAMP." rep_origin complement(1473..2061) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" CDS complement(2235..3092) /codon_start=1 /label=AmpR /note="beta-lactamase" /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW EPELNEAIPNDERDTTMPVAMPTTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS LIKHW" promoter complement(3093..3197) /label=AmpR promoter primer_bind 3671..3687 /label=M13 fwd /note="common sequencing primer, one of multiple similar variants" promoter 3690..3708 /label=SP6 promoter /note="promoter for bacteriophage SP6 RNA polymerase"
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