Basic Vector Information
- Vector Name:
- pMQ75
- Antibiotic Resistance:
- Gentamicin
- Length:
- 7353 bp
- Type:
- Shuttle/allelic-replacement vector
- Replication origin:
- p15A ori
- Source/Author:
- Shanks RM, Caiazza NC, Hinsa SM, Toutain CM, O'toole GA.
- Promoter:
- sacB
pMQ75 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 ℃.
pMQ75 vector Sequence
LOCUS 40924_32145 7353 bp DNA circular SYN 18-DEC-2018 DEFINITION Shuttle/allelic-replacement vector pMQ75, complete sequence. ACCESSION . VERSION . KEYWORDS . SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 7353) AUTHORS Shanks RM, Caiazza NC, Hinsa SM, Toutain CM, O'toole GA. TITLE Saccharomyces cerevisiae-Based Molecular Tool Kit for Manipulation of Genes from Gram-Negative Bacteria JOURNAL Appl. Environ. Microbiol. 72 (7), 5027-5036 (2006) PUBMED 16820502 REFERENCE 2 (bases 1 to 7353) AUTHORS Shanks RMQ., Caiazza NC, Hinsa SM, Toutain CM, O'Toole GA. TITLE Direct Submission JOURNAL Submitted (15-SEP-2005) Microbiology and Immunology, Dartmouth Medical School, Vail Building Room 505, Hanover, NH 03755, USA REFERENCE 3 (bases 1 to 7353) TITLE Direct Submission REFERENCE 4 (bases 1 to 7353) AUTHORS . TITLE Direct Submission COMMENT SGRef: number: 1; type: "Journal Article"; journalName: "Appl. Environ. Microbiol."; date: "2006"; volume: "72"; issue: "7"; pages: "5027-5036" COMMENT SGRef: number: 2; type: "Journal Article"; journalName: "Submitted (15-SEP-2005) Microbiology and Immunology, Dartmouth Medical School, Vail Building Room 505, Hanover, NH 03755, USA" COMMENT SGRef: number: 3; type: "Journal Article" FEATURES Location/Qualifiers source 1..7353 /mol_type="other DNA" /organism="synthetic DNA construct" CDS complement(212..1630) /codon_start=1 /label=SacB /note="secreted levansucrase that renders bacterial growth sensitive to sucrose" /translation="MNIKKFAKQATVLTFTTALLAGGATQAFAKETNQKPYKETYGISH ITRHDMLQIPEQQKNEKYQVPEFDSSTIKNISSAKGLDVWDSWPLQNADGTVANYHGYH IVFALAGDPKNADDTSIYMFYQKVGETSIDSWKNAGRVFKDSDKFDANDSILKDQTQEW SGSATFTSDGKIRLFYTDFSGKHYGKQTLTTAQVNVSASDSSLNINGVEDYKSIFDGDG KTYQNVQQFIDEGNYSSGDNHTLRDPHYVEDKGHKYLVFEANTGTEDGYQGEESLFNKA YYGKSTSFFRQESQKLLQSDKKRTAELANGALGMIELNDDYTLKKVMKPLIASNTVTDE IERANVFKMNGKWYLFTDSRGSKMTIDGITSNDIYMLGYVSNSLTGPYKPLNKTGLVLK MDLDPNDVTFTYSHFAVPQAKGNNVVITSYMTNRGFYADKQSTFAPSFLLNIKGKKTSV VKDSILEQGQLTVNK" promoter complement(1631..2076) /label=sacB promoter /note="sacB promoter and control region" oriT complement(2450..2559) /direction=LEFT /label=oriT /note="incP origin of transfer" terminator complement(2980..3007) /label=rrnB T2 terminator /note="transcription terminator T2 from the E. coli rrnB gene" terminator complement(3099..3185) /label=rrnB T1 terminator /note="transcription terminator T1 from the E. coli rrnB gene" primer_bind 3535..3551 /label=M13 fwd /note="common sequencing primer, one of multiple similar variants" misc_feature complement(3555..3611) /label=MCS /note="pUC18/19 multiple cloning site" primer_bind complement(3621..3637) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" protein_bind complement(3645..3661) /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(3669..3699) /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind complement(3714..3735) /label=CAP binding site /note="CAP binding activates transcription in the presence of cAMP." rep_origin 3862..4406 /label=p15A ori /note="Plasmids containing the medium-copy-number p15A origin of replication can be propagated in E. coli cells that contain a second plasmid with the ColE1 origin." CDS complement(4766..5296) /codon_start=1 /label=GmR /note="gentamycin acetyltransferase" /translation="MLRSSNDVTQQGSRPKTKLGGSSMGIIRTCRLGPDQVKSMRAALD LFGREFGDVATYSQHQPDSDYLGNLLRSKTFIALAAFDQEAVVGALAAYVLPRFEQPRS EIYIYDLAVSGEHRRQGIATALINLLKHEANALGAYVIYVQADYGDDPAVALYTKLGIR EEVMHFDIDPSTAT" promoter complement(5485..5513) /label=Pc promoter /note="class 1 integron promoter" promoter complement(5531..5635) /label=AmpR promoter CDS complement(5735..6535) /codon_start=1 /label=URA3 /note="orotidine-5'-phosphate decarboxylase, required for uracil biosynthesis" /translation="MSKATYKERAATHPSPVAAKLFNIMHEKQTNLCASLDVRTTKELL ELVEALGPKICLLKTHVDILTDFSMEGTVKPLKALSAKYNFLLFEDRKFADIGNTVKLQ YSAGVYRIAEWADITNAHGVVGPGIVSGLKQAAEEVTKEPRGLLMLAELSCKGSLSTGE YTKGTVDIAKSDKDFVIGFIAQRDMGGRDEGYDWLIMTPGVGLDDKGDALGQQYRTVDD VVSTGSDIIIVGRGLFAKGRDAKVEGERYRKAGWEAYLRRCGQQN" promoter complement(6536..6756) /label=URA3 promoter misc_feature 6784..7287 /label=CEN/ARS /note="S. cerevisiae CEN6 centromere fused to an autonomously replicating sequence"
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