Basic Vector Information
- Vector Name:
- pNOV076
- Antibiotic Resistance:
- Gentamycin
- Length:
- 8181 bp
- Type:
- Cloning vector
- Replication origin:
- ori
- Source/Author:
- Jones AK, Woods AL, Takeoka KT, Shen X, Wei J-R., Caughlan RE, Dean CR.
pNOV076 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 ℃.
pNOV076 vector Sequence
LOCUS 40924_33442 8181 bp DNA circular SYN 18-DEC-2018 DEFINITION Cloning vector pNOV076, complete sequence. ACCESSION . VERSION . KEYWORDS . SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 8181) AUTHORS Jones AK, Woods AL, Takeoka KT, Shen X, Wei J-R., Caughlan RE, Dean CR. TITLE Determinants of antibacterial spectrum and resistance potential of the elongation factor G inhibitor argyrin B in key Gram negative pathogens JOURNAL Unpublished REFERENCE 2 (bases 1 to 8181) AUTHORS Wei J-R. TITLE Direct Submission JOURNAL Submitted (12-OCT-2016) Infectious Diseases, Bacteriology, Novartis Institutes for Biomedical Research, 5300 Chiron Way, Emeryville, CA 94608, USA REFERENCE 3 (bases 1 to 8181) TITLE Direct Submission REFERENCE 4 (bases 1 to 8181) AUTHORS . TITLE Direct Submission COMMENT SGRef: number: 1; type: "Journal Article"; journalName: "Unpublished" COMMENT SGRef: number: 2; type: "Journal Article"; journalName: "Submitted (12-OCT-2016) Infectious Diseases, Bacteriology, Novartis Institutes for Biomedical Research, 5300 Chiron Way, Emeryville, CA 94608, USA" COMMENT SGRef: number: 3; type: "Journal Article" FEATURES Location/Qualifiers source 1..8181 /mol_type="other DNA" /organism="synthetic DNA construct" rep_origin 7..1337 CDS complement(1474..2553) /label=lacI /note="lac repressor" promoter complement(2554..2631) /label=lacIq promoter /note="In the lacIq allele, a single base change in the promoter boosts expression of the lacI gene about 10-fold." promoter 2861..2889 /label=tac promoter /note="strong E. coli promoter; hybrid between the trp and lac UV5 promoters" protein_bind 2897..2913 /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)." CDS 2936..5056 /codon_start=1 /gene="fusA" /product="elongation factor G" /label=fusA /note="S459F" /protein_id="APS85742.1" /translation="MARTTPINRYRNIGICAHVDAGKTTTTERVLFYTGVNHKLGEVHD GAATTDWMVQEQERGITITSAAVTTFWKGSRGQYDNYRVNVIDTPGHVDFTIEVERSLR VLDGAVVVFCGTSGVEPQSETVWRQANKYGVPRIVYVNKMDRQGANFLRVVEQIKKRLG HTPVPVQLAIGAEENFVGQVDLIKMKAIYWNDDDKGMTYREEEIPAELKDLAEEWRSSM VEAAAEANEELMNKYLEEGELSEAEIKEGLRLRTLACEIVPAVCGSSFKNKGVPLVLDA VIDYLPAPTEIPAIKGVSPDDETVEDERHADDNEPFSSLAFKIATDPFVGTLTFARVYS GVLSSGDSVLNSVKGKKERVGRMVQMHANQREEIKEVRAGDIAALIGMKDVTTGDTLCS IEKPIILERMDFPEPVISVAVEPKTKADQEKMGIALGKLAQEDPSFRVKTDEESGQTII FGMGELHLDIIVDRMKREFGVEANIGKPQVAYRETITKDNVEIEGKFVRQSGGRGQFGH CWIRFSAADVDEKGNITEGLVFENEVVGGVVPKEYIPAIQKGIEEQMKNGVVAGYPLIG LKATVFDGSYHDVDSNEMAFKIAASMATKQLAQKGGGKVLEPIMKVEVVTPEDYMGDVM GDLNRRRGLIQGMEDTVSGKVIRAEVPLGEMFGYATDVRSMSQGRASYSMEFSKYAEAP SNIVEALVKKQG" gene 2936..5056 /gene="fusA" /label=fusA /note="S459F" primer_bind complement(5066..5082) /label=SK primer /note="common sequencing primer, one of multiple similar variants" terminator 5342..5428 /label=rrnB T1 terminator /note="transcription terminator T1 from the E. coli rrnB gene" terminator 5520..5547 /label=rrnB T2 terminator /note="transcription terminator T2 from the E. coli rrnB gene" promoter 5579..5607 /label=Pc promoter /note="class 1 integron promoter" CDS 5796..6326 /label=GmR /note="gentamycin acetyltransferase" rep_origin 6351..6806 /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" rep_origin 6917..7505 /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" misc_feature complement(7691..7831) /label=bom /note="basis of mobility region from pBR322"
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