Basic Vector Information
- Vector Name:
- pNM262
- Antibiotic Resistance:
- Ampicillin
- Length:
- 8497 bp
- Type:
- Cloning vector
- Replication origin:
- ori
- Source/Author:
- Gonzalez LM, Mukhitov N, Voigt CA.
pNM262 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 ℃.
pNM262 vector Sequence
LOCUS 62056_18095 8497 bp DNA circular SYN 16-DEC-2019 DEFINITION Cloning vector pNM262, complete sequence. ACCESSION MN443785 VERSION . KEYWORDS . SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 8497) AUTHORS Gonzalez LM, Mukhitov N, Voigt CA. TITLE Resilient living materials built by printing bacterial spores JOURNAL Nat. Chem. Biol. (2019) In press PUBMED 31792444 REFERENCE 2 (bases 1 to 8497) AUTHORS Mukhitov N, Gonzalez LM, Voigt CA. TITLE Direct Submission JOURNAL Submitted (11-SEP-2019) Biological Engineering, MIT, NE47-140, 500 Tech Square, Cambridge, MA 02139, USA REFERENCE 3 (bases 1 to 8497) AUTHORS . TITLE Direct Submission COMMENT SGRef: number: 1; type: "Journal Article"; journalName: "Nat. Chem. Biol. (2019) In press" COMMENT SGRef: number: 2; type: "Journal Article"; journalName: "Submitted (11-SEP-2019) Biological Engineering, MIT, NE47-140, 500 Tech Square, Cambridge, MA 02139, USA" COMMENT ##Assembly-Data-START## Sequencing Technology :: Sanger dideoxy sequencing ##Assembly-Data-END## FEATURES Location/Qualifiers source 1..8497 /mol_type="other DNA" /organism="synthetic DNA construct" promoter 106..210 /label=AmpR promoter CDS 211..1068 /label=AmpR /note="beta-lactamase" rep_origin 1242..1830 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" misc_feature 2963..4138 /label=similar to AmyE /note="similar to AmyE" gene complement(4263..5045) /gene="specR" /label=specR terminator 5341..5427 /label=rrnB T1 terminator /note="transcription terminator T1 from the E. coli rrnB gene" terminator 5446..5473 /label=rrnB T2 terminator /note="transcription terminator T2 from the E. coli rrnB gene" misc_binding 5593..5604 /label=VanO binding site /bound_moiety="VanO" regulatory 5618..5647 /label=Pspank /note="Pspank" /regulatory_class="promoter" misc_binding 5647..5658 /label=VanO binding site /bound_moiety="VanO" misc_RNA 5660..5710 /label=sTRSV HHRz /note="hammerhead ribozyme from the tobacco ringspot virus satellite RNA (Khvorova et al., 2003)" regulatory 5741..5757 /label=gsiB /note="gsiB" /regulatory_class="ribosome_binding_site" CDS 5758..6597 /codon_start=1 /transl_table=11 /product="AmyE SP-lysostaphin" /label=AmyE SP-lysostaphin /protein_id="QGV56747.1" /translation="MFAKRFKTSLLPLFAGFLLLFHLVLAGPAAASAGSTHEHSAQWLN NYKKGYGYGPYPLGINGGMHYGVDFFMNIGTPVKAISSGKIVEAGWSNYGGGNQIGLIE NDGVHRQWYMHLSKYNVKVGDYVKAGQIIGWSGSTGYSTAPHLHFQRMVNSFSNSTAQD PMPFLKSAGYGKAGGTVTPTPNTGWKTNKYGTLYKSESASFTPNTDIITRTTGPFRSMP QSGVLKAGQTIHYDEVMKQDGHVWVGYTGNSGQRIYLPVRTWNKSTNTLGVLWGTIK" terminator 6633..6727 /label=lambda t0 terminator /note="transcription terminator from phage lambda" regulatory 6835..6905 /label=Ppen /note="Ppen" /regulatory_class="promoter" regulatory 6913..6924 /label=spoVG /note="spoVG" /regulatory_class="ribosome_binding_site" CDS 6930..7664 /codon_start=1 /transl_table=11 /product="VanR-IVS" /label=VanR-IVS /protein_id="QGV56748.1" /translation="MDMPRIKPGQRVMMALRKMIASGEIKSGERIAEIPTAAALGVSRM PVRIALRSLEQEGLVVRLGARGYAARGVSSDQIRDAIEVRGVLEGFAARRLAERGMTAE THARFVVLIAEGEALFAAGRLNGEDLDRYAAYNQAFHDTLVSAAGNGAVESALARNGFE PFAAAGALALDLMDLSAEYEHLLAAHRQHQAVLDAVSCGDAEGAERIMRDHALAAIRNA KVFEAAASAGAPLGAAWSIRAD" regulatory 7788..7808 /label=hairpin terminator /note="hairpin terminator" /regulatory_class="terminator" misc_feature 7830..8497 /label=similar to AmyE /note="similar to AmyE"
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