Basic Vector Information
- Vector Name:
- pNM261
- Antibiotic Resistance:
- Ampicillin
- Length:
- 8395 bp
- Type:
- Cloning vector
- Replication origin:
- ori
- Source/Author:
- Gonzalez LM, Mukhitov N, Voigt CA.
pNM261 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 ℃.
pNM261 vector Sequence
LOCUS 62056_18090 8395 bp DNA circular SYN 16-DEC-2019
DEFINITION Cloning vector pNM261, complete sequence.
ACCESSION MN443784
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 8395)
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 8395)
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 8395)
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..8395
/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..6495
/codon_start=1
/transl_table=11
/product="lysostaphin"
/label=lysostaphin
/protein_id="QGV56745.1"
/translation="MTHEHSAQWLNNYKKGYGYGPYPLGINGGMHYGVDFFMNIGTPVK
AISSGKIVEAGWSNYGGGNQIGLIENDGVHRQWYMHLSKYNVKVGDYVKAGQIIGWSGS
TGYSTAPHLHFQRMVNSFSNSTAQDPMPFLKSAGYGKAGGTVTPTPNTGWKTNKYGTLY
KSESASFTPNTDIITRTTGPFRSMPQSGVLKAGQTIHYDEVMKQDGHVWVGYTGNSGQR
IYLPVRTWNKSTNTLGVLWGTIK"
terminator 6531..6625
/label=lambda t0 terminator
/note="transcription terminator from phage lambda"
regulatory 6733..6803
/label=Ppen
/note="Ppen"
/regulatory_class="promoter"
regulatory 6811..6827
/label=spoVG
/note="spoVG"
/regulatory_class="ribosome_binding_site"
CDS 6828..7562
/codon_start=1
/transl_table=11
/product="VanR-IVS"
/label=VanR-IVS
/protein_id="QGV56746.1"
/translation="MDMPRIKPGQRVMMALRKMIASGEIKSGERIAEIPTAAALGVSRM
PVRIALRSLEQEGLVVRLGARGYAARGVSSDQIRDAIEVRGVLEGFAARRLAERGMTAE
THARFVVLIAEGEALFAAGRLNGEDLDRYAAYNQAFHDTLVSAAGNGAVESALARNGFE
PFAAAGALALDLMDLSAEYEHLLAAHRQHQAVLDAVSCGDAEGAERIMRDHALAAIRNA
KVFEAAASAGAPLGAAWSIRAD"
regulatory 7686..7706
/label=hairpin terminator
/note="hairpin terminator"
/regulatory_class="terminator"
misc_feature 7728..8395
/label=similar to AmyE
/note="similar to AmyE"
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