Basic Vector Information
- Vector Name:
- pToxin-mfLON
- Antibiotic Resistance:
- Ampicillin
- Length:
- 4692 bp
- Type:
- Cloning vector
- Replication origin:
- p15A ori
- Source/Author:
- Chan CT, Lee JW, Cameron DE, Bashor CJ, Collins JJ.
pToxin-mfLON 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 ℃.
pToxin-mfLON vector Sequence
LOCUS 40924_43813 4692 bp DNA circular SYN 18-DEC-2018
DEFINITION Cloning vector pToxin-mfLON, complete sequence.
ACCESSION .
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 4692)
AUTHORS Chan CT, Lee JW, Cameron DE, Bashor CJ, Collins JJ.
TITLE 'Deadman' and 'Passcode' microbial kill switches for bacterial
containment
JOURNAL Nat. Chem. Biol. 12 (2), 82-86 (2016)
PUBMED 26641934
REFERENCE 2 (bases 1 to 4692)
AUTHORS Chan CTY., Lee JW, Cameron DE, Bashor CJ, Collins JJ.
TITLE Direct Submission
JOURNAL Submitted (12-OCT-2015) Department of Biological Engineering,
Massachusetts Institute of Technology, 77 Massachusetts Ave.,
E25-302b, Cambridge, MA 02139, USA
REFERENCE 3 (bases 1 to 4692)
TITLE Direct Submission
REFERENCE 4 (bases 1 to 4692)
AUTHORS .
TITLE Direct Submission
COMMENT SGRef: number: 1; type: "Journal Article"; journalName: "Nat. Chem.
Biol."; date: "2016"; volume: "12"; issue: "2"; pages: "82-86"
COMMENT SGRef: number: 2; type: "Journal Article"; journalName: "Submitted
(12-OCT-2015) Department of Biological Engineering, Massachusetts
Institute of Technology, 77 Massachusetts Ave., E25-302b, Cambridge,
MA 02139, USA"
COMMENT SGRef: number: 3; type: "Journal Article"
COMMENT ##Assembly-Data-START##
Sequencing Technology :: Sanger dideoxy sequencing
##Assembly-Data-END##
FEATURES Location/Qualifiers
source 1..4692
/mol_type="other DNA"
/organism="synthetic DNA construct"
regulatory 7..80
/label=pLscrO-2
/note="pLscrO-2"
/regulatory_class="promoter"
regulatory 81..102
/label=RBS
/note="RBS"
/regulatory_class="ribosome_binding_site"
CDS 103..2547
/codon_start=1
/product="mfLON protease"
/label=mfLON protease
/protein_id="ALP32109.1"
/translation="MSKKIKLPIFQIRGSFIVPGIKENLEVGRKNTLASVNYAIKNSNN
QMIAIPQIDASVEKPEFSDLHEFGILIDFEVIKEWKDNSLTISTNPIQRCKVISFFENE
DQVPYAEVELIESINDFSDEELKELIEKISDAIKTKASLVTKQIKQLISGESDDLSLAF
DSIMFKLAPSKILTNPEYITSPSLKTRWSIIEKIIFAEDGIITRNAESIDAARQKNEIE
QELNHKLKEKMDKQQKEYYLREKMRIIKDELEDEDDSDDSSLEKYKERLAKEPFPEEVK
RKIMASIKRVEALQSGTPEWNTEKNYIDWMMSIPWWEETEDLTDLKYAKKILDKHHYGM
KKVKERIIEYLAVKTKTKSLKAPIITLVGPPGVGKTSLAKSIAEAVGKNFVKVSLGGVK
DESEIRGHRKTYVGSMPGRIIQTMKRAKVKNPLFLLDEIDKMASDHRGDPASAMLEVLD
PEQNKEFSDHYIEEPYDLSQVMFIATANYPEDIPEALYDRMEIINLSSYTEIEKVKIAQ
DYLVPKAIEQHELTSEEISFTEGAINEIIKYYTREAGVRQLERHINSIIRKYIVKNLNG
EMDKIVIDEKQVNDLLGKRIFDHTEKQEESQIGVVTGLAYTQFGGDILPIEVSLYPGKG
NLILTGKLGEVMKESATIALTYVKSNFEKFGVDKKVFEENDIHVHVPEGAVPKDGPSAG
ITITTALISALSDKPVSKEIGMTGEITLRGNVLPIGGLREKSISASRSGLKTIIIPKKN
ERDLDEIPDEVKAKLKIIPAEKYEEVFAIVFKTKAANKNEENTNEVPTFMLNAGQANRR
RV"
terminator 2589..2675
/label=rrnB T1 terminator
/note="transcription terminator T1 from the E. coli rrnB
gene"
rep_origin 2796..3341
/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."
terminator complement(3503..3597)
/label=lambda t0 terminator
/note="transcription terminator from phage lambda"
CDS complement(3630..4487)
/label=AmpR
/note="beta-lactamase"
promoter complement(4488..4592)
/label=AmpR promoter
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