Basic Vector Information
| Basic Vector Information | |||
|---|---|---|---|
| Vector Name | pToxin-mfLON-ecoRI | Length | 5729 bp |
| Type | Cloning vector | Source | Chan CT, Lee JW, Cameron DE, Bashor CJ, Collins JJ. |
pToxin-mfLON-ecoRI 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-ecoRI vector Sequence
LOCUS Exported 5729 bp ds-DNA circular SYN 18-DEC-2018
DEFINITION Cloning vector pToxin-mfLON-ecoRI, complete sequence.
ACCESSION KT895277
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 5729)
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 5729)
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 5729)
AUTHORS .
TITLE Direct Submission
COMMENT ##Assembly-Data-START##
Sequencing Technology :: Sanger dideoxy sequencing
##Assembly-Data-END##
FEATURES Location/Qualifiers
source 1..5729
/organism="Cloning vector pToxin-mfLON-ecoRI"
/mol_type="other DNA"
/db_xref="taxon:1758747"
regulatory 7..54
/regulatory_class="promoter"
/note="pLscrO-2"
regulatory 81..101
/regulatory_class="ribosome_binding_site"
/note="RBS"
CDS 103..2547
/codon_start=1
/product="mfLON protease"
/label=mfLON protease
/protein_id="ALP32113.1"
/translation="MSKKIKLPIFQIRGSFIVPGIKENLEVGRKNTLASVNYAIKNSNN
QMIAIPQIDASVEKPEFSDLHEFGILIDFEVIKEWKDNSLTISTNPIQRCKVISFFENE
DQVPYAEVELIESINDFSDEELKELIEKISDAIKTKASLVTKQIKQLISGESDDLSLAF
DSIMFKLAPSKILTNPEYITSPSLKTRWSIIEKIIFAEDGIITRNAESIDAARQKNEIE
QELNHKLKEKMDKQQKEYYLREKMRIIKDELEDEDDSDDSSLEKYKERLAKEPFPEEVK
RKIMASIKRVEALQSGTPEWNTEKNYIDWMMSIPWWEETEDLTDLKYAKKILDKHHYGM
KKVKERIIEYLAVKTKTKSLKAPIITLVGPPGVGKTSLAKSIAEAVGKNFVKVSLGGVK
DESEIRGHRKTYVGSMPGRIIQTMKRAKVKNPLFLLDEIDKMASDHRGDPASAMLEVLD
PEQNKEFSDHYIEEPYDLSQVMFIATANYPEDIPEALYDRMEIINLSSYTEIEKVKIAQ
DYLVPKAIEQHELTSEEISFTEGAINEIIKYYTREAGVRQLERHINSIIRKYIVKNLNG
EMDKIVIDEKQVNDLLGKRIFDHTEKQEESQIGVVTGLAYTQFGGDILPIEVSLYPGKG
NLILTGKLGEVMKESATIALTYVKSNFEKFGVDKKVFEENDIHVHVPEGAVPKDGPSAG
ITITTALISALSDKPVSKEIGMTGEITLRGNVLPIGGLREKSISASRSGLKTIIIPKKN
ERDLDEIPDEVKAKLKIIPAEKYEEVFAIVFKTKAANKNEENTNEVPTFMLNAGQANRR
RV"
regulatory 2561..2655
/regulatory_class="terminator"
/note="T0"
terminator 2561..2655
/label=lambda t0 terminator
/note="transcription terminator from phage lambda"
regulatory 2662..2709
/regulatory_class="promoter"
/note="pLscrO-2"
regulatory 2742..2761
/regulatory_class="ribosome_binding_site"
/note="RBS"
CDS 2768..3601
/codon_start=1
/product="type-2 restriction enzyme EcoRI"
/label=type-2 restriction enzyme EcoRI
/protein_id="ALP32114.1"
/translation="MSNKKQSNRLTEQHKLSQGVIGIFGDYAKAHDLAVGEVSKLVKKA
LSNEYPQLSFRYRDSIKKTEINEALKKIDPDLGGTLFVSNSSIKPDGGIVEVKDDYGEW
RVVLVAEAKHQGKDIINIRNGLLVGKRGDQDLMAAGNAIERSHKNISEIANFMLSESHF
PYVLFLEGSNFLTENISITRPDGRVVNLEYNSGILNRLDRLTAANYGMPINSNLCINKF
VNHKDKSIMLQAASIYTQGDGREWDSKIMFEIMFDISTTSLRVLGRDLFEQLTSK"
regulatory 3621..3680
/regulatory_class="terminator"
/note="T1"
terminator 3626..3712
/gene="Escherichia coli rrnB"
/label=rrnB T1 terminator
/note="transcription terminator T1 from the E. coli rrnB
gene"
rep_origin 3732..4528
/label=p15A
/note="p15A"
rep_origin 3833..4378
/direction=RIGHT
/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 4540..4634
/label=lambda t0 terminator
/note="transcription terminator from phage lambda"
CDS complement(4664..5524)
/codon_start=1
/product="beta-lactmase"
/label=beta-lactmase
/protein_id="ALP32115.1"
/translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
LIKHW"
promoter complement(5525..5629)
/gene="bla"
/label=AmpR promoter
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