LOCUS       Exported                3182 bp ds-DNA     circular SYN 18-DEC-2018
DEFINITION  Cloning vector pToxin-ecoRI, complete sequence.
ACCESSION   KT895276
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3182)
  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 3182)
  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 3182)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     ##Assembly-Data-START##
            Sequencing Technology :: Sanger dideoxy sequencing 
            ##Assembly-Data-END##
FEATURES             Location/Qualifiers
     source          1..3182
                     /organism="Cloning vector pToxin-ecoRI"
                     /mol_type="other DNA"
                     /db_xref="taxon:1758745"
     terminator      14..108
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     regulatory      115..189
                     /regulatory_class="promoter"
                     /note="T0"
     regulatory      195..214
                     /regulatory_class="ribosome_binding_site"
                     /note="RBS"
     CDS             221..1054
                     /codon_start=1
                     /product="type-2 restriction enzyme EcoRI"
                     /label=type-2 restriction enzyme EcoRI
                     /protein_id="ALP32111.1"
                     /translation="MSNKKQSNRLTEQHKLSQGVIGIFGDYAKAHDLAVGEVSKLVKKA
                     LSNEYPQLSFRYRDSIKKTEINEALKKIDPDLGGTLFVSNSSIKPDGGIVEVKDDYGEW
                     RVVLVAEAKHQGKDIINIRNGLLVGKRGDQDLMAAGNAIERSHKNISEIANFMLSESHF
                     PYVLFLEGSNFLTENISITRPDGRVVNLEYNSGILNRLDRLTAANYGMPINSNLCINKF
                     VNHKDKSIMLQAASIYTQGDGREWDSKIMFEIMFDISTTSLRVLGRDLFEQLTSK"
     regulatory      1074..1133
                     /regulatory_class="terminator"
                     /note="T1"
     terminator      1079..1165
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     rep_origin      1185..1981
                     /label=p15A
                     /note="p15A"
     rep_origin      1286..1831
                     /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      1993..2087
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             complement(2117..2977)
                     /codon_start=1
                     /product="beta-lactmase"
                     /label=beta-lactmase
                     /protein_id="ALP32112.1"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     promoter        complement(2978..3082)
                     /gene="bla"
                     /label=AmpR promoter