LOCUS       Exported                5886 bp ds-DNA     circular SYN 18-DEC-2018
DEFINITION  Cloning vector pPasscode3, complete sequence.
ACCESSION   KT895274
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5886)
  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 5886)
  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 5886)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     ##Assembly-Data-START##
            Sequencing Technology :: Sanger dideoxy sequencing 
            ##Assembly-Data-END##
FEATURES             Location/Qualifiers
     source          1..5886
                     /organism="Cloning vector pPasscode3"
                     /mol_type="other DNA"
                     /db_xref="taxon:1758744"
     promoter        42..115
                     /label=PLtetO-1 promoter
                     /note="modified phage lambda PL promoter with tet operator 
                     sites (Lutz and Bujard, 1997)"
     regulatory      42..89
                     /regulatory_class="promoter"
                     /note="pLtetO-1"
     protein_bind    42..60
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    67..85
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     regulatory      96..121
                     /regulatory_class="ribosome_binding_site"
                     /note="RBS"
     CDS             122..1159
                     /codon_start=1
                     /product="GalR-LacI"
                     /label=GalR-LacI
                     /protein_id="ALP32105.1"
                     /translation="MKPVTLYDVAEYAGVSYQTVSRVVNQASHVSAKTREKVEAAMESL
                     SYHPNANARALAQQTTETVGLVVGDVSDPFFGAMVKAVEQVAYHTGNFLLIGNGYHNEQ
                     KERQAIEQLIRHRCAALVVHAKMIPDADLASLMKQMPGMVLINRILPGFENRCIALDDR
                     YGAWLATRHLIQQGHTRIGYLCSNHSISDAEDRLQGYYDALAESGIAANDRLVTFGEPD
                     ESGGEQAMTELLGRGRNFTAVACYNDSMAAGAMGVLNDNGIDVPGEISLIGFDDVLVSR
                     YVRPRLTTVRYPIVTMATQAAELALALADNRPLPEITNVFSPTLVRRHSVSTPSLEASH
                     HATSD"
     regulatory      1180..1274
                     /regulatory_class="terminator"
                     /note="T0"
     terminator      1180..1274
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     promoter        1323..1396
                     /label=PLtetO-1 promoter
                     /note="modified phage lambda PL promoter with tet operator 
                     sites (Lutz and Bujard, 1997)"
     regulatory      1323..1370
                     /regulatory_class="promoter"
                     /note="pLtetO-1"
     protein_bind    1323..1341
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    1348..1366
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     regulatory      1377..1402
                     /regulatory_class="ribosome_binding_site"
                     /note="RBS"
     CDS             1403..2485
                     /codon_start=1
                     /product="LacI"
                     /label=LacI
                     /protein_id="ALP32106.1"
                     /translation="MKPVTLYDVAEYAGVSYQTVSRVVNQASHVSAKTREKVEAAMAEL
                     NYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSRADQLGASVVVSMVERSGV
                     EACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSIIFSH
                     EDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREGDWSA
                     MSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTEDSSC
                     YIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR
                     ALADSLMQLARQVSRLESGQ"
     regulatory      2506..2600
                     /regulatory_class="terminator"
                     /note="T0"
     terminator      2506..2600
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     regulatory      complement(2640..2734)
                     /regulatory_class="terminator"
                     /note="T0"
     terminator      2640..2734
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             complement(2755..3783)
                     /codon_start=1
                     /product="CelR-ScrR"
                     /label=CelR-ScrR
                     /protein_id="ALP32107.1"
                     /translation="MKTKRVTIKDIAELAGVSKATASLVLNGRGKELRVAQETRERVLA
                     IIKELGYVPNRAARTLVTRRTDTVALVVSENNQKLFAEPFYAGIVLGVGVALSERGFQF
                     VLATGRSGIEHERLGGYLAGQHVDGVLLLSLHRDDPLPQMLDEAGVPYVYGGRPLGVPE
                     EQVSYVDIDNIGGGRQATQRLIETGHRRIATIAGPQDMVAGVERLQGYREALLAAGMEY
                     DETLVSYGDFTYDSGVAAMRELLDRAPDVDAVFAASDLMGLAALRVLRASGRRVPEDVA
                     VVGYDDSTVAEHAEPPMTSVNQPTELMGREMARLLVDRITGETTEPVRLVLETHLMVRE
                     SG"
     regulatory      complement(3784..3809)
                     /regulatory_class="ribosome_binding_site"
                     /note="RBS"
     regulatory      complement(3841..3869)
                     /regulatory_class="promoter"
                     /note="pLlacO"
     regulatory      3900..4004
                     /regulatory_class="terminator"
                     /note="T1"
     terminator      3905..3991
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     rep_origin      4011..4818
                     /label=ColE1
                     /note="ColE1"
     rep_origin      complement(4155..4743)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     regulatory      complement(4825..4930)
                     /regulatory_class="terminator"
                     /note="T0"
     terminator      4831..4925
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             complement(4956..5750)
                     /codon_start=1
                     /product="kanamycin resistance protein"
                     /label=kanamycin resistance protein
                     /protein_id="ALP32108.1"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"