LOCUS       Exported                5887 bp ds-DNA     circular SYN 18-DEC-2018
DEFINITION  Cloning vector pPasscode2, complete sequence.
ACCESSION   KT895273
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5887)
  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 5887)
  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 5887)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     ##Assembly-Data-START##
            Sequencing Technology :: Sanger dideoxy sequencing 
            ##Assembly-Data-END##
FEATURES             Location/Qualifiers
     source          1..5887
                     /organism="Cloning vector pPasscode2"
                     /mol_type="other DNA"
                     /db_xref="taxon:1758743"
     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..1135
                     /codon_start=1
                     /product="CelR-LacI"
                     /label=CelR-LacI
                     /protein_id="ALP32101.1"
                     /translation="MKPVTLYDVAEYAGVSYQTVSRVVNQASHVSAKTREKVEAAIKEL
                     GYVPNRAARTLVTRRTDTVALVVSENNQKLFAEPFYAGIVLGVGVALSERGFQFVLATG
                     RSGIEHERLGGYLAGQHVDGVLLLSLHRDDPLPQMLDEAGVPYVYGGRPLGVPEEQVSY
                     VDIDNIGGGRQATQRLIETGHRRIATIAGPQDMVAGVERLQGYREALLAAGMEYDETLV
                     SYGDFTYDSGVAAMRELLDRAPDVDAVFAASDLMGLAALRVLRASGRRVPEDVAVVGYD
                     DSTVAEHAEPPMTSVNQPTELMGREMARLLVDRITGETTEPVRLVLETHLMVRESG"
     regulatory      1156..1250
                     /regulatory_class="terminator"
                     /note="T0"
     terminator      1156..1250
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     promoter        1299..1372
                     /label=PLtetO-1 promoter
                     /note="modified phage lambda PL promoter with tet operator 
                     sites (Lutz and Bujard, 1997)"
     regulatory      1299..1346
                     /regulatory_class="promoter"
                     /note="pLtetO-1"
     protein_bind    1299..1317
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    1324..1342
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     regulatory      1353..1378
                     /regulatory_class="ribosome_binding_site"
                     /note="RBS"
     CDS             1379..2461
                     /codon_start=1
                     /product="LacI"
                     /label=LacI
                     /protein_id="ALP32102.1"
                     /translation="MKPVTLYDVAEYAGVSYQTVSRVVNQASHVSAKTREKVEAAMAEL
                     NYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSRADQLGASVVVSMVERSGV
                     EACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSIIFSH
                     EDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREGDWSA
                     MSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTEDSSC
                     YIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR
                     ALADSLMQLARQVSRLESGQ"
     regulatory      2482..2576
                     /regulatory_class="terminator"
                     /note="T0"
     terminator      2482..2576
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     regulatory      complement(2616..2710)
                     /regulatory_class="terminator"
                     /note="T0"
     terminator      2616..2710
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             complement(2731..3783)
                     /codon_start=1
                     /product="GalR-ScrR"
                     /label=GalR-ScrR
                     /protein_id="ALP32103.1"
                     /translation="MKTKRVTIKDIAELAGVSKATASLVLNGRGKELRVAQETRERVLA
                     IMESLSYHPNANARALAQQTTETVGLVVGDVSDPFFGAMVKAVEQVAYHTGNFLLIGNG
                     YHNEQKERQAIEQLIRHRCAALVVHAKMIPDADLASLMKQMPGMVLINRILPGFENRCI
                     ALDDRYGAWLATRHLIQQGHTRIGYLCSNHSISDAEDRLQGYYDALAESGIAANDRLVT
                     FGEPDESGGEQAMTELLGRGRNFTAVACYNDSMAAGAMGVLNDNGIDVPGEISLIGFDD
                     VLVSRYVRPRLTTVRYPIVTMATQAAELALALADNRPLPEITNVFSPTLVRRHSVSTPS
                     LEASHHATSD"
     regulatory      complement(3784..3809)
                     /regulatory_class="ribosome_binding_site"
                     /note="RBS"
     regulatory      complement(3842..3870)
                     /regulatory_class="promoter"
                     /note="pLlacO"
     regulatory      3901..4005
                     /regulatory_class="terminator"
                     /note="T1"
     terminator      3906..3992
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     rep_origin      4012..4819
                     /label=ColE1
                     /note="ColE1"
     rep_origin      complement(4156..4744)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     regulatory      complement(4826..4931)
                     /regulatory_class="terminator"
                     /note="T0"
     terminator      4832..4926
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             complement(4957..5751)
                     /codon_start=1
                     /product="kanamycin resistance protein"
                     /label=kanamycin resistance protein
                     /protein_id="ALP32104.1"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"