pPasscode2 vector (V004176)

LOCUS       40924_34096        5887 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Cloning vector pPasscode2, complete sequence.
ACCESSION   .
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)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 5887)
  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..5887
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        42..115
                     /label=PLtetO-1 promoter
                     /note="modified phage lambda PL promoter with tet operator
                     sites (Lutz and Bujard, 1997)"
     CDS             122..1135
                     /codon_start=1
                     /product="CelR-LacI"
                     /label=CelR-LacI
                     /protein_id="ALP32101.1"
                     /translation="MKPVTLYDVAEYAGVSYQTVSRVVNQASHVSAKTREKVEAAIKEL
                     GYVPNRAARTLVTRRTDTVALVVSENNQKLFAEPFYAGIVLGVGVALSERGFQFVLATG
                     RSGIEHERLGGYLAGQHVDGVLLLSLHRDDPLPQMLDEAGVPYVYGGRPLGVPEEQVSY
                     VDIDNIGGGRQATQRLIETGHRRIATIAGPQDMVAGVERLQGYREALLAAGMEYDETLV
                     SYGDFTYDSGVAAMRELLDRAPDVDAVFAASDLMGLAALRVLRASGRRVPEDVAVVGYD
                     DSTVAEHAEPPMTSVNQPTELMGREMARLLVDRITGETTEPVRLVLETHLMVRESG"
     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
                     /label=pLtetO-1
                     /note="pLtetO-1"
                     /regulatory_class="promoter"
     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
                     /label=RBS
                     /note="RBS"
                     /regulatory_class="ribosome_binding_site"
     CDS             1379..2458
                     /label=lacI
                     /note="lac repressor"
     regulatory      2482..2576
                     /label=T0
                     /note="T0"
                     /regulatory_class="terminator"
     terminator      2482..2576
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     regulatory      complement(2616..2710)
                     /label=T0
                     /note="T0"
                     /regulatory_class="terminator"
     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)
                     /label=RBS
                     /note="RBS"
                     /regulatory_class="ribosome_binding_site"
     regulatory      complement(3842..3870)
                     /label=pLlacO
                     /note="pLlacO"
                     /regulatory_class="promoter"
     terminator      3906..3992
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB
                     gene"
     rep_origin      complement(4156..4744)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     terminator      complement(4832..4926)
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             complement(4960..5751)
                     /label=NeoR/KanR
                     /note="aminoglycoside phosphotransferase"