pTOX10-yhaV vector (V015445)

Basic Vector Information

Vector Name:
pTOX10-yhaV
Antibiotic Resistance:
Gentamycin
Length:
5849 bp
Type:
UNVERIFIED: Cloning vector
Replication origin:
R6K γ ori
Source/Author:
Lazarus JE, Warr AR, Kuehl CJ, Giorgio RT
Promoter:
rhaB

pTOX10-yhaV vector Map

pTOX10-yhaV5849 bp60012001800240030003600420048005400GmRrrnB T1 terminatorrrnB T2 terminatorrhaB promoterRibonuclease toxin YhaVlambda tL3 terminatorlac operatorR6K gamma oritraJoriTAmpR promoterAmpR promoterrhaSstrong forward terminator

pTOX10-yhaV vector Sequence

LOCUS       V015445                 5849 bp    DNA     circular SYN 30-JUN-2019
DEFINITION  Exported.
ACCESSION   V015445
VERSION     V015445
KEYWORDS    UNVERIFIED
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
            .
REFERENCE   1  (bases 1 to 5849)
  AUTHORS   Lazarus JE, Warr AR, Kuehl CJ, Giorgio RT, Davis BM, Waldor MK.
  TITLE     A New Suite of Allelic Exchange Vectors for the Scarless
            Modification of Proteobacterial Genomes
  JOURNAL   Appl. Environ. Microbiol. (2019) In press
   PUBMED   31201277
REFERENCE   2  (bases 1 to 5849)
  AUTHORS   Lazarus JE, Warr AR, Kuehl CJ, Giorgio RT, Davis BM, Waldor MK.
  TITLE     Direct Submission
  JOURNAL   Submitted (24-MAY-2019) Medicine, Massachusetts General Hospital, 55
            Fruit St, Jackson 5, Boston, MA 02114, USA
REFERENCE   3  (bases 1 to 5849)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Appl.
            Environ. Microbiol. (2019) In press"
            SGRef: number: 2; type: "Journal Article"; journalName: "Submitted
            (24-MAY-2019) Medicine, Massachusetts General Hospital, 55 Fruit St,
            Jackson 5, Boston, MA 02114, USA"
            GenBank staff is unable to verify sequence and/or annotation
            provided by the submitter.
FEATURES             Location/Qualifiers
     source          1..5849
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(4..534)
                     /label="GmR"
                     /note="gentamycin acetyltransferase"
     terminator      751..837
                     /label="rrnB T1 terminator"
                     /note="transcription terminator T1 from the E. coli rrnB
                     gene"
     terminator      929..956
                     /label="rrnB T2 terminator"
                     /note="transcription terminator T2 from the E. coli rrnB
                     gene"
     promoter        1159..1277
                     /label="rhaB promoter"
                     /note="promoter of the E. coli rhaBAD operon, conferring
                     tight induction with L-rhamnose and repression with
                     D-glucose in the presence of RhaR and RhaS (Giacalone et
                     al., 2006)"
     CDS             1301..1762
                     /gene="yhaV"
                     /label="Ribonuclease toxin YhaV"
                     /note="Ribonuclease toxin YhaV from Escherichia coli
                     (strain K12). Accession#: P64594"
     terminator      1825..2071
                     /label="lambda tL3 terminator"
                     /note="transcription terminator tL3 from phage lambda"
     protein_bind    complement(2107..2123)
                     /label="lac operator"
                     /note="The lac repressor binds to the lac operator to
                     inhibit transcription in E. coli. This inhibition can be
                     relieved by adding lactose or
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     rep_origin      complement(2158..2546)
                     /direction=LEFT
                     /label="R6K gamma ori"
                     /note="gamma replication origin from E. coli plasmid R6K;
                     requires the R6K initiator protein pi for replication"
     CDS             complement(3167..3535)
                     /label="traJ"
                     /note="oriT-recognizing protein"
     oriT            complement(3568..3677)
                     /direction=LEFT
                     /label="oriT"
                     /note="incP origin of transfer"
     regulatory      4590..4607
                     /label="AmpR promoter"
                     /note="AmpR promoter"
                     /regulatory_class="promoter"
     regulatory      4608..4689
                     /label="AmpR promoter"
                     /note="AmpR promoter"
                     /regulatory_class="promoter"
     CDS             4717..5550
                     /label="rhaS"
                     /note="positive regulator of the rhaB promoter"
     regulatory      5638..5671
                     /label="strong forward terminator"
                     /note="strong forward terminator"
                     /regulatory_class="terminator"

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