pTOX4-yhaV-amilCP vector (V016130)

Basic Vector Information

Vector Name:
pTOX4-yhaV-amilCP
Antibiotic Resistance:
Chloramphenicol
Length:
6808 bp
Type:
UNVERIFIED: Cloning vector
Replication origin:
R6K γ ori
Source/Author:
Lazarus JE, Warr AR, Kuehl CJ, Giorgio RT
Promoter:
rhaB

pTOX4-yhaV-amilCP vector Map

pTOX4-yhaV-amilCP6808 bp3006009001200150018002100240027003000330036003900420045004800510054005700600063006600strong forward terminatorcat promoterCmRrrnB T1 terminatorrrnB T2 terminatorrhaB promoterRibonuclease toxin YhaVlambda tL3 terminatorlac operatorR6K gamma oritraJoriTAmpR promoterAmpR promoterrhaStac promoteramilCP

pTOX4-yhaV-amilCP vector Sequence

LOCUS       V016130                 6808 bp    DNA     circular SYN 30-JUN-2019
DEFINITION  Exported.
ACCESSION   V016130
VERSION     V016130
KEYWORDS    UNVERIFIED
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
            .
REFERENCE   1  (bases 1 to 6808)
  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 6808)
  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 6808)
  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..6808
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     regulatory      62..95
                     /label="strong forward terminator"
                     /note="strong forward terminator"
                     /regulatory_class="terminator"
     promoter        346..448
                     /label="cat promoter"
                     /note="promoter of the E. coli cat gene encoding
                     chloramphenicol acetyltransferase"
     CDS             449..1105
                     /label="CmR"
                     /note="chloramphenicol acetyltransferase"
     terminator      1209..1295
                     /label="rrnB T1 terminator"
                     /note="transcription terminator T1 from the E. coli rrnB
                     gene"
     terminator      1387..1414
                     /label="rrnB T2 terminator"
                     /note="transcription terminator T2 from the E. coli rrnB
                     gene"
     promoter        1617..1735
                     /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             1759..2220
                     /gene="yhaV"
                     /label="Ribonuclease toxin YhaV"
                     /note="Ribonuclease toxin YhaV from Escherichia coli
                     (strain K12). Accession#: P64594"
     terminator      2283..2529
                     /label="lambda tL3 terminator"
                     /note="transcription terminator tL3 from phage lambda"
     protein_bind    complement(2565..2581)
                     /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(2616..3004)
                     /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(3625..3993)
                     /label="traJ"
                     /note="oriT-recognizing protein"
     oriT            complement(4026..4135)
                     /direction=LEFT
                     /label="oriT"
                     /note="incP origin of transfer"
     regulatory      5048..5065
                     /label="AmpR promoter"
                     /note="AmpR promoter"
                     /regulatory_class="promoter"
     regulatory      5066..5147
                     /label="AmpR promoter"
                     /note="AmpR promoter"
                     /regulatory_class="promoter"
     CDS             5175..6008
                     /label="rhaS"
                     /note="positive regulator of the rhaB promoter"
     promoter        6046..6074
                     /label="tac promoter"
                     /note="strong E. coli promoter; hybrid between the trp and
                     lac UV5 promoters"
     CDS             6141..6803
                     /label="amilCP"
                     /note="amilCP is a fluorescent protein published in 2008,
                     derived from Acropora millepora. It is reported to be a
                     somewhat slowly-maturing tetramer."

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