Product Data Sheet

Recovery

1.Obtain an LB agar plate with the appropriate antibiotic.

2.Using a sterile pipette tip, touch the bacteria growing within the punctured area of the stab culture. (A sterilized wire loop or sterile toothpick can be used in place of a sterile pipette tip.)

3.Run this tip lightly over a section of the plate, as shown in the figure, to create streak #1.

4.Using another sterile pipette tip, pass through streak #1 and spread the bacteria over a second section of the plate, to create streak #2.

5.Using a third sterile pipette tip, pass through streak #2 and spread the bacteria over the last section of the plate, to create streak #3.

6.Grow overnight in a 37 ^o C incubator (unless a different growth temperature is indicated on the plasmid datasheet).

7.In the morning, single colonies should be visible. If the bacterial growth is too dense, re-streak onto a new agar plate to obtain single colonies.

LOCUS       XXU25061                4768 bp    DNA     linear   SYN 10-JAN-2005

DEFINITION  Cloning vector pBBR1MCS-5 REP, LacZ alpha peptide (lacZ alpha), and

            gentamicin-3-acetyltransferase genes, complete cds.

ACCESSION   U25061

VERSION     U25061.1  GI:833825

KEYWORDS    .

SOURCE      Cloning vector pBBR1MCS-5

  ORGANISM  Cloning vector pBBR1MCS-5

            other sequences; artificial sequences; vectors.

REFERENCE   1  (bases 1 to 4768)

  AUTHORS   Kovach,M.E., Phillips,R.W., Elzer,P.H., Roop,R.M. II and

            Peterson,K.M.

  TITLE     pBBR1MCS: a broad-host-range cloning vector

  JOURNAL   BioTechniques 16 (5), 800-802 (1994)

   PUBMED   8068328

REFERENCE   2  (bases 1 to 4768)

  AUTHORS   Kovach,M.E., Elzer,P.H., Hill,D.S., Robertson,G.T., Farris,M.A.,

            Roop,R.M. II and Peterson,K.M.

  TITLE     Four new derivatives of the broad-host-range cloning vector

            pBBR1MCS, carrying different antibiotic-resistance cassettes

  JOURNAL   Gene 166 (1), 175-176 (1995)

   PUBMED   8529885

REFERENCE   3  (bases 1 to 4768)

  AUTHORS   Kovach,M.E.

  TITLE     Direct Submission

  JOURNAL   Submitted (13-APR-1995) Michael E. Kovach, Microbiology, Louisiana

            State University, 1501 Kings Highway, Shreveport, LA 71130, USA

FEATURES             Location/Qualifiers

     source          1..4768

                     /organism="Cloning vector pBBR1MCS-5"

                     /mol_type="genomic DNA"

                     /db_xref="taxon:51200"

                     /lab_host="Bordetella bronchiseptica"

     CDS             1793..2455

                     /note="required for plasmid replication"

                     /codon_start=1

                     /transl_table=11

                     /product="REP"

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