pLL39 vector (V004989)

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Price Information

Cat No.	Plasmid Name	Availability	Add to cart
V004989	pLL39	In stock, 1 week for quality controls	Buy one, get one free!

Two tubes of lyophilized plasmid will be delivered, each tube is about 5µg.

Basic Vector Information

Vector Name:: pLL39

Antibiotic Resistance:: Streptomycin

Length:: 8019 bp

Type:: Single-copy integration vector

Replication origin:: pSC101 ori

Source/Author:: Luong TT, Lee CY.

pLL39 vector Map

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Plasmid Protocol

1. Centrifuge at 5,000×g for 5 min.

2. Carefully open the tube and add 20 μl of sterile water to dissolve the DNA.

3. Close the tube and incubate for 10 minutes at room temperature.

4. Briefly vortex the tube and then do a quick spin to concentrate the liquid at the bottom. Speed is less than 5000×g.

5. Store the plasmid at -20 ℃.

6. The concentration of plasmid re-measurement sometimes differs from the nominal value, which may be due to the position of the lyophilized plasmid in the tube, the efficiency of the re-dissolution, the measurement bias, and adsorption on the wall of the tube, therefore, it is recommended to transform and extract the plasmid before using it

General Plasmid Transform Protocol

1. Take one 100μl of the competent cells and thaw it on ice for 10min, add 2μl of plasmid, then ice bath for 30min, then heat-shock it at 42℃ for 60s, do not stir, and then ice bath for 2min.

2. Add 900μl of LB liquid medium without antibiotics, and incubate at 37℃ for 45min (30℃ for 1-1.5 hours) with 180rpm shaking.

3. Centrifuge at 6000rpm for 5min, leave only 100μl of supernatant to resuspend the bacterial precipitate and spread it onto the target plasmid-resistant LB plate.

4. Invert the plate and incubate at 37℃ for 14h, or at 30℃ for 20h.

5. Pick a single colony into LB liquid medium, add the corresponding antibiotics, incubate at 220rpm for 14h, and extract the plasmid according to the experimental needs and the instructions of the plasmid extraction kit.

pLL39 vector Sequence

LOCUS       V004989                 8019 bp    DNA     circular SYN 18-DEC-2018
DEFINITION  Exported.
ACCESSION   V004989
VERSION     V004989
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
            .
REFERENCE   1  (bases 1 to 8019)
  AUTHORS   Luong TT, Lee CY.
  TITLE     Improved single-copy integration vectors for Staphylococcus aureus
  JOURNAL   J. Microbiol. Methods 70 (1), 186-190 (2007)
   PUBMED   17512993
REFERENCE   2  (bases 1 to 8019)
  AUTHORS   Luong TT, Lee CY.
  TITLE     Direct Submission
  JOURNAL   Submitted (06-APR-2007) Microbiology and Immunology, Univ of
            Arkansas for Medical Sciences, 4301 W. Markham St. Mail Slot 511,
            Little Rock, AR 72205, USA
REFERENCE   3  (bases 1 to 8019)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 8019)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "J.
            Microbiol. Methods"; date: "2007"; volume: "70"; issue: "1"; pages:
            "186-190"
            SGRef: number: 2; type: "Journal Article"; journalName: "Submitted
            (06-APR-2007) Microbiology and Immunology, Univ of Arkansas for
            Medical Sciences, 4301 W. Markham St. Mail Slot 511, Little Rock, AR
            72205, USA"
            SGRef: number: 3; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..8019
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    1..57
                     /label="MCS"
                     /note="pUC18/19 multiple cloning site"
     CDS             complement(715..2091)
                     /gene="tet"
                     /label="Tetracycline resistance protein"
                     /note="Tetracycline resistance protein from Staphylococcus
                     aureus. Accession#: P02983"
     CDS             complement(3288..4076)
                     /label="SmR"
                     /note="aminoglycoside adenylyltransferase (Murphy, 1985)"
     CDS             complement(5331..6278)
                     /label="Rep101"
                     /note="RepA protein needed for replication with the pSC101
                     origin"
     rep_origin      complement(6326..6548)
                     /direction=LEFT
                     /label="pSC101 ori"
                     /note="low-copy replication origin that requires the Rep101
                     protein"
     terminator      7166..7209
                     /label="rrnB T1 terminator"
                     /note="transcription terminator T1 from the E. coli rrnB
                     gene"
     terminator      7309..7395
                     /gene="Escherichia coli rrnB"
                     /label="rrnB T1 terminator"
                     /note="transcription terminator T1 from the E. coli rrnB
                     gene"
     terminator      7492..7578
                     /gene="Escherichia coli rrnB"
                     /label="rrnB T1 terminator"
                     /note="transcription terminator T1 from the E. coli rrnB
                     gene"
     terminator      7675..7761
                     /gene="Escherichia coli rrnB"
                     /label="rrnB T1 terminator"
                     /note="transcription terminator T1 from the E. coli rrnB
                     gene"
     terminator      complement(7858..7944)
                     /label="rrnB T1 terminator"
                     /note="transcription terminator T1 from the E. coli rrnB
                     gene"