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pAM1 vector (V009520)

Price Information

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V009520 pAM1 In stock, instant shipping

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

Basic Vector Information

The pAM1 is a Shuttle vector.

Vector Name:
pAM1
Antibiotic Resistance:
Ampicillin
Length:
6245 bp
Type:
Shuttle vector
Replication origin:
ori
Source/Author:
De Schamphelaire W, Olbrechts A, Meert J, Verhelst K, Roggeman Fonseca M, Vanhoucke M, Beyaert R.
Growth Strain(s):
DH10B
Growth Temperature:
37℃

pAM1 vector Map

Copy Sequence View Map
pAM16245 bp30060090012001500180021002400270030003300360039004200450048005100540057006000M13 fwdrRNA adenine N-6-methyltransferaseunnamed protein product; copG-likeunnamed protein product; repBM13 revlac operatorlac promoterCAP binding siteoriAmpRAmpR promoter

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.

pAM1 vector Sequence

LOCUS       V009520                 6245 bp    DNA     circular SYN 17-DEC-2018
DEFINITION  Exported.
ACCESSION   V009520
VERSION     V009520
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
            .
REFERENCE   1  (bases 1 to 6245)
  AUTHORS   De Schamphelaire W, Olbrechts A, Meert J, Verhelst K, Roggeman
            Fonseca M, Vanhoucke M, Beyaert R.
  TITLE     BCCM/LMBP Plasmid collection
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 6245)
  AUTHORS   De Schamphelaire W.
  TITLE     Direct Submission
  JOURNAL   Submitted (02-FEB-2017) BCCM/LMBP, Universiteit Gent,
            Technologiepark 927, 9052, BELGIUM
REFERENCE   3  (bases 1 to 6245)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 6245)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName:
            "Unpublished"
            SGRef: number: 2; type: "Journal Article"; journalName: "Submitted
            (02-FEB-2017) BCCM/LMBP, Universiteit Gent, Technologiepark 927,
            9052, BELGIUM"
            SGRef: number: 3; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..6245
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     379..395
                     /label="M13 fwd"
                     /note="common sequencing primer, one of multiple similar
                     variants"
     CDS             complement(502..1233)
                     /gene="ermC"
                     /label="rRNA adenine N-6-methyltransferase"
                     /note="rRNA adenine N-6-methyltransferase from
                     Staphylococcus aureus. Accession#: P02979"
     CDS             1698..1982
                     /codon_start=1
                     /note="unnamed protein product; copG-like"
                     /protein_id="SJL86728.1"
                     /translation="MSDFSERDRELLAMFGMGEDQVRDDVERMESETAGHGITGPVHYA
                     LHMLPEHGEEMVSMTVKLPKSQLEKVTETARRYHISRSEYVRRQLAGAV"
     CDS             complement(2174..2464)
                     /codon_start=1
                     /note="unnamed protein product; orfX-like"
                     /protein_id="SJL86729.1"
                     /translation="MTIQTIRKKRPLPAKELAAMYDVSVRTIQRWASQTRKDWIDEQAT
                     LRESIRAYHDDEGHTWPQTAEHFGMSQDAVRSRCYRARKERAAEAKAARPE"
     CDS             complement(2461..3408)
                     /codon_start=1
                     /note="unnamed protein product; repB"
                     /protein_id="SJL86730.1"
                     /translation="MSDEYSQPTLELSRTFEGWWLPRRPLCCDDDYSQLVRRSRTDALR
                     CKHIEANPSALVNTIVVDIDDANAKAMALWGHRGMLPNWIAENPANGHAHAGWVLTYPV
                     PRTDMARLKPLKLLHAVTEGLRRSVDGDEGYSGLLMKNPLSDAWDSDLCREDTYDLPDL
                     VAALEEHGDMPPKSWTRTKRAREVGVGRNCTLFDEARTLAYRQVRRLPDRTPASSDLLR
                     EYVRRTCHEINASFPDPLPVREVNDTAKSIHKWITTRSRMWRDGAVANAATFVAIQSAR
                     GRKGGRGNKRDKQGNVQNAFKQKAELFGKEMMGQ"
     primer_bind     complement(4024..4040)
                     /label="M13 rev"
                     /note="common sequencing primer, one of multiple similar
                     variants"
     protein_bind    complement(4048..4064)
                     /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)."
     promoter        complement(4072..4102)
                     /label="lac promoter"
                     /note="promoter for the E. coli lac operon"
     protein_bind    complement(4117..4138)
                     /label="CAP binding site"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     rep_origin      complement(4426..5014)
                     /direction=LEFT
                     /label="ori"
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
     CDS             complement(5188..6045)
                     /label="AmpR"
                     /note="beta-lactamase"
     promoter        complement(6046..6150)
                     /label="AmpR promoter"