pMG14 vector (V004747)

MSDS

Basic Vector Information

Vector Name:: pMG14

Antibiotic Resistance:: Ampicillin

Length:: 5169 bp

Type:: Shuttle vector

Replication origin:: ori

Source/Author:: Gimpel M, Brantl S.

pMG14 vector Vector Map

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Plasmid Resuspension 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 ℃.

pMG14 vector Sequence

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LOCUS       40924_30765        5169 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Shuttle vector pMG14, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5169)
  AUTHORS   Gimpel M, Brantl S.
  TITLE     Construction of a modular plasmid family for chromosomal integration
            in Bacillus subtilis
  JOURNAL   J. Microbiol. Methods 91 (2), 312-317 (2012)
  PUBMED    22982324
REFERENCE   2  (bases 1 to 5169)
  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  JOURNAL   Unpublished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   3  (bases 1 to 5169)
  AUTHORS   De Schamphelaire W.
  TITLE     Direct Submission
  JOURNAL   Submitted (02-FEB-2017) BCCM/LMBP, Universiteit Gent, 
            Technologiepark 927, 9052, BELGIUM
REFERENCE   4  (bases 1 to 5169)
  TITLE     Direct Submission
REFERENCE   5  (bases 1 to 5169)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "J. 
            Microbiol. Methods"; date: "2012"; volume: "91"; issue: "2"; pages: 
            "312-317"
COMMENT     SGRef: number: 2; type: "Journal Article"; journalName: 
            "Unpublished"
COMMENT     SGRef: number: 3; type: "Journal Article"; journalName: "Submitted 
            (02-FEB-2017) BCCM/LMBP, Universiteit Gent, Technologiepark 927, 
            9052, BELGIUM"
COMMENT     SGRef: number: 4; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..5169
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     regulatory      5..234
                     /label=cggR promoter
                     /note="cggR promoter"
                     /regulatory_class="promoter"
     misc_feature    262..302
                     /label=5' UTR of gapA
                     /note="5' UTR of gapA"
     CDS             311..334
                     /label=Strep-Tag II
                     /note="peptide that binds Strep-Tactin(R), an engineered 
                     form
                     of streptavidin"
     CDS             complement(470..1009)
                     /codon_start=1
                     /note="unnamed protein product; amyE fragment"
                     /protein_id="SJL86547.1"
                     /translation="MFAKRFKTSLLPLFAGFLLLFHLVLAGPAAASAETANKSNELTAP
                     SIKSGTILHAWNWSFNTLKHNMKDIHDAGYTAIQTSPINQVKEGNQGDKSMSNWYWLYQ
                     PTSYQIGNRYLGTEQEFKEMCAAAEEYGIKVIVDAVINHTTSDYAAISNEVKSIPNWTH
                     GNTQIKNWSDRWDVTQN"
     misc_feature    complement(1010..1125)
                     /label=5' UTR of amyE
                     /note="5' UTR of amyE"
     promoter        1229..1333
                     /label=AmpR promoter
     CDS             1334..2191
                     /label=AmpR
                     /note="beta-lactamase"
     rep_origin      2365..2953
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     protein_bind    3241..3262
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     promoter        3277..3307
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3315..3331
                     /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)."
     primer_bind     3339..3355
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     CDS             complement(3396..4199)
                     /codon_start=1
                     /note="unnamed protein product; amyE fragment"
                     /protein_id="SJL86549.1"
                     /translation="STWMSDDDIRLGWAVIASRSGSTPLFFSRPEGGGNGVRFPGKSQI
                     GDRGSALFEDQAITAVNRFHNVMAGQPEELSNPNGNNQIFMNQRGSHGVVLANAGSSSV
                     SINTATKLPDGRYDNKAGAGSFQVNDGKLTGTINARSVAVLYPDDIAKAPHVFLENYKT
                     GVTHSFNDQLTITLRADANTTKAVYQINNGPETAFKDGDQFTIGKGDPFGKTYTIMLKG
                     TNSDGVTRTEKYSFVKRDPASAKTIGYQNPNHWSQVNAYIYKHDGS"
     CDS             complement(4289..5059)
                     /codon_start=1
                     /note="unnamed protein product; kanamycin/tobramycin
                     resistance (aadD)"
                     /protein_id="SJL86550.1"
                     /translation="MRIVNGPIIMTREERMKIVHEIKERILDKYGDDVKAIGVYGSLGR
                     QTDGPYSDIEMMCVMSTEEAEFSHEWTTGEWKVEVNFDSEEILLDYASQVESDWPLTHG
                     QFFSILPIYDSGGYLEKVYQTAKSVEAQTFHDAICALIVEELFEYAGKWRNIRVQGPTT
                     FLPSLTVQVAMAGAMLIGLHHRICYTTSASVLTEAVKQSDLPSGYDHLCQFVMSGQLSD
                     SEKLLESLENFWNGIQEWTERHGYIVDVSKRIPF"

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