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pMHL120 vector (V004731)

Basic Vector Information

Vector Name:
pMHL120
Antibiotic Resistance:
Kanamycin
Length:
13561 bp
Type:
TnphoZ mutagenesis vector
Replication origin:
ori
Source/Author:
Lee MH, Nittayajarn A, Ross RP, Rothschild CB, Parsonage D, Claiborne A, Rubens CE.
Promoter:
tet

pMHL120 vector Vector Map

pMHL12013561 bp600120018002400300036004200480054006000660072007800840090009600102001080011400120001260013200RBSermBoribomropTcRtet promoterphoZKanRrepAtsFactor Xa site

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 ℃.

pMHL120 vector Sequence

Copy Sequence

Download GeneBank File(.gb)

LOCUS       40924_30860       13561 bp DNA     circular SYN 18-DEC-2018
DEFINITION  TnphoZ mutagenesis vector pMHL120, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 13561)
  AUTHORS   Lee MH, Nittayajarn A, Ross RP, Rothschild CB, Parsonage D, 
            Claiborne A, Rubens CE.
  TITLE     Characterization of Enterococcus faecalis alkaline phosphatase and 
            use in identifying Streptococcus agalactiae secreted proteins
  JOURNAL   J. Bacteriol. 181 (18), 5790-5799 (1999)
  PUBMED    10482522
REFERENCE   2  (bases 1 to 13561)
  AUTHORS   Clancy A, Lee MH, Jones AL, Rubens CE.
  TITLE     Construction and characterization of transposon TnphoZ for the 
            identification of genes encoding exported proteins in Streptococcus 
            agalactiae
  JOURNAL   FEMS Microbiol. Lett. 241 (2), 257-264 (2004)
  PUBMED    15598541
REFERENCE   3  (bases 1 to 13561)
  AUTHORS   Lee MH, Clancy A, Jones AL, Nittayajarn A, Rubens CE.
  TITLE     Direct Submission
  JOURNAL   Submitted (19-MAR-2001) Infectious Disease, Children's Hospital and 
            Regional Medical Center, 4800 Sand Point Way NE, Seattle, WA 98105, 
            USA
REFERENCE   4  (bases 1 to 13561)
  TITLE     Direct Submission
REFERENCE   5  (bases 1 to 13561)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "J. 
            Bacteriol."; date: "1999"; volume: "181"; issue: "18"; pages: 
            "5790-5799"
COMMENT     SGRef: number: 2; type: "Journal Article"; journalName: "FEMS 
            Microbiol. Lett."; date: "2004"; volume: "241"; issue: "2"; pages: 
            "257-264"
COMMENT     SGRef: number: 3; type: "Journal Article"; journalName: "Submitted 
            (19-MAR-2001) Infectious Disease, Children's Hospital and Regional 
            Medical Center, 4800 Sand Point Way NE, Seattle, WA 98105, USA"
COMMENT     SGRef: number: 4; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..13561
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     RBS             3821..3829
                     /label=Shine-Dalgarno sequence
                     /note="full consensus sequence for ribosome-binding sites 
                     upstream of start codons in E. coli; complementary to a 
                     region in the 3' end of the 16S rRNA (Chen et al., 1994)"
     CDS             complement(4170..4904)
                     /gene="ermB"
                     /label=ermB
                     /note="rRNA adenine N-6-methyltransferase from Enterococcus
                     faecalis (strain ATCC 700802 / V583). Accession#: P20173"
     rep_origin      5356..5944
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     misc_feature    complement(6130..6270)
                     /label=bom
                     /note="basis of mobility region from pBR322"
     CDS             complement(6375..6563)
                     /label=rop
                     /note="Rop protein, which maintains plasmids at low copy
                     number"
     CDS             complement(7204..8391)
                     /label=TcR
                     /note="tetracycline efflux protein"
     promoter        complement(8439..8467)
                     /label=tet promoter
                     /note="E. coli promoter for tetracycline efflux protein
                     gene"
     gene            complement(8683..10032)
                     /gene="phoZ"
                     /allele="phoZ2"
                     /label=phoZ
                     /note="alkaline phosphatase; derived from Enterococcus
                     faecalis; expression is dependent upon integration event to
                     create in-frame start codon"
     CDS             10619..11410
                     /label=KanR
                     /note="aminoglycoside phosphotransferase"
     CDS             complement(11817..12515)
                     /codon_start=1
                     /gene="repAts"
                     /product="temperature-sensitive RepA"
                     /label=repAts
                     /protein_id="AAK27832.1"
                     /translation="MAIKNTKARNFGFLLYPDSIPNDWKEKLESLGVSMAVSPLHDMDE
                     KKDKDTWNNSNIIQNGKHYKKPHYHVIYIARNPVTIESVRNKIKRKLGNSSVAHVEILD
                     YIKGSYEYLTHESKDAIAKNKHIYDKKDILNINDFDIDRYITLDESQKRELKNLLLDIV
                     DDYNLVNTKDLMAFIRLRGAEFGILNTNDVKDIVSTNSSAFRLWFEGNYQCGYRASYAK
                     VLDAETGEIK"
     gene            complement(11817..12515)
                     /gene="repAts"
                     /label=repAts
     CDS             13127..13138
                     /label=Factor Xa site
                     /note="Factor Xa recognition and cleavage site"

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