pMHC9-1 vector (V004742)

MSDS

Basic Vector Information

Vector Name:: pMHC9-1

Antibiotic Resistance:: Ampicillin

Length:: 7104 bp

Type:: Cloning vector

Replication origin:: ori

Source/Author:: Maglennon GA, Cook BS, Deeney AS, Bosse JT, Peters SE, Langford PR, Maskell DJ, Tucker AW, Wren BW, Rycroft AN.

pMHC9-1 vector Vector Map

View Map in NovoBuilder

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 ℃.

pMHC9-1 vector Sequence

Copy Sequence

Download GeneBank File(.gb)

LOCUS       40924_30805        7104 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Cloning vector pMHC9-1, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7104)
  AUTHORS   Maglennon GA, Cook BS, Deeney AS, Bosse JT, Peters SE, Langford PR, 
            Maskell DJ, Tucker AW, Wren BW, Rycroft AN.
  TITLE     Transposon mutagenesis in Mycoplasma hyopneumoniae using a novel 
            mariner-based system for generating random mutations
  JOURNAL   Vet. Res. 44 (1), 124 (2013)
  PUBMED    24359443
REFERENCE   2  (bases 1 to 7104)
  AUTHORS   Maglennon GA, Cook BS, Deeney AS, Bosse JT, Langford PR, Maskell DJ,
            Tucker AW, Wren BW, Rycroft AN.
  TITLE     Direct Submission
  JOURNAL   
REFERENCE   3  (bases 1 to 7104)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 7104)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Vet. Res.";
            date: "2013"; volume: "44"; issue: "1"; pages: "124"
COMMENT     SGRef: number: 2; type: "Journal Article"; journalName: "Submitted 
            (16-NOV-2013) Pathology "
COMMENT     SGRef: number: 3; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..7104
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             805..1851
                     /codon_start=1
                     /product="transposase"
                     /label=transposase
                     /note="C9 mutant transposase from Himar1"
                     /protein_id="AHE63356.1"
                     /translation="MEKKEFRVLIKYCFLKGKNTVEAKTWLDNEFPDSAPGKSTIIDWY
                     AKFKRGEMSTEDGERSGRPKEVVTDENIKKIHKMILNDRKMKLIEIAEALKISKERVGH
                     IIHQYLDMRKLCAKWVPRELTFDQKQRRVDDSKRCLQLLTRNTPEFFRRYVTMDETWLH
                     HYTPESNRQSAEWTATGEPSPKRGKTQKSAGKVMASVFWDAHGIIFIDYLEKGKTINSD
                     YYMALLERLKVEIAAKRPHMKKKKVLFHQDNAPCHKSLRTMAKIHELGFELLPHPPYSP
                     DLAPSDFFLFSDIKRMLAGKKFGCNEEVIAETEAYFEAKPKEYYQNGIKKLEGRYNRCI
                     ALEGNYVE"
     CDS             2622..4541
                     /codon_start=1
                     /gene="tetM"
                     /product="tetracycline resistance protein"
                     /label=tetM
                     /protein_id="AHE63355.1"
                     /translation="MKIINIGVLAHVDAGKTTLTESLLYNSGAITELGSVDKGTTRTDN
                     TLLERQRGITIQTGITSFQWENTKVNIIDTPGHMDFLAEVYRSLSVLDGAILLISAKDG
                     VQAQTRILFHALRKMGIPTIFFINKIDQNGIDLSTVYQDIKEKLSAEIVIKQKVELYPN
                     VCVTNFTESEQWDTVIEGNDDLLEKYMSGKSLEALELEQEESIRFQNCSLFPLYHGSAK
                     SNIGIDNLIEVITNKFYSSTHRGPSELCGNVFKIEYTKKRQRLAYIRLYSGVLHLRDSV
                     RVSEKEKIKVTEMYTSINGELCKIDRAYSGEIVILQNEFLKLNSVLGDTKLLPQRKKIE
                     NPHPLLQTTVEPSKPEQREMLLDALLEISDSDPLLRYYVDSTTHEIILSFLGKVQMEVI
                     SALLQEKYHVEIEITEPTVIYMERPLKNAEYTIHIEVPPNPFWASIGLSVSPLPLGSGM
                     QYESSVSLGYLNQSFQNAVMEGIRYGCEQGLYGWNVTDCKICFKYGLYYSPVSTPADFR
                     MLAPIVLEQVLKKAGTELLEPYLSFKIYAPQEYLSRAYNDAPKYCANIVDTQLKNNEVI
                     LSGEIPARCIQEYRSDLTFFTNGRSVCLTELKGYHVTTGEPVCQPRRPNSRIDKVRYMF
                     NKIT"
     gene            2622..4541
                     /gene="tetM"
                     /label=tetM
     primer_bind     complement(4883..4899)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    complement(4907..4923)
                     /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(4931..4961)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    complement(4976..4997)
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     rep_origin      complement(5285..5873)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(6047..6904)
                     /codon_start=1
                     /label=AmpR
                     /note="beta-lactamase"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     promoter        complement(6905..7009)
                     /label=AmpR promoter

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