pUCP20T-mKO1 vector (V002368)

MSDS

Basic Vector Information

Vector Name:: pUCP20T-mKO1

Antibiotic Resistance:: Ampicillin

Length:: 4831 bp

Type:: Cloning vector

Replication origin:: ori

Source/Author:: Barbier M, Damron FH.

pUCP20T-mKO1 vector Vector Map

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 ℃.

pUCP20T-mKO1 vector Sequence

Copy Sequence

Download GeneBank File(.gb)

LOCUS       40924_45333        4831 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Cloning vector pUCP20T-mKO1, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4831)
  AUTHORS   Barbier M, Damron FH.
  TITLE     Rainbow Vectors for Broad-Range Bacterial Fluorescence Labeling
  JOURNAL   PLoS ONE 11 (3), E0146827 (2016)
  PUBMED    26937640
REFERENCE   2  (bases 1 to 4831)
  AUTHORS   Barbier M, Damron FH.
  TITLE     Direct Submission
  JOURNAL   Submitted (01-DEC-2015) Microbiology, Immunology and Cell Biology, 
            West Virginia University, School of Medicine, One Medical Center 
            Drive, Morgantown, WV 26506, USA
REFERENCE   3  (bases 1 to 4831)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 4831)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "PLoS ONE"; 
            date: "2016"; volume: "11"; issue: "3"; pages: "E0146827"
COMMENT     SGRef: number: 2; type: "Journal Article"; journalName: "Submitted 
            (01-DEC-2015) Microbiology, Immunology and Cell Biology, West 
            Virginia University, School of Medicine, One Medical Center Drive, 
            Morgantown, WV 26506, USA"
COMMENT     SGRef: number: 3; type: "Journal Article"
COMMENT     ##Assembly-Data-START##
            Sequencing Technology :: Sanger dideoxy sequencing 
            ##Assembly-Data-END##
FEATURES             Location/Qualifiers
     source          1..4831
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      complement(232..583)
                     /direction=LEFT
                     /label=pRO1600 oriV
                     /note="broad-host-range origin of replication from
                     Pseudomonas aeruginosa plasmid pRO1600; requires the 
                     pRO1600 Rep protein for replication (West et al., 1994)"
     CDS             597..1427
                     /codon_start=1
                     /label=pRO1600 Rep
                     /note="replication protein for the broad-host-range plasmid
                     pRO1600 from Pseudomonas aeruginosa"
                     /translation="VASPPMVYKSNALVEAAYRLSVQEQRIVLACISQVKRSEPVTDEV
                     MYSVTAEDIATMAGVPIESSYNQLKEAALRLKRREVRLTQEPNGKGKRPSVMITGWVQT
                     IIYREGEGRVELRFTKDMLPYLTELTKQFTKYALADVAKMDSTHAIRLYELLMQWDSIG
                     QREIEIDQLRKWFQLEGRYPSIKDFKLRVLDPAVTQINEHSPLQVEWAQRKTGRKVTHL
                     LFSFGPKKPAKAVGKAPAKRKAGKISDAEIAKQARPGETWEAARARLTQMPLDLA"
     primer_bind     1591..1607
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     CDS             complement(1650..2303)
                     /codon_start=1
                     /label=Kusabira-Orange
                     /note="Kusabira-Orange fluorescent protein (Karasawa et
                     al., 2004)"
                     /translation="MVSVIKPEMKMKYFMDGSVNGHEFTVEGEGTGKPYEGHQEMTLRV
                     TMAKGGPMPFSFDLVSHTFCYGHRPFTKYPEEIPDYFKQAFPEGLSWERSLQFEDGGFA
                     AVSAHISLRGNCFEHKSKFVGVNFPADGPVMQNQSSDWEPSTEKITTCDGVLKGDVTMY
                     LKLAGGGNHKCQFKTTYKAAKKILKMPQSHFIGHRLVRKTEGNITELVEDAVAHC"
     primer_bind     complement(2334..2350)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    complement(2358..2374)
                     /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(2382..2412)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    complement(2427..2448)
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     oriT            2832..2940
                     /label=oriT
                     /note="incP origin of transfer"
     rep_origin      complement(3012..3600)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(3774..4631)
                     /codon_start=1
                     /label=AmpR
                     /note="beta-lactamase"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     promoter        complement(4632..4736)
                     /label=AmpR promoter

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