pTRK1-bglT2mcsHEstop vector (V002525)

Basic Vector Information

Vector Name:
pTRK1-bglT2mcsHEstop
Antibiotic Resistance:
Ampicillin
Length:
9207 bp
Type:
Reporter vector
Replication origin:
ori
Source/Author:
Vian A, Carrascosa AV, Garcia JL, Cortes E.

pTRK1-bglT2mcsHEstop vector Vector Map

pTRK1-bglT2mcsHEstop9207 bp400800120016002000240028003200360040004400480052005600600064006800720076008000840088009200polylinker HindIII-NruI-SalI-NotI-PstI-SwaI-EcoRV-PvuII-EcoRIbgaArepAKNTAmpRori

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 ℃.

pTRK1-bglT2mcsHEstop vector Sequence

Copy Sequence

Download GeneBank File(.gb)

LOCUS       40924_44272        9207 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Reporter vector pTRK1-bglT2mcsHEstop DNA, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9207)
  AUTHORS   Vian A, Carrascosa AV, Garcia JL, Cortes E.
  TITLE     Structure of the beta-galactosidase gene from Thermus sp. strain T2:
            expression in Escherichia coli and purification in a single step of 
            an active fusion protein
  JOURNAL   Appl. Environ. Microbiol. 64 (6), 2187-2191 (1998)
  PUBMED    9603833
REFERENCE   2  (bases 1 to 9207)
  AUTHORS   Fujita A, Misumi Y, Koyama Y.
  TITLE     Two versatile shuttle vectors for Thermus thermophilus-Escherichia 
            coli containing multiple cloning sites, lacZalpha gene and kanamycin
            or hygromycin resistance marker
  JOURNAL   Plasmid 67 (3), 272-275 (2012)
  PUBMED    22252135
REFERENCE   3  (bases 1 to 9207)
  AUTHORS   Fujita A, Misumi Y, Honda S, Sato T, Koyama Y.
  TITLE     Construction of new cloning vectors that employ the phytoene 
            synthase encoding gene for color screening of cloned DNA inserts in 
            Thermus thermophilus
  JOURNAL   Gene 527 (2), 655-662 (2013)
  PUBMED    23845779
REFERENCE   4  (bases 1 to 9207)
  AUTHORS   Fujita A, Sato T, Koyama Y, Misumi Y.
  TITLE     Development of a novel reporter gene system incorporating a gene 
            encoding a thermostable beta-galactosidase for use in Thermus 
            thermophilus HB27
  JOURNAL   Unpublished
REFERENCE   5  (bases 1 to 9207)
  AUTHORS   Fujita A.
  TITLE     Direct Submission
  JOURNAL   Submitted (27-JAN-2015) Contact:Atsushi Fujita National Institute of
            Advanced Industrial Science and Technology, Health Research 
            Institute; 1-8-31 Midorigaoka, Ikeda, Osaka 563-8577, Japan URL 
            :https://unit.aist.go.jp/hri/
REFERENCE   6  (bases 1 to 9207)
  TITLE     Direct Submission
REFERENCE   7  (bases 1 to 9207)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Appl. 
            Environ. Microbiol."; date: "1998"; volume: "64"; issue: "6"; pages:
            "2187-2191"
COMMENT     SGRef: number: 2; type: "Journal Article"; journalName: "Plasmid"; 
            date: "2012"; volume: "67"; issue: "3"; pages: "272-275"
COMMENT     SGRef: number: 3; type: "Journal Article"; journalName: "Gene"; 
            date: "2013"; volume: "527"; issue: "2"; pages: "655-662"
COMMENT     SGRef: number: 4; type: "Journal Article"; journalName: 
            "Unpublished"
COMMENT     SGRef: number: 5; type: "Journal Article"; journalName: "Submitted 
            (27-JAN-2015) Contact:Atsushi Fujita National Institute of Advanced 
            Industrial Science and Technology, Health Research Institute; 1-8-31
            Midorigaoka, Ikeda, Osaka 563-8577, Japan URL 
            :https://unit.aist.go.jp/hri/"
COMMENT     SGRef: number: 6; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..9207
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    69..123
                     /note="polylinker 
                     HindIII-NruI-SalI-NotI-PstI-SwaI-EcoRV-PvuII-EcoRI"
     CDS             137..2071
                     /gene="bgaA"
                     /label=bgaA
                     /note="Beta-galactosidase BgaA from Thermus sp. Accession#:
                     O54315"
     CDS             complement(2922..4085)
                     /codon_start=1
                     /gene="repA"
                     /product="putative repA protein"
                     /label=repA
                     /protein_id="BAS05947.1"
                     /translation="MVLRAYAALRGLSPEALRAHLLAPPLRPERAREAFQRPYLAHFAQ
                     TLPRYPYATDDPKEGVRIYKRENALKRVHVQVGHYPHAVLRLVVDVDLPWPQVEERIHA
                     LPPSLVLVNPRSGHFHAWYELDPIPLTPPPGREGSLKGALALLAEVEALLEAYYGADPG
                     YNGLLSRNPFLHPPEWTWGGGKRWSLRDLHRELRGLLPSGTRRRVDPGLASYGRNNALF
                     DRLRAEAYAHVALFRGVPGGEEAFRAWVEQRAHALNQSLFRDHPKGPLDPREVHHTAKS
                     VAKWTYRNYRGARVYPVSSTGRPDRSRLSPQARALIPPLQGQELQEAVREGGRRRGSRR
                     RQEAEEKLTEALKRLQARGERVTARALAREAGVKPHTASKWLKRMRE"
     gene            complement(2922..4085)
                     /gene="repA"
                     /label=repA
     CDS             6577..7347
                     /codon_start=1
                     /gene="KNT"
                     /product="kanamycin nucleotidyltransferase"
                     /label=KNT
                     /protein_id="BAS05948.1"
                     /translation="MRIVNGPIIMTREERMKIVHEIKERILDKYGDDVKAIGVYGSLGR
                     QTDGPYSDIEMMCVMSTEEAEFSHEWTTGEWKAEVNFYSEEILLDYASRVEPDWPLTHG
                     RFFSILPIYDPGGYFEKVYQTAKSVEAQKFHDAICALIVEELFEYAGKWRNIRVQGPTT
                     FLPSLTVQVAMAGAMLIGLHHRICYTTSASVLTEAVKQPDLPPGYVQLCQLVMSGQLSD
                     PEKLLESLENFWNGIQEWTERHGYIVDVSKRIPF"
     gene            6577..7347
                     /gene="KNT"
                     /label=KNT
     CDS             7406..8263
                     /label=AmpR
                     /note="beta-lactamase"
     rep_origin      8437..9025
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"

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