Basic Vector Information
- Vector Name:
- pEKH-rsgI6
- Antibiotic Resistance:
- Ampicillin
- Length:
- 10707 bp
- Type:
- Cloning vector
- Replication origin:
- ori
- Host:
- Yeast
- Source/Author:
- Sand A, Holwerda EK, Ruppertsberger NM, Maloney M, Olson DG, Nataf Y, Borovok I, Sonenshein AL, Bayer EA, Lamed R, Lynd LR, Shoham Y.
- Promoter:
- URA3
pEKH-rsgI6 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 ℃.
pEKH-rsgI6 vector Sequence
LOCUS 40924_17304 10707 bp DNA circular SYN 17-DEC-2018
DEFINITION Cloning vector pEKH-rsgI6, complete sequence.
ACCESSION .
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 10707)
AUTHORS Sand A, Holwerda EK, Ruppertsberger NM, Maloney M, Olson DG, Nataf
Y, Borovok I, Sonenshein AL, Bayer EA, Lamed R, Lynd LR, Shoham Y.
TITLE Three cellulosomal xylanase genes in Clostridium thermocellum are
regulated by both vegetative SigA (sigma(A)) and alternative SigI6
(sigma(I6)) factors
JOURNAL FEBS Lett. 589 (20), 3133-3140 (2015)
PUBMED 26320414
REFERENCE 2 (bases 1 to 10707)
AUTHORS Holwerda EK.
TITLE Direct Submission
JOURNAL Submitted (24-JUL-2015) Thayer School of Engineering, Dartmouth
College, 14 Engineering drive, Hanover, NH 03755, USA
REFERENCE 3 (bases 1 to 10707)
TITLE Direct Submission
REFERENCE 4 (bases 1 to 10707)
AUTHORS .
TITLE Direct Submission
COMMENT SGRef: number: 1; type: "Journal Article"; journalName: "FEBS
Lett."; date: "2015"; volume: "589"; issue: "20"; pages: "3133-3140"
COMMENT SGRef: number: 2; type: "Journal Article"; journalName: "Submitted
(24-JUL-2015) Thayer School of Engineering, Dartmouth College, 14
Engineering drive, Hanover, NH 03755, 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..10707
/mol_type="other DNA"
/organism="synthetic DNA construct"
misc_feature 11..514
/label=CEN/ARS
/note="S. cerevisiae CEN6 centromere fused to an
autonomously replicating sequence"
CDS complement(1202..1960)
/gene="sigI6"
/label=sigI6
/note="RNA polymerase sigma factor SigI6 from Acetivibrio
thermocellus (strain ATCC 27405 / DSM 1237 / JCM 9322 /
NBRC 103400 / NCIMB 10682 / NRRL B-4536 / VPI 7372).
Accession#: A3DH98"
CDS 2594..3241
/gene="cat"
/label=cat
/note="Chloramphenicol acetyltransferase from
Staphylococcus aureus. Accession#: P00485"
CDS 3262..3807
/codon_start=1
/product="hypoxanthine phosphoribosyltransferase"
/EC_number="2.4.2.8"
/label=hypoxanthine phosphoribosyltransferase
/protein_id="ALI61571.1"
/translation="MENLSKDIDEILITEEELKEKIKELGRQITKDYKGKNLMLVGVLK
GALMFMADLSRHIDLPLSLDFMAVSSYGSSTHSSGIVKIIKDLDISIEGKDVLIVEDII
DSGLTLSYLRETLLGRKPKSLKICTILDKPERREASVKVDYVGFKIPDKFVVGYGLDFD
EKYRNLPFIGVLKPEMYS"
terminator 4703..4950
/label=CYC1 terminator
/note="transcription terminator for CYC1"
rep_origin complement(5198..5786)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
CDS complement(5873..6451)
/codon_start=1
/product="thymidine kinase"
/EC_number="2.7.1.21"
/label=thymidine kinase
/protein_id="ALI61572.1"
/translation="MYGPKDHGYIEVVTGPMFSGKSEELIRRIKRAKIARQKVQVFKPA
IDDRYSIDKVVSHNGDNMHAIAIVKASDILAYAEEDTDVFAIDEVQFFDSEIVDIVKEI
ADSGKRVICAGLDMDFRGEPFGPTPELMAIAEFVDKLTAICMKCGNPATRTQRLINGKP
ANYDDPIIMVGAKESYEARCRKCHEVPRT"
CDS complement(7151..8152)
/label=repB
/note="RepB replication protein"
CDS complement(8713..9570)
/label=AmpR
/note="beta-lactamase"
CDS complement(9669..10469)
/label=URA3
/note="orotidine-5'-phosphate decarboxylase, required for
uracil biosynthesis"
promoter complement(10470..10685)
/label=URA3 promoter
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