Basic Vector Information
- Vector Name:
- pMC212
- Antibiotic Resistance:
- Kanamycin
- Length:
- 13012 bp
- Type:
- Cloning vector
- Replication origin:
- p15A ori
- Host:
- Yeast
- Source/Author:
- Currie DH, Herring CD, Guss AM, Olson DG, Hogsett DA, Lynd LR.
- Promoter:
- URA3
pMC212 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 ℃.
pMC212 vector Sequence
LOCUS 40924_29931 13012 bp DNA circular SYN 18-DEC-2018
DEFINITION Cloning vector pMC212, complete sequence.
ACCESSION .
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 13012)
AUTHORS Currie DH, Herring CD, Guss AM, Olson DG, Hogsett DA, Lynd LR.
TITLE Functional heterologous expression of an engineered full length CipA
from Clostridium thermocellum in Thermoanaerobacterium
saccharolyticum
JOURNAL Biotechnol Biofuels 6 (1), 32 (2013)
PUBMED 23448319
REFERENCE 2 (bases 1 to 13012)
AUTHORS Currie DH, Herring CD, Guss AM, Olson DG, Hogsett DA, Lynd LR.
TITLE Direct Submission
JOURNAL Submitted (22-FEB-2013) Thayer School of Engineering, Dartmouth
College, 14 Engineering Drive, Hanover, NH 03755, USA
REFERENCE 3 (bases 1 to 13012)
TITLE Direct Submission
REFERENCE 4 (bases 1 to 13012)
AUTHORS .
TITLE Direct Submission
COMMENT SGRef: number: 1; type: "Journal Article"; journalName: "Biotechnol
Biofuels"; date: "2013"; volume: "6"; issue: "1"; pages: "32"
COMMENT SGRef: number: 2; type: "Journal Article"; journalName: "Submitted
(22-FEB-2013) Thayer School of Engineering, Dartmouth College, 14
Engineering Drive, Hanover, NH 03755, USA"
COMMENT SGRef: number: 3; type: "Journal Article"
FEATURES Location/Qualifiers
source 1..13012
/mol_type="other DNA"
/organism="synthetic DNA construct"
misc_feature 1..1500
/label=similar to xynA upstream region
/note="similar to xynA upstream region"
CDS 1501..3723
/gene="celS"
/label=celS
/note="Cellulose 1,4-beta-cellobiosidase (reducing end)
CelS from Acetivibrio thermocellus (strain ATCC 27405 / DSM
1237 / JCM 9322 / NBRC 103400 / NCIMB 10682 / NRRL B-4536 /
VPI 7372). Accession#: A3DH67"
CDS 3739..3765
/label=9xHis
/note="9xHis affinity tag"
CDS 3966..4952
/codon_start=1
/product="pta"
/label=pta
/protein_id="AGG53972.1"
/translation="MSIIQNIIEKAKSDKKKIVLPEGAEPRTLKAAEIVLKEGIADLVL
LGNEDEIRNAAKDLDISKAEIIDPVKSEMFDRYANDFYELRKNKGITLEKARETIKDNI
YFGCMMVKEGYADGLVSGAIHATADLLRPAFQIIKTAPGAKIVSSFFIMEVPNCEYGEN
GVFLFADCAVNPSPNAEELASIAVQSANTAKNLLGFEPKVAMLSFSTKGSASHELVDKV
RKATEIAKELMPDVAIDGELQLDAALVKEVAELKAPGSKVAGCANVLIFPDLQAGNIGY
KLVQRLAKANAIGPITQGMGAPVNDLSRGCSYRDIVDVIATTAVQAQ"
CDS 4970..6181
/codon_start=1
/product="ack"
/label=ack
/protein_id="AGG53973.1"
/translation="MKIMKILVINCGSSSLKYQLIESTDGNVLAKGLAERIGINDSMLT
HNANGEKIKIKKDMKDHKDAIKLVLDALVNSDYGVIKDMSEIDAVGHRVVHGGESFTSS
VLINDEVLKAITDCIELAPLHNPANIEGIKACQQIMPNVPMVAVFDTAFHQTMPDYAYL
YPIPYEYYTKYRIRRYGFHGTSHKYVSNRAAEILNKPIEDLKIITCHLGNGSSIAAVKY
GKSIDTSMGFTPLEGLAMGTRSGSIDPSIISYLMEKENISAEEVVNILNKKSGVYGISG
ISSDFRDLEDAAFKNGDERAQLALNVFAYRVKKTIGAYAAAMGGVDVIVFTAGVGENGP
EIREFILDGLEFLGFSLDKEKNKVRGKETIISTPNSKVSVMVVPTNEEYMIAKDTEKIV
KSIK"
CDS 6840..7631
/label=KanR
/note="aminoglycoside phosphotransferase"
misc_feature 7780..9217
/label=similar to xynA downstream region
/note="similar to xynA downstream region"
rep_origin complement(9556..10101)
/direction=LEFT
/label=p15A ori
/note="Plasmids containing the medium-copy-number p15A
origin of replication can be propagated in E. coli cells
that contain a second plasmid with the ColE1 origin."
promoter 10213..10241
/label=tet promoter
/note="E. coli promoter for tetracycline efflux protein
gene"
CDS complement(10318..11118)
/label=URA3
/note="orotidine-5'-phosphate decarboxylase, required for
uracil biosynthesis"
promoter complement(11119..11339)
/label=URA3 promoter
misc_feature 11367..11870
/label=CEN/ARS
/note="S. cerevisiae CEN6 centromere fused to an
autonomously replicating sequence"
oriT complement(12269..12378)
/direction=LEFT
/label=oriT
/note="incP origin of transfer"
terminator complement(12799..12826)
/label=rrnB T2 terminator
/note="transcription terminator T2 from the E. coli rrnB
gene"
terminator complement(12918..13004)
/label=rrnB T1 terminator
/note="transcription terminator T1 from the E. coli rrnB
gene"
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