Basic Vector Information
PSM1 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 ℃.
PSM1 vector Sequence
LOCUS 40924_40702 6321 bp DNA circular SYN 18-DEC-2018
DEFINITION Cloning vector PSM1, complete sequence.
ACCESSION .
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 6321)
AUTHORS Rahnama M, Forester N, Ariyawansa S, Voisey CR, Johnson LJ, Johnson
RD, Fleetwood DJ.
TITLE Efficient targeted mutagenesis in Epichloe festucae using a split
marker system
JOURNAL Unpublished
REFERENCE 2 (bases 1 to 6321)
AUTHORS Rahnama M, Fleetwood D.
TITLE Direct Submission
JOURNAL Submitted (27-SEP-2016) School of Biological Sciences, University of
Auckland, 3A Symonds Street, Auckland, Auckland 1010, New Zealand
REFERENCE 3 (bases 1 to 6321)
TITLE Direct Submission
REFERENCE 4 (bases 1 to 6321)
AUTHORS .
TITLE Direct Submission
COMMENT SGRef: number: 1; type: "Journal Article"; journalName:
"Unpublished"
COMMENT SGRef: number: 2; type: "Journal Article"; journalName: "Submitted
(27-SEP-2016) School of Biological Sciences, University of Auckland,
3A Symonds Street, Auckland, Auckland 1010, New Zealand"
COMMENT SGRef: number: 3; type: "Journal Article"
FEATURES Location/Qualifiers
source 1..6321
/mol_type="other DNA"
/organism="synthetic DNA construct"
terminator complement(268..295)
/label=rrnB T2 terminator
/note="transcription terminator T2 from the E. coli rrnB
gene"
terminator complement(387..473)
/label=rrnB T1 terminator
/note="transcription terminator T1 from the E. coli rrnB
gene"
CDS complement(513..1124)
/codon_start=1
/gene="hph"
/product="hygromycin B phosphotransferase"
/label=hph
/protein_id="APD72149.1"
/translation="MPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRGYV
LRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLPET
ELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQTV
MDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLT"
gene complement(513..1124)
/gene="hph"
/label=hph
intron complement(1133..1248)
/label=gpdA intron
/note="intron from the Aspergillus nidulans
glyceraldehyde-3-phosphate dehydrogenase gene (Punt et al.,
1990)"
primer_bind 2096..2112
/label=M13 fwd
/note="common sequencing primer, one of multiple similar
variants"
protein_bind 2129..2360
/label=attP1
/note="recombination site for the Gateway(R) BP reaction
(pDONR(TM)221 version)"
CDS complement(2759..3061)
/codon_start=1
/label=ccdB
/note="CcdB, a bacterial toxin that poisons DNA gyrase"
/translation="MQFKVYTYKRESRYRLFVDVQSDIIDTPGRRMVIPLASARLLSDK
VSRELYPVVHIGDESWRMMTTDMASVPVSVIGEEVADLSHRENDIKNAINLMFWGI"
CDS complement(3409..4065)
/codon_start=1
/label=CmR
/note="chloramphenicol acetyltransferase"
/translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
promoter complement(4066..4168)
/label=cat promoter
/note="promoter of the E. coli cat gene encoding
chloramphenicol acetyltransferase"
protein_bind complement(4313..4544)
/label=attP2
/note="recombination site for the Gateway(R) BP reaction
(pDONR(TM)221 version)"
promoter complement(4563..4581)
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
primer_bind complement(4586..4602)
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
CDS 4715..5521
/codon_start=1
/label=KanR
/note="aminoglycoside phosphotransferase"
/translation="MSHIQRETSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYGKP
DAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGKTA
FQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDASD
FDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGIAD
RYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
rep_origin 5671..6259
/direction=RIGHT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
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