PSM2 vector (V003137)

Basic Vector Information

      • Vector Name:
      • PSM2
      • Antibiotic Resistance:
      • Chloramphenicol
      • Length:
      • 6504 bp
      • Type:
      • Cloning vector
      • Source/Author:
      • Rahnama M, Forester N, Ariyawansa S, Voisey CR, Johnson LJ, Johnson RD, Fleetwood DJ.

PSM2 vector Vector Map

PSM26504 bp300600900120015001800210024002700300033003600390042004500480051005400570060006300rrnB T2 terminatorrrnB T1 terminatorhphTtrpCM13 fwdattP1ccdBcatcat promoterattP2T7 promoterM13 revnptIIpUCOri

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 ℃.

PSM2 vector Sequence

Copy Sequence

Download GeneBank File(.gb)

LOCUS       Exported                6504 bp ds-DNA     circular SYN 18-DEC-2018
DEFINITION  Cloning vector PSM2, complete sequence.
ACCESSION   KX904529
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6504)
  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  JOURNAL   Unpublished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 6504)
  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 6504)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..6504
                     /organism="Cloning vector PSM2"
                     /mol_type="other DNA"
                     /db_xref="taxon:1920725"
     terminator      268..295
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      387..473
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     CDS             513..1454
                     /codon_start=1
                     /gene="hph"
                     /product="hygromycin B phosphotransferase"
                     /label=hph
                     /protein_id="APD72153.1"
                     /translation="SEGEESRAFSFDVGGRGYVLRVNSCADGFYKDRYVYRHFASAALP
                     IPEVLDIGEFSESLTYCISRRAQGVTLQDLPETELPAVLQPVAEAMDAIAAADLSQTSG
                     FGPFGPQGIGQYTTWRDFICAIADPHVYHWQTVMDDTVSASVAQALDELMLWAEDCPEV
                     RHLVHADFGSNNVLTDNGRITAVIDWSEAMFGDSQYEVANIFFWRPWLACMEQQTRYFE
                     RRHPELAGSPRLRAYMLRIGLDQLYQSLVDGNFDDAAWAQGRCDAIVRSGAGTVGRTQI
                     ARRSAAVWTDGCVEVLADSGNRRPSTRPRAKE"
     gene            513..1454
                     /gene="hph"
                     /label=hph
     regulatory      1472..2240
                     /regulatory_class="terminator"
                     /note="TtrpC"
     primer_bind     2279..2295
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     misc_recomb     2312..2543
                     /label=attP1
                     /note="attP1"
     protein_bind    2312..2543
                     /gene="mutant version of attP"
                     /label=attP1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction 
                     (pDONR(TM)201 version)"
     CDS             complement(2939..3244)
                     /codon_start=1
                     /gene="ccdB"
                     /product="plasmid maintenance protein CcdB"
                     /label=ccdB
                     /protein_id="APD72154.1"
                     /translation="MQFKVYTYKRESRYRLFVDVQSDIIDTPGRRMVIPLASARLLSDK
                     VSRELYPVVHIGDESWRMMTTDMASVPVSVIGEEVADLSHRENDIKNAINLMFWGI"
     gene            complement(2939..3244)
                     /gene="ccdB"
                     /label=ccdB
     CDS             complement(3589..4248)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=cat
                     /protein_id="APD72155.1"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     gene            complement(3589..4248)
                     /gene="cat"
                     /label=cat
     promoter        complement(4249..4351)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     misc_recomb     complement(4493..4724)
                     /label=attP2
                     /note="attP2"
     protein_bind    complement(4496..4727)
                     /gene="mutant version of attP"
                     /label=attP2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction 
                     (pDONR(TM)201 version)"
     promoter        complement(4746..4764)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(4769..4785)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     CDS             4898..5707
                     /codon_start=1
                     /gene="nptII"
                     /product="kanamycin resistance protein"
                     /label=nptII
                     /protein_id="APD72156.1"
                     /translation="MSHIQRETSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYGKP
                     DAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGKTA
                     FQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDASD
                     FDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGIAD
                     RYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     gene            4898..5707
                     /gene="nptII"
                     /label=nptII
     rep_origin      5825..6498
                     /label=pUCOri
                     /note="pUCOri"
     rep_origin      5854..6442
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"

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