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  JOURNAL   Unpublished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"