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  JOURNAL   Unpublished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            "Unpublished"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"
ORIGIN
        1 ctttcctgcg ttatcccctg attctgtgga taaccgtatt accgcctttg agtgagctga
       61 taccgctcgc cgcagccgaa cgaccgagcg cagcgagtca gtgagcgagg aagcggaaga
      121 gcgcccaata cgcaaaccgc ctctccccgc gcgttggccg attcattaat gcagctggca
      181 cgacaggttt cccgactgga aagcgggcag tgagcgcaac gcaattaata cgcgtaccgc
      241 tagccaggaa gagtttgtag aaacgcaaaa aggccatccg tcaggatggc cttctgctta
      301 gtttgatgcc tggcagttta tggcgggcgt cctgcccgcc accctccggg ccgttgcttc
      361 acaacgttca aatccgctcc cggcggattt gtcctactca ggagagcgtt caccgacaaa
      421 caacagataa aacgaaaggc ccagtcttcc gactgagcct ttcgttttat ttgatgcctg
      481 gcagttccct actctcgcgt taacgcggcc gccgtcagga cattgttgga gccgaaatcc
      541 gcgtgcacga ggtgccggac ttcggggcag tcctcggccc aaagcatcag ctcatcgaga
      601 gcctgcgcga cggacgcact gacggtgtcg tccatcacag tttgccagtg atacacatgg
      661 ggatcagcaa tcgcgcatat gaaatcacgc catgtagtgt attgaccgat tccttgcggt
      721 ccgaatgggc cgaacccgct cgtctggcta agatcggccg cagcgatcgc atccatggcc
      781 tccgcgaccg gctgcagaac agcgggcagt tcggtttcag gcaggtcttg caacgtgaca
      841 ccctgtgcac ggcgggagat gcaataggtc aggctctcgc tgaattcccc aatgtcaagc
      901 acttccggaa tcgggagcgc ggccgatgca aagtgccgat aaacataacg atctttgtag
      961 aaaccatcgg cgcagctatt tacccgcagg acatatccac gccctcctac atcgaagctg
     1021 aaagcacgag attcttcgcc ctccgagagc tgcatcaggt cggagacgct gtcgaacttt
     1081 tcgatcagaa acttctcgac agacgtcgcg gtgagttcag gcatggtgat gtctgctcaa
     1141 gcggggtagc tgttagtcaa gctgcgatga agtgggaaag ctcgaactga aaggttcaaa
     1201 ggaataaggg atgggaagga tggagtatgg atgtagcaaa gtacttactt aggggaaata
     1261 aaggttcttg gatgggaaga tgaatatact gaagatggga aaagaaagag aaaagaaaag
     1321 agcagctggt ggggagagca ggaaaatatg gcaacaaatg ttggactgac gcaacgacct
     1381 tgtcaacccc gccgacacac cgggcggaca gacggggcaa agctgcctac cagggactga
     1441 gggacctcag caggtcgagt gcagagcacc ggatgggtcg actgccagct tgtgttcccg
     1501 gtctgcgccg ctggccagct cctgagcggc ctttccggtt tcatacaccg ggcaaagcag
     1561 gagaggcacg atatttggac gccctacaga tgccggatgg gccaattagg gagcttacgc
     1621 gccgggtact cgctctacct acttcggaga aggtactatc tcgtgaatct tttaccagat
     1681 cggaagcaat tggacttctg tacctaggtt aatggcatgc tatttcgccg acggctatac
     1741 acccctggct tcacattctc cttcgcttac tgccggtgat tcgatgaagc tccatattct
     1801 ccgatgatgc aatagattct tggtcaacga ggggcacacc agcctttcca cttcggggcg
     1861 gaggggcggc cggtcccgga ttaataatca tccactgcac ctcagagccg ccagagctgt
     1921 ctggccagtg gcttattact cagcccttct ctctgcgtcc gtccgtctct ccgcatgcca
     1981 gaaagagtca ccggtcactg tacagagctc acgagttcgt cacatttttc tacaaatggt
     2041 ggaggcggcg gattttagtt aacgctagca tggatgtttt cccagtcacg acgttgtaaa
     2101 acgacggcca gtcttaagct cgggccccaa ataatgattt tattttgact gatagtgacc
     2161 tgttcgttgc aacacattga tgagcaatgc ttttttataa tgccaacttt gtacaaaaaa
     2221 gctgaacgag aaacgtaaaa tgatataaat atcaatatat taaattagat tttgcataaa
     2281 aaacagacta cataatactg taaaacacaa catatccagt cactatgaat caactactta
     2341 gatggtatta gtgacctgta gtcgaccgac agccttccaa atgttcttcg ggtgatgctg
     2401 ccaacttagt cgaccgacag ccttccaaat gttcttctca aacggaatcg tcgtatccag
     2461 cctactcgct attgtcctca atgccgtatt aaatcataaa aagaaataag aaaaagaggt
     2521 gcgagcctct tttttgtgtg acaaaataaa aacatctacc tattcatata cgctagtgtc
     2581 atagtcctga aaatcatctg catcaagaac aatttcacaa ctcttatact tttctcttac
     2641 aagtcgttcg gcttcatctg gattttcagc ctctatactt actaaacgtg ataaagtttc
     2701 tgtaatttct actgtatcga cctgcagact ggctgtgtat aagggagcct gacatttata
     2761 ttccccagaa catcaggtta atggcgtttt tgatgtcatt ttcgcggtgg ctgagatcag
     2821 ccacttcttc cccgataacg gagaccggca cactggccat atcggtggtc atcatgcgcc
     2881 agctttcatc cccgatatgc accaccgggt aaagttcacg ggagacttta tctgacagca
     2941 gacgtgcact ggccaggggg atcaccatcc gtcgcccggg cgtgtcaata atatcactct
     3001 gtacatccac aaacagacga taacggctct ctcttttata ggtgtaaacc ttaaactgca
     3061 tttcaccagc ccctgttctc gtcagcaaaa gagccgttca tttcaataaa ccgggcgacc
     3121 tcagccatcc cttcctgatt ttccgctttc cagcgttcgg cacgcagacg acgggcttca
     3181 ttctgcatgg ttgtgcttac cagaccggag atattgacat catatatgcc ttgagcaact
     3241 gatagctgtc gctgtcaact gtcactgtaa tacgctgctt catagcatac ctctttttga
     3301 catacttcgg gtatacatat cagtatatat tcttataccg caaaaatcag cgcgcaaata
     3361 cgcatactgt tatctggctt ttagtaagcc ggatccacgc ggcgtttacg ccccgccctg
     3421 ccactcatcg cagtactgtt gtaattcatt aagcattctg ccgacatgga agccatcaca
     3481 gacggcatga tgaacctgaa tcgccagcgg catcagcacc ttgtcgcctt gcgtataata
     3541 tttgcccatg gtgaaaacgg gggcgaagaa gttgtccata ttggccacgt ttaaatcaaa
     3601 actggtgaaa ctcacccagg gattggctga gacgaaaaac atattctcaa taaacccttt
     3661 agggaaatag gccaggtttt caccgtaaca cgccacatct tgcgaatata tgtgtagaaa
     3721 ctgccggaaa tcgtcgtggt attcactcca gagcgatgaa aacgtttcag tttgctcatg
     3781 gaaaacggtg taacaagggt gaacactatc ccatatcacc agctcaccgt ctttcattgc
     3841 catacggaat tccggatgag cattcatcag gcgggcaaga atgtgaataa aggccggata
     3901 aaacttgtgc ttatttttct ttacggtctt taaaaaggcc gtaatatcca gctgaacggt
     3961 ctggttatag gtacattgag caactgactg aaatgcctca aaatgttctt tacgatgcca
     4021 ttgggatata tcaacggtgg tatatccagt gatttttttc tccattttag cttccttagc
     4081 tcctgaaaat ctcgataact caaaaaatac gcccggtagt gatcttattt cattatggtg
     4141 aaagttggaa cctcttacgt gccgatcaac gtctcatttt cgccaaaagt tggcccaggg
     4201 cttcccggta tcaacaggga caccaggatt tatttattct gcgaagtgat cttccgtcac
     4261 aggtatttat tcggcgcaaa gtgcgtcggg tgatgctgcc aacttagtcg actacaggtc
     4321 actaatacca tctaagtagt tgattcatag tgactggata tgttgtgttt tacagtatta
     4381 tgtagtctgt tttttatgca aaatctaatt taatatattg atatttatat cattttacgt
     4441 ttctcgttca gctttcttgt acaaagttgg cattataaga aagcattgct tatcaatttg
     4501 ttgcaacgaa caggtcacta tcagtcaaaa taaaatcatt atttgccatc cagctgatat
     4561 cccctatagt gagtcgtatt acatggtcat agctgtttcc tggcagctct ggcccgtgtc
     4621 tcaaaatctc tgatgttaca ttgcacaaga taaaataata tcatcatgaa caataaaact
     4681 gtctgcttac ataaacagta atacaagggg tgttatgagc catattcaac gggaaacgtc
     4741 gaggccgcga ttaaattcca acatggatgc tgatttatat gggtataaat gggctcgcga
     4801 taatgtcggg caatcaggtg cgacaatcta tcgcttgtat gggaagcccg atgcgccaga
     4861 gttgtttctg aaacatggca aaggtagcgt tgccaatgat gttacagatg agatggtcag
     4921 actaaactgg ctgacggaat ttatgcctct tccgaccatc aagcatttta tccgtactcc
     4981 tgatgatgca tggttactca ccactgcgat ccccggaaaa acagcattcc aggtattaga
     5041 agaatatcct gattcaggtg aaaatattgt tgatgcgctg gcagtgttcc tgcgccggtt
     5101 gcattcgatt cctgtttgta attgtccttt taacagcgat cgcgtatttc gtctcgctca
     5161 ggcgcaatca cgaatgaata acggtttggt tgatgcgagt gattttgatg acgagcgtaa
     5221 tggctggcct gttgaacaag tctggaaaga aatgcataaa cttttgccat tctcaccgga
     5281 ttcagtcgtc actcatggtg atttctcact tgataacctt atttttgacg aggggaaatt
     5341 aataggttgt attgatgttg gacgagtcgg aatcgcagac cgataccagg atcttgccat
     5401 cctatggaac tgcctcggtg agttttctcc ttcattacag aaacggcttt ttcaaaaata
     5461 tggtattgat aatcctgata tgaataaatt gcagtttcat ttgatgctcg atgagttttt
     5521 ctaatcagaa ttggttaatt ggttgtaaca ctggcagagc attacgctga cttgacggga
     5581 cggcgcaagc tcatgaccaa aatcccttaa cgtgagttac gcgtcgttcc actgagcgtc
     5641 agaccccgta gaaaagatca aaggatcttc ttgagatcct ttttttctgc gcgtaatctg
     5701 ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt tgtttgccgg atcaagagct
     5761 accaactctt tttccgaagg taactggctt cagcagagcg cagataccaa atactgttct
     5821 tctagtgtag ccgtagttag gccaccactt caagaactct gtagcaccgc ctacatacct
     5881 cgctctgcta atcctgttac cagtggctgc tgccagtggc gataagtcgt gtcttaccgg
     5941 gttggactca agacgatagt taccggataa ggcgcagcgg tcgggctgaa cggggggttc
     6001 gtgcacacag cccagcttgg agcgaacgac ctacaccgaa ctgagatacc tacagcgtga
     6061 gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg
     6121 cagggtcgga acaggagagc gcacgaggga gcttccaggg ggaaacgcct ggtatcttta
     6181 tagtcctgtc gggtttcgcc acctctgact tgagcgtcga tttttgtgat gctcgtcagg
     6241 ggggcggagc ctatggaaaa acgccagcaa cgcggccttt ttacggttcc tggccttttg
     6301 ctggcctttt gctcacatgt t
//
