LOCUS       40924_40707        6504 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Cloning vector PSM2, complete sequence.
ACCESSION   .
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  JOURNAL   Unpublished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)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 6504)
  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            "Unpublished"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..6504
                     /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             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
     terminator      1472..2240
                     /label=trpC terminator
                     /note="transcription terminator from the Aspergillus
                     nidulans trpC gene (Punt et al., 1987)"
     primer_bind     2279..2295
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    2312..2543
                     /label=attP1
                     /note="recombination site for the Gateway(R) BP reaction 
                     (pDONR(TM)221 version)"
     CDS             complement(2942..3244)
                     /codon_start=1
                     /label=ccdB
                     /note="CcdB, a bacterial toxin that poisons DNA gyrase"
                     /translation="MQFKVYTYKRESRYRLFVDVQSDIIDTPGRRMVIPLASARLLSDK
                     VSRELYPVVHIGDESWRMMTTDMASVPVSVIGEEVADLSHRENDIKNAINLMFWGI"
     CDS             complement(3592..4248)
                     /codon_start=1
                     /label=CmR
                     /note="chloramphenicol acetyltransferase"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     promoter        complement(4249..4351)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding
                     chloramphenicol acetyltransferase"
     protein_bind    complement(4496..4727)
                     /label=attP2
                     /note="recombination site for the Gateway(R) BP reaction 
                     (pDONR(TM)221 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..5704
                     /codon_start=1
                     /label=KanR
                     /note="aminoglycoside phosphotransferase"
                     /translation="MSHIQRETSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYGKP
                     DAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGKTA
                     FQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDASD
                     FDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGIAD
                     RYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     rep_origin      5854..6442
                     /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 gctcggaggg cgaagaatct cgtgctttca
      541 gcttcgatgt aggagggcgt ggatatgtcc tgcgggtaaa tagctgcgcc gatggtttct
      601 acaaagatcg ttatgtttat cggcactttg catcggccgc gctcccgatt ccggaagtgc
      661 ttgacattgg ggaattcagc gagagcctga cctattgcat ctcccgccgt gcacagggtg
      721 tcacgttgca agacctgcct gaaaccgaac tgcccgctgt tctgcagccg gtcgcggagg
      781 ccatggatgc gatcgctgcg gccgatctta gccagacgag cgggttcggc ccattcggac
      841 cgcaaggaat cggtcaatac actacatggc gtgatttcat atgcgcgatt gctgatcccc
      901 atgtgtatca ctggcaaact gtgatggacg acaccgtcag tgcgtccgtc gcgcaggctc
      961 tcgatgagct gatgctttgg gccgaggact gccccgaagt ccggcacctc gtgcacgcgg
     1021 atttcggctc caacaatgtc ctgacggaca atggccgcat aacagcggtc attgactgga
     1081 gcgaggcgat gttcggggat tcccaatacg aggtcgccaa catcttcttc tggaggccgt
     1141 ggttggcttg tatggagcag cagacgcgct acttcgagcg gaggcatccg gagcttgcag
     1201 gatcgccgcg gctccgggcg tatatgctcc gcattggtct tgaccaactc tatcagagct
     1261 tggttgacgg caatttcgat gatgcagctt gggcgcaggg tcgatgcgac gcaatcgtcc
     1321 gatccggagc cgggactgtc gggcgtacac aaatcgcccg cagaagcgcg gccgtctgga
     1381 ccgatggctg tgtagaagta ctcgccgata gtggaaaccg acgccccagc actcgtccga
     1441 gggcaaagga atagagtaga tgccgaccgc gggatccact taacgttact gaaatcatca
     1501 aacagcttga cgaatctgga tataagatcg ttggtgtcga tgtcagctcc ggagttgaga
     1561 caaatggtgt tcaggatctc gataagatac gttcatttgt ccaagcagca aagagtgcct
     1621 tctagtgatt taatagctcc atgtcaacaa gaataaaacg cgttttcggg tttacctctt
     1681 ccagatacag ctcatctgca atgcattaat gcattgactg caacctagta acgccttnca
     1741 ggctccggcg aagagaagaa tagcttagca gagctatttt cattttcggg agacgagatc
     1801 aagcagatca acggtcgtca agagacctac gagactgagg aatccgctct tggctccacg
     1861 cgactatata tttgtctcta attgtacttt gacatgctcc tcttctttac tctgatagct
     1921 tgactatgaa aattccgtca ccagcncctg ggttcgcaaa gataattgca tgtttcttcc
     1981 ttgaactctc aagcctacag gacacacatt catcgtaggt ataaacctcg aaatcanttc
     2041 ctactaagat ggtatacaat agtaaccatg catggttgcc tagtgaatgc tccgtaacac
     2101 ccaatacgcc ggccgaaact tttttacaac tctcctatga gtcgtttacc cagaatgcac
     2161 aggtacactt gtttagaggt aatccttctt tctagaagtc ctcgtgtact gtgtaagcgc
     2221 ccactccaca tctccactcg gttaacgcta gcatggatgt tttcccagtc acgacgttgt
     2281 aaaacgacgg ccagtcttaa gctcgggccc caaataatga ttttattttg actgatagtg
     2341 acctgttcgt tgcaacacat tgatgagcaa tgctttttta taatgccaac tttgtacaaa
     2401 aaagctgaac gagaaacgta aaatgatata aatatcaata tattaaatta gattttgcat
     2461 aaaaaacaga ctacataata ctgtaaaaca caacatatcc agtcactatg aatcaactac
     2521 ttagatggta ttagtgacct gtagtcgacc gacagccttc caaatgttct tcgggtgatg
     2581 ctgccaactt agtcgaccga cagccttcca aatgttcttc tcaaacggaa tcgtcgtatc
     2641 cagcctactc gctattgtcc tcaatgccgt attaaatcat aaaaagaaat aagaaaaaga
     2701 ggtgcgagcc tcttttttgt gtgacaaaat aaaaacatct acctattcat atacgctagt
     2761 gtcatagtcc tgaaaatcat ctgcatcaag aacaatttca caactcttat acttttctct
     2821 tacaagtcgt tcggcttcat ctggattttc agcctctata cttactaaac gtgataaagt
     2881 ttctgtaatt tctactgtat cgacctgcag actggctgtg tataagggag cctgacattt
     2941 atattcccca gaacatcagg ttaatggcgt ttttgatgtc attttcgcgg tggctgagat
     3001 cagccacttc ttccccgata acggagaccg gcacactggc catatcggtg gtcatcatgc
     3061 gccagctttc atccccgata tgcaccaccg ggtaaagttc acgggagact ttatctgaca
     3121 gcagacgtgc actggccagg gggatcacca tccgtcgccc gggcgtgtca ataatatcac
     3181 tctgtacatc cacaaacaga cgataacggc tctctctttt ataggtgtaa accttaaact
     3241 gcatttcacc agcccctgtt ctcgtcagca aaagagccgt tcatttcaat aaaccgggcg
     3301 acctcagcca tcccttcctg attttccgct ttccagcgtt cggcacgcag acgacgggct
     3361 tcattctgca tggttgtgct taccagaccg gagatattga catcatatat gccttgagca
     3421 actgatagct gtcgctgtca actgtcactg taatacgctg cttcatagca tacctctttt
     3481 tgacatactt cgggtataca tatcagtata tattcttata ccgcaaaaat cagcgcgcaa
     3541 atacgcatac tgttatctgg cttttagtaa gccggatcca cgcggcgttt acgccccgcc
     3601 ctgccactca tcgcagtact gttgtaattc attaagcatt ctgccgacat ggaagccatc
     3661 acagacggca tgatgaacct gaatcgccag cggcatcagc accttgtcgc cttgcgtata
     3721 atatttgccc atggtgaaaa cgggggcgaa gaagttgtcc atattggcca cgtttaaatc
     3781 aaaactggtg aaactcaccc agggattggc tgagacgaaa aacatattct caataaaccc
     3841 tttagggaaa taggccaggt tttcaccgta acacgccaca tcttgcgaat atatgtgtag
     3901 aaactgccgg aaatcgtcgt ggtattcact ccagagcgat gaaaacgttt cagtttgctc
     3961 atggaaaacg gtgtaacaag ggtgaacact atcccatatc accagctcac cgtctttcat
     4021 tgccatacgg aattccggat gagcattcat caggcgggca agaatgtgaa taaaggccgg
     4081 ataaaacttg tgcttatttt tctttacggt ctttaaaaag gccgtaatat ccagctgaac
     4141 ggtctggtta taggtacatt gagcaactga ctgaaatgcc tcaaaatgtt ctttacgatg
     4201 ccattgggat atatcaacgg tggtatatcc agtgattttt ttctccattt tagcttcctt
     4261 agctcctgaa aatctcgata actcaaaaaa tacgcccggt agtgatctta tttcattatg
     4321 gtgaaagttg gaacctctta cgtgccgatc aacgtctcat tttcgccaaa agttggccca
     4381 gggcttcccg gtatcaacag ggacaccagg atttatttat tctgcgaagt gatcttccgt
     4441 cacaggtatt tattcggcgc aaagtgcgtc gggtgatgct gccaacttag tcgactacag
     4501 gtcactaata ccatctaagt agttgattca tagtgactgg atatgttgtg ttttacagta
     4561 ttatgtagtc tgttttttat gcaaaatcta atttaatata ttgatattta tatcatttta
     4621 cgtttctcgt tcagctttct tgtacaaagt tggcattata agaaagcatt gcttatcaat
     4681 ttgttgcaac gaacaggtca ctatcagtca aaataaaatc attatttgcc atccagctga
     4741 tatcccctat agtgagtcgt attacatggt catagctgtt tcctggcagc tctggcccgt
     4801 gtctcaaaat ctctgatgtt acattgcaca agataaaata atatcatcat gaacaataaa
     4861 actgtctgct tacataaaca gtaatacaag gggtgttatg agccatattc aacgggaaac
     4921 gtcgaggccg cgattaaatt ccaacatgga tgctgattta tatgggtata aatgggctcg
     4981 cgataatgtc gggcaatcag gtgcgacaat ctatcgcttg tatgggaagc ccgatgcgcc
     5041 agagttgttt ctgaaacatg gcaaaggtag cgttgccaat gatgttacag atgagatggt
     5101 cagactaaac tggctgacgg aatttatgcc tcttccgacc atcaagcatt ttatccgtac
     5161 tcctgatgat gcatggttac tcaccactgc gatccccgga aaaacagcat tccaggtatt
     5221 agaagaatat cctgattcag gtgaaaatat tgttgatgcg ctggcagtgt tcctgcgccg
     5281 gttgcattcg attcctgttt gtaattgtcc ttttaacagc gatcgcgtat ttcgtctcgc
     5341 tcaggcgcaa tcacgaatga ataacggttt ggttgatgcg agtgattttg atgacgagcg
     5401 taatggctgg cctgttgaac aagtctggaa agaaatgcat aaacttttgc cattctcacc
     5461 ggattcagtc gtcactcatg gtgatttctc acttgataac cttatttttg acgaggggaa
     5521 attaataggt tgtattgatg ttggacgagt cggaatcgca gaccgatacc aggatcttgc
     5581 catcctatgg aactgcctcg gtgagttttc tccttcatta cagaaacggc tttttcaaaa
     5641 atatggtatt gataatcctg atatgaataa attgcagttt catttgatgc tcgatgagtt
     5701 tttctaatca gaattggtta attggttgta acactggcag agcattacgc tgacttgacg
     5761 ggacggcgca agctcatgac caaaatccct taacgtgagt tacgcgtcgt tccactgagc
     5821 gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat
     5881 ctgctgcttg caaacaaaaa aaccaccgct accagcggtg gtttgtttgc cggatcaaga
     5941 gctaccaact ctttttccga aggtaactgg cttcagcaga gcgcagatac caaatactgt
     6001 tcttctagtg tagccgtagt taggccacca cttcaagaac tctgtagcac cgcctacata
     6061 cctcgctctg ctaatcctgt taccagtggc tgctgccagt ggcgataagt cgtgtcttac
     6121 cgggttggac tcaagacgat agttaccgga taaggcgcag cggtcgggct gaacgggggg
     6181 ttcgtgcaca cagcccagct tggagcgaac gacctacacc gaactgagat acctacagcg
     6241 tgagctatga gaaagcgcca cgcttcccga agggagaaag gcggacaggt atccggtaag
     6301 cggcagggtc ggaacaggag agcgcacgag ggagcttcca gggggaaacg cctggtatct
     6361 ttatagtcct gtcgggtttc gccacctctg acttgagcgt cgatttttgt gatgctcgtc
     6421 aggggggcgg agcctatgga aaaacgccag caacgcggcc tttttacggt tcctggcctt
     6481 ttgctggcct tttgctcaca tgtt
//
