LOCUS       40924_17104        9267 bp DNA     circular SYN 17-DEC-2018
DEFINITION  Cloning vector pEG(KT), complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9267)
  AUTHORS   Pierce BD, Wendland B.
  TITLE     Sequence of the yeast protein expression plasmid pEG(KT)
  JOURNAL   Yeast 26 (6), 349-353 (2009)
  PUBMED    19350530
REFERENCE   2  (bases 1 to 9267)
  AUTHORS   Pierce BD, Wendland B.
  TITLE     Direct Submission
  JOURNAL   Submitted (03-DEC-2008) Biology, The Johns Hopkins University, 3400 
            N. Charles St., Mudd Hall, Baltimore, MD 21218, USA
REFERENCE   3  (bases 1 to 9267)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 9267)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Yeast"; 
            date: "2009"; volume: "26"; issue: "6"; pages: "349-353"
COMMENT     SGRef: number: 2; type: "Journal Article"; journalName: "Submitted 
            (03-DEC-2008) Biology, The Johns Hopkins University, 3400 N. Charles
            St., Mudd Hall, Baltimore, MD 21218, USA"
COMMENT     SGRef: number: 3; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..9267
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             22..39
                     /label=thrombin site
                     /note="thrombin recognition and cleavage site"
     rep_origin      327..1309
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     CDS             1676..2767
                     /label=LEU2
                     /note="3-isopropylmalate dehydrogenase, required for
                     leucine biosynthesis"
     primer_bind     complement(3021..3037)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     rep_origin      3477..3990
                     /label=M13 ori
                     /note="M13 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     promoter        4735..4839
                     /label=AmpR promoter
     CDS             4840..5697
                     /label=AmpR
                     /note="beta-lactamase"
     rep_origin      5871..6459
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     protein_bind    6746..6767
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     promoter        6782..6812
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6820..6836
                     /label=lac operator
                     /note="The lac repressor binds to the lac operator to
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     primer_bind     6844..6860
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        6880..7091
                     /label=URA3 promoter
     CDS             7092..7892
                     /label=URA3
                     /note="orotidine-5'-phosphate decarboxylase, required for
                     uracil biosynthesis"
     protein_bind    complement(8152..8269)
                     /label=UAS
                     /note="upstream activating sequence mediating
                     Gal4-dependent induction"
     regulatory      8344..8589
                     /label=CYC1
                     /note="CYC1"
                     /regulatory_class="promoter"
     CDS             8608..9261
                     /label=GST
                     /note="glutathione S-transferase from Schistosoma
                     japonicum"
ORIGIN
        1 ctgtctggtg gtggtggtgg tctggttccg cgtggatccc cgggaattct agacaccatg
       61 ggtcgactcg agctcaagct taattcatcg tgactgactg acgatctgcc tcgcgcgttt
      121 cggtgatgac ggtgaaaacc tctgacacat gcagctcccg gagacggtca cagcttgtct
      181 gtaagcggat gccgggagca gacaagcccg tcagggcgcg tcagcgggtg ttggcgggtg
      241 tcggggcgca gctcgcgttg cattttcgga agcgctcgtt ttcggaaacg ctttgaagtt
      301 cctattccga agttcctatt ctctagctag aaagtatagg aacttcagag cgcttttgaa
      361 aaccaaaagc gctctgaaga cgcactttca aaaaaccaaa aacgcaccgg actgtaacga
      421 gctactaaaa tattgcgaat accgcttcca caaacattgc tcaaaagtat ctctttgcta
      481 tatatctctg tgctatatcc ctatataacc tacccatcca cctttcgctc cttgaacttg
      541 catctaaact cgacctctac attttttatg tttatctcta gtattactct ttagacaaaa
      601 aaattgtagt aagaactatt catagagtga atcgaaaaca atacgaaaat gtaaacattt
      661 cctatacgta gtatatagag acaaaataga agaaaccgtt cataattttc tgaccaatga
      721 agaatcatca acgctatcac tttctgttca caaagtatgc gcaatccaca tcggtataga
      781 atataatcgg ggatgccttt atcttgaaaa aatgcacccg cagcttcgct agtaatcagt
      841 aaacgcggga agtggagtca ggcttttttt atggaagaga aaatagacac caaagtagcc
      901 ttcttctaac cttaacggac ctacagtgca aaaagttatc aagagactgc attatagagc
      961 gcacaaagga gaaaaaaagt aatctaagat gctttgttag aaaaatagcg ctctcgggat
     1021 gcatttttgt agaacaaaaa agaagtatag attctttgtt ggtaaaatag cgctctcgcg
     1081 ttgcatttct gttctgtaaa aatgcagctc agattctttg tttgaaaaat tagcgctctc
     1141 gcgttgcatt tttgttttac aaaaatgaag cacagattct tcgttggtaa aatagcgctt
     1201 tcgcgttgca tttctgttct gtaaaaatgc agctcagatt ctttgtttga aaaattagcg
     1261 ctctcgcgtt gcatttttgt tctacaaaat gaagcacaga tgcttcgtta acaaagatat
     1321 gctattgaag tgcaagatgg aaacgcagaa aatgaaccgg ggatgcgacg tgcaagatta
     1381 cctatgcaat agatgcaata gtttctccag gaaccgaaat acatacattg tcttccgtaa
     1441 agcgctagac tatatattat tatacaggtt caaatatact atctgtttca gggaaaactc
     1501 ccaggttcgg atgttcaaaa ttcaatgatg ggtaacaagt acgatcgtaa atctgtaaaa
     1561 cagtttgtcg gatattaggc tgtatctcct caaagcgtat tcgaatatca ttgagaagct
     1621 gcattttttt tttttttttt tttttttata tatatttcaa ggatatacca ttgtaatgtc
     1681 tgcccctaag aagatcgtcg ttttgccagg tgaccacgtt ggtcaagaaa tcacagccga
     1741 agccattaag gttcttaaag ctatttctga tgttcgttcc aatgtcaagt tcgatttcga
     1801 aaatcattta attggtggtg ctgctatcga tgctacaggt gttccacttc cagatgaggc
     1861 gctggaagcc tccaagaagg ctgatgccgt tttgttaggt gctgtgggtg gtcctaaatg
     1921 gggtaccggt agtgttagac ctgaacaagg tttactaaaa atccgtaaag aacttcaatt
     1981 gtacgccaac ttaagaccat gtaactttgc atccgactct cttttagact tatctccaat
     2041 caagccacaa tttgctaaag gtactgactt cgttgttgtt agagaattag tgggaggtat
     2101 ttactttggt aagagaaagg aagacgatgg tgatggtgtc gcttgggata gtgaacaata
     2161 caccgttcca gaagtgcaaa gaatcacaag aatggccgct ttcatggccc tacaacatga
     2221 gccaccattg cctatttggt ccttggataa agctaatgtt ttggcctctt caagattatg
     2281 gagaaaaact gtggaggaaa ccatcaagaa cgaattccct acattgaaag ttcaacatca
     2341 attgattgat tctgccgcca tgatcctagt taagaaccca acccacctaa atggtattat
     2401 aatcaccagc aacatgtttg gtgatatcat ctccgatgaa gcctccgtta tcccaggctc
     2461 cttgggtttg ttgccatctg cgtccttggc ctctttgcca gacaagaaca ccgcatttgg
     2521 tttgtacgaa ccatgccatg gttccgctcc agatttgcca aagaataagg tcaaccctat
     2581 cgccactatc ttgtctgctg caatgatgtt gaaattgtca ttgaacttgc ctgaagaagg
     2641 taaagccatt gaagatgcag ttaaaaaggt tttggatgca ggtatcagaa ctggtgattt
     2701 aggtggttcc aacagtacca ccgaagtcgg tgatgctgtc gccgaagaag ttaagaaaat
     2761 ccttgcttaa aaagattctc tttttttatg atatttgtac aaaaaaaaaa aaaaaaaaaa
     2821 aaaaaaaaaa aaaaaaaaaa aaaaaatgca gcgtcacatc ggataataat gatggcagcc
     2881 attgtagaag tgccttttgc atttctagtc tctttctcgg tctagctagt tttactacat
     2941 cgcgaagata gaatcttaga tcacactgcc tttgctgagc tggatcaata gagtaacaaa
     3001 agagtggtaa ggagcttggc actggccgtc gttttacaac gtcgtgactg ggaaaaccct
     3061 ggcgttaccc aacttaatcg ccttgcagca catccccctt tcgccagtgg cgtaatagcg
     3121 aagaggcccg caccgatcgc ccttcccaac agttgcgcag cctgaatggc gaatggcgat
     3181 tccgttgcaa tggctggcgg taatattgtt ctggatatta ccagcaaggc cgatagtttg
     3241 agttcttcta ctcaggcaag tgatgttatt actaatcaaa gaagtattgc gacaacggtt
     3301 aatttgcgtg atggacagac tcttttactc ggtggcctca ctgattataa aaacacttct
     3361 caggattctg gcgtaccgtt cctgtctaaa atccctttaa tcggcctcct gtttagctcc
     3421 cgctctgatt ctaacgagga aagcacgtta tacgtgctcg tcaaagcaac catagtacgc
     3481 gccctgtagc ggcgcattaa gcgcggcggg tgtggtggtt acgcgcagcg tgaccgctac
     3541 acttgccagc gccctagcgc ccgctccttt cgctttcttc ccttcctttc tcgccacgtt
     3601 cgccggcttt ccccgtcaag ctctaaatcg ggggctccct ttagggttcc gatttagtgc
     3661 tttacggcac ctcgacccca aaaaacttga ttagggtgat ggttcacgta gtgggccatc
     3721 gccctgatag acggtttttc gccctttgac gttggagtcc acgttcttta atagtggact
     3781 cttgttccaa actggaacaa cactcaaccc tatctcggtc tattcttttg atttataagg
     3841 gattttgccg atttcggcct attggttaaa aaatgagctg atttaacaaa aatttaacgc
     3901 gaattttaac aaaatattaa cgtttacaat ttaaatattt gcttatacaa tcttcctgtt
     3961 tttggggctt ttctgattat caaccggggt acatatgatt gacatgctag ttttacgatt
     4021 accgttcatc gattctcttg tttgctccag actctcaggc aatgacctga tagcctttgt
     4081 agacctctca aaaatagcta ccctctccgg catgaattta tcagctagaa cggttgaata
     4141 tcatattgat ggtgatttga ctgtctccgg cctttctcac ccgtttgaat ctttacctac
     4201 acattactca ggcattgcat ttaaaatata tgagggttct aaaaattttt atccttgcgt
     4261 tgaaataaag gcttctcccg caaaagtatt acagggtcat aatgtttttg gtacaaccga
     4321 tttagcttta tgctctgagg ctttattgct taattttgct aattctttgc cttgcctgta
     4381 tgatttattg gatgttggaa tcgcctgatg cggtattttc tccttacgca tctgtgcggt
     4441 atttcacacc gcatatggtg cactctcagt acaatctgct ctgatgccgc atagttaagc
     4501 cagccccgac acccgccaac acccgctgac gcgccctgac gggcttgtct gctcccggca
     4561 tccgcttaca gacaagctgt gaccgtctcc gggagctgca tgtgtcagag gttttcaccg
     4621 tcatcaccga aacgcgcgag acgaaagggc ctcgtgatac gcctattttt ataggttaat
     4681 gtcatgataa taatggtttc ttagacgtca ggtggcactt ttcggggaaa tgtgcgcgga
     4741 acccctattt gtttattttt ctaaatacat tcaaatatgt atccgctcat gagacaataa
     4801 ccctgataaa tgcttcaata atattgaaaa aggaagagta tgagtattca acatttccgt
     4861 gtcgccctta ttcccttttt tgcggcattt tgccttcctg tttttgctca cccagaaacg
     4921 ctggtgaaag taaaagatgc tgaagatcag ttgggtgcac gagtgggtta catcgaactg
     4981 gatctcaaca gcggtaagat ccttgagagt tttcgccccg aagaacgttt tccaatgatg
     5041 agcactttta aagttctgct atgtggcgcg gtattatccc gtattgacgc cgggcaagag
     5101 caactcggtc gccgcataca ctattctcag aatgacttgg ttgagtactc accagtcaca
     5161 gaaaagcatc ttacggatgg catgacagta agagaattat gcagtgctgc cataaccatg
     5221 agtgataaca ctgcggccaa cttacttctg acaacgatcg gaggaccgaa ggagctaacc
     5281 gcttttttgc acaacatggg ggatcatgta actcgccttg atcgttggga accggagctg
     5341 aatgaagcca taccaaacga cgagcgtgac accacgatgc ctgtagcaat ggcaacaacg
     5401 ttgcgcaaac tattaactgg cgaactactt actctagctt cccggcaaca attaatagac
     5461 tggatggagg cggataaagt tgcaggacca cttctgcgct cggcccttcc ggctggctgg
     5521 tttattgctg ataaatctgg agccggtgag cgtgggtctc gcggtatcat tgcagcactg
     5581 gggccagatg gtaagccctc ccgtatcgta gttatctaca cgacggggag tcaggcaact
     5641 atggatgaac gaaatagaca gatcgctgag ataggtgcct cactgattaa gcattggtaa
     5701 ctgtcagacc aagtttactc atatatactt tagattgatt taaaacttca tttttaattt
     5761 aaaaggatct aggtgaagat cctttttgat aatctcatga ccaaaatccc ttaacgtgag
     5821 ttttcgttcc actgagcgtc agaccccgta gaaaagatca aaggatcttc ttgagatcct
     5881 ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt
     5941 tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt cagcagagcg
     6001 cagataccaa atactgttct tctagtgtag ccgtagttag gccaccactt caagaactct
     6061 gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc tgccagtggc
     6121 gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa ggcgcagcgg
     6181 tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac ctacaccgaa
     6241 ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg
     6301 gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga gcttccaggg
     6361 ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact tgagcgtcga
     6421 tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa cgcggccttt
     6481 ttacggttcc tggccttttg ctggcctttt gctcacatgt tctttcctgc gttatcccct
     6541 gattctgtgg ataaccgtat taccgccttt gagtgagctg ataccgctcg ccgcagccga
     6601 acgaccgagc gcagcgagtc agtgagcgag gaagcggaag agcgcccaat acgcaaaccg
     6661 cctctccccg cgcgttggcc gattcattaa tgcagtggca cgacaggttt cccgactgga
     6721 aagcgggcag tgagcgcaac gcaattaatg tgagttagct cactcattag gcaccccagg
     6781 ctttacactt tatgcttccg gctcgtatgt tgtgtggaat tgtgagcgga taacaatttc
     6841 acacaggaaa cagctatgac catgattacg aattagcttt tcaattcatc attttttttt
     6901 tattcttttt tttgatttcg gtttccttga aatttttttg attcggtaat ctccgaacag
     6961 aaggaagaac gaaggaagga gcacagactt agattggtat atatacgcat atgtagtgtt
     7021 gaagaaacat gaaattgccc agtattctta acccaactgc acagaacaaa aaccggaaac
     7081 gaagataaat catgtcgaaa gctacatata aggaacgtgc tgctactcat cctagtcctg
     7141 ttgctgccaa gctatttaat atcatgcacg aaaagcaaac aaacttgtgt gcttcattgg
     7201 atgttcgtac caccaaggaa ttactggagt tagttgaagc attaggtccc aaaatttgtt
     7261 tactaaaaac acatgtggat atcttgactg atttttccat ggagggcaca gttaagccgc
     7321 taaaggcatt atccgccaag tacaattttt tactcttcga agacagaaaa tttgctgaca
     7381 ttggtaatac agtcaaattg cagtactctg cgggtgtata cagaatagca gaatgggcag
     7441 acattacgaa tgcacacggt gtggtgggcc caggtattgt tagcggtttg aagcaggcgg
     7501 cggaagaagt aacaaaggaa cctagaggcc ttttgatgtt agcagaattg tcatgcaagg
     7561 gctccctagc tactggagaa tatactaagg gtactgttga cattgcgaag agcgacaaag
     7621 attttgttat cggctttatt gctcaaagag acatgggtgg aagagatgaa ggttacgatt
     7681 ggttgattat gacacccggt gtgggtttag atgacaaggg agacgcattg ggtcaacagt
     7741 atagaaccgt ggatgatgtg gtctctacag gatctgacat tattattgtt ggaagaggac
     7801 tatttgcaaa gggaagggat gctaaggtag agggtgaacg ttacagaaaa gcaggctggg
     7861 aagcatattt gagaagatgc ggccagcaaa actaaaaaac tgtattataa gtaaatgcat
     7921 gtatactaaa ctcacaaatt agagcttcaa tttaattata tcagttatta cccgatcaaa
     7981 aatcatcgct tcgctgatta attaccccag aaataaggct aaaaaactaa tcgcattatc
     8041 atcctatggt tgttaatttg attcgttcat ttgaaggttt gtggggccag gttactgcca
     8101 atttttcctc ttcataacca taaaagctag tattgtagaa tctttattgt tcggagcagt
     8161 gcggcgcgag gcacatctgc gtttcaggaa cgcgaccggt gaagacgagg acgcacggag
     8221 gagagtcttc cttcggaggg ctgtcacccg ctcggcggct tctaatccgt acttcaatat
     8281 agcaatgagc agttaagcgt attactgaaa gttccaaaga gaaggttttt ttaggctaat
     8341 cgacctcgag cagatccgcc aggcgtgtat atagcgtgga tggccaggca actttagtgc
     8401 tgacacatac aggcatatat atatgtgtgc gacgacacat gatcatatgg catgcatgtg
     8461 ctctgtatgt atataaaact cttgttttct tcttttctct aaatattctt tccttataca
     8521 ttaggtcctt tgtagcataa attactatac ttctatagac acgcaaacac aaatacacac
     8581 actaaattac cggatcaatt cgagctcatg tcccctatac taggttattg gaaaattaag
     8641 ggccttgtgc aacccactcg acttcttttg gaatatcttg aagaaaaata tgaagagcat
     8701 ttgtatgagc gcgatgaagg tgataaatgg cgaaacaaaa agtttgaatt gggtttggag
     8761 tttcccaatc ttccttatta tattgatggt gatgttaaat taacacagtc tatggccatc
     8821 atacgttata tagctgacaa gcacaacatg ttgggtggtt gtccaaaaga gcgtgcagag
     8881 atttcaatgc ttgaaggagc ggttttggat attagatacg gtgtttcgag aattgcatat
     8941 agtaaagact ttgaaactct caaagttgat tttcttagca agctacctga aatgctgaaa
     9001 atgttcgaag atcgtttatg tcataaaaca tatttaaatg gtgatcatgt aacccatcct
     9061 gacttcatgt tgtatgacgc tcttgatgtt gttttataca tggacccaat gtgcctggat
     9121 gcgttcccaa aattagtttg ttttaaaaaa cgtattgaag ctatcccaca aattgataag
     9181 tacttgaaat ccagcaagta tatagcatgg cctttgcagg gctggcaagc cacgtttggt
     9241 ggtggcgacc atcctccaaa atcggat
//
