LOCUS       V005319                 7456 bp    DNA     circular SYN 18-DEC-2018
DEFINITION  Exported.
ACCESSION   V005319
VERSION     V005319
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
            .
REFERENCE   1  (bases 1 to 7456)
  AUTHORS   Gong Z, Lv H.
  TITLE     Vector pJKPN for targeted integration at the Leu1-32 of
            Schizosaccharomyces pombe
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 7456)
  AUTHORS   Gong Z, Lv H.
  TITLE     Direct Submission
  JOURNAL   Submitted (28-SEP-2011) School of Life Science, Fudan University,
            Room 618, Science Building, Fudan University, Handan Road 220,
            Shanghai, Shanghai 200433, China
REFERENCE   3  (bases 1 to 7456)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 7456)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName:
            "Unpublished"
            SGRef: number: 2; type: "Journal Article"; journalName: "Submitted
            (28-SEP-2011) School of Life Science, Fudan University, Room 618,
            Science Building, Fudan University, Handan Road 220, Shanghai,
            Shanghai 200433, China"
            SGRef: number: 3; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..7456
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     5'UTR           187..306
                     /gene="Leu1"
     CDS             307..1419
                     /gene="leu1"
                     /label="3-isopropylmalate dehydrogenase"
                     /note="3-isopropylmalate dehydrogenase from
                     Schizosaccharomyces pombe (strain 972 / ATCC 24843).
                     Accession#: P18869"
     3'UTR           1423..2258
                     /gene="Leu1"
     rep_origin      complement(2308..2763)
                     /direction=LEFT
                     /label="f1 ori"
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     primer_bind     2910..2926
                     /label="M13 fwd"
                     /note="common sequencing primer, one of multiple similar
                     variants"
     promoter        2936..2954
                     /label="T7 promoter"
                     /note="promoter for bacteriophage T7 RNA polymerase"
     promoter        2995..4159
                     /label="nmt1 promoter"
                     /note="wild-type nmt1 promoter from Schizosaccharomyces
                     pombe, conferring strong thiamine-repressible expression"
     regulatory      4181..5177
                     /label="NMT1 terminator"
                     /note="NMT1 terminator"
                     /regulatory_class="terminator"
     promoter        complement(5196..5214)
                     /label="T3 promoter"
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(5235..5251)
                     /label="M13 rev"
                     /note="common sequencing primer, one of multiple similar
                     variants"
     protein_bind    complement(5259..5275)
                     /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)."
     promoter        complement(5283..5313)
                     /label="lac promoter"
                     /note="promoter for the E. coli lac operon"
     protein_bind    complement(5328..5349)
                     /label="CAP binding site"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     rep_origin      complement(5637..6225)
                     /direction=LEFT
                     /label="ori"
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
     CDS             complement(6399..7256)
                     /label="AmpR"
                     /note="beta-lactamase"
     promoter        complement(7257..7361)
                     /label="AmpR promoter"
ORIGIN
        1 tcgcgcgttt cggtgatgac ggtgaaaacc tctgacacat gcagctcccg gagacggtca
       61 cagcttgtct gtaagcggat gccgggagca gacaagcccg tcagggcgcg tcagcgggtg
      121 ttggcgggtg tcggggctgg cttaactatg cggcatcaga gcagattgta ctgagagtgc
      181 accatacgat atcccaatct gtagtataaa aagtgcaaca gtgttgagtt tcccgaaacc
      241 aggcattttt gcggtagggt ctgaaacttt cacgaaacca tttcacaaca aaacttcttc
      301 aacaacatgt gtgcaaagaa aatcgtcgtc ttaccaggag accatattgg ccctgaaatt
      361 gttgcttctg ccttggaggt tttgaaagtc gttgagaaga agcgacctga gttaaaactc
      421 gagtttgaag aacacaagat tggaggtgcc tctattgatg cctatggaac ccctttgact
      481 gacgagactg tgaaggcttg tttggaagct gacggtgttc ttttgggtgc cgttggtggt
      541 cctgaatgga ccaaccccaa ttgtcgtcct gagcaaggtt tattgaagct tcgtaagagt
      601 atgggtgttt gggccaacct tcgaccttgc aactttgcca gcaagtcttt agtcaagtac
      661 agccctttga agcctgaaat cgttgaaggt gtcgattttt gtgttgtacg agaacttact
      721 ggaggttgtt actttggtga gcgcactgag gacaacggat cgggttatgc tatggacact
      781 tggccttaca gtttggaaga agtttctcgt attgctcgtt tggctgcttg gttagctgaa
      841 acttccaacc ctcctgctcc cgtcacatta ctcgacaaag ctaatgtttt ggcaacttct
      901 cgtctttggc gcaagactgt agctaagatt tttaaggagg aatatcctca ccttacctta
      961 aaaaaccagc tcattgactc cgcagccatg cttttggtca agagccctcg tacacttaac
     1021 ggtgttgttt tgactgacaa cttgtttggt gacattattt cagatgaggc ttctgtcatt
     1081 cctggtagct tgggcctttt gccttctgcc tccctttccg gtgtggtagg aaaatcagaa
     1141 gaaaaggttc attgtttggt tgagcccatt cacggtagcg ctcccgatat cgctggtaag
     1201 ggcattgtta atcctgttgg tacaatttta tctgcttccc ttctccttcg ttatggtttg
     1261 aatgctccta aggaggctga agctatcgaa gccgccgtac gcaaggtctt ggatgatact
     1321 tcaattggtg gacgtggtct ttatactcgc gatttgggag gtgaggcttc taccgctgat
     1381 attactaagg ctgttgttga agaacttgaa aaaattttgt aatagggttt aatgtagaat
     1441 aaattcatat gatttgattt gacctaattt ttattgatta attgtgtcgc atatttatta
     1501 tcgttaaaaa tgcaattgac aaaagcgtaa attttaaggc tctaatgttt tttcttttgg
     1561 tatgttatag tcaagcaaag atgtattaag ttgcaaaagc atggaattga tttacgacat
     1621 ccgttcaact ttatgaccga gttgtcgctg tgtacttact ctttttttac tatgaaaagt
     1681 ttcaaacata atatcattaa ttatttcttt ttaaacgaag taacctgtaa cataaataat
     1741 aggggaaaag aagaaaaaaa ccataaaaat tcgagaatcg taacaattat tattgtaacg
     1801 atgacaaccc gattctttga gagagccaga gacatgactt ttgcattacc gtaattggtt
     1861 tctacttatg aattgtaaac ttgtgtaggg aatttcatcc accagctatc gcgggctgct
     1921 ctctgcatca gtgttagcac attttatatt aatgaagatt ttgctatgca aaatacgtaa
     1981 atttctggcg cgatatattt ttatcaaacg aacatgacta tcctttccac tttgatggtt
     2041 cgcaagtatt ttgatttgta ttaaatgcac atctaacaag ccatttcgcc caaaatcacc
     2101 aactggcaca tgtacccaac cagttaggtt cctcgacttg aactgttgta acaatttcca
     2161 aatcttgaaa ccctgtccca gctgatattc gcaaagtact gggagtataa gattcatcaa
     2221 gggtgtattg aaggtacata ctcacatact taatcgatgc ggtgtgaaat accgcacaga
     2281 tgcgtaagga gaaaataccg catcaggaaa ttgtaaacgt taatattttg ttaaaattcg
     2341 cgttaaattt ttgttaaatc agctcatttt ttaaccaata ggccgaaatc ggcaaaatcc
     2401 cttataaatc aaaagaatag accgagatag ggttgagtgt tgttccagtt tggaacaaga
     2461 gtccactatt aaagaacgtg gactccaacg tcaaagggcg aaaaaccgtc tatcagggcg
     2521 atggcccact acgtgaacca tcaccctaat caagtttttt ggggtcgagg tgccgtaaag
     2581 cactaaatcg gaaccctaaa gggagccccc gatttagagc ttgacgggga aagccggcga
     2641 acgtggcgag aaaggaaggg aagaaagcga aaggagcggg cgctagggcg ctggcaagtg
     2701 tagcggtcac gctgcgcgta accaccacac ccgccgcgct taatgcgccg ctacagggcg
     2761 cgtcgcgcca ttcgccattc aggctgcgca actgttggga agggcgatcg cggtgcgggc
     2821 ctcttcgcta ttacgccagc tggcgaaagg gggatgtgct gcaaggcgat taagttgggt
     2881 aacgccaggg ttttcccagt cacgacgttg taaaacgacg gccagtgagc gcgcgtaata
     2941 cgactcacta tagggcgaat tggggtcgat cgactctaga ggatcagaaa attatcgcca
     3001 taaaagacag aataagtcat cagcggttgt ttcatttcct atattttttt tttatttttt
     3061 tattttttaa taagggaaaa tttaacgtct aaggatacag aagattgtta gcacattaaa
     3121 gtaataaagg cttaagtagt aagtgcctta gcatgttatt gtatttcaaa ggacataatc
     3181 taaaataata acaatatcat ttctcacaag ttattcaatt ttcttttttt tttctaataa
     3241 tatcaagaat gtattatttg tttgacataa gtcaactaat ttatttaata tgctggatta
     3301 atcttgcaga catgtaaatt aacaagtttt agtcaaataa cgttgaagtt tcaatgaact
     3361 caaataattt ctcttttttt ttatataacc ataagtctaa tctgatttat attttccgca
     3421 gggatcaact gaagttatga catttggatt ggatcactta taaccttggt cgccaaataa
     3481 tacaaaaatc agcgttataa aacaaagaag gtttttgtta agaaattaat cctctttctt
     3541 gataagaaag ttgaaccgaa attgcagata ctgatatatg aaaataatac ccacaatttt
     3601 gggaatagcg caagcctcaa tttaaacaat aggtgaggac acatgataat gacctcaatg
     3661 attgttagaa gaaaagagcc tcattacaaa atcgaaaaat gaatggttgg gtacaagttt
     3721 ccaaaacatg gtaaagtgga ctttgcgtat gagacgtaaa tagaaaaaaa cacttgttat
     3781 atgttttcta gaattattgt tgtctcttta tggttggatg atgcaaaata gtaatttcgg
     3841 ttagttgctg taaaacacca cgagacaaat agatatggat atttattaaa tcaggaaaaa
     3901 cgtaactctc ggctactgga tggttcagtc acccaacgat tactggggag agaaaacagg
     3961 gcaaaagcaa agcttaaagg aatccgattg tcattcggca atgtgcagcg aaactaaaaa
     4021 ccggataatg gacctgttaa tcgaaacatt gaagatatat aaaggaagag gaatcctggc
     4081 atatcatcaa ttgaataagt tgaattaatt atttcaatct cattctcact ttctgactta
     4141 tagtcgcttt gttaaatcat atgtcgactc tagaggatcc ccgggtaaaa ggaatgtctc
     4201 ccttgccagt actgctaggg tttttctttc aaactatgga agcccattca agctgcatat
     4261 tacgattttg tttttcgctt ttagaaagtg gtttagatga gataatagaa aaattcttga
     4321 tctccgacaa cgagtacttt tatttttttt gctaatcact ttactcaata ttagctcgaa
     4381 atcgtagaaa cgtagacggg tgcgggatac cgagtggtgt agttaagaat ttttataaac
     4441 cacgtggccc aaaaatatga acccaaaacg tttatacatg agtatacttt aagaaggcta
     4501 taccccttcg tgttagatgt agttttagct acccaacccg agtctatgag cttgacttca
     4561 gatgtagaag gcattaaatc gttttgaata ttaattaaaa aacgatgaaa attaaatatt
     4621 taaaagcaat catacgctga aaatttagtg ctgtggctaa tccttcaaca tggaaatgcc
     4681 ataaaagtga ctttgacaaa aaaaaaagta tatacaggta gtaaactcat ctacttcatt
     4741 gactttgttt acagcatgtg gaaggaggaa tatttattgc taaatcgtag tttaacattc
     4801 aataagtaat actattgaaa ttcgacaaga ttggccgcat ggatgaaaaa gaggcatttt
     4861 gctttgggag aattagttca aattagaact gaaaaaaaaa actttacgag gcaaaaatgt
     4921 cggattgaga tcgtaaaagt tcgctcgtcg tcttttgctt tgtgattgtt ttcatggata
     4981 catcttgctg gatatttaaa ttttagtact atgtataaga tattctataa atgttttatc
     5041 acccaaacct gttagcgcct tcttaattct attcaatctg gcttttgctc tgagactact
     5101 tcttggactt tcactacttg ttagttatac ggaatttgtg taattagaag tgaaataatc
     5161 ctttctatta gtaatgcgag ctccagcttt tgttcccttt agtgagggtt aattgcgcgc
     5221 ttggcgtaat catggtcata gctgtttcct gtgtgaaatt gttatccgct cacaattcca
     5281 cacaacatag gagccggaag cataaagtgt aaagcctggg gtgcctaatg agtgaggtaa
     5341 ctcacattaa ttgcgttgcg ctcactgccc gctttccagt cgggaaacct gtcgtgccag
     5401 ctgcattaat gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg gcgctcttcc
     5461 gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct
     5521 cactcaaagg cggtaatacg gttatccaca gaatcagggg ataacgcagg aaagaacatg
     5581 tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc
     5641 cataggctcg gcccccctga cgagcatcac aaaaatcgac gctcaagtca gaggtggcga
     5701 aacccgacag gactataaag ataccaggcg ttcccccctg gaagctccct cgtgcgctct
     5761 cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc gggaagcgtg
     5821 gcgctttctc aatgctcacg ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag
     5881 ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc cggtaactat
     5941 cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc cactggtaac
     6001 aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg gtggcctaac
     6061 tacggctaca ctagaaggac agtatttggt atctgcgctc tgctgaagcc agttaccttc
     6121 ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag cggtggtttt
     6181 tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc
     6241 ttttctacgg ggtctgacgc tcagtggaac gaaaactcac gttaagggat tttggtcatg
     6301 agattatcaa aaaggatctt cacctagatc cttttaaatt aaaaatgaag ttttaaatca
     6361 atctaaagta tatatgagta aacttggtct gacagttacc aatgcttaat cagtgaggca
     6421 cctatctcag cgatctgtct atttcgttca tccatagttg cctgactgcc cgtcgtgtag
     6481 ataactacga tacgggaggg cttaccatct ggccccagtg ctgcaatgat accgcgagac
     6541 ccacgctcac cggctccaga tttatcagca ataaaccagc cagccggaag ggccgagcgc
     6601 agaagtggtc ctgcaacttt atccgcctcc atccagtcta ttaattgttg ccgggaagct
     6661 agagtaagta gttcgccagt taatagtttg cgcaacgttg ttgccattgc tacaggcatc
     6721 gtggtgtcac gctcgtcgtt tggtatggct tcattcagct ccggttccca acgatcaagg
     6781 cgagttacat gatcccccat gttgtgaaaa aaagcggtta gctccttcgg tcctccgatc
     6841 gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg ttatggcagc actgcataat
     6901 tctcttactg tcatgccatc cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag
     6961 tcattctgag aatagtgtat gcggcgaccg agttgctctt gcccggcgtc aatacgggat
     7021 aataccgcgc cacatagcag aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg
     7081 cgaaaactct caaggatctt accgctgttg agatccagtt cgatgtaacc cactcgtgca
     7141 cccaactgat cttcagcatc ttttactttc accagcgttt ctgggtgagc aaaaacagga
     7201 aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat actcatactc
     7261 ttcctttttc aatattattg aagcatttat cagggttatt gtctcatgag cggatacata
     7321 tttgaatgta tttagaaaaa taaacaaata ggggttccgc gcacatttcc ccgaaaagtg
     7381 ccacctgacg tctaagaaac cattattatc atgacattaa cctataaaaa taggcgtatc
     7441 acgaggccct ttcgtc
//
