LOCUS       40924_34366        5559 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Expression vector pPeCre DNA, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5559)
  AUTHORS   Kasai Y, Tsukahara T, Ikeda F, Ide Y, Harayama S.
  TITLE     Improving triacylglycerol production in the unicellular green alga 
            Pseudochoricystis ellipsoidea by metabolic engineering using 
            iterative self-cloning approaches
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 5559)
  AUTHORS   Kasai Y, Harayama S.
  TITLE     Direct Submission
  JOURNAL   Submitted (27-OCT-2017) Contact:Yuki Kasai Chuo University, Research
            and Development Initiative; 1-13-27 Kasuga, Bunkyo-ku, Tokyo 
            112-8551, Japan
REFERENCE   3  (bases 1 to 5559)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 5559)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: 
            "Unpublished"
COMMENT     SGRef: number: 2; type: "Journal Article"; journalName: "Submitted 
            (27-OCT-2017) Contact:Yuki Kasai Chuo University, Research and 
            Development Initiative; 1-13-27 Kasuga, Bunkyo-ku, Tokyo 112-8551, 
            Japan"
COMMENT     SGRef: number: 3; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..5559
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     regulatory      1..814
                     /note="includes promoter region of Pseudochoricystis
                     ellipsoidea nuclear gene RBCS for ribulose bisphosphate 
                     carboxylase/oxygenase small subunit"
                     /regulatory_class="promoter"
     CDS             1105..2133
                     /label=Cre
                     /note="site-specific recombinase"
     regulatory      2137..2674
                     /note="includes terminator region of Pseudochoricystis 
                     ellipsoidea nuclear gene RBCS for ribulose bisphosphate 
                     carboxylase/oxygenase small subunit"
                     /regulatory_class="terminator"
     promoter        complement(2699..2717)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2727..2743)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     rep_origin      complement(2884..3339)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     promoter        3366..3470
                     /label=AmpR promoter
     CDS             3471..4328
                     /label=AmpR
                     /note="beta-lactamase"
     rep_origin      4502..5090
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     protein_bind    5378..5399
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     promoter        5414..5444
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5452..5468
                     /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     5476..5492
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        5513..5531
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
ORIGIN
        1 gcggccgctc tagaactagt atgccctcaa tcggccgatt tcatgcatga agaagaagat
       61 gatgattaca catcattcca gccatctagg gctgccatca gtaattgcat gacccgcaac
      121 ttcccagtca gtcaaatgca caatagaagc atagttgcat gggaatcaac ccagcagcct
      181 cccgcagagc tagcctaggt acgtgtacgc agccacataa tcatcatgaa tgattctgca
      241 cggtgctatg atagcaacag atactcgcca gtgcgcacgc aagcaatctc atcacaccac
      301 tgtggtcagg caagcacgat atagatgtca aatgacagct ccattgggag tgcaagtcta
      361 ctagagtgca agcctactag tagtaggaag gtccagcgca tgccaaaact cctgctaagg
      421 ggttgcccaa gccccttggc tggccaaaag tccttaatat tttgctaggt aaaggggatc
      481 ttcccggagg tggtgttgat gaacatttgt acaccagcat ccgggtcctt ttggactctg
      541 aaaacgtgag cgagcagcga ctgcttgcag cttccaaacg aaaggtctgg aagcgctgga
      601 gtggtgagtc aggggtgact cagcaaacac caaggacaca cgttttcacc ctaccctcag
      661 aacccttgaa ataggggtcg atttccctct aacttaagtg gttatccaca aactcacgac
      721 gataagagcc agcgccagca ccatttcgtt cctcattttc agctgaaagc aaagcctttg
      781 catacttatc gctctcagat tacaatcgca atgggtacgc agctttctca ctccctcacc
      841 gcgtgtggtt cactcaggtt tctggctaag gaaagtctga tgtgactttc cttagacgtt
      901 aaagcagaat caccccatgc tttaagcagt tttatgtgtc ttgcccttca tgtgaagggt
      961 ctgtactgtt gcagagtgct ctcttgtgtt cttgagtgtt cttgtctcca gtgtgcccat
     1021 taactttatc catgtcttat gtgtccagga ctgcgccaaa gaaactgacg ctgcgctgtt
     1081 ttgctttgtg cagccatggg atccatgagc aacctgctga ccgtgcacca gaacctgccc
     1141 gccctgcccg tggacgcaac aagcgacgag gtgcgcaaga acctgatgga catgttccgc
     1201 gaccgccagg ccttctcaga gcacacctgg aagatgctgc tgagcgtgtg ccgctcatgg
     1261 gccgcatggt gcaagctgaa caaccgcaag tggttccccg cagagccaga ggacgtgcgc
     1321 gactacctgc tgtacctgca ggcccgcggc ctggcagtga agaccatcca gcagcacctg
     1381 ggccagctga acatgctgca ccgccgcagc ggcctgcccc gcccaagcga ctccaacgcc
     1441 gtgtcactgg tcatgcgccg catccgcaag gagaacgtgg acgccggcga gcgcgccaag
     1501 caggccctgg cattcgagcg caccgacttc gaccaggtgc gctccctgat ggagaacagc
     1561 gaccgctgcc aggacatccg caacctggcc ttcctgggca tcgcatacaa caccctgctg
     1621 cgcatcgccg agatcgcacg catccgcgtg aaggacatca gccgcacaga cggcggccgc
     1681 atgctgatcc acatcggccg caccaagaca ctggtgtcaa ccgccggcgt ggagaaggca
     1741 ctgtccctgg gcgtgaccaa gctggtggag cgctggatct ccgtgagcgg cgtggccgac
     1801 gaccccaaca actacctgtt ctgccgcgtg cgcaagaacg gcgtggccgc accatcagca
     1861 acctcccagc tgagcacacg cgccctggag ggcatcttcg aggcaaccca ccgcctgatc
     1921 tacggcgcca aggacgactc cggccagcgc tacctggcct ggtcaggcca ctccgcacgc
     1981 gtgggcgccg cacgcgacat ggcccgcgca ggcgtgagca tccccgagat catgcaggcc
     2041 ggcggctgga ccaacgtgaa catcgtgatg aactacatcc gcaacctgga ctccgagaca
     2101 ggcgccatgg tgcgcctgct ggaggacggc gactgaaagc ttatcgatac cgtcgacagg
     2161 ttctaagcaa tgacctgggc tgtacatcat tagcaaactg aatctgcagc tttctaggct
     2221 cttgctgttc ctaggaactt ggggttcaag actgggtttt tgctgtggtt cctgactgca
     2281 gaccatcact gtctgggttg tcgtgtctaa tgtccatagg tgtgagcaca tgctccactt
     2341 caattcctac tgctgggtaa ctggcgctgt ctgagggact tctgagaaga atgccgagaa
     2401 gaacttgagc gatttgtaag gggatatttt gggtctgtac caaaggccca gactgggctt
     2461 ccagtgcatc gcacatatgc atctgcatgt ccattcatac aatcaacctg ttgactgcat
     2521 gacccagatc acaacctcct tgagttggaa gtagcttctt ctacagatag aaaatggtgc
     2581 atgtcagctg agggttgcag gcgtttaaaa gtgcatcaag tttgcagaac acatggtgat
     2641 tcatgactgc atggaaacag accagcacac tcgagggggg gcccggtacc caattcgccc
     2701 tatagtgagt cgtattacgc gcgctcactg gccgtcgttt tacaacgtcg tgactgggaa
     2761 aaccctggcg ttacccaact taatcgcctt gcagcacatc cccctttcgc cagctggcgt
     2821 aatagcgaag aggcccgcac cgatcgccct tcccaacagt tgcgcagcct gaatggcgaa
     2881 tggaaattgt aagcgttaat attttgttaa aattcgcgtt aaatttttgt taaatcagct
     2941 cattttttaa ccaataggcc gaaatcggca aaatccctta taaatcaaaa gaatagaccg
     3001 agatagggtt gagtgttgtt ccagtttgga acaagagtcc actattaaag aacgtggact
     3061 ccaacgtcaa agggcgaaaa accgtctatc agggcgatgg cccactacgt gaaccatcac
     3121 cctaatcaag ttttttgggg tcgaggtgcc gtaaagcact aaatcggaac cctaaaggga
     3181 gcccccgatt tagagcttga cggggaaagc cggcgaacgt ggcgagaaag gaagggaaga
     3241 aagcgaaagg agcgggcgct agggcgctgg caagtgtagc ggtcacgctg cgcgtaacca
     3301 ccacacccgc cgcgcttaat gcgccgctac agggcgcgtc aggtggcact tttcggggaa
     3361 atgtgcgcgg aacccctatt tgtttatttt tctaaataca ttcaaatatg tatccgctca
     3421 tgagacaata accctgataa atgcttcaat aatattgaaa aaggaagagt atgagtattc
     3481 aacatttccg tgtcgccctt attccctttt ttgcggcatt ttgccttcct gtttttgctc
     3541 acccagaaac gctggtgaaa gtaaaagatg ctgaagatca gttgggtgca cgagtgggtt
     3601 acatcgaact ggatctcaac agcggtaaga tccttgagag ttttcgcccc gaagaacgtt
     3661 ttccaatgat gagcactttt aaagttctgc tatgtggcgc ggtattatcc cgtattgacg
     3721 ccgggcaaga gcaactcggt cgccgcatac actattctca gaatgacttg gttgagtact
     3781 caccagtcac agaaaagcat cttacggatg gcatgacagt aagagaatta tgcagtgctg
     3841 ccataaccat gagtgataac actgcggcca acttacttct gacaacgatc ggaggaccga
     3901 aggagctaac cgcttttttg cacaacatgg gggatcatgt aactcgcctt gatcgttggg
     3961 aaccggagct gaatgaagcc ataccaaacg acgagcgtga caccacgatg cctgtagcaa
     4021 tggcaacaac gttgcgcaaa ctattaactg gcgaactact tactctagct tcccggcaac
     4081 aattaataga ctggatggag gcggataaag ttgcaggacc acttctgcgc tcggcccttc
     4141 cggctggctg gtttattgct gataaatctg gagccggtga gcgtgggtct cgcggtatca
     4201 ttgcagcact ggggccagat ggtaagccct cccgtatcgt agttatctac acgacgggga
     4261 gtcaggcaac tatggatgaa cgaaatagac agatcgctga gataggtgcc tcactgatta
     4321 agcattggta actgtcagac caagtttact catatatact ttagattgat ttaaaacttc
     4381 atttttaatt taaaaggatc taggtgaaga tcctttttga taatctcatg accaaaatcc
     4441 cttaacgtga gttttcgttc cactgagcgt cagaccccgt agaaaagatc aaaggatctt
     4501 cttgagatcc tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac
     4561 cagcggtggt ttgtttgccg gatcaagagc taccaactct ttttccgaag gtaactggct
     4621 tcagcagagc gcagatacca aatactgtcc ttctagtgta gccgtagtta ggccaccact
     4681 tcaagaactc tgtagcaccg cctacatacc tcgctctgct aatcctgtta ccagtggctg
     4741 ctgccagtgg cgataagtcg tgtcttaccg ggttggactc aagacgatag ttaccggata
     4801 aggcgcagcg gtcgggctga acggggggtt cgtgcacaca gcccagcttg gagcgaacga
     4861 cctacaccga actgagatac ctacagcgtg agctatgaga aagcgccacg cttcccgaag
     4921 ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg
     4981 agcttccagg gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc cacctctgac
     5041 ttgagcgtcg atttttgtga tgctcgtcag gggggcggag cctatggaaa aacgccagca
     5101 acgcggcctt tttacggttc ctggcctttt gctggccttt tgctcacatg ttctttcctg
     5161 cgttatcccc tgattctgtg gataaccgta ttaccgcctt tgagtgagct gataccgctc
     5221 gccgcagccg aacgaccgag cgcagcgagt cagtgagcga ggaagcggaa gagcgcccaa
     5281 tacgcaaacc gcctctcccc gcgcgttggc cgattcatta atgcagctgg cacgacaggt
     5341 ttcccgactg gaaagcgggc agtgagcgca acgcaattaa tgtgagttag ctcactcatt
     5401 aggcacccca ggctttacac tttatgcttc cggctcgtat gttgtgtgga attgtgagcg
     5461 gataacaatt tcacacagga aacagctatg accatgatta cgccaagcgc gcaattaacc
     5521 ctcactaaag ggaacaaaag ctggagctcc accgcggtg
//
