LOCUS       40924_6676        5357 bp DNA     circular SYN 17-DEC-2018
DEFINITION  Expression vector pbleCrCre DNA, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5357)
  AUTHORS   Kasai Y, Harayama S.
  TITLE     Construction of marker-free transgenic strains of Chlamydomonas 
            reinhardtii using a Cre/loxP-mediated recombinase system
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 5357)
  AUTHORS   Kasai Y, Harayama S.
  TITLE     Direct Submission
  JOURNAL   Submitted (06-MAY-2016) Contact:Yuki Kasai Chuo University, 
            Department of Biological sciences; Kasuga 1-13-27, Bunkyo-ku, Tokyo 
            112-8551, Japan
REFERENCE   3  (bases 1 to 5357)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 5357)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: 
            "Unpublished"
COMMENT     SGRef: number: 2; type: "Journal Article"; journalName: "Submitted 
            (06-MAY-2016) Contact:Yuki Kasai Chuo University, Department of 
            Biological sciences; Kasuga 1-13-27, Bunkyo-ku, Tokyo 112-8551, 
            Japan"
COMMENT     SGRef: number: 3; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..5357
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     18..34
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     regulatory      43..317
                     /note="includes promoter region of Chlamydomonas
                     reinhardtii nuclear gene hsp70A for cytosolic HSP70"
                     /regulatory_class="promoter"
     regulatory      325..540
                     /note="includes promoter region and start codon of
                     Chlamydomonas reinhardtii nuclear gene rbcS2 for ribulose 
                     bisphosphate carboxylase/oxygenase small subunit"
                     /regulatory_class="promoter"
     intron          541..685
                     /gene="ble::CrCRE"
                     /note="includes intron 1 from Chlamydomonas reinhardtii
                     rbcS2 gene"
     exon            686..854
                     /gene="ble::CrCRE"
                     /note="includes a fragment of Streptoalloteichus
                     hindustanus Ble gene"
     intron          855..999
                     /gene="ble::CrCRE"
                     /note="includes intron 1 from Chlamydomonas reinhardtii
                     rbcS2 gene"
     CDS             1210..2232
                     /label=Cre
                     /note="site-specific recombinase"
     regulatory      2236..2476
                     /note="includes terminator region from Chlamydomonas
                     reinhardtii rbcS2 gene"
                     /regulatory_class="terminator"
     promoter        complement(2513..2531)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2541..2557)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     rep_origin      complement(2698..3153)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     promoter        3180..3284
                     /label=AmpR promoter
     CDS             3285..4142
                     /label=AmpR
                     /note="beta-lactamase"
     rep_origin      4316..4904
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     protein_bind    5192..5213
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     promoter        5228..5258
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5266..5282
                     /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     5290..5306
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        5327..5345
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
ORIGIN
        1 ggtaccgggc cccccctcga ggtcgacggt atcgataagc ttgacggcgg ggagctcgct
       61 gaggcttgac atgattggtg cgtatgtttg tatgaagcta caggactgat ttggcgggct
      121 atgagggcgg gggaagctct ggaagggccg cgatggggcg cgcggcgtcc agaaggcgcc
      181 atacggcccg ctggcggcac ccatccggta taaaagcccg cgaccccgaa cggtgacctc
      241 cactttcagc gacaaacgag cacttataca tacgcgacta ttctgccgct atacataacc
      301 actcagctag cttaagatcc caatgccaga aggagcgcag ccaaaccagg atgatgtttg
      361 atggggtatt tgagcacttg caacccttat ccggaagccc cctggcccac aaaggctagg
      421 cgccaatgca agcagttcgc atgcagcccc tggagcggtg ccctcctgat aaaccggcca
      481 gggggcctat gttctttact tttttacaag agaagtcact caacatctta aaatggccag
      541 gtgagtcgac gagcaagccc ggcggatcag gcagcgtgct tgcagatttg acttgcaacg
      601 cccgcattgt gtcgacgaag gcttttggct cctctgtcgc tgtctcaagc agcatctaac
      661 cctgcgtcgc cgtttccatt tgcaggatgg ccaagctgac cagcgccgtt ccggtgctca
      721 ccgcgcgcga cgtcgccgga gcggtcgagt tctggaccga ccggctcggg ttctcccggg
      781 acttcgtgga ggacgacttc gccggtgtgg tccgggacga cgtgaccctg ttcatcagcg
      841 cggtccagga ccaggtgagt cgacgagcaa gcccggcgga tcaggcagcg tgcttgcaga
      901 tttgacttgc aacgcccgca ttgtgtcgac gaaggctttt ggctcctctg tcgctgtctc
      961 aagcagcatc taaccctgcg tcgccgtttc catttgcagg accaggtggt gccggacaac
     1021 accctggcct gggtgtgggt gcgcggcctg gacgagctgt acgccgagtg gtcggaggtc
     1081 gtgtccacga acttccggga cgcctccggg ccggccatga ccgagatcgg cgagcagccg
     1141 tgggggcggg agttcgccct gcgcgacccg gccggcaact gcgtgcactt cgtggccgag
     1201 gagcaggaca acctgctgac cgtgcaccag aacctgcccg ccctgcccgt ggacgcgacg
     1261 agcgacgagg tgcgcaagaa cctgatggac atgttccgcg accgccaggc cttctcggag
     1321 cacacctgga agatgctgct gagcgtgtgc cgctcgtggg ccgcgtggtg caagctgaac
     1381 aaccgcaagt ggttccccgc ggagccggag gacgtgcgcg actacctgct gtacctgcag
     1441 gcccgcggcc tggcggtgaa gacgatccag cagcacctgg gccagctgaa catgctgcac
     1501 cgccgcagcg gcctgccccg cccgagcgac tcgaacgccg tgtcgctggt catgcgccgc
     1561 atccgcaagg agaacgtgga cgccggcgag cgcgcgaagc aggccctggc gttcgagcgc
     1621 accgacttcg accaggtgcg ctccctgatg gagaacagcg accgctgcca ggacatccgc
     1681 aacctggcct tcctgggcat cgcgtacaac accctgctgc gcatcgccga gatcgcgcgc
     1741 atccgcgtga aggacatcag ccgcacggac ggcggccgca tgctgatcca catcggccgc
     1801 accaagacgc tggtgtcgac cgccggcgtg gagaaggcgc tgtccctggg cgtgacgaag
     1861 ctggtggagc gctggatctc cgtgagcggc gtggccgacg accccaacaa ctacctgttc
     1921 tgccgcgtgc gcaagaacgg cgtggccgcg ccgtcggcga cctcccagct gagcacgcgc
     1981 gccctggagg gcatcttcga ggcgacccac cgcctgatct acggcgccaa ggacgactcc
     2041 ggccagcgct acctggcctg gtcgggccac tccgcgcgcg tgggcgccgc gcgcgacatg
     2101 gcccgcgcgg gcgtgagcat ccccgagatc atgcaggccg gcggctggac caacgtgaac
     2161 atcgtgatga actacatccg caacctggac tccgagacgg gcgcgatggt gcgcctgctg
     2221 gaggacggcg actaaggatc cccgctccgt gtaaatggag gcgctcgttg atctgagcct
     2281 tgccccctga cgaacggcgg tggatggaag atactgctct caagtgctga agcggtagct
     2341 tagctccccg tttcgtgctg atcagtcttt ttcaacacgt aaaaagcgga ggagttttgc
     2401 aattttgttg gttgtaacga tcctccgttg attttggcct ctttctccat gggcgggctg
     2461 ggcgtatttg aagctctaga gcggccgcca ccgcggtgga gctccaattc gccctatagt
     2521 gagtcgtatt acgcgcgctc actggccgtc gttttacaac gtcgtgactg ggaaaaccct
     2581 ggcgttaccc aacttaatcg ccttgcagca catccccctt tcgccagctg gcgtaatagc
     2641 gaagaggccc gcaccgatcg cccttcccaa cagttgcgca gcctgaatgg cgaatggaaa
     2701 ttgtaagcgt taatattttg ttaaaattcg cgttaaattt ttgttaaatc agctcatttt
     2761 ttaaccaata ggccgaaatc ggcaaaatcc cttataaatc aaaagaatag accgagatag
     2821 ggttgagtgt tgttccagtt tggaacaaga gtccactatt aaagaacgtg gactccaacg
     2881 tcaaagggcg aaaaaccgtc tatcagggcg atggcccact acgtgaacca tcaccctaat
     2941 caagtttttt ggggtcgagg tgccgtaaag cactaaatcg gaaccctaaa gggagccccc
     3001 gatttagagc ttgacgggga aagccggcga acgtggcgag aaaggaaggg aagaaagcga
     3061 aaggagcggg cgctagggcg ctggcaagtg tagcggtcac gctgcgcgta accaccacac
     3121 ccgccgcgct taatgcgccg ctacagggcg cgtcaggtgg cacttttcgg ggaaatgtgc
     3181 gcggaacccc tatttgttta tttttctaaa tacattcaaa tatgtatccg ctcatgagac
     3241 aataaccctg ataaatgctt caataatatt gaaaaaggaa gagtatgagt attcaacatt
     3301 tccgtgtcgc ccttattccc ttttttgcgg cattttgcct tcctgttttt gctcacccag
     3361 aaacgctggt gaaagtaaaa gatgctgaag atcagttggg tgcacgagtg ggttacatcg
     3421 aactggatct caacagcggt aagatccttg agagttttcg ccccgaagaa cgttttccaa
     3481 tgatgagcac ttttaaagtt ctgctatgtg gcgcggtatt atcccgtatt gacgccgggc
     3541 aagagcaact cggtcgccgc atacactatt ctcagaatga cttggttgag tactcaccag
     3601 tcacagaaaa gcatcttacg gatggcatga cagtaagaga attatgcagt gctgccataa
     3661 ccatgagtga taacactgcg gccaacttac ttctgacaac gatcggagga ccgaaggagc
     3721 taaccgcttt tttgcacaac atgggggatc atgtaactcg ccttgatcgt tgggaaccgg
     3781 agctgaatga agccatacca aacgacgagc gtgacaccac gatgcctgta gcaatggcaa
     3841 caacgttgcg caaactatta actggcgaac tacttactct agcttcccgg caacaattaa
     3901 tagactggat ggaggcggat aaagttgcag gaccacttct gcgctcggcc cttccggctg
     3961 gctggtttat tgctgataaa tctggagccg gtgagcgtgg gtctcgcggt atcattgcag
     4021 cactggggcc agatggtaag ccctcccgta tcgtagttat ctacacgacg gggagtcagg
     4081 caactatgga tgaacgaaat agacagatcg ctgagatagg tgcctcactg attaagcatt
     4141 ggtaactgtc agaccaagtt tactcatata tactttagat tgatttaaaa cttcattttt
     4201 aatttaaaag gatctaggtg aagatccttt ttgataatct catgaccaaa atcccttaac
     4261 gtgagttttc gttccactga gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag
     4321 atcctttttt tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg
     4381 tggtttgttt gccggatcaa gagctaccaa ctctttttcc gaaggtaact ggcttcagca
     4441 gagcgcagat accaaatact gtccttctag tgtagccgta gttaggccac cacttcaaga
     4501 actctgtagc accgcctaca tacctcgctc tgctaatcct gttaccagtg gctgctgcca
     4561 gtggcgataa gtcgtgtctt accgggttgg actcaagacg atagttaccg gataaggcgc
     4621 agcggtcggg ctgaacgggg ggttcgtgca cacagcccag cttggagcga acgacctaca
     4681 ccgaactgag atacctacag cgtgagctat gagaaagcgc cacgcttccc gaagggagaa
     4741 aggcggacag gtatccggta agcggcaggg tcggaacagg agagcgcacg agggagcttc
     4801 cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc
     4861 gtcgattttt gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg
     4921 cctttttacg gttcctggcc ttttgctggc cttttgctca catgttcttt cctgcgttat
     4981 cccctgattc tgtggataac cgtattaccg cctttgagtg agctgatacc gctcgccgca
     5041 gccgaacgac cgagcgcagc gagtcagtga gcgaggaagc ggaagagcgc ccaatacgca
     5101 aaccgcctct ccccgcgcgt tggccgattc attaatgcag ctggcacgac aggtttcccg
     5161 actggaaagc gggcagtgag cgcaacgcaa ttaatgtgag ttagctcact cattaggcac
     5221 cccaggcttt acactttatg cttccggctc gtatgttgtg tggaattgtg agcggataac
     5281 aatttcacac aggaaacagc tatgaccatg attacgccaa gcgcgcaatt aaccctcact
     5341 aaagggaaca aaagctg
//
