LOCUS       40924_6681        5381 bp DNA     circular SYN 17-DEC-2018
DEFINITION  Expression vector pbleLCrCre DNA, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5381)
  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  JOURNAL   Unpublished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 5381)
  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 5381)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 5381)
  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            "Unpublished"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..5381
                     /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="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::linker::CrCRE"
                     /note="includes intron 1 from Chlamydomonas reinhardtii
                     rbcS2 gene"
     exon            686..854
                     /gene="ble::linker::CrCRE"
     intron          855..999
                     /gene="ble::linker::CrCRE"
                     /note="includes intron 1 from Chlamydomonas reinhardtii
                     rbcS2 gene"
     misc_feature    1210..1227
                     /gene="ble::linker::CrCRE"
                     /label=artificial linker
                     /note="artificial linker"
     CDS             1228..2256
                     /label=Cre
                     /note="site-specific recombinase"
     regulatory      2260..2498
                     /note="includes terminator region of Chlamydomonas
                     reinhardtii rbcS2 gene"
                     /regulatory_class="terminator"
     promoter        complement(2537..2555)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2565..2581)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     rep_origin      complement(2722..3177)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     promoter        3204..3308
                     /label=AmpR promoter
     CDS             3309..4166
                     /label=AmpR
                     /note="beta-lactamase"
     rep_origin      4340..4928
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     protein_bind    5216..5237
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     promoter        5252..5282
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5290..5306
                     /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     5314..5330
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        5351..5369
                     /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 gagcaggacg gcggctccgg cggccgcatg agcaacctgc tgaccgtgca ccagaacctg
     1261 cccgccctgc ccgtggacgc gacgagcgac gaggtgcgca agaacctgat ggacatgttc
     1321 cgcgaccgcc aggccttctc ggagcacacc tggaagatgc tgctgagcgt gtgccgctcg
     1381 tgggccgcgt ggtgcaagct gaacaaccgc aagtggttcc ccgcggagcc ggaggacgtg
     1441 cgcgactacc tgctgtacct gcaggcccgc ggcctggcgg tgaagacgat ccagcagcac
     1501 ctgggccagc tgaacatgct gcaccgccgc agcggcctgc cccgcccgag cgactcgaac
     1561 gccgtgtcgc tggtcatgcg ccgcatccgc aaggagaacg tggacgccgg cgagcgcgcg
     1621 aagcaggccc tggcgttcga gcgcaccgac ttcgaccagg tgcgctccct gatggagaac
     1681 agcgaccgct gccaggacat ccgcaacctg gccttcctgg gcatcgcgta caacaccctg
     1741 ctgcgcatcg ccgagatcgc gcgcatccgc gtgaaggaca tcagccgcac ggacggcggc
     1801 cgcatgctga tccacatcgg ccgcaccaag acgctggtgt cgaccgccgg cgtggagaag
     1861 gcgctgtccc tgggcgtgac gaagctggtg gagcgctgga tctccgtgag cggcgtggcc
     1921 gacgacccca acaactacct gttctgccgc gtgcgcaaga acggcgtggc cgcgccgtcg
     1981 gcgacctccc agctgagcac gcgcgccctg gagggcatct tcgaggcgac ccaccgcctg
     2041 atctacggcg ccaaggacga ctccggccag cgctacctgg cctggtcggg ccactccgcg
     2101 cgcgtgggcg ccgcgcgcga catggcccgc gcgggcgtga gcatccccga gatcatgcag
     2161 gccggcggct ggaccaacgt gaacatcgtg atgaactaca tccgcaacct ggactccgag
     2221 acgggcgcga tggtgcgcct gctggaggac ggcgactaag gatccccgct ccgtgtaaat
     2281 ggaggcgctc gttgatctga gccttgcccc ctgacgaacg gcggtggatg gaagatactg
     2341 ctctcaagtg ctgaagcggt agcttagctc cccgtttcgt gctgatcagt ctttttcaac
     2401 acgtaaaaag cggaggagtt ttgcaatttt gttggttgta acgatcctcc gttgattttg
     2461 gcctctttct ccatgggcgg gctgggcgta tttgaagctc tagagcggcc gccaccgcgg
     2521 tggagctcca attcgcccta tagtgagtcg tattacgcgc gctcactggc cgtcgtttta
     2581 caacgtcgtg actgggaaaa ccctggcgtt acccaactta atcgccttgc agcacatccc
     2641 cctttcgcca gctggcgtaa tagcgaagag gcccgcaccg atcgcccttc ccaacagttg
     2701 cgcagcctga atggcgaatg gaaattgtaa gcgttaatat tttgttaaaa ttcgcgttaa
     2761 atttttgtta aatcagctca ttttttaacc aataggccga aatcggcaaa atcccttata
     2821 aatcaaaaga atagaccgag atagggttga gtgttgttcc agtttggaac aagagtccac
     2881 tattaaagaa cgtggactcc aacgtcaaag ggcgaaaaac cgtctatcag ggcgatggcc
     2941 cactacgtga accatcaccc taatcaagtt ttttggggtc gaggtgccgt aaagcactaa
     3001 atcggaaccc taaagggagc ccccgattta gagcttgacg gggaaagccg gcgaacgtgg
     3061 cgagaaagga agggaagaaa gcgaaaggag cgggcgctag ggcgctggca agtgtagcgg
     3121 tcacgctgcg cgtaaccacc acacccgccg cgcttaatgc gccgctacag ggcgcgtcag
     3181 gtggcacttt tcggggaaat gtgcgcggaa cccctatttg tttatttttc taaatacatt
     3241 caaatatgta tccgctcatg agacaataac cctgataaat gcttcaataa tattgaaaaa
     3301 ggaagagtat gagtattcaa catttccgtg tcgcccttat tccctttttt gcggcatttt
     3361 gccttcctgt ttttgctcac ccagaaacgc tggtgaaagt aaaagatgct gaagatcagt
     3421 tgggtgcacg agtgggttac atcgaactgg atctcaacag cggtaagatc cttgagagtt
     3481 ttcgccccga agaacgtttt ccaatgatga gcacttttaa agttctgcta tgtggcgcgg
     3541 tattatcccg tattgacgcc gggcaagagc aactcggtcg ccgcatacac tattctcaga
     3601 atgacttggt tgagtactca ccagtcacag aaaagcatct tacggatggc atgacagtaa
     3661 gagaattatg cagtgctgcc ataaccatga gtgataacac tgcggccaac ttacttctga
     3721 caacgatcgg aggaccgaag gagctaaccg cttttttgca caacatgggg gatcatgtaa
     3781 ctcgccttga tcgttgggaa ccggagctga atgaagccat accaaacgac gagcgtgaca
     3841 ccacgatgcc tgtagcaatg gcaacaacgt tgcgcaaact attaactggc gaactactta
     3901 ctctagcttc ccggcaacaa ttaatagact ggatggaggc ggataaagtt gcaggaccac
     3961 ttctgcgctc ggcccttccg gctggctggt ttattgctga taaatctgga gccggtgagc
     4021 gtgggtctcg cggtatcatt gcagcactgg ggccagatgg taagccctcc cgtatcgtag
     4081 ttatctacac gacggggagt caggcaacta tggatgaacg aaatagacag atcgctgaga
     4141 taggtgcctc actgattaag cattggtaac tgtcagacca agtttactca tatatacttt
     4201 agattgattt aaaacttcat ttttaattta aaaggatcta ggtgaagatc ctttttgata
     4261 atctcatgac caaaatccct taacgtgagt tttcgttcca ctgagcgtca gaccccgtag
     4321 aaaagatcaa aggatcttct tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa
     4381 caaaaaaacc accgctacca gcggtggttt gtttgccgga tcaagagcta ccaactcttt
     4441 ttccgaaggt aactggcttc agcagagcgc agataccaaa tactgtcctt ctagtgtagc
     4501 cgtagttagg ccaccacttc aagaactctg tagcaccgcc tacatacctc gctctgctaa
     4561 tcctgttacc agtggctgct gccagtggcg ataagtcgtg tcttaccggg ttggactcaa
     4621 gacgatagtt accggataag gcgcagcggt cgggctgaac ggggggttcg tgcacacagc
     4681 ccagcttgga gcgaacgacc tacaccgaac tgagatacct acagcgtgag ctatgagaaa
     4741 gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa
     4801 caggagagcg cacgagggag cttccagggg gaaacgcctg gtatctttat agtcctgtcg
     4861 ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc
     4921 tatggaaaaa cgccagcaac gcggcctttt tacggttcct ggccttttgc tggccttttg
     4981 ctcacatgtt ctttcctgcg ttatcccctg attctgtgga taaccgtatt accgcctttg
     5041 agtgagctga taccgctcgc cgcagccgaa cgaccgagcg cagcgagtca gtgagcgagg
     5101 aagcggaaga gcgcccaata cgcaaaccgc ctctccccgc gcgttggccg attcattaat
     5161 gcagctggca cgacaggttt cccgactgga aagcgggcag tgagcgcaac gcaattaatg
     5221 tgagttagct cactcattag gcaccccagg ctttacactt tatgcttccg gctcgtatgt
     5281 tgtgtggaat tgtgagcgga taacaatttc acacaggaaa cagctatgac catgattacg
     5341 ccaagcgcgc aattaaccct cactaaaggg aacaaaagct g
//
