pbleCrCre vector (V009158)

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  JOURNAL   Unpublished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            "Unpublished"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"