LOCUS       Exported                4881 bp ds-DNA     circular SYN 17-DEC-2018
DEFINITION  Expression vector pCrCre DNA, complete sequence.
ACCESSION   LC150884
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4881)
  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  JOURNAL   Unpublished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 4881)
  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 4881)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..4881
                     /organism="Expression vector pCrCre"
                     /mol_type="other DNA"
                     /db_xref="taxon:1849578"
     primer_bind     18..34
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     regulatory      67..333
                     /regulatory_class="promoter"
                     /note="includes promoter region of Chlamydomonas 
                     reinhardtii nuclear gene hsp70A for cytosolic HSP70"
     regulatory      342..574
                     /regulatory_class="promoter"
                     /note="includes promoter region and start codon of 
                     Chlamydomonas reinhardtii nuclear gene rbcS2 for ribulose 
                     bisphosphate carboxylase/oxygenase small subunit"
     CDS             join(567..574,720..1758)
                     /codon_start=1
                     /gene="CrCRE"
                     /product="cre recombinase"
                     /label=CrCRE
                     /protein_id="BAV82214.1"
                     /translation="MARMAMSNLLTVHQNLPALPVDATSDEVRKNLMDMFRDRQAFSEH
                     TWKMLLSVCRSWAAWCKLNNRKWFPAEPEDVRDYLLYLQARGLAVKTIQQHLGQLNMLH
                     RRSGLPRPSDSNAVSLVMRRIRKENVDAGERAKQALAFERTDFDQVRSLMENSDRCQDI
                     RNLAFLGIAYNTLLRIAEIARIRVKDISRTDGGRMLIHIGRTKTLVSTAGVEKALSLGV
                     TKLVERWISVSGVADDPNNYLFCRVRKNGVAAPSATSQLSTRALEGIFEATHRLIYGAK
                     DDSGQRYLAWSGHSARVGAARDMARAGVSIPEIMQAGGWTNVNIVMNYIRNLDSETGAM
                     VRLLEDGD"
     gene            567..574
                     /gene="CrCRE"
                     /label=CrCRE
     intron          575..719
                     /note="includes intron 1 from Chlamydomonas reinhardtii 
                     rbcS2 gene"
     gene            720..1758
                     /gene="CrCRE"
                     /label=CrCRE
     regulatory      1759..1998
                     /regulatory_class="terminator"
                     /note="includes terminator region from Chlamydomonas 
                     reinhardtii rbcS2 gene"
     misc_feature    1999..4881
                     /label=derived from plasmid pBluescript SK(+)
                     /note="derived from plasmid pBluescript SK(+)"
     promoter        complement(2037..2055)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2065..2081)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     rep_origin      complement(2222..2677)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     promoter        2704..2808
                     /gene="bla"
                     /label=AmpR promoter
     CDS             2809..3669
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     rep_origin      3840..4428
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     protein_bind    4716..4737
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        4752..4782
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4790..4806
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /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     4814..4830
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        4851..4869
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"