LOCUS       Exported                5559 bp ds-DNA     circular SYN 18-DEC-2018
DEFINITION  Expression vector pPeCre DNA, complete sequence.
ACCESSION   LC332843
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  JOURNAL   Unpublished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)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..5559
                     /organism="Expression vector pPeCre"
                     /mol_type="other DNA"
                     /db_xref="taxon:2051842"
     regulatory      1..814
                     /regulatory_class="promoter"
                     /note="includes promoter region of Pseudochoricystis 
                     ellipsoidea nuclear gene RBCS for ribulose bisphosphate 
                     carboxylase/oxygenase small subunit"
     misc_feature    1105..2136
                     /gene="PeCre"
                     /note="include intron 1 of Pseudochoricystis ellipsoidea 
                     nuclear gene RBCS for ribulose bisphosphate 
                     carboxylase/oxygenase small subunit"
     CDS             1105..2136
                     /codon_start=1
                     /gene="PeCre"
                     /product="cre recombinase"
                     /label=PeCre
                     /protein_id="BBB04238.1"
                     /translation="MSNLLTVHQNLPALPVDATSDEVRKNLMDMFRDRQAFSEHTWKML
                     LSVCRSWAAWCKLNNRKWFPAEPEDVRDYLLYLQARGLAVKTIQQHLGQLNMLHRRSGL
                     PRPSDSNAVSLVMRRIRKENVDAGERAKQALAFERTDFDQVRSLMENSDRCQDIRNLAF
                     LGIAYNTLLRIAEIARIRVKDISRTDGGRMLIHIGRTKTLVSTAGVEKALSLGVTKLVE
                     RWISVSGVADDPNNYLFCRVRKNGVAAPSATSQLSTRALEGIFEATHRLIYGAKDDSGQ
                     RYLAWSGHSARVGAARDMARAGVSIPEIMQAGGWTNVNIVMNYIRNLDSETGAMVRLLE
                     DGD"
     gene            1105..2136
                     /gene="PeCre"
                     /label=PeCre
     regulatory      2137..2674
                     /regulatory_class="terminator"
                     /note="includes terminator region of Pseudochoricystis 
                     ellipsoidea nuclear gene RBCS for ribulose bisphosphate 
                     carboxylase/oxygenase small subunit"
     misc_feature    2675..5559
                     /gene="derived from plasmid pBluescript SK(+)"
     gene            2675..5559
                     /gene="derived from plasmid pBluescript SK(+)"
                     /label=derived from plasmid pBluescript SK(+)
     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
                     /gene="bla"
                     /label=AmpR promoter
     CDS             3471..4331
                     /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      4502..5090
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     protein_bind    5378..5399
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /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
                     /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     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"