LOCUS       Exported                6127 bp ds-DNA     circular SYN 18-DEC-2018
DEFINITION  Expression vector ploxP_cPeUMPS DNA, complete sequence.
ACCESSION   LC332841
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6127)
  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  JOURNAL   Unpublished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 6127)
  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 6127)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..6127
                     /organism="Expression vector ploxP_cPeUMPS"
                     /mol_type="other DNA"
                     /db_xref="taxon:2051840"
     misc_feature    1..706
                     /note="derived from plasmid pBluescript SK(+), 
                     Acc#:X52328.1"
     rep_origin      complement(3..458)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     600..616
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        626..644
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     670..686
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     misc_feature    707..740
                     /label=loxP
                     /note="loxP"
     protein_bind    707..740
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (GCATACAT)."
     regulatory      741..1530
                     /regulatory_class="promoter"
                     /note="includes promoter region of Pseudochoricystis 
                     ellipsoidea nuclear gene RBCS for ribulose bisphosphate 
                     carboxylase/oxygenase small subunit"
     misc_feature    1531..1815
                     /note="include intron 1 of Pseudochoricystis ellipsoidea 
                     nuclear gene RBCS for ribulose bisphosphate 
                     carboxylase/oxygenase small subunit"
     CDS             1816..3318
                     /codon_start=1
                     /gene="UMPS"
                     /product="uridine monophosphate synthase"
                     /label=UMPS
                     /protein_id="BBB04236.1"
                     /translation="MATSTPSVATAAELKPKEGPITSQEFEELVLRLHEIEAVKFGNFK
                     LKSGLMSPIYIDLRVIVSYPDVLRRVAEVMWHQVSGAAFDVMCGVPYTALPIATCMSLL
                     HGTPMLMRRKEVKEYGTKKAIEGAFKKGQTCLIVEDLVTSGASVMETVEPLEVEGLKVT
                     DVVVLIDREQGGAARMASNGLRLHSAFTLSFIIKTLQAHGLVSAKVADSVAAFIAANQT
                     FTPSAAAPTPSPAAPAQPKRLPFEERASLCQNAAGRKLLELMARKRTNLAVAADVATVE
                     EMLRIADAAGPHIAVFKTHVDIFDKWDDGIATQLRHLADKHEFLIFEDRKFADIGNTVV
                     SQYGGGIYKIADWSDITNAHLVPGPGIIDGLRKVGQEKGRGLLLLAEMSSKGALATGAY
                     TEKVAEAAAANQDFVMGFICQSPAKWATPVPPGLVHMTPGVQLASGSDALGQQYNTPAS
                     VIGQGGSDVIIVGRGIIKAADPAAAAAQYREAGWAAYEATLA"
     gene            1816..3318
                     /gene="UMPS"
                     /label=UMPS
     regulatory      3319..3850
                     /regulatory_class="terminator"
                     /note="includes terminator region of Pseudochoricystis 
                     ellipsoidea nuclear gene RBCS for ribulose bisphosphate 
                     carboxylase/oxygenase small subunit"
     misc_feature    3851..3884
                     /label=loxP
                     /note="loxP"
     protein_bind    3851..3884
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (GCATACAT)."
     misc_feature    3885..6127
                     /note="derived from plasmid pBluescript SK(+), 
                     Acc#:X52328.1"
     primer_bind     complement(3886..3902)
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        complement(3939..3957)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(3978..3994)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    4002..4018
                     /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)."
     promoter        complement(4026..4056)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4071..4092
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     rep_origin      complement(4380..4968)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(5139..5999)
                     /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"
     promoter        complement(6000..6104)
                     /gene="bla"
                     /label=AmpR promoter