LOCUS       Exported                8164 bp ds-DNA     circular SYN 18-DEC-2018
DEFINITION  Cloning vector pLM005-EPYC1-Venus, complete sequence.
ACCESSION   KX077944
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8164)
  AUTHORS   Mackinder LC, Meyer MT, Mettler-Altmann T, Chen VK, Mitchell MC, 
            Caspari O, Freeman Rosenzweig ES, Pallesen L, Reeves G, Itakura A, 
            Roth R, Sommer F, Geimer S, Muhlhaus T, Schroda M, Goodenough U, 
            Stitt M, Griffiths H, Jonikas MC.
  TITLE     A repeat protein links Rubisco to form the eukaryotic 
            carbon-concentrating organelle
  JOURNAL   Proc. Natl. Acad. Sci. U.S.A. (2016) In press
  PUBMED    27166422
REFERENCE   2  (bases 1 to 8164)
  AUTHORS   Mackinder LCM., Meyer MT, Mettler-Altmann T, Chen V, Mitchell M, 
            Caspari O, Freeman Rosenzweig E, Pallesen L, Reeves G, Itakura A, 
            Roth R, Sommer F, Geimer S, Muhlhaus T, Schroda M, Goodenough U, 
            Stitt M, Griffiths H, Jonikas MC.
  TITLE     Direct Submission
  JOURNAL   Submitted (13-APR-2016) Department of Plant Biology, Carnegie 
            Institution of Washington, 260 Panama Street, Stanford, CA 94305, 
            USA
REFERENCE   3  (bases 1 to 8164)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..8164
                     /organism="Cloning vector pLM005-EPYC1-Venus"
                     /mol_type="other DNA"
                     /db_xref="taxon:1850518"
     CDS             join(101..166,418..479,742..2389)
                     /codon_start=1
                     /product="EPYC1-VENUS"
                     /label=EPYC1-VENUS
                     /note="EPYC1 Venus 3xFLAG fusion protein"
                     /protein_id="ANF29831.1"
                     /translation="MATISSMRVGAASRVVVSGRVKTVKVAARGSWRESSTATVQASRA
                     SSATNRVSPTRSVLPANWRQELESLRNGNGSSSAASSAPAPARSSSASWRDAAPASSAP
                     ARSSSASKKAVTPSRSALPSNWKQELESLRSSSPAPASSAPAPARSSSASWRDAAPASS
                     APARSSSSKKAVTPSRSALPSNWKQELESLRSSSPAPASSAPAPARSSSASWRDAAPAS
                     SAPARSSSASKKAVTPSRSALPSNWKQELESLRSNSPAPASSAPAPARSSSASWRDAPA
                     SSSSSSADKAGTNPWTGKSKPEIKRTALPADWRKGLNGDLGGSGGRMVSKGEELFTGVV
                     PILVELDGDVNGHKFSVSGEGEGDATYGKLTLKLICTTGKLPVPWPTLVTTLGYGLQCF
                     ARYPDHMKQHDFFKSAMPEGYVQERTIFFKDDGNYKTRAEVKFEGDTLVNRIELKGIDF
                     KEDGNILGHKLEYNYNSHNVYITADKQKNGIKANFKIRHNIEDGGVQLADHYQQNTPIG
                     DGPVLLPDNHYLSYQSALSKDPNEKRDHMVLLEFVTAAGITLGMDELIKGGGDYKDHDG
                     DYKDHDIDYKDDDDK"
     CDS             1595..2113
                     /codon_start=1
                     /product="N-terminal fragment of mVenus for use in 
                     bimolecular fluorescence complementation (BiFC) (Kodama and
                     Hu, 2010)"
                     /label=VN173
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KLICTTGKLPVPWPTLVTTLGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYITADKQKNGIK
                     ANFKIRHNIE"
     CDS             2321..2386
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     rep_origin      complement(3747..4335)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(4506..5366)
                     /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(5367..5471)
                     /gene="bla"
                     /label=AmpR promoter
     CDS             6350..7153
                     /codon_start=1
                     /product="APHVIII"
                     /label=APHVIII
                     /note="paromomycin resistance marker"
                     /protein_id="ANF29832.1"
                     /translation="MDDALRALRGRYPGCEWVVVEDGASGAGVYRLRGGGRELFVKVAA
                     LGAGVGLLGEAERLVWLAEVGIPVPRVVEGGGDERVAWLVTEAVPGRPASARWPREQRL
                     DVAVALAGLARSLHALDWERCPFDRSLAVTVPQAARAVAEGSVDLEDLDEERKGWSGER
                     LLAELERTRPADEDLAVCHGDLCPDNVLLDPRTCEVTGLIDVGRVGRADRHSDLALVLR
                     ELAHEEDPWFGPECSAAFLREYGRGWDGAVSEEKLAFYRLLDEFF"
     promoter        complement(7478..7496)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"