LOCUS       Exported                7216 bp ds-DNA     circular SYN 18-DEC-2018
DEFINITION  Cloning vector pLM005-Venus, complete sequence.
ACCESSION   KX077945
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7216)
  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 7216)
  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 7216)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..7216
                     /organism="Cloning vector pLM005-Venus"
                     /mol_type="other DNA"
                     /db_xref="taxon:1850519"
     CDS             784..1587
                     /codon_start=1
                     /product="APHVIII"
                     /label=APHVIII
                     /protein_id="ANF29834.1"
                     /translation="MDDALRALRGRYPGCEWVVVEDGASGAGVYRLRGGGRELFVKVAA
                     LGAGVGLLGEAERLVWLAEVGIPVPRVVEGGGDERVAWLVTEAVPGRPASARWPREQRL
                     DVAVALAGLARSLHALDWERCPFDRSLAVTVPQAARAVAEGSVDLEDLDEERKGWSGER
                     LLAELERTRPADEDLAVCHGDLCPDNVLLDPRTCEVTGLIDVGRVGRADRHSDLALVLR
                     ELAHEEDPWFGPECSAAFLREYGRGWDGAVSEEKLAFYRLLDEFF"
     promoter        complement(1912..1930)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     CDS             complement(2752..3075)
                     /codon_start=1
                     /gene="lacZ fragment"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /label=lacZ-alpha
                     /translation="MTMITPSLHACRSTLEDPRVPSSNSLAVVLQRRDWENPGVTQLNR
                     LAAHPPFASWRNSEEARTDRPSQQLRSLNGEWRLMRYFLLTHLCGISHRIWCTLSTICS
                     DAA"
     primer_bind     2985..3001
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     misc_feature    3002..3058
                     /label=MCS
                     /note="pUC18/19 multiple cloning site"
     primer_bind     complement(3071..3087)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    3095..3111
                     /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(3119..3149)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3164..3185
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     CDS             3215..4039
                     /codon_start=1
                     /product="VENUS 3xFLAG"
                     /label=VENUS 3xFLAG
                     /note="VENUS 3xFLAG fusion protein"
                     /protein_id="ANF29833.1"
                     /translation="NGDLGGSGGRMVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGE
                     GDATYGKLTLKLICTTGKLPVPWPTLVTTLGYGLQCFARYPDHMKQHDFFKSAMPEGYV
                     QERTIFFKDDGNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYI
                     TADKQKNGIKANFKIRHNIEDGGVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALSKDP
                     NEKRDHMVLLEFVTAAGITLGMDELIKGGGDYKDHDGDYKDHDIDYKDDDDK"
     CDS             3245..3763
                     /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             3971..4036
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     rep_origin      complement(5397..5985)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(6156..7016)
                     /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(7017..7121)
                     /gene="bla"
                     /label=AmpR promoter