LOCUS       Exported                9997 bp ds-DNA     circular SYN 18-DEC-2018
DEFINITION  Expression vector pLM005-RCA1-Venus, complete sequence.
ACCESSION   KY550375
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9997)
  AUTHORS   Freeman Rosenzweig ES, Xu B, Kuhn Cuellar L, Marinez-Sanchez A, 
            Schaffer M, Strauss M, Cartwright HN, Forster F, Engel BD, Wingreen 
            NS, Mackinder LCM., Jonikas MC.
  TITLE     The pyrenoid is liquid-like and exhibits dynamic reorganization
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 9997)
  AUTHORS   Mackinder LCM., Jonikas MC.
  TITLE     Direct Submission
  JOURNAL   Submitted (29-JAN-2017) Department of Biology, University of York, 
            Wentworth Way, York, North Yorkshire YO10 5DD, United Kingdom
REFERENCE   3  (bases 1 to 9997)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..9997
                     /organism="Expression vector pLM005-RCA1-Venus"
                     /mol_type="other DNA"
                     /db_xref="taxon:2066026"
     CDS             join(101..106,197..213,336..393,478..564,1114..1314,
                     1511..1855,2084..2251,2502..2690,3033..3132,3345..4222)
                     /codon_start=1
                     /product="RCA1-Venus"
                     /label=RCA1-Venus
                     /note="RCA1 Venus 3xFLAG fusion protein"
                     /protein_id="AUR45266.1"
                     /translation="MQVTMKSSAVSGQRVGGARVATRSVRRAQLQVVASSRKQMGRWRS
                     IDAGVDASDDQQDITRGREMVDDLFQGGFGAGGTHNAVLSSQEYLSQSRASFNNIEDGF
                     YISPAFLDKMTIHIAKNFMDLPKIKVPLILGIWGGKGQGKTFQCALAYKKLGIAPIVMS
                     AGELESGNAGEPAKLIRTRYREASDIIKKGRMCSLFINDLDAGAGRMGDTTQYTVNNQM
                     VNATLMNIADNPTNVQLPGVYKNEEIPRVPIVCTGNDFSTLYAPLIRDGRMEKYYWNPT
                     REDRIGVCMGIFQEDNVQRREVENLVDTFPGQSIDFFGALRARVYDDMVRQWITDTGVD
                     KIGQQLVNARQKVAMPKVSMDLNVLIKYGKSLVDEQENVKRVQLADAYLSGAELAGHGG
                     SSLPEAYSRNGDLGGSGGRMVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATY
                     GKLTLKLICTTGKLPVPWPTLVTTLGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTI
                     FFKDDGNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYITADKQ
                     KNGIKANFKIRHNIEDGGVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALSKDPNEKRD
                     HMVLLEFVTAAGITLGMDELIKGGGDYKDHDGDYKDHDIDYKDDDDK"
     CDS             3428..3946
                     /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             4154..4219
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     rep_origin      complement(5580..6168)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(6339..7199)
                     /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(7200..7304)
                     /gene="bla"
                     /label=AmpR promoter
     CDS             8183..8986
                     /codon_start=1
                     /product="APHVIII"
                     /label=APHVIII
                     /note="Paromomycin resistance marker"
                     /protein_id="AUR45267.1"
                     /translation="MDDALRALRGRYPGCEWVVVEDGASGAGVYRLRGGGRELFVKVAA
                     LGAGVGLLGEAERLVWLAEVGIPVPRVVEGGGDERVAWLVTEAVPGRPASARWPREQRL
                     DVAVALAGLARSLHALDWERCPFDRSLAVTVPQAARAVAEGSVDLEDLDEERKGWSGER
                     LLAELERTRPADEDLAVCHGDLCPDNVLLDPRTCEVTGLIDVGRVGRADRHSDLALVLR
                     ELAHEEDPWFGPECSAAFLREYGRGWDGAVSEEKLAFYRLLDEFF"
     promoter        complement(9311..9329)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"