LOCUS       Exported                8025 bp ds-DNA     circular SYN 18-DEC-2018
DEFINITION  Cloning vector pLM006-EPYC1-mCherry, complete sequence.
ACCESSION   KX077947
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8025)
  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 8025)
  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 8025)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..8025
                     /organism="Cloning vector pLM006-EPYC1-mCherry"
                     /mol_type="other DNA"
                     /db_xref="taxon:1850521"
     CDS             join(423..509,655..1566)
                     /codon_start=1
                     /product="APHVII"
                     /label=APHVII
                     /note="hygromycin resistance marker"
                     /protein_id="ANF29838.1"
                     /translation="MTQESLLLLDRIDSDDSYASLRNDQEFWEPLARRALEELGLPVPP
                     VLRVPGESTNPVLVGEPGPVIKLFGEHWCGPESLASESEAYAVLADAPVPVPRLLGRGE
                     LRPGTGAWPWPYLVMSRMTGTTWRSAMDGTTDRNALLALARELGRVLGRLHRVPLTGNT
                     VLTPHSEVFPELLRERRAATVEDHRGWGYLSPRLLDRLEDWLPDVDTLLAGREPRFVHG
                     DLHGTNIFVDLAATEVTGIVDFTDVYAGDSRYSLVQLHLNAFRGDREILAALLDGAQWK
                     RTEDFARELLAFTFLHDFEVFEETPLDLSGFTDPEELAQFLWGPPDTAPGA"
     promoter        complement(1827..1845)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     CDS             join(2614..2679,2931..2992,3255..4857)
                     /codon_start=1
                     /product="EPYC1-mCherry"
                     /label=EPYC1-mCherry
                     /note="EPYC1 mCherry 6xHIS fusion protein"
                     /protein_id="ANF29837.1"
                     /translation="MATISSMRVGAASRVVVSGRVKTVKVAARGSWRESSTATVQASRA
                     SSATNRVSPTRSVLPANWRQELESLRNGNGSSSAASSAPAPARSSSASWRDAAPASSAP
                     ARSSSASKKAVTPSRSALPSNWKQELESLRSSSPAPASSAPAPARSSSASWRDAAPASS
                     APARSSSSKKAVTPSRSALPSNWKQELESLRSSSPAPASSAPAPARSSSASWRDAAPAS
                     SAPARSSSASKKAVTPSRSALPSNWKQELESLRSNSPAPASSAPAPARSSSASWRDAPA
                     SSSSSSADKAGTNPWTGKSKPEIKRTALPADWRKGLNGDLGGSGGRMVSKGEEDNMAII
                     KEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEGTQTAKLKVTKGGPLPFAWDILSPQFMY
                     GSKAYVKHPADIPDYLKLSFPEGFKWERVMNFEDGGVVTVTQDSSLQDGEFIYKVKLRG
                     TNFPSDGPVMQKKTMGWEASSERMYPEDGALKGEIKQRLKLKDGGHYDAEVKTTYKAKK
                     PVQLPGAYNVNIKLDITSHNEDYTIVEQYERAEGRHSTGGMDELYRSGGGHHHHHHGDL
                     "
     CDS             4108..4812
                     /codon_start=1
                     /product="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
                     /label=mCherry
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEG
                     TQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNF
                     EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK
                     GEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERA
                     EGRHSTGGMDELY"
     CDS             4828..4845
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     rep_origin      complement(6206..6794)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(6965..7825)
                     /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(7826..7930)
                     /gene="bla"
                     /label=AmpR promoter