pLM005-Venus vector (V004983)

Basic Vector Information

Vector Name:
pLM005-Venus
Antibiotic Resistance:
Ampicillin
Length:
7216 bp
Type:
Cloning vector
Replication origin:
ori
Source/Author:
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, Sti
Promoter:
T3

pLM005-Venus vector Map

pLM005-Venus7216 bp300600900120015001800210024002700300033003600390042004500480051005400570060006300660069007200APHVIIIT3 promoterM13 fwdMCSM13 revlac operatorlac promoterCAP binding siteVenusFLAGoriAmpRAmpR promoter

pLM005-Venus vector Sequence

LOCUS       40924_28436        7216 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Cloning vector pLM005-Venus, complete sequence.
ACCESSION   .
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)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 7216)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Proc. Natl.
            Acad. Sci. U.S.A. (2016) In press"
COMMENT     SGRef: number: 2; type: "Journal Article"; journalName: "Submitted 
            (13-APR-2016) Department of Plant Biology, Carnegie Institution of 
            Washington, 260 Panama Street, Stanford, CA 94305, USA"
COMMENT     SGRef: number: 3; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..7216
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     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"
     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    complement(3095..3111)
                     /label=lac operator
                     /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    complement(3164..3185)
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     CDS             3245..3961
                     /codon_start=1
                     /label=Venus
                     /note="yellow fluorescent protein (YFP) with fast and
                     efficient maturation (Nagai et al., 2002)"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KLICTTGKLPVPWPTLVTTLGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYITADKQKNGIK
                     ANFKIRHNIEDGGVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELIK"
     CDS             4013..4036
                     /codon_start=1
                     /label=FLAG
                     /note="FLAG(R) epitope tag, followed by an enterokinase
                     cleavage site"
                     /translation="DYKDDDDK"
     rep_origin      complement(5397..5985)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(6159..7016)
                     /codon_start=1
                     /label=AmpR
                     /note="beta-lactamase"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     promoter        complement(7017..7121)
                     /label=AmpR promoter

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