LOCUS       Exported                8732 bp ds-DNA     circular SYN 17-DEC-2018
DEFINITION  Binary vector pCRB_PMpHSP17.8, complete sequence.
ACCESSION   MF939096
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8732)
  AUTHORS   Yu Z, Boehm CR, Hibberd JM, Abell C, Haseloff J, Burgess SJ, 
            Reyna-Llorens I.
  TITLE     Droplet-based microfluidic analysis and screening of single plant 
            cells
  JOURNAL   PLoS ONE 13 (5), e0196810 (2018)
  PUBMED    29723275
REFERENCE   2  (bases 1 to 8732)
  AUTHORS   Boehm CR.
  TITLE     Direct Submission
  JOURNAL   Submitted (10-SEP-2017) Plant Sciences, University of Cambridge, 
            Downing Street, Cambridge, Cambridgeshire CB2 3EA, United Kingdom
REFERENCE   3  (bases 1 to 8732)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     ##Assembly-Data-START##
            Sequencing Technology :: Sanger dideoxy sequencing 
            ##Assembly-Data-END##
FEATURES             Location/Qualifiers
     source          1..8732
                     /organism="Binary vector pCRB_PMpHSP17.8"
                     /mol_type="other DNA"
                     /db_xref="taxon:2200773"
     misc_feature    1..32
                     /label=LB
                     /note="LB"
     misc_feature    6..28
                     /label=LB T-DNA repeat
                     /note="left border repeat from nopaline C58 T-DNA 
                     (truncated)"
     regulatory      33..1761
                     /regulatory_class="promoter"
                     /note="MpEF1a promoter & 5UTR"
     CDS             1767..2882
                     /codon_start=1
                     /gene="hygR"
                     /product="hygromycin phosphotransferase"
                     /label=hygR
                     /protein_id="AWK40017.1"
                     /translation="MTHNPTILRKTLPLYKEVHFIWRGQPKLIPMKKPELTATSVEKFL
                     IEKFDSVSDLMQLSEGEESRAFSFDVGGRGYVLRVNSCADGFYKDRYVYRHFASAALPI
                     PEVLDIGEFSESLTYCISRRAQGVTLQDLPETELPAVLQPVAEAMDAIAAADLSQTSGF
                     GPFGPQGIGQYTTWRDFICAIADPHVYHWQTVMDDTVSASVAQALDELMLWAEDCPEVR
                     HLVHADFGSNNVLTDNGRITAVIDWSEAMFGDSQYEVANIFFWRPWLACMEQQTRYFER
                     RHPELAGSPRLRAYMLRIGLDQLYQSLVDGNFDDAAWAQGRCDAIVRSGAGTVGRTQIA
                     RRSAAVWTDGCVEVLADSGNRRPSTRPRAKE"
     gene            1767..2882
                     /gene="hygR"
                     /label=hygR
     promoter        1792..1838
                     /label=minimal CaMV 35S promoter
                     /note="minimal 35S promoter from cauliflower mosaic virus"
     CDS             1857..2882
                     /codon_start=1
                     /gene="aph(4)-Ia"
                     /product="aminoglycoside phosphotransferase from E. coli"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
                     YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP
                     ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
                     TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
                     GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
                     NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK
                     E"
     regulatory      2883..3292
                     /regulatory_class="terminator"
                     /note="35ST"
     polyA_signal    3113..3289
                     /label=CaMV poly(A) signal
                     /note="cauliflower mosaic virus polyadenylation signal"
     primer_bind     3295..3311
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     regulatory      3417..3687
                     /regulatory_class="terminator"
                     /note="NosT"
     terminator      3427..3679
                     /label=NOS terminator
                     /note="nopaline synthase terminator and poly(A) signal"
     regulatory      3724..4925
                     /regulatory_class="promoter"
                     /note="MpHSP17.8 promoter & 5UTR"
     CDS             4926..5645
                     /codon_start=1
                     /gene="mVenus"
                     /product="mVenus"
                     /label=mVenus
                     /protein_id="AWK40018.1"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KLICTTGKLPVPWPTLVTTLGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYITADKQKNGIK
                     ANFKIRHNIEDGGVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     gene            4926..5645
                     /gene="mVenus"
                     /label=mVenus
     regulatory      5723..6027
                     /regulatory_class="terminator"
                     /note="NosT"
     terminator      5740..5992
                     /label=NOS terminator
                     /note="nopaline synthase terminator and poly(A) signal"
     primer_bind     complement(6040..6056)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     misc_feature    6114..6166
                     /label=RB
                     /note="RB"
     misc_feature    6117..6141
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     misc_feature    6167..8732
                     /label=pGreenII backbone
                     /note="pGreenII backbone"
     rep_origin      complement(6232..6820)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     rep_origin      8170..8605
                     /label=pSa ori
                     /note="origin of replication from bacterial plasmid pSa"