LOCUS       40924_13365        8732 bp DNA     circular SYN 17-DEC-2018
DEFINITION  Binary vector pCRB_PMpHSP17.8, complete sequence.
ACCESSION   .
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)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 8732)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "PLoS ONE"; 
            date: "2018"; volume: "13"; issue: "5"; pages: "e0196810"
COMMENT     SGRef: number: 2; type: "Journal Article"; journalName: "Submitted 
            (10-SEP-2017) Plant Sciences, University of Cambridge, Downing 
            Street, Cambridge, Cambridgeshire CB2 3EA, United Kingdom"
COMMENT     SGRef: number: 3; type: "Journal Article"
COMMENT     ##Assembly-Data-START##
            Sequencing Technology :: Sanger dideoxy sequencing 
            ##Assembly-Data-END##
FEATURES             Location/Qualifiers
     source          1..8732
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    6..28
                     /label=LB T-DNA repeat
                     /note="left border repeat from nopaline C58 T-DNA
                     (truncated)"
     regulatory      33..1761
                     /label=MpEF1a promoter & 5'UTR
                     /note="MpEF1a promoter & 5'UTR"
                     /regulatory_class="promoter"
     promoter        1792..1838
                     /label=minimal CaMV 35S promoter
                     /note="minimal 35S promoter from cauliflower mosaic virus"
     CDS             1857..2879
                     /label=HygR
                     /note="aminoglycoside phosphotransferase from E. coli"
     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"
     terminator      3427..3679
                     /label=NOS terminator
                     /note="nopaline synthase terminator and poly(A) signal"
     regulatory      3724..4925
                     /label=MpHSP17.8 promoter & 5'UTR
                     /note="MpHSP17.8 promoter & 5'UTR"
                     /regulatory_class="promoter"
     CDS             4926..5642
                     /label=Venus
                     /note="yellow fluorescent protein (YFP) with fast and
                     efficient maturation (Nagai et al., 2002)"
     regulatory      5723..6027
                     /label=NosT
                     /note="NosT"
                     /regulatory_class="terminator"
     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    6117..6141
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     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"
ORIGIN
        1 gatcttggca ggatatattg tggtgtaacg ttcaaatgag tcacacacat tgttgagaga
       61 catatcaaaa taactcaatt gtaaggtaag ttgatagatt tcccagttaa atgcattatt
      121 ttaacaatat ttgtatctca aattttcaaa atcatatcaa atttgcagat cttgatgatg
      181 attcacaact ctgtcttcga tttttgattg ttaacgggta tcgatatttg ggtaacttaa
      241 aaagaatttg ctaaagatct taaaaaacag attaaaaaaa cctttcaaaa tagctaatcc
      301 tttaaatccg tttcacaata taatgctcat tgatgataat cgaaaagatt ccattttaaa
      361 gtgcatataa attgcttaat tatctctttc tgtctcgtaa tattttgttc tctcctattg
      421 gatcggaccc gcatcttctc gatcagttca ctatactaat atttggtcaa cttcaagaga
      481 gattagaact tcgttctcca aagtgtaaag gccgacaagc agaaaaggaa acttcaatca
      541 gaagccttct aggttttccc gcgtcccaaa aagcgaaggg aggacgggga ggacggtgag
      601 gccagaagaa gaaagcaaca gtaaaatctc atgtgattgc gtgcatgtca tggatctatg
      661 ggatgctagt ccaccgtttg gaagtatgtg cagaaggaga aaggcgcggg aggccaaaag
      721 cgacttcggc cacctctcta ctttctagaa agtttgctcg cagcttgtaa taagcgcgga
      781 aaaggtactc gaaaaaccgg agcaatcggt gtggtatcag cagtcgggtt cgctaaacct
      841 acgtcttctt attttggaat gttaaattga aagggcgtgc cggccctaag cacggacgat
      901 gtgacagacg gaggagtgtg agcaattcgg atgacgatgc tggccggcag tcacgacaaa
      961 gagtccgaag gggcatggac agggcaatat ataaatggaa aagaggcttt gaatattcct
     1021 aacccttcgt cgtcggcctc ccaacttgcc ctcgcatgct gaattggccg cctatctgcg
     1081 aatggccggc cgccgaccct aagtcgcaat aacttgaacg gagccgatgc aaagtcctta
     1141 tcgaacgctc gttattataa acggggcaga cctaggtatt taattatatc aaaaccctcc
     1201 gcaatttcgc taatcattcc ctaatttctt ctactttggc gtctagtgct gtggtcgcgc
     1261 ttgctctcaa agctcgctgg gaggcagtgg agcgtctggc ttaaaatcga tcgccgcaac
     1321 cctagagttc ctttcacttc tttctctctt gcgtggagag catcgacatc gaagagctgc
     1381 atcttgtctt ctggtatgcg ccttgtgccc tcgtctctgt acgtcgttac gattgctcga
     1441 tattgtgatt gtctagacct gaaacccgac tggagctttt acgtggtgtc gattggggcc
     1501 tcgagcgagt gattttttag gttcgtttta gccaagtaca tcccactttg gtcagtcctg
     1561 ttatctcggt aaggtgttca gttcggtctc tggtgcggag gtgcctcgta agtttcctgc
     1621 atgatagatt tgtctatcct tgacttgaag aggagtcgaa ggtcagcgag gttggctgtg
     1681 tgctatggac gtcggtgctc cttggttccg tgcttctatg ggtgttgcct caacagtatt
     1741 gatgtgcctg cagaaaggtt gtcaccatga cgcacaatcc cactatcctt cgcaagaccc
     1801 ttcctctata taaggaagtt catttcattt ggagaggaca gcccaagctg atccctatga
     1861 aaaagcctga actcaccgcg acgtctgtcg agaagtttct gatcgaaaag ttcgacagcg
     1921 tctccgacct gatgcagctc tcggagggcg aagaatctcg tgctttcagc ttcgatgtag
     1981 gagggcgtgg atatgtcctg cgggtaaata gctgcgccga tggtttctac aaagatcgtt
     2041 atgtttatcg gcactttgca tcggccgcgc tcccgattcc ggaagtgctt gacattgggg
     2101 agttcagcga gagcctgacc tattgcatct cccgccgtgc acagggtgtc acgttgcaag
     2161 acctgcctga aaccgaactg cccgctgttc ttcagccggt cgcggaggct atggatgcga
     2221 tcgctgcggc cgatcttagc cagacgagcg ggttcggccc attcggaccg caaggaatcg
     2281 gtcaatacac tacatggcgt gatttcatat gcgcgattgc tgatccccat gtgtatcact
     2341 ggcaaactgt gatggacgac accgtcagtg cgtccgtcgc gcaggctctc gatgagctga
     2401 tgctttgggc cgaggactgc cccgaagtcc ggcacctcgt gcacgcggat ttcggctcca
     2461 acaatgtcct gacggacaat ggccgcataa cagcggtcat tgactggagc gaggcgatgt
     2521 tcggggattc ccaatacgag gtcgccaaca tcttcttctg gaggccgtgg ttggcttgta
     2581 tggagcagca gacgcgctac ttcgagcgga ggcatccgga gcttgcagga tcgccacgcc
     2641 tccgggcgta tatgctccgc attggtcttg accaactcta tcagagcttg gttgacggca
     2701 atttcgatga tgcagcttgg gcgcagggtc gatgcgacgc aatcgtccga tccggagccg
     2761 ggactgtcgg gcgtacacaa atcgcccgca gaagcgcggc cgtctggacc gatggctgtg
     2821 tagaagtact cgccgatagt ggaaaccgac gccccagcac tcgtccgagg gcaaaggaat
     2881 agagtagatg ccgaccgaac aagagctgat ttcgagaacg cctcagccag caactcgcgc
     2941 gagcctagca aggcaaatgc gagagaacgg ccttacgctt ggtggcacag ttctcgtcca
     3001 cagttcgcta agctcgctcg gctggtcgcg ggagggccgg tcgcaatgat tcaggaatta
     3061 attcggtacg ctgaaatcac cagtctctct ctacaaatct atctctctct attttctcca
     3121 taaataatgt gtgagtagtt tcccgataag ggaaattagg gttcttatag ggtttcgctc
     3181 atgtgttgag catataagaa acccttagta tgtatttgta tttgtaaaat acttctatca
     3241 ataaaatttc taattcctaa aaccaaaatc cagtactaaa atccagatcg atcacaggaa
     3301 acagctatga ccatgattac gcccgggctt gacctgaagc agatcaacaa ctggttcatc
     3361 aaccagcgga agcggcactg gaagccatcc gagtaacatc aaaaatgaat ctgcccgaat
     3421 ttccccgatc gttcaaacat ttggcaataa agtttcttaa gattgaatcc tgttgccggt
     3481 cttgcgatga ttatcatata atttctgttg aattacgtta agcatgtaat aattaacatg
     3541 taatgcatga cgttatttat gagatgggtt tttatgatta gagtcccgca attatacatt
     3601 taatacgcga tagaaaacaa aatatagcgc gcaaactagg ataaattatc gcgcgcggtg
     3661 tcatctatgt tactagatcg ggaattcata gtttggccgc gaattcgccc ttgatatcca
     3721 cgtccagcca ttatagcaac tatgttaaaa gtgacagttt tctctaatcg ggaatcttgt
     3781 ccgtattgaa agttccctgt tcagtgactg agccgcttaa taatattttc agtttcgagt
     3841 acgagggaga gattgaacaa attttggggt ttcagggcta aattctgcag gggctttagt
     3901 tttgagtcgg gtgaaacttt cctaagtctt ggcaatgaac tcacattcgc cattgtacct
     3961 attaaatgac ccaaacatcc ttgacaacaa attgacaagt acgggaacat gatcttcaat
     4021 atgaaattta atatgtcctt tatgttggct ttcctcatca atgattggaa actcacatac
     4081 aaagaaaagc gtgatataat aaatcactag aatcttctcc gctttctaag acagtaatga
     4141 tggatccacg tgagtgtgaa acatcaatat caacatcggt aggagtagag aagtttctaa
     4201 atcagaaaaa gtgaatagac aagacaactt actccttgaa cgagaatatt cctcgagaaa
     4261 gagggaaaag agtaaaaaga gtaaaaagcc atggtacaga atagcccatt ccatggttat
     4321 tgcagtcaaa ccgctacgcc agcaaaaacg gaacgaaact aaacccgttc cacaaattgc
     4381 gaaagatgga atgtattcgc catgtatatt tcgagattat tatgagactt ccggtcccgg
     4441 gaagatggcc tgggttcacc gggatcagcc gaattccgac gtgatggagc acggccatgc
     4501 atacttctag agaagtcggc caccgtgaaa ttcgacgacg agatggtact agaatcaccg
     4561 ggacagtaca cacctcgttc atggcctcga gcattcgatg tcgctctcac ctgcacagcc
     4621 tcgaggtatc gagagggttc taggatggac cagcgatgtc tcgaggaatt tagaaagatt
     4681 tcatgcgctc gttgtgtgag agccaaggaa gcacattatg gggcctataa gtaccacgca
     4741 agggcctggg gttcccatca atcaaatcag tgttcagacg atttcctcaa gcgatcattg
     4801 caccgtgatc agtgaagcac agacttcaag cttttggctt gcctacttcc acgttcccat
     4861 ccgagatcga caccgaattg ttgagttctg tctgatcttc ctgcagtcga gagcgaagtt
     4921 ccaccatggt gagcaagggc gaggagctgt tcaccggggt ggtgcccatc ctggtcgagc
     4981 tggacggcga cgtaaacggc cacaagttca gcgtgtccgg cgagggcgag ggcgatgcca
     5041 cctacggcaa gctgaccctg aagctgatct gcaccaccgg caagctgccc gtgccctggc
     5101 ccaccctcgt gaccaccctg ggctacggcc tgcagtgctt cgcccgctac cccgaccaca
     5161 tgaagcagca cgacttcttc aagtccgcca tgcccgaagg ctacgtccag gagcgcacca
     5221 tcttcttcaa ggacgacggc aactacaaga cccgcgccga ggtgaagttc gagggcgaca
     5281 ccctggtgaa ccgcatcgag ctgaagggca tcgacttcaa ggaggacggc aacatcctgg
     5341 ggcacaagct ggagtacaac tacaacagcc acaacgtcta tatcaccgcc gacaagcaga
     5401 agaacggcat caaggccaac ttcaagatcc gccacaacat cgaggacggc ggcgtgcagc
     5461 tcgccgacca ctaccagcag aacaccccca tcggcgacgg ccccgtgctg ctgcccgaca
     5521 accactacct gagctaccag tccgccctga gcaaagaccc caacgagaag cgcgatcaca
     5581 tggtcctgct ggagttcgtg accgccgccg ggatcactct cggcatggac gagctgtaca
     5641 agtaataaag ggccgccact ccaccggcgc ctaagagctc gaatttcccc gatcggcgtc
     5701 ttggattcgc ttgaattgct accgagctcg aatttccccg atcgttcaaa catttggcaa
     5761 taaagtttct taagattgaa tcctgttgcc ggtcttgcga tgattatcat ataatttctg
     5821 ttgaattacg ttaagcatgt aataattaac atgtaatgca tgacgttatt tatgagatgg
     5881 gtttttatga ttagagtccc gcaattatac atttaatacg cgatagaaaa caaaatatag
     5941 cgcgcaaact aggataaatt atcgcgcgcg gtgtcatcta tgttactaga tcgggaattc
     6001 tccagctttt gttcccttta gtgagggggg aattaattca ctggccgtcg ttttacaacg
     6061 tcgtgactgg gaaaaccctg gcgttaggga taacgcagga aagaacatga aggccttgac
     6121 aggatatatt ggcgggtaaa ctaagtcgct gtatgtgttt gtttgagatc tcatgtgagc
     6181 aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag
     6241 gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc
     6301 gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt
     6361 tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct
     6421 ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg
     6481 ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct
     6541 tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat
     6601 tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg
     6661 ctacactaga agaacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaag
     6721 aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt
     6781 ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc
     6841 tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt
     6901 atcaaaaagg atcttcacct agatcctttt aaattaaaaa tgaagtttta aatcaatcta
     6961 aagtatatat gtgtaacatt ggtctagtga ttagaaaaac tcatcgagca tcaaatgaaa
     7021 ctgcaattta ttcatatcag gattatcaat accatatttt tgaaaaagcc gtttctgtaa
     7081 tgaaggagaa aactcaccga ggcagttcca taggatggca agatcctggt atcggtctgc
     7141 gattccgact cgtccaacat caatacaacc tattaatttc ccctcgtcaa aaataaggtt
     7201 atcaagtgag aaatcaccat gagtgacgac tgaatccggt gagaatggca aaagtttatg
     7261 catttctttc cagacttgtt caacaggcca gccattacgc tcgtcatcaa aatcactcgc
     7321 atcaaccaaa ccgttattca ttcgtgattg cgcctgagcg agacgaaata cgcgatcgct
     7381 gttaaaagga caattacaaa caggaatcga atgcaaccgg cgcagcctga gcgagacgaa
     7441 atacgcgatc gctgttaaaa ggacaattac aaacaggaat cgaatgcaac cggcgcagga
     7501 acactgccag cgcatcaaca atattttcac ctgaatcagg atattcttct aatacctgga
     7561 atgctgtttt ccctgggatc gcagtggtga gtaaccatgc atcatcagga gtacggataa
     7621 aatgcttgat ggtcggaaga ggcataaatt ccgtcagcca gtttagtctg accatctcat
     7681 ctgtaacaac attggcaacg ctacctttgc catgtttcag aaacaactct ggcgcatcgg
     7741 gcttcccata caatcggtag attgtcgcac ctgattgccc gacattatcg cgagcccatt
     7801 tatacccata taaatcagca tccatgttgg aatttaatcg cggccttgag caagacgttt
     7861 cccgttgaat atggctcata acaccccttg tattactgtt tatgtaagca gacagtttta
     7921 ttgttcatga tgatatattt ttatcttgtg caatgtaaca tcagagattt tgagacacaa
     7981 cgtggctttg ttgaataaat cgaacttttg ctgagttgaa ggatcagatc acgcatcttc
     8041 ccgacaacgc agaccgttcc gtggcaaagc aaaagttcaa aatcaccaac tggtccacct
     8101 acaacaaagc tctcatcaac cgtggctccc tcactttctg gctggatgat ggggcgattc
     8161 aggcgatccc catccaacag cccgccgtcg agcgggcttt tttatccccg gaagcctgtg
     8221 gatagagggt agttatccac gtgaaaccgc taatgccccg caaagccttg attcacgggg
     8281 ctttccggcc cgctccaaaa actatccacg tgaaatcgct aatcagggta cgtgaaatcg
     8341 ctaatcggag tacgtgaaat cgctaataag gtcacgtgaa atcgctaatc aaaaaggcac
     8401 gtgagaacgc taatagccct ttcagatcaa cagcttgcaa acacccctcg ctccggcaag
     8461 tagttacagc aagtagtatg ttcaattagc ttttcaatta tgaatatata tatcaattat
     8521 tggtcgccct tggcttgtgg acaatgcgct acgcgcaccg gctccgcccg tggacaaccg
     8581 caagcggttg cccaccgtcg agcgccagcg cctttgccca caacccggcg gccggccgca
     8641 acagatcgtt ttataaattt ttttttttga aaaagaaaaa gcccgaaagg cggcaacctc
     8701 tcgggcttct ggatttccga tccccggaat ta
//
