LOCUS       40924_2239        9789 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Cloning vector nLucFlucGW, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9789)
  AUTHORS   Reis RS.
  TITLE     nLuc/Fluc dual luciferase for protoplast transformation
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 9789)
  AUTHORS   Reis RS.
  TITLE     Direct Submission
  JOURNAL   Submitted (29-JUN-2018) Department of Plant Molecular Biology, 
            University of Lausanne, Quartier UNIL-Sorge, Batiment Biophore, 
            Lausanne, Vaud 1015, Switzerland
REFERENCE   3  (bases 1 to 9789)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 9789)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: 
            "Unpublished"
COMMENT     SGRef: number: 2; type: "Journal Article"; journalName: "Submitted 
            (29-JUN-2018) Department of Plant Molecular Biology, University of 
            Lausanne, Quartier UNIL-Sorge, Batiment Biophore, Lausanne, Vaud 
            1015, Switzerland"
COMMENT     SGRef: number: 3; type: "Journal Article"
COMMENT     ##Assembly-Data-START##
            Sequencing Technology :: Sanger dideoxy sequencing 
            ##Assembly-Data-END##
FEATURES             Location/Qualifiers
     source          1..9789
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      103..189
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB
                     gene"
     terminator      281..308
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB
                     gene"
     regulatory      324..1449
                     /label=RHIP1pro
                     /note="RHIP1pro"
                     /regulatory_class="promoter"
     protein_bind    1470..1594
                     /label=attR1
                     /note="recombination site for the Gateway(R) LR reaction"
     promoter        1619..1649
                     /label=lac UV5 promoter
                     /note="E. coli lac promoter with an 'up' mutation"
     CDS             1703..2359
                     /label=CmR
                     /note="chloramphenicol acetyltransferase"
     CDS             2704..3006
                     /label=ccdB
                     /note="CcdB, a bacterial toxin that poisons DNA gyrase"
     protein_bind    complement(3050..3174)
                     /label=attR2
                     /note="recombination site for the Gateway(R) LR reaction"
     CDS             3188..3253
                     /label=F2A
                     /note="2A peptide from foot-and-mouth disease virus
                     polyprotein"
     CDS             3272..3784
                     /label=Nluc
                     /note="NanoLuc(R) luciferase"
     CDS             3785..3811
                     /label=HA
                     /note="HA (human influenza hemagglutinin) epitope tag"
     terminator      3834..4082
                     /label=HSP terminator
                     /note="efficient transcription terminator from the
                     Arabidopsis thaliana heat shock protein 18.2 gene (Nagaya 
                     et al., 2010)"
     protein_bind    complement(4091..4115)
                     /label=attB2
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             5850..7499
                     /label=luciferase
                     /note="firefly luciferase"
     terminator      7529..7781
                     /label=NOS terminator
                     /note="nopaline synthase terminator and poly(A) signal"
     promoter        8118..8188
                     /label=AmpR promoter
     CDS             8189..9046
                     /label=AmpR
                     /note="beta-lactamase"
     rep_origin      9139..9727
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
ORIGIN
        1 ctttcctgcg ttatcccctg attctgtgga taaccgtatt accgctagca tggatctcgg
       61 ggacgtctaa ctactaagcg agagtaggga actgccaggc atcaaataaa acgaaaggct
      121 cagtcggaag actgggcctt tcgttttatc tgttgtttgt cggtgaacgc tctcctgagt
      181 aggacaaatc cgccgggagc ggatttgaac gttgtgaagc aacggcccgg agggtggcgg
      241 gcaggacgcc cgccataaac tgccaggcat caaactaagc agaaggccat cctgacggat
      301 ggcctttttg cgtttctacc attgctctca tactcagaag caatccctgc acgaataccc
      361 tgaagcaact gcaatatagt agctgagctt ctccgtctcc tcctaaggat agactcaaac
      421 tctctgtcta aatcaatctc tctaccttgt tgcctgtggc gatgacgacg atgatgatgt
      481 cgatcaatat cgacgctgtc atttccatca acatcatcag atacattgac ctcatcatta
      541 tccacaccaa actctgtcct cctaaacctt ctacgtctac gaggattact catcattcca
      601 agcaagattg gaccccatag agacaatgca cggtctgtac caaaatcaat ctcagggtct
      661 tgaacctccc ttagactgct actgctgccg ccattaatct cacggctcat ttcctcgaca
      721 aacccgcttt ggcaaaaggg gcatttgatc tcggcaccaa taactggatt caccatttgt
      781 gaacacatgt gacaccaata cctagctgac attgcctctt cctccttctt tccttcccca
      841 cccaaacctc tcgtttctga aacaaccaga agaacaacaa aaaaaaaaac atgaagaagc
      901 aaacctataa agtttcgacc tttgtttcat cagatctaaa taaagagaat ggtattccca
      961 aaagtttcac taaaaagagg ataatatcat cttcaccaac atcattagct caagtttcag
     1021 acaacaaaac gcaagctttc tatgcaaatg cgaataaaca gatcaaccaa aatttgcctc
     1081 tataaagaga taacaataga aatcagggct aatccatgaa aagaaaactc tcaattctca
     1141 tggagtaatt aaaacaaaac aaatacaaaa acaaaccttg taaatgttct tacttttgga
     1201 gatccagagg gaagaaatgt cttcctcctt cttttagatc tccacgaaga taaatgagag
     1261 cttttttatt tgtcttcttc tgcagaaatg gtgggagacg aagaaaagag gaatgaagga
     1321 ggaaaggaag acgaagatga agacgaagaa aagggaatgt aatgtatagt tcgcagaccc
     1381 aatcttaaag cccattaacc gcaaactgat atataaagga gccgggtcta tcttaaccat
     1441 ttaaaaccac tccccgagct cgaattatca caagtttgta caaaaaagct gaacgagaaa
     1501 cgtaaaatga tataaatatc aatatattaa attagatttt gcataaaaaa cagactacat
     1561 aatactgtaa aacacaacat atccagtcac tatggcggcc gcattaggca ccccaggctt
     1621 tacactttat gcttccggct cgtataatgt gtggattttg agttaggatc cggcgagatt
     1681 ttcaggagct aaggaagcta aaatggagaa aaaaatcact ggatatacca ccgttgatat
     1741 atcccaatgg catcgtaaag aacattttga ggcatttcag tcagttgctc aatgtaccta
     1801 taaccagacc gttcagctgg atattacggc ctttttaaag accgtaaaga aaaataagca
     1861 caagttttat ccggccttta ttcacattct tgcccgcctg atgaatgctc atccggaatt
     1921 ccgtatggca atgaaagacg gtgagctggt gatatgggat agtgttcacc cttgttacac
     1981 cgttttccat gagcaaactg aaacgttttc atcgctctgg agtgaatacc acgacgattt
     2041 ccggcagttt ctacacatat attcgcaaga tgtggcgtgt tacggtgaaa acctggccta
     2101 tttccctaaa gggtttattg agaatatgtt tttcgtctca gccaatccct gggtgagttt
     2161 caccagtttt gatttaaacg tggccaatat ggacaacttc ttcgcccccg ttttcaccat
     2221 gggcaaatat tatacgcaag gcgacaaggt gctgatgccg ctggcgattc aggttcatca
     2281 tgccgtctgt gatggcttcc atgtcggcag aatgcttaat gaattacaac agtactgcga
     2341 tgagtggcag ggcggggcgt aaacgcgtgg atccggctta ctaaaagcca gataacagta
     2401 tgcgtatttg cgcgctgatt tttgcggtat aagaatatat actgatatgt atacccgaag
     2461 tatgtcaaaa agaggtgtgc tatgaagcag cgtattacag tgacagttga cagcgacagc
     2521 tatcagttgc tcaaggcata tatgatgtca atatctccgg tctggtaagc acaaccatgc
     2581 agaatgaagc ccgtcgtctg cgtgccgaac gctggaaagc ggaaaatcag gaagggatgg
     2641 ctgaggtcgc ccggtttatt gaaatgaacg gctcttttgc tgacgagaac agggactggt
     2701 gaaatgcagt ttaaggttta cacctataaa agagagagcc gttatcgtct gtttgtggat
     2761 gtacagagtg atattattga cacgcccggg cgacggatgg tgatccccct ggccagtgca
     2821 cgtctgctgt cagataaagt ctcccgtgaa ctttacccgg tggtgcatat cggggatgaa
     2881 agctggcgca tgatgaccac cgatatggcc agtgtgccgg tctccgttat cggggaagaa
     2941 gtggctgatc tcagccaccg cgaaaatgac atcaaaaacg ccattaacct gatgttctgg
     3001 ggaatataaa tgtcaggctc ccttatacac agccagtctg caggtcgacc atagtgactg
     3061 gatatgttgt gttttacagt attatgtagt ctgtttttta tgcaaaatct aatttaatat
     3121 attgatattt atatcatttt acgtttctcg ttcagctttc ttgtacaaag tggtgatagc
     3181 ttggccggtg aaacagctgt tgaattttga ccttcttaag cttgcgggag acgtcgagtc
     3241 caaccccggg ccctcgatcg aggcacgggc aatggtcttc acactcgaag atttcgttgg
     3301 ggactggcga cagacagccg gctacaacct ggaccaagtc cttgaacagg gaggtgtgtc
     3361 cagtttgttt cagaatctcg gggtgtccgt aactccgatc caaaggattg tcctgagcgg
     3421 tgaaaatggg ctgaagatcg acatccatgt catcatcccg tatgaaggtc tgagcggcga
     3481 ccaaatgggc cagatcgaaa aaatttttaa ggtggtgtac cctgtggatg atcatcactt
     3541 taaggtgatc ctgcactatg gcacactggt aatcgacggg gttacgccga acatgatcga
     3601 ctatttcgga cggccgtatg aaggcatcgc cgtgttcgac ggcaaaaaga tcactgtaac
     3661 agggaccctg tggaacggca acaaaattat cgacgagcgc ctgatcaacc ccgacggctc
     3721 cctgctgttc cgagtaacca tcaacggagt gaccggctgg cggctgtgcg aacgcattct
     3781 ggcgtaccca tacgatgttc cagattacgc ttgaccttag taaaagggtg ggatatgaag
     3841 atgaagatga aatatttggt gtgtcaaata aaaagcttgt gtgcttaagt ttgtgttttt
     3901 ttcttggctt gttgtgttat gaatttgtgg ctttttctaa tattaaatga atgtaagatc
     3961 tcattataat gaataaacaa atgtttctat aatccattgt gaatgttttg ttggatctct
     4021 tctgcagcat ataactactg tatgtgctat ggtatggact atggaatatg attaaagata
     4081 aggcgcgccg acccagcttt cttgtacaaa gtggtgatat cactagtgta tgtaggtata
     4141 gtaacatgaa caagaacaga ctcaagtaca agatcgccag ggatccctca acttttgatt
     4201 cgctatttgc agtgcacctg tggcgttcat cacatctttt gtgacactgt ttgcactggt
     4261 cattgctatt acaaaggacc ttcctgatgt tgaaggagat cgaaagtaag taactgcacg
     4321 cataaccatt ttctttccgc tctttggctc aatccatttg acagtcaaag acaatgttta
     4381 accagctccg tttgatatat tgtctttatg tgtttgttca agcatgttta gttaatcatg
     4441 cctttgattg atcttgaata ggttccaaat atcaaccctg gcaacaaaac ttggagtgag
     4501 aaacattgca ttcctcggtt ctggacttct gctagtaaat tatgtttcag ccatatcact
     4561 agctttctac atgcctcagg tgaattcatc tatttccgtc ttaactattt cggttaatta
     4621 aagcacgaac accattactg catgtagaag cttgataaac tatcgccacc aatttatttt
     4681 tgttgcgata ttgttacttt cctcagtatg cagctttgaa aagaccaacc ctcttatcct
     4741 ttaacaatga acaggttttt agaggtagct tgatgattcc tgcacatgtg atcttggctt
     4801 caggcttaat tttccaggta aagcattatg agatactctt atatctctta catacttttg
     4861 agataatgca caagaacttc ataactatat gctttagttt ctgcatttga cactgccaaa
     4921 ttcattaatc tctaatatct ttgttgttga tctttggtag acatgggtac tagaaaaagc
     4981 aaactacacc aaggtaaaat acttttgtac aaacataaac tcgttatcac ggaacatcaa
     5041 tggagtgtat atctaacgga gtgtagaaac atttgattat tgcaggaagc tatctcagga
     5101 tattatcggt ttatatggaa tctcttctac gcagagtatc tgttattccc cttcctctag
     5161 ctttcaattt catggtgagg atatgcagtt ttctttgtat atcattcttc ttcttctttg
     5221 tagcttggag tcaaaatcgg ttccttcatg tacatacatc aaggatatgt ccttctgaat
     5281 ttttatatct tgcaataaaa atgcttgtac caattgaaac accagctttt tgagttctat
     5341 gatcactgac ttggttctaa ccaaaaaaaa aaaatgttta atttacatat ctaaaagtag
     5401 gtttagggaa acctaaacag taaaatattt gtatattatt cgaatttcac tcatcataaa
     5461 aacttaaatt gcaccataaa attttgtttt actattaatg atgtaatttg tgtaacttaa
     5521 gataaaaata atattccgta agttaaccgg ctaaaaccac gtataaacca gggaacctgt
     5581 taaaccggtt ctttactgga taaagaaatg aaagcccatg tagacagctc cattagagcc
     5641 caaaccctaa atttctcatc tatataaaag gagtgacatt agggtttttg ttcgtcctct
     5701 taaagcttct cgttttctct gccgtctctc tcattcgcgc gacgcaaacg atcttcaggt
     5761 gatcttcttt ctccaaatcc tctctcataa ctctgatttc gtacttgtgt atttgagctc
     5821 acgctctgtt tctctcacca cagcccacca tggaagacgc caaaaacata aagaaaggcc
     5881 cggcgccatt ctatccgctg gaagatggaa ccgctggaga gcaactgcat aaggctatga
     5941 agagatacgc cctggttcct ggaacaattg cttttacaga tgcacatatc gaggtggaca
     6001 tcacttacgc tgagtacttc gaaatgtccg ttcggttggc agaagctatg aaacgatatg
     6061 ggctgaatac aaatcacaga atcgtcgtat gcagtgaaaa ctctcttcaa ttctttatgc
     6121 cggtgttggg cgcgttattt atcggagttg cagttgcgcc cgcgaacgac atttataatg
     6181 aacgtgaatt gctcaacagt atgggcattt cgcagcctac cgtggtgttc gtttccaaaa
     6241 aggggttgca aaaaattttg aacgtgcaaa aaaagctccc aatcatccaa aaaattatta
     6301 tcatggattc taaaacggat taccagggat ttcagtcgat gtacacgttc gtcacatctc
     6361 atctacctcc cggttttaat gaatacgatt ttgtgccaga gtccttcgat agggacaaga
     6421 caattgcact gatcatgaac tcctctggat ctactggtct gcctaaaggt gtcgctctgc
     6481 ctcatagaac tgcctgcgtg agattctcgc atgccagaga tcctattttt ggcaatcaaa
     6541 tcattccgga tactgcgatt ttaagtgttg ttccattcca tcacggtttt ggaatgttta
     6601 ctacactcgg atatttgata tgtggatttc gagtcgtctt aatgtataga tttgaagaag
     6661 agctgtttct gaggagcctt caggattaca agattcaaag tgcgctgctg gtgccaaccc
     6721 tattctcctt cttcgccaaa agcactctga ttgacaaata cgatttatct aatttacacg
     6781 aaattgcttc tggtggcgct cccctctcta aggaagtcgg ggaagcggtt gccaagaggt
     6841 tccatctgcc aggtatcagg caaggatatg ggctcactga gactacatca gctattctga
     6901 ttacacccga gggggatgat aaaccgggcg cggtcggtaa agttgttcca ttttttgaag
     6961 cgaaggttgt ggatctggat accgggaaaa cgctgggcgt taatcaaaga ggcgaactgt
     7021 gtgtgagagg tcctatgatt atgtccggtt atgtaaacaa tccggaagcg accaacgcct
     7081 tgattgacaa ggatggatgg ctacattctg gagacatagc ttactgggac gaagacgaac
     7141 acttcttcat cgttgaccgc ctgaagtctc tgattaagta caaaggctat caggtggctc
     7201 ccgctgaatt ggaatccatc ttgctccaac accccaacat cttcgacgca ggtgtcgcag
     7261 gtcttcccga cgatgacgcc ggtgaacttc ccgccgccgt tgttgttttg gagcacggaa
     7321 agacgatgac ggaaaaagag atcgtggatt acgtcgccag tcaagtaaca accgcgaaaa
     7381 agttgcgcgg aggagttgtg tttgtggacg aagtaccgaa aggtcttacc ggaaaactcg
     7441 acgcaagaaa aatcagagag atcctcataa aggccaagaa gggcggaaag atcgccgtgt
     7501 aatgagcttg gaatggatct tcgatcccga tcgttcaaac atttggcaat aaagtttctt
     7561 aagattgaat cctgttgccg gtcttgcgac gattatcata taatttctgt tgaattacgt
     7621 taagcatgta ataattaaca tgtaatgcat gacgttattt atgagatggg tttttatgat
     7681 tagagtcccg caattataca tttaatacgc gatagaaaac aaaatatagc gcgcaaacta
     7741 ggataaatta tcgcgcgcgg tgtcatctat gttactagat cgggaattgc caagctaatt
     7801 cttgaagacg aaagggcctc gtgatacgcc tatttttata ggttaatgtc atgataataa
     7861 tggtttctta gacgtcaggt ggcacttttc ggggaaatgt gcgcggaacc cctatttgtt
     7921 tatttttcta aatacattca aatatgtatc cgctcatgag acaataaccc tgataaatgc
     7981 ttcaataatg ggaccgactc gcgctgtcag gagcatgccg tgtctcaaaa tctctgatgt
     8041 tacattgcac aagataaaaa tatatcatca tgaacaataa aactgtctgc ttacataaac
     8101 agtaatacaa ggggtgtttt caaatatgta tccgctcatg agacaataac cctgataaat
     8161 gcttcaataa tattgaaaaa ggaagagtat gagtattcaa catttccgtg tcgcccttat
     8221 tccctttttt gcggcatttt gccttcctgt ttttgctcac ccagaaacgc tggtgaaagt
     8281 aaaagatgct gaagatcagt tgggtgcacg agtgggttac atcgaactgg atctcaacag
     8341 cggtaagatc cttgagagtt ttcgccccga agaacgtttt ccaatgatga gcacttttaa
     8401 agttctgcta tgtggcgcgg tattatcccg tgttgacgcc gggcaagagc aactcggtcg
     8461 ccgcatacac tattctcaga atgacttggt tgagtactca ccagtcacag aaaagcatct
     8521 tacggatggc atgacagtaa gagaattatg cagtgctgcc ataaccatga gtgataacac
     8581 tgcggccaac ttacttctga caacgatcgg aggaccgaag gagctaaccg cttttttgca
     8641 caacatgggg gatcatgtaa ctcgccttga tcgttgggaa ccggagctga atgaagccat
     8701 accaaacgac gagcgtgaca ccacgatgcc tgcagcaatg gcaacaacgt tgcgcaaact
     8761 attaactggc gaactactta ctctagcttc ccggcaacaa ttaatagact ggatggaggc
     8821 ggataaagtt gcaggaccac ttctgcgctc ggcccttccg gctggctggt ttattgctga
     8881 taaatctgga gccggtgagc gtgggtctcg cggtatcatt gcagcactgg ggccagatgg
     8941 taagccctcc cgtatcgtag ttatctacac gacggggagt caggcaacta tggatgaacg
     9001 aaatagacag atcgctgaga taggtgcctc actgattaag cattggtaat cagaattggt
     9061 taattggttg taacattatt cagattgggc cccgttccac tgagcgtcag accccgtaga
     9121 aaagatcaaa ggatcttctt gagatccttt ttttctgcgc gtaatctgct gcttgcaaac
     9181 aaaaaaacca ccgctaccag cggtggtttg tttgccggat caagagctac caactctttt
     9241 tccgaaggta actggcttca gcagagcgca gataccaaat actgttcttc tagtgtagcc
     9301 gtagttaggc caccacttca agaactctgt agcaccgcct acatacctcg ctctgctaat
     9361 cctgttacca gtggctgctg ccagtggcga taagtcgtgt cttaccgggt tggactcaag
     9421 acgatagtta ccggataagg cgcagcggtc gggctgaacg gggggttcgt gcacacagcc
     9481 cagcttggag cgaacgacct acaccgaact gagataccta cagcgtgagc tatgagaaag
     9541 cgccacgctt cccgaaggga gaaaggcgga caggtatccg gtaagcggca gggtcggaac
     9601 aggagagcgc acgagggagc ttccaggggg aaacgcctgg tatctttata gtcctgtcgg
     9661 gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct
     9721 atggaaaaac gccagcaacg cggccttttt acggttcctg gccttttgct ggccttttgc
     9781 tcacatgtt
//
