LOCUS       40924_44923        4199 bp DNA     circular SYN 18-DEC-2018
DEFINITION  BiFC expression vector pUC-triVR, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4199)
  AUTHORS   Offenborn JN, Waadt R, Kudla J.
  TITLE     Visualization and translocation of ternary 
            Calcineurin-A/Calcineurin-B/Calmodulin-2 protein complexes by 
            dual-color trimolecular fluorescence complementation
  JOURNAL   New Phytol. (2015) In press
  PUBMED    25919910
REFERENCE   2  (bases 1 to 4199)
  AUTHORS   Offenborn JN, Waadt R, Kudla J.
  TITLE     Direct Submission
  JOURNAL   Submitted (22-MAY-2015) University of Muenster, Institute for 
            Biology and Plant Biotechnology, Schlossplatz 7, Muenster 48149, 
            Germany
REFERENCE   3  (bases 1 to 4199)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 4199)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "New Phytol.
            (2015) In press"
COMMENT     SGRef: number: 2; type: "Journal Article"; journalName: "Submitted 
            (22-MAY-2015) University of Muenster, Institute for Biology and 
            Plant Biotechnology, Schlossplatz 7, Muenster 48149, Germany"
COMMENT     SGRef: number: 3; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..4199
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        96..200
                     /label=AmpR promoter
     CDS             201..1058
                     /codon_start=1
                     /label=AmpR
                     /note="beta-lactamase"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     rep_origin      1232..1820
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     protein_bind    2108..2129
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     promoter        2144..2174
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2182..2198
                     /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)."
     primer_bind     2206..2222
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        2751..3096
                     /label=CaMV 35S promoter
                     /note="strong constitutive promoter from cauliflower mosaic
                     virus"
     CDS             3114..3365
                     /codon_start=1
                     /label=VC155
                     /note="C-terminal fragment of mVenus for use in bimolecular
                     fluorescence complementation (BiFC) (Kodama and Hu, 2010)"
                     /translation="DKQKNGIKANFKIRHNIEDGGVQLADHYQQNTPIGDGPVLLPDNH
                     YLSYQSKLSKDPNEKRDHMVLLEFVTAAGITLGMDELYK"
     CDS             3366..3392
                     /codon_start=1
                     /label=HA
                     /note="HA (human influenza hemagglutinin) epitope tag"
                     /translation="YPYDVPDYA"
     misc_feature    3393..3443
                     /label=MCS2
                     /note="MCS2"
     CDS             3447..3473
                     /codon_start=1
                     /label=HA
                     /note="HA (human influenza hemagglutinin) epitope tag"
                     /translation="YPYDVPDYA"
     misc_feature    3474..3704
                     /label=mCherry 160-236
                     /note="mCherry 160-236"
     terminator      3724..3976
                     /label=NOS terminator
                     /note="nopaline synthase terminator and poly(A) signal"
     primer_bind     complement(3985..4001)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
ORIGIN
        1 gacgaaaggg cctcgtgata cgcctatttt tataggttaa tgtcatgata ataatggttt
       61 cttagacgtc aggtggcact tttcggggaa atgtgcgcgg aacccctatt tgtttatttt
      121 tctaaataca ttcaaatatg tatccgctca tgagacaata accctgataa atgcttcaat
      181 aatattgaaa aaggaagagt atgagtattc aacatttccg tgtcgccctt attccctttt
      241 ttgcggcatt ttgccttcct gtttttgctc acccagaaac gctggtgaaa gtaaaagatg
      301 ctgaagatca gttgggtgca cgagtgggtt acatcgaact ggatctcaac agcggtaaga
      361 tccttgagag ttttcgcccc gaagaacgtt ttccaatgat gagcactttt aaagttctgc
      421 tatgtggcgc ggtattatcc cgtattgacg ccgggcaaga gcaactcggt cgccgcatac
      481 actattctca gaatgacttg gttgagtact caccagtcac agaaaagcat cttacggatg
      541 gcatgacagt aagagaatta tgcagtgctg ccataaccat gagtgataac actgcggcca
      601 acttacttct gacaacgatc ggaggaccga aggagctaac cgcttttttg cacaacatgg
      661 gggatcatgt aactcgcctt gatcgttggg aaccggagct gaatgaagcc ataccaaacg
      721 acgagcgtga caccacgatg cctgtagcaa tggcaacaac gttgcgcaaa ctattaactg
      781 gcgaactact tactctagct tcccggcaac aattaataga ctggatggag gcggataaag
      841 ttgcaggacc acttctgcgc tcggcccttc cggctggctg gtttattgct gataaatctg
      901 gagccggtga gcgtgggtct cgcggtatca ttgcagcact ggggccagat ggtaagccct
      961 cccgtatcgt agttatctac acgacgggga gtcaggcaac tatggatgaa cgaaatagac
     1021 agatcgctga gataggtgcc tcactgatta agcattggta actgtcagac caagtttact
     1081 catatatact ttagattgat ttaaaacttc atttttaatt taaaaggatc taggtgaaga
     1141 tcctttttga taatctcatg accaaaatcc cttaacgtga gttttcgttc cactgagcgt
     1201 cagaccccgt agaaaagatc aaaggatctt cttgagatcc tttttttctg cgcgtaatct
     1261 gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg gatcaagagc
     1321 taccaactct ttttccgaag gtaactggct tcagcagagc gcagatacca aatactgtcc
     1381 ttctagtgta gccgtagtta ggccaccact tcaagaactc tgtagcaccg cctacatacc
     1441 tcgctctgct aatcctgtta ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg
     1501 ggttggactc aagacgatag ttaccggata aggcgcagcg gtcgggctga acggggggtt
     1561 cgtgcacaca gcccagcttg gagcgaacga cctacaccga actgagatac ctacagcgtg
     1621 agcattgaga aagcgccacg cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg
     1681 gcagggtcgg aacaggagag cgcacgaggg agcttccagg gggaaacgcc tggtatcttt
     1741 atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg atttttgtga tgctcgtcag
     1801 gggggcggag cctatggaaa aacgccagca acgcggcctt tttacggttc ctggcctttt
     1861 gctggccttt tgctcacatg ttctttcctg cgttatcccc tgattctgtg gataaccgta
     1921 ttaccgcctt tgagtgagct gataccgctc gccgcagccg aacgaccgag cgcagcgagt
     1981 cagtgagcga ggaagcggaa gagcgcccaa tacgcaaacc gcctctcccc gcgcgttggc
     2041 cgattcatta atgcagctgg cacgacaggt ttcccgactg gaaagcgggc agtgagcgca
     2101 acgcaattaa tgtgagttag ctcactcatt aggcacccca ggctttacac tttatgcttc
     2161 cggctcgtat gttgtgtgga attgtgagcg gataacaatt tcacacagga aacagctatg
     2221 accatgatta cgccaagctt agcttgcatg cctgcaggtc cccagattag ccttttcaat
     2281 ttcagaaaga atgctaaccc acagatggtt agagaggctt acgcagcagg tctcatcaag
     2341 acgatctacc cgagcaataa tctccaggaa atcaaatacc ttcccaagaa ggttaaagat
     2401 gcagtcaaaa gattcaggac taactgcatc aagaacacag agaaagatat atttctcaag
     2461 atcagaagta ctattccagt atggacgatt caaggcttgc ttcacaaacc aaggcaagta
     2521 atagagattg gagtctctaa aaaggtagtt cccactgaat caaaggccat ggagtcaaag
     2581 attcaaatag aggacctaac agaactcgcc gtaaagactg gcgaacagtt catacagagt
     2641 ctcttacgac tcaatgacaa gaagaaaatc ttcgtcaaca tggtggagca cgacacactt
     2701 gtctactcca aaaatatcaa agatacagtc tcagaagacc aaagggcaat tgagactttt
     2761 caacaaaggg taatatccgg aaacctcctc ggattccatt gcccagctat ctgtcacttt
     2821 attgtgaaga tagtggaaaa ggaaggtggc tcctacaaat gccatcattg cgataaagga
     2881 aaggccatcg ttgaagatgc ctctgccgac agtggtccca aagatggacc cccacccacg
     2941 aggagcatcg tggaaaaaga agacgttcca accacgtctt caaagcaagt ggattgatgt
     3001 gatatctcca ctgacgtaag ggatgacgca caatcccact atccttcgca agacccttcc
     3061 tctatataag gaagttcatt tcatttggag agaacacggg ggactctaga atggacaagc
     3121 agaagaacgg catcaaggcc aacttcaaga tccgccacaa catcgaggac ggcggcgtgc
     3181 agctcgccga ccactaccag cagaacaccc ccatcggcga cggccccgtg ctgctgcccg
     3241 acaaccacta cctgagctac cagtccaagc tgagcaaaga ccccaacgag aagcgcgatc
     3301 acatggtcct gctggagttc gtgaccgccg ccgggatcac tctcggcatg gacgagctgt
     3361 acaagtaccc atacgatgtt ccagattacg ctgggcccag gcctactagt ggatccgtcg
     3421 acctcgaggg taccgctccc gggatgtacc catacgatgt tccagattac gctggcgccc
     3481 tgaagggcga gatcaagcag aggctgaagc tgaaggacgg cggccactac gacgctgagg
     3541 tcaagaccac ctacaaggcc aagaagcccg tgcagctgcc cggcgcctac aacgtcaaca
     3601 tcaagttgga catcacctcc cacaacgagg actacaccat cgtggaacag tacgaacgcg
     3661 ccgagggccg ccactccacc ggcggcatgg acgagctgta caagtaggag ctcgaatttc
     3721 cccgatcgtt caaacatttg gcaataaagt ttcttaagat tgaatcctgt tgccggtctt
     3781 gcgatgatta tcatataatt tctgttgaat tacgttaagc atgtaataat taacatgtaa
     3841 tgcatgacgt tatttatgag atgggttttt atgattagag tcccgcaatt atacatttaa
     3901 tacgcgatag aaaacaaaat atagcgcgca aactaggata aattatcgcg cgcggtgtca
     3961 tctatgttac tagatcggga attcactggc cgtcgtttta caacgtcgtg actgggaaaa
     4021 ccctggcgtt acccaactta atcgccttgc agcacatccc cctttcgcca gctggcgtaa
     4081 tagcgaagag gcccgcaccg atcgcccttc ccaacagttg cgcagcctga atggcgaatg
     4141 gcgcctgatg cggtattttc tccttacgca tctgtgcggt atttcacacc gcatatggt
//
