LOCUS       40924_44938        4235 bp DNA     circular SYN 18-DEC-2018
DEFINITION  BiFC expression vector pUC-VYNE, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4235)
  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 4235)
  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 4235)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 4235)
  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..4235
                     /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"
     misc_feature    3105..3188
                     /label=MCS
                     /note="MCS"
     misc_feature    3192..3221
                     /label=myc-Tag
                     /note="myc-Tag"
     CDS             3192..3221
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             3222..3740
                     /codon_start=1
                     /label=VN173
                     /note="N-terminal fragment of mVenus for use in bimolecular
                     fluorescence complementation (BiFC) (Kodama and Hu, 2010)"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KLICTTGKLPVPWPTLVTTLGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYITADKQKNGIK
                     ANFKIRHNIE"
     terminator      3760..4012
                     /label=NOS terminator
                     /note="nopaline synthase terminator and poly(A) signal"
     primer_bind     complement(4021..4037)
                     /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 gttaaccggg
     3121 ctcaggcctg gcgcgccact agtggatcca tcgatagtac tgtcgacctc gagggtaccg
     3181 ctcccgggat ggagcaaaag ttgatttctg aggaggatct tatggtgagc aagggcgagg
     3241 agctgttcac cggggtggtg cccatcctgg tcgagctgga cggcgacgta aacggccaca
     3301 agttcagcgt gtccggcgag ggcgagggcg atgccaccta cggcaagctg accctgaagc
     3361 tgatctgcac caccggcaag ctgcccgtgc cctggcccac cctcgtgacc accctgggct
     3421 acggcctgca gtgcttcgcc cgctaccccg accacatgaa gcagcacgac ttcttcaagt
     3481 ccgccatgcc cgaaggctac gtccaggagc gcaccatctt cttcaaggac gacggcaact
     3541 acaagacccg cgccgaggtg aagttcgagg gcgacaccct ggtgaaccgc atcgagctga
     3601 agggcatcga cttcaaggag gacggcaaca tcctggggca caagctggag tacaactaca
     3661 acagccacaa cgtctatatc accgccgaca agcagaagaa cggcatcaag gccaacttca
     3721 agatccgcca caacatcgag taggagctcg aatttccccg atcgttcaaa catttggcaa
     3781 taaagtttct taagattgaa tcctgttgcc ggtcttgcga tgattatcat ataatttctg
     3841 ttgaattacg ttaagcatgt aataattaac atgtaatgca tgacgttatt tatgagatgg
     3901 gtttttatga ttagagtccc gcaattatac atttaatacg cgatagaaaa caaaatatag
     3961 cgcgcaaact aggataaatt atcgcgcgcg gtgtcatcta tgttactaga tcgggaattc
     4021 actggccgtc gttttacaac gtcgtgactg ggaaaaccct ggcgttaccc aacttaatcg
     4081 ccttgcagca catccccctt tcgccagctg gcgtaatagc gaagaggccc gcaccgatcg
     4141 cccttcccaa cagttgcgca gcctgaatgg cgaatggcgc ctgatgcggt attttctcct
     4201 tacgcatctg tgcggtattt cacaccgcat atggt
//
