LOCUS       Exported                4813 bp ds-DNA     circular SYN 18-DEC-2018
DEFINITION  Cloning vector pNB1, complete sequence.
ACCESSION   KU705395
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4813)
  AUTHORS   Budhagatapalli N, Schedel S, Gurushidze M, Pencs S, Hiekel S, Rutten
            T, Kusch S, Morbitzer R, Lahaye T, Panstruga R, Kumlehn J, Hensel G.
  TITLE     A simple test for the cleavage activity of customized endonucleases 
            in plants
  JOURNAL   Plant Methods 12, 18 (2016)
  PUBMED    26962325
REFERENCE   2  (bases 1 to 4813)
  AUTHORS   Budhagatapalli N, Kumlehn J, Hensel G.
  TITLE     Direct Submission
  JOURNAL   Submitted (10-FEB-2016) Physiology and Cell Biology, Leibniz 
            Institute of Plant Genetics and Crop Plant Research (IPK), 
            Corrensstr. 3, Stadt Seeland/OT Gatersleben 06466, Germany
REFERENCE   3  (bases 1 to 4813)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     ##Assembly-Data-START##
            Sequencing Technology :: Sanger dideoxy sequencing 
            ##Assembly-Data-END##
FEATURES             Location/Qualifiers
     source          1..4813
                     /organism="Cloning vector pNB1"
                     /mol_type="other DNA"
                     /db_xref="taxon:1814154"
     regulatory      24..813
                     /regulatory_class="promoter"
                     /note="CaMV 35S"
     promoter        99..776
                     /label=CaMV 35S promoter (enhanced)
                     /note="cauliflower mosaic virus 35S promoter with a 
                     duplicated enhancer region"
     misc_feature    828..901
                     /note="sig peptide; derived from Vicia faba legumin B gene 
                     LeB4"
     misc_feature    903..926
                     /label=multiple cloning site
                     /note="multiple cloning site"
     misc_feature    926..1661
                     /label=synthetic yfp
                     /note="synthetic yfp"
     CDS             1389..1640
                     /codon_start=1
                     /product="C-terminal fragment of mVenus for use in 
                     bimolecular fluorescence complementation (BiFC) (Kodama and
                     Hu, 2010)"
                     /label=VC155
                     /translation="DKQKNGIKVNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNH
                     YLSYQSALSKDPNEKRDHMVLLEFVTAAGITLGMDELYK"
     regulatory      1672..1925
                     /regulatory_class="terminator"
                     /note="nos"
     terminator      1672..1924
                     /label=NOS terminator
                     /note="nopaline synthase terminator and poly(A) signal"
     promoter        complement(1972..1990)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1997..2013)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     rep_origin      complement(2154..2609)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     promoter        2636..2740
                     /gene="bla"
                     /label=AmpR promoter
     CDS             2741..3601
                     /codon_start=1
                     /gene="amp"
                     /product="beta-lactamase"
                     /label=amp
                     /note="derived from Escherichia coli"
                     /protein_id="AMQ36131.1"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     gene            2741..3601
                     /gene="amp"
                     /label=amp
     rep_origin      complement(3644..4360)
                     /direction=LEFT
                     /label=ColE1
                     /note="ColE1"
     rep_origin      3772..4360
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     protein_bind    4648..4669
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        4684..4714
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4722..4738
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /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     4746..4762
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        4783..4801
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"