LOCUS       40924_33008        4813 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Cloning vector pNB1, complete sequence.
ACCESSION   .
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)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 4813)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Plant 
            Methods 12, 18 (2016)"
COMMENT     SGRef: number: 2; type: "Journal Article"; journalName: "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"
COMMENT     SGRef: number: 3; type: "Journal Article"
COMMENT     ##Assembly-Data-START##
            Sequencing Technology :: Sanger dideoxy sequencing 
            ##Assembly-Data-END##
FEATURES             Location/Qualifiers
     source          1..4813
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     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"
     CDS             927..1640
                     /label=EGFP
                     /note="enhanced GFP"
     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
                     /label=AmpR promoter
     CDS             2741..3598
                     /label=AmpR
                     /note="beta-lactamase"
     rep_origin      3772..4360
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     protein_bind    4648..4669
                     /label=CAP binding site
                     /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
                     /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"
ORIGIN
        1 gagctcatgt tactagatcg ggccaacatg gtggagcacg acactctcgt ctactccaag
       61 aatatcaaag atacagtctc agaagaccaa agggctattg agacttttca acaaagggta
      121 atatcgggaa acctcctcgg attccattgc ccagctatct gtcacttcat caaaaggaca
      181 gtagaaaagg aaggtggcac ctacaaatgc catcattgcg ataaaggaaa ggctatcgtt
      241 caagatgcct ctgccgacag tggtcccaaa gatggacccc cacccacgag gagcatcgtg
      301 gaaaaagaag acgttccaac cacgtcttca aagcaagtgg attgatgtga taacatggtg
      361 gagcacgaca ctctcgtcta ctccaagaat atcaaagata cagtctcaga agaccaaagg
      421 gctattgaga cttttcaaca aagggtaata tcgggaaacc tcctcggatt ccattgccca
      481 gctatctgtc acttcatcaa aaggacagta gaaaaggaag gtggcaccta caaatgccat
      541 cattgcgata aaggaaaggc tatcgttcaa gatgcctctg ccgacagtgg tcccaaagat
      601 ggacccccac ccacgaggag catcgtggaa aaagaagacg ttccaaccac gtcttcaaag
      661 caagtggatt gatgtgatat ctccactgac gtaagggatg acgcacaatc ccactatcct
      721 tcgcaagacc ttcctctata taaggaagtt catttcattt ggagaggaca cgctgaaatc
      781 accagtctct ctctacaaat ctatctctct cgagcactag tggatctatg gcttccaaac
      841 cttttctatc tttgctttca ctttccttgc ttctctttac aagcacatgt ttagcaggct
      901 ctggatcccc cgggctgcag gaattcgtga gcaagggcga ggagctgttc accggggtgg
      961 tgcccatcct ggtcgagctg gacggcgacg taaacggcca caagttcagc gtgtccggcg
     1021 agggcgaggg cgatgccacc tacggcaagc tgaccctgaa gttcatctgc accaccggca
     1081 agctgcccgt gccctggccc accctcgtga ccaccctgac ctacggcgtg cagtgcttca
     1141 gccgctaccc cgaccacatg aagcagcacg acttcttcaa gtccgccatg cccgaaggct
     1201 acgtccagga gcgcaccatc ttcttcaagg acgacggcaa ctacaagacc cgcgccgagg
     1261 tgaagttcga gggcgacacc ctggttaata gaattgaatt aaagggcatc gactttaaag
     1321 aagatggtaa catcctgggg cacaagctgg agtacaacta caacagccac aacgtctata
     1381 tcatggccga caagcagaag aacggcatca aggtgaactt caagatccgc cacaacatcg
     1441 aggacggcag cgtgcagctc gccgaccact accagcagaa cacccccatc ggcgacggcc
     1501 ccgtgctgct gcccgacaac cactacctga gctaccagtc cgccctgagc aaagacccca
     1561 acgagaagcg cgatcacatg gtcctgctgg agttcgtgac cgccgccggg atcactctcg
     1621 gcatggacga gctgtacaag tctgagaaag atgagctata agcggccgcg ggatcgttca
     1681 aacatttggc aataaagttt cttaagattg aatcctgttg ccggtcttgc gatgattatc
     1741 atataatttc tgttgaatta cgttaagcat gtaataatta acatgtaatg catgacgtta
     1801 tttatgagat gggtttttat gattagagtc ccgcaattat acatttaata cgcgatagaa
     1861 aacaaaatat agcgcgcaaa ctaggataaa ttatcgcgcg cggtgtcatc tatgttacta
     1921 gatcgggcca acatggtgga tctagatgca ttcgcgaggt acccaattcg ccctatagtg
     1981 agtcgtatta caattcactg gccgtcgttt tacaacgtcg tgactgggaa aaccctggcg
     2041 ttacccaact taatcgcctt gcagcacatc cccctttcgc cagctggcgt aatagcgaag
     2101 aggcccgcac cgatcgccct tcccaacagt tgcgcagcct gaatggcgaa tggaaattgt
     2161 aagcgttaat attttgttaa aattcgcgtt aaatttttgt taaatcagct cattttttaa
     2221 ccaataggcc gaaatcggca aaatccctta taaatcaaaa gaatagaccg agatagggtt
     2281 gagtgttgtt ccagtttgga acaagagtcc actattaaag aacgtggact ccaacgtcaa
     2341 agggcgaaaa accgtctatc agggcgatgg cccactacgt gaaccatcac cctaatcaag
     2401 ttttttgggg tcgaggtgcc gtaaagcact aaatcggaac cctaaaggga gcccccgatt
     2461 tagagcttga cggggaaagc cggcgaacgt ggcgagaaag gaagggaaga aagcgaaagg
     2521 agcgggcgct agggcgctgg caagtgtagc ggtcacgctg cgcgtaacca ccacacccgc
     2581 cgcgcttaat gcgccgctac agggcgcgtc aggtggcact tttcggggaa atgtgcgcgg
     2641 aacccctatt tgtttatttt tctaaataca ttcaaatatg tatccgctca tgagacaata
     2701 accctgataa atgcttcaat aatattgaaa aaggaagagt atgagtattc aacatttccg
     2761 tgtcgccctt attccctttt ttgcggcatt ttgccttcct gtttttgctc acccagaaac
     2821 gctggtgaaa gtaaaagatg ctgaagatca gttgggtgca cgagtgggtt acatcgaact
     2881 ggatctcaac agcggtaaga tccttgagag ttttcgcccc gaagaacgtt ttccaatgat
     2941 gagcactttt aaagttctgc tatgtggcgc ggtattatcc cgtattgacg ccgggcaaga
     3001 gcaactcggt cgccgcatac actattctca gaatgacttg gttgagtact caccagtcac
     3061 agaaaagcat cttacggatg gcatgacagt aagagaatta tgcagtgctg ccataaccat
     3121 gagtgataac actgcggcca acttacttct gacaacgatc ggaggaccga aggagctaac
     3181 cgcttttttg cacaacatgg gggatcatgt aactcgcctt gatcgttggg aaccggagct
     3241 gaatgaagcc ataccaaacg acgagcgtga caccacgatg cctgtagcaa tggcaacaac
     3301 gttgcgcaaa ctattaactg gcgaactact tactctagct tcccggcaac aattaataga
     3361 ctggatggag gcggataaag ttgcaggacc acttctgcgc tcggcccttc cggctggctg
     3421 gtttattgct gataaatctg gagccggtga gcgtgggtct cgcggtatca ttgcagcact
     3481 ggggccagat ggtaagccct cccgtatcgt agttatctac acgacgggga gtcaggcaac
     3541 tatggatgaa cgaaatagac agatcgctga gataggtgcc tcactgatta agcattggta
     3601 actgtcagac caagtttact catatatact ttagattgat ttaaaacttc atttttaatt
     3661 taaaaggatc taggtgaaga tcctttttga taatctcatg accaaaatcc cttaacgtga
     3721 gttttcgttc cactgagcgt cagaccccgt agaaaagatc aaaggatctt cttgagatcc
     3781 tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt
     3841 ttgtttgccg gatcaagagc taccaactct ttttccgaag gtaactggct tcagcagagc
     3901 gcagatacca aatactgtcc ttctagtgta gccgtagtta ggccaccact tcaagaactc
     3961 tgtagcaccg cctacatacc tcgctctgct aatcctgtta ccagtggctg ctgccagtgg
     4021 cgataagtcg tgtcttaccg ggttggactc aagacgatag ttaccggata aggcgcagcg
     4081 gtcgggctga acggggggtt cgtgcacaca gcccagcttg gagcgaacga cctacaccga
     4141 actgagatac ctacagcgtg agctatgaga aagcgccacg cttcccgaag ggagaaaggc
     4201 ggacaggtat ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg agcttccagg
     4261 gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg
     4321 atttttgtga tgctcgtcag gggggcggag cctatggaaa aacgccagca acgcggcctt
     4381 tttacggttc ctggcctttt gctggccttt tgctcacatg ttctttcctg cgttatcccc
     4441 tgattctgtg gataaccgta ttaccgcctt tgagtgagct gataccgctc gccgcagccg
     4501 aacgaccgag cgcagcgagt cagtgagcga ggaagcggaa gagcgcccaa tacgcaaacc
     4561 gcctctcccc gcgcgttggc cgattcatta atgcagctgg cacgacaggt ttcccgactg
     4621 gaaagcgggc agtgagcgca acgcaattaa tgtgagttag ctcactcatt aggcacccca
     4681 ggctttacac tttatgcttc cggctcgtat gttgtgtgga attgtgagcg gataacaatt
     4741 tcacacagga aacagctatg accatgatta cgccaagctc gaaattaacc ctcactaaag
     4801 ggaacaaaag ctg
//
