LOCUS       40924_26989        4231 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Cloning vector pKS plasmid pKS Chlamy dual, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4231)
  AUTHORS   Hahn F, Mantegazza O, Greiner A, Hegemann P, Eisenhut M, Weber AP.
  TITLE     An Efficient Visual Screen for CRISPR/Cas9 Activity in Arabidopsis 
            thaliana
  JOURNAL   Front Plant Sci 8, 39 (2017)
  PUBMED    28174584
REFERENCE   2  (bases 1 to 4231)
  AUTHORS   Hahn F, Mantegazza O, Eisenhut M, Greiner A, Hegemann P, Weber APM.
  TITLE     Direct Submission
  JOURNAL   Submitted (03-NOV-2016) Plant Biochemistry, Heinrich Heine 
            University Dusseldorf, Universitatsstr. 1, Dusseldorf 40225, Germany
REFERENCE   3  (bases 1 to 4231)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 4231)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Front Plant
            Sci 8, 39 (2017)"
COMMENT     SGRef: number: 2; type: "Journal Article"; journalName: "Submitted 
            (03-NOV-2016) Plant Biochemistry, Heinrich Heine University 
            Dusseldorf, Universitatsstr. 1, Dusseldorf 40225, Germany"
COMMENT     SGRef: number: 3; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..4231
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     regulatory      1..463
                     /label=U6/C8
                     /note="U6/C8"
                     /regulatory_class="promoter"
     misc_feature    465..470
                     /label=BsmBI site
                     /note="BsmBI site"
     misc_feature    483..488
                     /label=BsmBI site
                     /note="BsmBI site"
     misc_RNA        490..565
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     primer_bind     576..592
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(626..642)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        complement(672..690)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(711..727)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    complement(735..751)
                     /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)."
     promoter        complement(759..789)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    complement(804..825)
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     rep_origin      complement(1113..1701)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(1875..2732)
                     /label=AmpR
                     /note="beta-lactamase"
     promoter        complement(2733..2837)
                     /label=AmpR promoter
     rep_origin      2864..3319
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     primer_bind     3460..3476
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        3486..3504
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      3572..4090
                     /label=U6/C15
                     /note="U6/C15"
                     /regulatory_class="promoter"
     misc_feature    4093..4098
                     /label=BbsI site
                     /note="BbsI site"
     misc_feature    4101..4106
                     /label=BbsI site
                     /note="BbsI site"
     misc_structure  4109..4188
     misc_RNA        4109..4184
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     regulatory      4195..4231
                     /label=U6/C8
                     /note="U6/C8"
                     /regulatory_class="promoter"
ORIGIN
        1 cctggcagca gggcagcgcg cggtgggtag ctgctcccct cgcgcattgc atgggtgcaa
       61 attcatgctt ttgcgcaggg agcaggatgc cctcataaat cctacgcgcg caaggttggg
      121 agtcattgcc cacccgtacc ccaacaagct atgcataccc aagctgccac accactgggc
      181 tgtcgccggc aaccaatggc gcgaaagctg gcttgctgtc acaatgggcc tttttcccag
      241 cctcgcagcg tgggcgtggc tacctagaac ccgcatgcac gctggagagc gcccaggaac
      301 atctgcgcta tggaaggggc cttggggcta ctgttccacg atgggactta ccgggtcggg
      361 atcaacgcgc tcccgcgcgg gaaaggtcgg gctggcgccg gtgtctgttt ggacgcgact
      421 tttgcgccct agtgatatgt atattacacc gacgagcact gtctgagacg taccatatga
      481 accgtctccg ttttagagct agaaatagca agttaaaata aggctagtcc gttatcaact
      541 tgaaaaagtg gcaccgagtc ggtgcaaatt ttttttctag aactagtgga tcccccgggc
      601 tgcaggaatt cgatatcaag cttatcgata ccgtcgacct cgaggggggg cccggtaccc
      661 agcttttgtt ccctttagtg agggttaatt gcgcgcttgg cgtaatcatg gtcatagctg
      721 tttcctgtgt gaaattgtta tccgctcaca attccacaca acatacgagc cggaagcata
      781 aagtgtaaag cctggggtgc ctaatgagtg agctaactca cattaattgc gttgcgctca
      841 ctgcccgctt tccagtcggg aaacctgtcg tgccagctgc attaatgaat cggccaacgc
      901 gcggggagag gcggtttgcg tattgggcgc tcttccgctt cctcgctcac tgactcgctg
      961 cgctcggtcg ttcggctgcg gcgagcggta tcagctcact caaaggcggt aatacggtta
     1021 tccacagaat caggggataa cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc
     1081 aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc ccctgacgag
     1141 catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact ataaagatac
     1201 caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct gccgcttacc
     1261 ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcatag ctcacgctgt
     1321 aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc
     1381 gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggtaaga
     1441 cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc gaggtatgta
     1501 ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag aaggacagta
     1561 tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg tagctcttga
     1621 tccggcaaac aaaccaccgc tggtagcggt ggtttttttg tttgcaagca gcagattacg
     1681 cgcagaaaaa aaggatctca agaagatcct ttgatctttt ctacggggtc tgacgctcag
     1741 tggaacgaaa actcacgtta agggattttg gtcatgagat tatcaaaaag gatcttcacc
     1801 tagatccttt taaattaaaa atgaagtttt aaatcaatct aaagtatata tgagtaaact
     1861 tggtctgaca gttaccaatg cttaatcagt gaggcaccta tctcagcgat ctgtctattt
     1921 cgttcatcca tagttgcctg actccccgtc gtgtagataa ctacgatacg ggagggctta
     1981 ccatctggcc ccagtgctgc aatgataccg cgagacccac gctcaccggc tccagattta
     2041 tcagcaataa accagccagc cggaagggcc gagcgcagaa gtggtcctgc aactttatcc
     2101 gcctccatcc agtctattaa ttgttgccgg gaagctagag taagtagttc gccagttaat
     2161 agtttgcgca acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt
     2221 atggcttcat tcagctccgg ttcccaacga tcaaggcgag ttacatgatc ccccatgttg
     2281 tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg tcagaagtaa gttggccgca
     2341 gtgttatcac tcatggttat ggcagcactg cataattctc ttactgtcat gccatccgta
     2401 agatgctttt ctgtgactgg tgagtactca accaagtcat tctgagaata gtgtatgcgg
     2461 cgaccgagtt gctcttgccc ggcgtcaata cgggataata ccgcgccaca tagcagaact
     2521 ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa aactctcaag gatcttaccg
     2581 ctgttgagat ccagttcgat gtaacccact cgtgcaccca actgatcttc agcatctttt
     2641 actttcacca gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga
     2701 ataagggcga cacggaaatg ttgaatactc atactcttcc tttttcaata ttattgaagc
     2761 atttatcagg gttattgtct catgagcgga tacatatttg aatgtattta gaaaaataaa
     2821 caaatagggg ttccgcgcac atttccccga aaagtgccac ctgacgcgcc ctgtagcggc
     2881 gcattaagcg cggcgggtgt ggtggttacg cgcagcgtga ccgctacact tgccagcgcc
     2941 ctagcgcccg ctcctttcgc tttcttccct tcctttctcg ccacgttcgc cggctttccc
     3001 cgtcaagctc taaatcgggg gctcccttta gggttccgat ttagtgcttt acggcacctc
     3061 gaccccaaaa aacttgatta gggtgatggt tcacgtagtg ggccatcgcc ctgatagacg
     3121 gtttttcgcc ctttgacgtt ggagtccacg ttctttaata gtggactctt gttccaaact
     3181 ggaacaacac tcaaccctat ctcggtctat tcttttgatt tataagggat tttgccgatt
     3241 tcggcctatt ggttaaaaaa tgagctgatt taacaaaaat ttaacgcgaa ttttaacaaa
     3301 atattaacgc ttacaatttc cattcgccat tcaggctgcg caactgttgg gaagggcgat
     3361 cggtgcgggc ctcttcgcta ttacgccagc tggcgaaagg gggatgtgct gcaaggcgat
     3421 taagttgggt aacgccaggg ttttcccagt cacgacgttg taaaacgacg gccagtgagc
     3481 gcgcgtaata cgactcacta tagggcgaat tggagctcca ccgcggtgcg gccgccagat
     3541 cttccggatg gctcgagttt ttcagcaaga tggccacccc gacaccacag ccaagcgcag
     3601 gtctgctggg ctctgcgaaa tcgaatgcaa gtttaactgg tgcgcgcgtt ccctggaccc
     3661 tgattggccc attttagaac attcatgaag ttgatatact gcagatatgc gagagataat
     3721 aaactaagca gtcggaggtc ggccatgcct cgtgacagcg gcgtcaactc cgagttcctg
     3781 cggttcctgt agcgcatgag catgggtcgc gtgtggtgat gaccaggaac ggcggcggca
     3841 ggaagcggaa gaggaggagg tcaggagcaa gagccaagca ggtaggagaa cgagggagca
     3901 gagtacttgt ggctgggagg ccacgggtgc cctagcaggg gcacgcgccg gtgacgtgag
     3961 caggaggaga aggcagtata cgcacttgac acggcttcag gaggcgcaca ggaggcggag
     4021 gccgccacga accagcagag gaagcaataa attgtcttac tatatatata tttggcctat
     4081 gagaattgac gggtcttcga gaagacctgt tttagagcta gaaatagcaa gttaaaataa
     4141 ggctagtccg ttatcaactt gaaaaagtgg caccgagtcg gtgcaaattt ttttaaatgc
     4201 ctgggtattg ggattgataa tccagttgat a
//
