LOCUS       62056_11890        5799 bp DNA     circular SYN 09-AUG-2020
DEFINITION  Cloning vector pgRNA-ccdB-1, complete sequence.
ACCESSION   MN934323
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5799)
  AUTHORS   Zhang K, Zhang Z, Kang J, Chen J, Liu J, Gao N, Fan L, Zheng P, Wang
            Y, Sun J.
  TITLE     CRISPR/Cas13d-Mediated Microbial RNA Knockdown
  JOURNAL   Front Bioeng Biotechnol 8, 856 (2020)
REFERENCE   2  (bases 1 to 5799)
  AUTHORS   Wang Y.
  TITLE     Direct Submission
  JOURNAL   Submitted (13-JAN-2020) Key Laboratory of Systems Microbial 
            Biotechnology, Tianjin Institute of Industrial Biotechnology, 
            Chinese Academy of Sciences, 32 West 7th Avenue, Tianjin Airport 
            Economic Area, Tianjin 300308, China
REFERENCE   3  (bases 1 to 5799)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Front 
            Bioeng Biotechnol 8, 856 (2020)"
COMMENT     SGRef: number: 2; type: "Journal Article"; journalName: "Submitted 
            (13-JAN-2020) Key Laboratory of Systems Microbial Biotechnology, 
            Tianjin Institute of Industrial Biotechnology, Chinese Academy of 
            Sciences, 32 West 7th Avenue, Tianjin Airport Economic Area, Tianjin
            300308, China"
COMMENT     ##Assembly-Data-START##
            Sequencing Technology :: Sanger dideoxy sequencing 
            ##Assembly-Data-END##
FEATURES             Location/Qualifiers
     source          1..5799
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     regulatory      1..53
                     /regulatory_class="promoter"
     misc_RNA        54..89
                     /label=RfxCas13d DR36
                     /note="36-base direct repeat for an RfxCas13d pre-gRNA 
                     (Konermann et al., 2018)"
     CDS             438..740
                     /label=ccdB
                     /note="CcdB, a bacterial toxin that poisons DNA gyrase"
     terminator      795..881
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB
                     gene"
     terminator      973..1000
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB
                     gene"
     rep_origin      2097..3560
     CDS             4084..4875
                     /label=NeoR/KanR
                     /note="aminoglycoside phosphotransferase"
     rep_origin      4978..5566
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
ORIGIN
        1 ttttctccac ataagctggc aatgttgcga cgcaacaggt acagtgtaat tcacaagtaa
       61 acccctacca actggtcggg gtttgaaact gagaccacgc gtggatccgg cttactaaaa
      121 gccagataac agtatgcgta tttgcgcgct gatttttgcg gtataagaat atatactgat
      181 atgtataccc gaagtatgtc aaaaagaggt atgctatgaa gcagcgtatt acagtgacag
      241 ttgacagcga cagctatcag ttgctcaagg catatatgat gtcaatatct ccggtctggt
      301 aagcacaacc atgcagaatg aagcccgtcg tctgcgtgcc gaacgctgga aagcggaaaa
      361 tcaggaaggg atggctgagg tcgcccggtt tattgaaatg aacggctctt ttgctgacga
      421 gaacaggggc tggtgaaatg cagtttaagg tttacaccta taaaagagag agccgttatc
      481 gtctgtttgt ggatgtacag agtgatatta ttgacacgcc cgggcgacgg atggtgatcc
      541 ccctggccag tgcacgtctg ctgtcagata aagtctcccg tgaactttac ccggtggtgc
      601 atatcgggga tgaaagctgg cgcatgatga ccaccgatat ggccagtgtg ccggtatccg
      661 ttatcgggga agaagtggct gatctcagcc accgcgaaaa tgacatcaaa aacgccatta
      721 acctgatgtt ctggggaata taaggtctca caagtaaacc cctaccaact ggtcggggtt
      781 tgaaacccag gcatcaaata aaacgaaagg ctcagtcgaa agactgggcc tttcgtttta
      841 tctgttgttt gtcggtgaac gctctcctga gtaggacaaa tccgccggga gcggatttga
      901 acgttgcgaa gcaacggccc ggagggtggc gggcaggacg cccgccataa actgccaggc
      961 atcaaattaa gcagaaggcc atcctgacgg atggcctttt tgcgtttcta caaactcttt
     1021 tgtttatttt tctaaataca ttcaaatatg tatccgctca tgaattaatt ccgctagatg
     1081 acgtgcggct tcgacctcct gggcgtggcg cttgttggcg cgctcgcggc tggctgcggc
     1141 acgacacgcg tctgagcagt attttgcgcg ccgtcctcgt gggtcaggcc ggggtgggat
     1201 caggccaccg cagtaggcgc agctgatgcg atcctccact actgcgcgtc ctcctggcgc
     1261 tgccgagcac gcagctcgtc ggccagctct tcaaggtcgg ccacaagcgt ttctaggtcg
     1321 ctcgcggcac ttgcccagtc gcgtgatgct ggcgcgtctg tcgtatcgag ggcgcggaaa
     1381 aatccgatca ccgtttttaa atcgacggcg gcatcgagtg cgtcggactc cagcgcgaca
     1441 tcggagagat ccaccgctga tgcttcaggc cagttttggt acttcgtcgt gaaggtcatg
     1501 acaccattat aacgaacgtt cgttaaaaat tctagcccca attctgataa tttcttccgg
     1561 cactcctgcg aaaacctgcg agacttcttg cccagaaaaa acgccaagcg cagcggttac
     1621 cgcacttttt ttccaggtga tttcaccctg accagcgaag cggcacttta gtgcatgagg
     1681 tgtgcccctg gtttcccctc tttggagggt tcaacccaaa aaagcacaca agcaaaaatg
     1741 aaaatcatca tgagcaagtt ggtgcgaagc agcaacgcgc tagctccaaa aaggtctgca
     1801 ggatctcgag gagatttttg agggggaggg agtcgaggaa gagccagagc agaaggcggg
     1861 gaaccgttct ctgccgacag cgtgagcccc ccttaaaaat caggccgggg aggaaccggg
     1921 gagggatcag agctaggagc gagacaccct aaaggggggg aaccgttttc tgctgacggt
     1981 gtttcgttta ttagttttca gcccgtggat agcggagggt gagggcaagt gagagccaga
     2041 gcaaggacgg gacccctaaa ggggggaacc gttttctgct gacggtgttt cgtttattag
     2101 ttttcagccc gtggacggcc gcgtttagct tccattccaa gtgcctttct gacttgttgg
     2161 atgcgccttt cactgacacc tagttcgcct gcaagctcac gagtcgaggg atcagcaacc
     2221 gattgagaac gggcatccag gatcgcagtt ttgacgcgaa gttcgagcaa ctcgcctgtc
     2281 atttctcggc gtttgtttgc ttccgctaat cgctgtcgcg tctcctgcgc atacttactt
     2341 tctgggtcag cccatctgcg tgcattcgat gtagctgcgc cccgtcgccc catcgtcgct
     2401 agagctttcc gccctcggct gctctgcgtt tccacccgac gagcagggac gactggctgg
     2461 cctttagcca cgtagccgcg cacacgacgc gccatcgtca ggcgatcacg catggcggga
     2521 agatccggct cccggccgtc tgcaccgacc gcctgggcaa cgttgtacgc cacttcatac
     2581 gcgtcgatga tcttggcatc ttttaggcgc tcaccagcag ctttgagctg gtatcccacg
     2641 gtcaacgcgt ggcgaaacgc ggtttcgtcg cgcgctcgct ctggatttgt ccagagcact
     2701 cgcacgccgt cgatcaggtc gccggacgcg tccagggcgc tcggcaggct cgcgtccaaa
     2761 atcgctagcg ccttggcttc tgcggtggcg cgttgtgccg cttcaatgcg ggcgcgtccg
     2821 ctggaaaagt cctgctcaat gtactttttc ggcttctgtg atccggtcat cgttcgagca
     2881 atctccatta ggtcggccag ccgatccaca cgatcatgct ggcagtgcca tttataggct
     2941 gtcggatcgt ctgagacgtg cagcggccac cggctcagcc tatgcgaaaa agcctggtca
     3001 gcgccgaaaa cacgagtcat ttcttccgtc gttgcagcca gcaggcgcat atttgggctg
     3061 gttttacctg ctgcggcata caccgggtca atgagccaga tgagctggca tttcccgctc
     3121 agcggattca cgccgatcca agccggcgct ttttctaggc gtgcccattt ctctaaaatc
     3181 gcgtagacct gcgggtttac gtgctcaatc ttcccgccgg cctggtggct gggcacatcg
     3241 atgtcaagca cgatcaccgc ggcatgttgc gcgtgcgtca gcgcaacgta ctggcaccgc
     3301 gtcagcgctt ttgagccagc ccggtagagc tttggttggg tttcgccggt atccgggttt
     3361 ttaatccagg cgctcgcgaa atctcttgtc ttgctgccct ggaagctttc gcgtcccagg
     3421 tgagcgagca gttcgcggcg atcttctgcc gtccagccgc gtgagccgca gcgcatagct
     3481 tcggggtggg tgtcgaacag atcggcggac aatttccacg cgctagctgt gactgtgtcc
     3541 tgcggatcgg ctagagtcat gtcttgagtg ctttctccca gctgatgact gggggttagc
     3601 cgacgccctg tgagttcccg ctcacggggc gttcaacttt ttcaggtatt tgtgcagctt
     3661 atcgtgtttt cttcgtaaat gaacgcttaa ctaccttgtt aaacgtggca aataggcagg
     3721 attgatgggg atctagcttc acgctgccgc aagcactcag ggcgcaaggg ctgctaaagg
     3781 aagcggtaga aagccagtcc gcagaaacgg tgctgacccc ggatgaatgt cagctactgg
     3841 gctatctgga caagggaaaa cgcaagcgca aagagaaagc aggtagcttg cagtgggctt
     3901 acatggcgat agctagactg ggcggtttta tggacagcaa gcgaaccgga attgccagct
     3961 ggggcgccct ctggtaaggt tgggaagccc tgcaaagtaa actggatggc tttcttgccg
     4021 ccaaggatct gatggcgcag gggatcaaga tctgatcaag agacaggatg aggatcgttt
     4081 cgcatgattg aacaagatgg attgcacgca ggttctccgg ccgcttgggt ggagaggcta
     4141 ttcggctatg actgggcaca acagacaatc ggctgctctg atgccgccgt gttccggctg
     4201 tcagcgcagg ggcgcccggt tctttttgtc aagaccgacc tgtccggtgc cctgaatgaa
     4261 ctccaagacg aggcagcgcg gctatcgtgg ctggccacga cgggcgttcc ttgcgcagct
     4321 gtgctcgacg ttgtcactga agcgggaagg gactggctgc tattgggcga agtgccgggg
     4381 caggatctcc tgtcatctca ccttgctcct gccgagaaag tatccatcat ggctgatgca
     4441 atgcggcggc tgcatacgct tgatccggct acctgcccat tcgaccacca agcgaaacat
     4501 cgcatcgagc gagcacgtac tcggatggaa gccggtcttg tcgatcagga tgatctggac
     4561 gaagagcatc aggggctcgc gccagccgaa ctgttcgcca ggctcaaggc gcggatgccc
     4621 gacggcgagg atctcgtcgt gacccatggc gatgcctgct tgccgaatat catggtggaa
     4681 aatggccgct tttctggatt catcgactgt ggccggctgg gtgtggcgga ccgctatcag
     4741 gacatagcgt tggctacccg tgatattgct gaagagcttg gcggcgaatg ggctgaccgc
     4801 ttcctcgtgc tttacggtat cgccgctccc gattcgcagc gcatcgcctt ctatcgcctt
     4861 cttgacgagt tcttctgagc gggactctgg ggttcgcgga atcatgacca aaatccctta
     4921 acgtgagttt tcgttccact gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg
     4981 agatcctttt tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac cgctaccagc
     5041 ggtggtttgt ttgccggatc aagagctacc aactcttttt ccgaaggtaa ctggcttcag
     5101 cagagcgcag ataccaaata ctgtccttct agtgtagccg tagttaggcc accacttcaa
     5161 gaactctgta gcaccgccta catacctcgc tctgctaatc ctgttaccag tggctgctgc
     5221 cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac cggataaggc
     5281 gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc agcttggagc gaacgaccta
     5341 caccgaactg agatacctac agcgtgagct atgagaaagc gccacgcttc ccgaagggag
     5401 aaaggcggac aggtatccgg taagcggcag ggtcggaaca ggagagcgca cgagggagct
     5461 tccaggggga aacgcctggt atctttatag tcctgtcggg tttcgccacc tctgacttga
     5521 gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc
     5581 ggccttttta cggttcctgg ccttttgctg gccttttgct cacatgttct ttcctgcgtt
     5641 atcccctgat tctgtggata accgtattac cgcctttgag tgagctgata ccgctcgccg
     5701 cagccgaacg accgagcgca gcgagtcagt gagcgaggaa gcggaagagc gcctgatgcg
     5761 gtattttctc cttacgcatc tgtgcggtat ttcacaccg
//
