LOCUS       62056_13615        3180 bp DNA     circular SYN 30-AUG-2019
DEFINITION  Cloning vector pKW5_MB1Kan_TracrK_Spacer, complete sequence.
ACCESSION   MN226642
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3180)
  AUTHORS   Wang K, de la Torre D, Robertson WE, Chin JW.
  TITLE     Programmed chromosome fission and fusion enable precise large-scale 
            genome rearrangement and assembly
  JOURNAL   Science 365 (6456), 922-926 (2019)
REFERENCE   2  (bases 1 to 3180)
  AUTHORS   Wang K, de la Torre D, Robertson WE, Chin JW.
  TITLE     Direct Submission
  JOURNAL   Submitted (24-JUL-2019) Protein and Nucleic Acid Chemistry, MRC 
            Laboratory of Molecular Biology, Francis Crick Avenue, Cambridge, 
            Cambridgeshire CB2 0QH, United Kingdom
REFERENCE   3  (bases 1 to 3180)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Science"; 
            date: "2019"; volume: "365"; issue: "6456"; pages: "922-926"
COMMENT     SGRef: number: 2; type: "Journal Article"; journalName: "Submitted 
            (24-JUL-2019) Protein and Nucleic Acid Chemistry, MRC Laboratory of 
            Molecular Biology, Francis Crick Avenue, Cambridge, Cambridgeshire 
            CB2 0QH, United Kingdom"
COMMENT     ##Assembly-Data-START##
            Sequencing Technology :: Sanger dideoxy sequencing 
            ##Assembly-Data-END##
FEATURES             Location/Qualifiers
     source          1..3180
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        104..208
                     /label=AmpR promoter
     CDS             209..1000
                     /label=NeoR/KanR
                     /note="aminoglycoside phosphotransferase"
     rep_origin      1174..1762
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     terminator      complement(1926..2012)
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB
                     gene"
     regulatory      2168..2202
                     /label=putative Cas9 promoter
                     /note="putative Cas9 promoter"
                     /regulatory_class="promoter"
     regulatory      2234..2261
                     /label=putative tracrRNA promoter
                     /note="putative tracrRNA promoter"
                     /regulatory_class="promoter"
     misc_RNA        2315..2393
                     /label=tracrRNA
                     /note="trans-activating CRISPR RNA for the Streptococcus 
                     pyogenes CRISPR/Cas9 system"
     misc_feature    2505..2636
                     /label=crRNA leader
                     /note="crRNA leader sequence for the Streptococcus pyogenes
                     CRISPR/Cas system"
     repeat_region   2637..2672
                     /label=DR
                     /note="direct repeat for the Streptococcus pyogenes
                     CRISPR/Cas system"
     gap             2673..2702
                     /estimated_length=30
     gap             2739..2768
                     /estimated_length=30
     gap             2805..2834
                     /estimated_length=30
     gap             2871..2900
                     /estimated_length=30
     gap             2937..2966
                     /estimated_length=30
     gap             3003..3032
                     /estimated_length=30
     promoter        complement(3101..3174)
                     /label=PLtetO-1 promoter
                     /note="modified phage lambda PL promoter with tet operator
                     sites (Lutz and Bujard, 1997)"
ORIGIN
        1 ttcttgaaga cgaaagggcc tcgtgatacg cctattttta taggttaatg tcatgataat
       61 aatggtttct tagacgtcag gtggcacttt tcggggaaat gtgcgcggaa cccctatttg
      121 tttatttttc taaatacatt caaatatgta tccgctcatg agacaataac cctgataaat
      181 gcttcaataa tattgaaaaa ggaagagtat gattgaacaa gatggattgc acgcaggttc
      241 tccggccgct tgggtggaga ggctattcgg ctatgactgg gcacaacaga caatcggctg
      301 ctctgatgcc gccgtgttcc ggctgtcagc gcaggggcgc ccggttcttt ttgtcaagac
      361 cgacctgtcc ggtgccctga atgaactgca ggacgaggca gcgcggctat cgtggctggc
      421 cacgacgggc gttccttgcg cagctgtgct cgacgttgtc actgaagcgg gaagggactg
      481 gctgctattg ggcgaagtgc cggggcagga tctcctgtca tctcaccttg ctcctgccga
      541 gaaagtatcc atcatggctg atgcaatgcg gcggctgcat acgcttgatc cggctacctg
      601 cccattcgac caccaagcga aacatcgcat cgagcgagca cgtactcgga tggaagccgg
      661 tcttgtcgat caggatgatc tggacgaaga gcatcagggg ctcgcgccag ccgaactgtt
      721 cgccaggctc aaggcgcgca tgcccgacgg cgaggatctc gtcgtgaccc atggcgatgc
      781 ctgcttgccg aatatcatgg tggaaaatgg ccgcttttct ggattcatcg actgtggccg
      841 gctgggtgtg gcggaccgct atcaggacat agcgttggct acccgtgata ttgctgaaga
      901 gcttggcggc gaatgggctg accgcttcct cgtgctttac ggtatcgccg ctcccgattc
      961 gcagcgcatc gccttctatc gccttcttga cgagttcttc tgactgtcag accaagttta
     1021 ctcatatata ctttagattg atttaaaact tcatttttaa tttaaaagga tctaggtgaa
     1081 gatccttttt gataatctca tgaccaaaat cccttaacgt gagttttcgt tccactgagc
     1141 gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat
     1201 ctgctgcttg caaacaaaaa aaccaccgct accagcggtg gtttgtttgc cggatcaaga
     1261 gctaccaact ctttttccga aggtaactgg cttcagcaga gcgcagatac caaatactgt
     1321 ccttctagtg tagccgtagt taggccacca cttcaagaac tctgtagcac cgcctacata
     1381 cctcgctctg ctaatcctgt taccagtggc tgctgccagt ggcgataagt cgtgtcttac
     1441 cgggttggac tcaagacgat agttaccgga taaggcgcag cggtcgggct gaacgggggg
     1501 ttcgtgcaca cagcccagct tggagcgaac gacctacacc gaactgagat acctacagcg
     1561 tgagctatga gaaagcgcca cgcttcccga agggagaaag gcggacaggt atccggtaag
     1621 cggcagggtc ggaacaggag agcgcacgag ggagcttcca gggggaaacg cctggtatct
     1681 ttatagtcct gtcgggtttc gccacctctg acttgagcgt cgatttttgt gatgctcgtc
     1741 aggggggcgg agcctatgga aaaacgccag caacgcggcc tttttacggt tcctggcctt
     1801 ttgctggcct tttgctcaca tgttctttcc tgcgttatcc cctgattctg tggataaccg
     1861 tattaccgcc tttgagtgag ctgataccgc tcgccgcagc cgaacgccta ggtctagggc
     1921 ggcggatttg tcctactcag gagagcgttc accgacaaac aacagataaa acgaaaggcc
     1981 cagtctttcg actgagcctt tcgttttatt tgatgcctct agcacgcgta ccatggaaaa
     2041 agtttaaatt aaatccataa tgatttgatg atttcaataa tagttttaat gacctccgaa
     2101 attagtttaa tatgctttaa tttttctttt tcaaaatatc tcttcaaaaa atattaccca
     2161 atacttaata ataaatagat tataacacaa aattctttta aaaagtagtt tattttgtta
     2221 tcattctata gtattaagta ttgttttatg gctgataaat ttctttgaat ttctccttga
     2281 ttatttgtta taaaagttat aaaataatct tgttggaacc attcaaaaca gcatagcaag
     2341 ttaaaataag gctagtccgt tatcaacttg aaaaagtggc accgagtcgg tgcttttttt
     2401 gatacttcta ttctactctg actgcaaacc aaaaaaacaa gcgctttcaa aacgcttgtt
     2461 ttatcatttt tagggaaatt aatctcttaa tccttttagg atcctatttc ttaataacta
     2521 aaaatatggt ataatactct taataaatgc agtaatacag gggcttttca agactgaagt
     2581 ctagctgaga caaatagtgc gattacgaaa ttttttagac aaaaatagtc tacgaggttt
     2641 tagagctatg ctgttttgaa tggtcccaaa acnnnnnnnn nnnnnnnnnn nnnnnnnnnn
     2701 nngttttaga gctatgctgt tttgaatggt cccaaaacnn nnnnnnnnnn nnnnnnnnnn
     2761 nnnnnnnngt tttagagcta tgctgttttg aatggtccca aaacnnnnnn nnnnnnnnnn
     2821 nnnnnnnnnn nnnngtttta gagctatgct gttttgaatg gtcccaaaac nnnnnnnnnn
     2881 nnnnnnnnnn nnnnnnnnnn gttttagagc tatgctgttt tgaatggtcc caaaacnnnn
     2941 nnnnnnnnnn nnnnnnnnnn nnnnnngttt tagagctatg ctgttttgaa tggtcccaaa
     3001 acnnnnnnnn nnnnnnnnnn nnnnnnnnnn nngttttaga gctatgctgt tttgaatggt
     3061 cccaaaactt cagcacactg agacttgttg agttgaattc ggtcagtgcg tcctgctgat
     3121 gtgctcagta tctctatcac tgatagggat gtcaatctct atcactgata gggactcgag
//
