LOCUS       62056_13610        3246 bp DNA     circular SYN 30-AUG-2019
DEFINITION  Cloning vector pKW3_MB1Amp_TracrK_Spacer, complete sequence.
ACCESSION   MN226641
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3246)
  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 3246)
  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 3246)
  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..3246
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        104..208
                     /label=AmpR promoter
     CDS             209..1066
                     /label=AmpR
                     /note="beta-lactamase"
     rep_origin      1240..1828
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     terminator      complement(1992..2078)
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB
                     gene"
     regulatory      2234..2268
                     /label=Cas9 promoter predicted
                     /note="Cas9 promoter predicted"
                     /regulatory_class="promoter"
     regulatory      2300..2327
                     /label=putative tracrRNA promoter
                     /note="putative tracrRNA promoter"
                     /regulatory_class="promoter"
     misc_RNA        2381..2459
                     /label=tracrRNA
                     /note="trans-activating CRISPR RNA for the Streptococcus 
                     pyogenes CRISPR/Cas9 system"
     misc_feature    2571..2702
                     /label=crRNA leader
                     /note="crRNA leader sequence for the Streptococcus pyogenes
                     CRISPR/Cas system"
     repeat_region   2703..2738
                     /label=DR
                     /note="direct repeat for the Streptococcus pyogenes
                     CRISPR/Cas system"
     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
     gap             3069..3098
                     /estimated_length=30
     promoter        complement(3167..3240)
                     /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 gagtattcaa catttccgtg tcgcccttat
      241 tccctttttt gcggcatttt gccttcctgt ttttgctcac ccagaaacgc tggtgaaagt
      301 aaaagatgct gaagatcagt tgggtgcacg agtgggttac atcgaactgg atctcaacag
      361 cggtaagatc cttgagagtt ttcgccccga agaacgtttt ccaatgatga gcacttttaa
      421 agttctgcta tgtggcgcgg tattatcccg tgttgacgcc gggcaagagc aactcggtcg
      481 ccgcatacac tattctcaga atgacttggt tgagtactca ccagtcacag aaaagcatct
      541 tacggatggc atgacagtaa gagaattatg cagtgctgcc ataaccatga gtgataacac
      601 tgcggccaac ttacttctga caacgatcgg aggaccgaag gagctaaccg cttttttgca
      661 caacatgggg gatcatgtaa ctcgccttga tcgttgggaa ccggagctga atgaagccat
      721 accaaacgac gagcgtgaca ccacgatgcc tgcagcaatg gcaacaacgt tgcgcaaact
      781 attaactggc gaactactta ctctagcttc ccggcaacaa ttaatagact ggatggaggc
      841 ggataaagtt gcaggaccac ttctgcgctc ggcccttccg gctggctggt ttattgctga
      901 taaatctgga gccggtgagc gtgggtctcg cggtatcatt gcagcactgg ggccagatgg
      961 taagccctcc cgtatcgtag ttatctacac gacggggagt caggcaacta tggatgaacg
     1021 aaatagacag atcgctgaga taggtgcctc actgattaag cattggtaac tgtcagacca
     1081 agtttactca tatatacttt agattgattt aaaacttcat ttttaattta aaaggatcta
     1141 ggtgaagatc ctttttgata atctcatgac caaaatccct taacgtgagt tttcgttcca
     1201 ctgagcgtca gaccccgtag aaaagatcaa aggatcttct tgagatcctt tttttctgcg
     1261 cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca gcggtggttt gtttgccgga
     1321 tcaagagcta ccaactcttt ttccgaaggt aactggcttc agcagagcgc agataccaaa
     1381 tactgtcctt ctagtgtagc cgtagttagg ccaccacttc aagaactctg tagcaccgcc
     1441 tacatacctc gctctgctaa tcctgttacc agtggctgct gccagtggcg ataagtcgtg
     1501 tcttaccggg ttggactcaa gacgatagtt accggataag gcgcagcggt cgggctgaac
     1561 ggggggttcg tgcacacagc ccagcttgga gcgaacgacc tacaccgaac tgagatacct
     1621 acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc
     1681 ggtaagcggc agggtcggaa caggagagcg cacgagggag cttccagggg gaaacgcctg
     1741 gtatctttat agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg
     1801 ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac gcggcctttt tacggttcct
     1861 ggccttttgc tggccttttg ctcacatgtt ctttcctgcg ttatcccctg attctgtgga
     1921 taaccgtatt accgcctttg agtgagctga taccgctcgc cgcagccgaa cgcctaggtc
     1981 tagggcggcg gatttgtcct actcaggaga gcgttcaccg acaaacaaca gataaaacga
     2041 aaggcccagt ctttcgactg agcctttcgt tttatttgat gcctctagca cgcgtaccat
     2101 ggaaaaagtt taaattaaat ccataatgat ttgatgattt caataatagt tttaatgacc
     2161 tccgaaatta gtttaatatg ctttaatttt tctttttcaa aatatctctt caaaaaatat
     2221 tacccaatac ttaataataa atagattata acacaaaatt cttttaaaaa gtagtttatt
     2281 ttgttatcat tctatagtat taagtattgt tttatggctg ataaatttct ttgaatttct
     2341 ccttgattat ttgttataaa agttataaaa taatcttgtt ggaaccattc aaaacagcat
     2401 agcaagttaa aataaggcta gtccgttatc aacttgaaaa agtggcaccg agtcggtgct
     2461 ttttttgata cttctattct actctgactg caaaccaaaa aaacaagcgc tttcaaaacg
     2521 cttgttttat catttttagg gaaattaatc tcttaatcct tttaggatcc tatttcttaa
     2581 taactaaaaa tatggtataa tactcttaat aaatgcagta atacaggggc ttttcaagac
     2641 tgaagtctag ctgagacaaa tagtgcgatt acgaaatttt ttagacaaaa atagtctacg
     2701 aggttttaga 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 cccaaaacnn nnnnnnnnnn nnnnnnnnnn nnnnnnnngt tttagagcta tgctgttttg
     3121 aatggtccca aaacttcagc acactgagac ttgttgagtt gaattcggtc agtgcgtcct
     3181 gctgatgtgc tcagtatctc tatcactgat agggatgtca atctctatca ctgataggga
     3241 ctcgag
//
