LOCUS       62056_18715        2578 bp DNA     circular SYN 06-JUN-2019
DEFINITION  Cloning vector pR6K-KCA, complete sequence.
ACCESSION   MK787297
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 2578)
  AUTHORS   Moore CL, Papa LJ III, Shoulders MD.
  TITLE     A Processive Protein Chimera Introduces Mutations across Defined DNA
            Regions In Vivo
  JOURNAL   J. Am. Chem. Soc. 140 (37), 11560-11564 (2018)
  PUBMED    29991261
REFERENCE   2  (bases 1 to 2578)
  AUTHORS   Moore CL, Papa LJ III, Shoulders MD.
  TITLE     Direct Submission
  JOURNAL   Submitted (11-APR-2019) Chemistry, Massachusetts Institute of 
            Technology, 77 Massachusetts Avenue, 16-573A, Cambridge, MA 02139, 
            USA
REFERENCE   3  (bases 1 to 2578)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "J. Am. 
            Chem. Soc."; date: "2018"; volume: "140"; issue: "37"; pages: 
            "11560-11564"
COMMENT     SGRef: number: 2; type: "Journal Article"; journalName: "Submitted 
            (11-APR-2019) Chemistry, Massachusetts Institute of Technology, 77 
            Massachusetts Avenue, 16-573A, Cambridge, MA 02139, USA"
FEATURES             Location/Qualifiers
     source          1..2578
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             23..325
                     /label=ccdB
                     /note="CcdB, a bacterial toxin that poisons DNA gyrase"
     promoter        374..404
                     /label=lac UV5 promoter
                     /note="E. coli lac promoter with an 'up' mutation"
     regulatory      440..451
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
                     /regulatory_class="ribosome_binding_site"
     CDS             458..1120
                     /label=amilCP
                     /note="amilCP is a fluorescent protein published in 2008, 
                     derived from Acropora millepora. It is reported to be a 
                     somewhat slowly-maturing tetramer."
     rep_origin      1180..1568
                     /label=R6K gamma ori
                     /note="gamma replication origin from E. coli plasmid R6K;
                     requires the R6K initiator protein pi for replication"
     promoter        1652..1699
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter"
     CDS             1718..2518
                     /label=NeoR/KanR
                     /note="aminoglycoside phosphotransferase from Tn5"
ORIGIN
        1 cacataacca ggaggtcaga ttatgcagtt taaggtttac acctataaaa gagagagccg
       61 ttatcgtctg tttgtggatg tacagagtga tattattgac acgcccgggc gacggatggt
      121 gatccccctg gccagtgcac gtctgctgtc agataaagtc tcccgtgaac tttacccggt
      181 ggtgcatatc ggggatgaaa gctggcgcat gatgaccacc gatatggcca gtgtgccggt
      241 ctccgttatc ggggaagaag tggctgatct cagccaccgc gaaaatgaca tcaaaaacgc
      301 cattaacctg atgttctggg gaatataacc cagaagctta gcaaaagcta aaaccaggag
      361 ctatttaggt acctttacac tttatgcttc cggctcgtat aatgtctttt tgtacctata
      421 atgtgtggat ctagtaatta aagaggagaa atactagatg agtgtgatcg ctaaacaaat
      481 gacctacaag gtttatatgt caggcacggt caatggacac tactttgagg tcgaaggcga
      541 tggaaaaggt aagccctacg agggggagca gacggtaaag ctcactgtca ccaagggcgg
      601 acctctgcca tttgcttggg atattttatc accacagtgt cagtacggaa gcataccatt
      661 caccaagtac cctgaagaca tccctgacta tgtaaagcag tcattcccgg agggctatac
      721 atgggagagg atcatgaact ttgaagatgg tgcagtgtgt actgtcagca atgattccag
      781 catccaaggc aactgtttca tctaccatgt caagttctct ggtttgaact ttcctcccaa
      841 tggacctgtc atgcagaaga agacacaggg ctgggaaccc aacactgagc gtctctttgc
      901 acgagatgga atgctgctag gaaacaactt tatggctctg aagttagaag gaggcggtca
      961 ctatttgtgt gaatttaaaa ctacttacaa ggcaaagaag cctgtgaaga tgccagggta
     1021 tcactatgtt gaccgcaaac tggatgtaac caatcacaac aaggattaca cttcggttga
     1081 gcagtgtgaa atttccattg cacgcaaacc tgtggtcgcc taaaccggtt ggttcgcccg
     1141 cctaatgagc gggctttttt ttgaattctt ttttaattcg atctgaagat cagcagttca
     1201 acctgttgat agtacgtact aagctctcat gtttcacgta ctaagctctc atgtttaacg
     1261 tactaagctc tcatgtttaa cgaactaaac cctcatggct aacgtactaa gctctcatgg
     1321 ctaacgtact aagctctcat gtttcacgta ctaagctctc atgtttgaac aataaaatta
     1381 atataaatca gcaacttaaa tagcctctaa ggttttaagt tttataagaa aaaaaagaat
     1441 atataaggct tttaaagctt ttaaggttta acggttgtgg acaacaagcc agggatgtaa
     1501 cgcactgaga agcccttaga gcctctcaaa gcaattttga gtgacacagg aacacttaac
     1561 ggctgacatg ggatccccct catcagtgcc aacatagtaa gccagtatac actccgctag
     1621 cgcggccgcc tcgagtttcg acctgcagcc tgttgacaat taatcatcgg catagtatat
     1681 cggcatagta taatacgaca aggtgaggaa ctaaaccatg ggatcggcca ttgaacaaga
     1741 tggattgcac gcaggttctc cggccgcttg ggtggagagg ctattcggct atgactgggc
     1801 acaacagaca atcggctgct ctgatgccgc cgtgttccgg ctgtcagcgc aggggcgccc
     1861 ggttcttttt gtcaagaccg acctgtccgg tgccctgaat gaactgcagg acgaggcagc
     1921 gcggctatcg tggctggcca cgacgggcgt tccttgcgca gctgtgctcg acgttgtcac
     1981 tgaagcggga agggactggc tgctattggg cgaagtgccg gggcaggatc tcctgtcatc
     2041 tcaccttgct cctgccgaga aagtatccat catggctgat gcaatgcggc ggctgcatac
     2101 gcttgatccg gctacctgcc cattcgacca ccaagcgaaa catcgcatcg agcgagcacg
     2161 tactcggatg gaagccggtc ttgtcgatca ggatgatctg gacgaagagc atcaggggct
     2221 cgcgccagcc gaactgttcg ccaggctcaa ggcgcgcatg cccgacggcg atgatctcgt
     2281 cgtgacccat ggcgatgcct gcttgccgaa tatcatggtg gaaaatggcc gcttttctgg
     2341 attcatcgac tgtggccggc tgggtgtggc ggaccgctat caggacatag cgttggctac
     2401 ccgtgatatt gctgaagagc ttggcggcga atgggctgac cgcttcctcg tgctttacgg
     2461 tatcgccgct cccgattcgc agcgcatcgc cttctatcgc cttcttgacg agttcttctg
     2521 aggggatcaa ttctctagag ctcgctgatc agcctcgact gtaccgttag ccctccca
//
