LOCUS       62056_22180        4506 bp DNA     circular SYN 13-JUN-2020
DEFINITION  Expression vector pUC57-Kan-mcr-6 DNA, complete seuence.
ACCESSION   LC548750
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4506)
  AUTHORS   Suzuki M.
  TITLE     Bacterial ARG expression libraries
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 4506)
  AUTHORS   Suzuki M.
  TITLE     Direct Submission
  JOURNAL   Submitted (19-MAY-2020) Contact:Masato Suzuki National Institute of 
            Infectious Diseases, AMR Research Center; 4-2-1 Aobacho, 
            Higashimurayama, Tokyo 189-0002, Japan
REFERENCE   3  (bases 1 to 4506)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: 
            "Unpublished"
COMMENT     SGRef: number: 2; type: "Journal Article"; journalName: "Submitted 
            (19-MAY-2020) Contact:Masato Suzuki National Institute of Infectious
            Diseases, AMR Research Center; 4-2-1 Aobacho, Higashimurayama, Tokyo
            189-0002, Japan"
FEATURES             Location/Qualifiers
     source          1..4506
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     379..395
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        466..494
                     /label=Pc promoter
                     /note="class 1 integron promoter"
     gene            742..2358
                     /gene="mcr-6"
                     /label=mcr-6
     CDS             742..2358
                     /codon_start=1
                     /transl_table=11
                     /gene="mcr-6"
                     /product="lipid A phosphoethanolamine transferase MCR-6"
                     /label=mcr-6
                     /protein_id="BCG50371.1"
                     /translation="MTQHSPWYRRPVNPYLLMSVVALFLSATANLTFFDKITNTYPMAQ
                     NAGFVISTALVLFGAMLLITVLLSYRYVLKPVLILLLIMGAVTSYFTDTYGTVYDTTML
                     QNALQTDQAESKDLMNMAFFVRIIGLGVLPSILVAWVKVDYPTLGKSLIQRAMTWGVAV
                     VMALVPILAFSSHYASFFREHKPLRSYVNPVMPIYSVGKLASIEYKKATAPKDTIYHAK
                     DAVQTTTPAERKPRLVVFVVGETARADHVQFNGYSRETFPQLAKIDNLANFSQVTSCGT
                     STAYSVPCMFSYLGQDDYDVDTAKYQENVLDTLDRLGVGILWRDNNSDSKGVMDKLPAS
                     QYFDYKSATNNTICNTNPYNECRDVGMLVGLDDYVSTNQGKDMLIMLHQMGNHGPAYFK
                     RYDEQFAKYTPVCEGNELAKCEHQSLINAYDNALLATDDFIAKSIDWLKTHQANYDVAM
                     LYVSDHGESLGENGVYLHGMPNAFAPKEQRAVPAFFWSNNPSFTPTASDTVLTHDAITP
                     TLLKLFDVTADKVKDRTAFIR"
     primer_bind     complement(2416..2432)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    complement(2440..2456)
                     /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(2464..2494)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    complement(2509..2530)
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     rep_origin      complement(2818..3406)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(3587..4393)
                     /label=KanR
                     /note="aminoglycoside phosphotransferase"
ORIGIN
        1 tcgcgcgttt cggtgatgac ggtgaaaacc tctgacacat gcagctcccg gagacggtca
       61 cagcttgtct gtaagcggat gccgggagca gacaagcccg tcagggcgcg tcagcgggtg
      121 ttggcgggtg tcggggctgg cttaactatg cggcatcaga gcagattgta ctgagagtgc
      181 accatatgcg gtgtgaaata ccgcacagat gcgtaaggag aaaataccgc atcaggcgcc
      241 attcgccatt caggctgcgc aactgttggg aagggcgatc ggtgcgggcc tcttcgctat
      301 tacgccagct ggcgaaaggg ggatgtgctg caaggcgatt aagttgggta acgccagggt
      361 tttcccagtc acgacgttgt aaaacgacgg ccagtgaatt cgagctcggt acctcgcgaa
      421 tgcatctaga tcacaccgtg gaaacggatg aaggcacgaa cccagttgac ataagcctgt
      481 tcggttcgta aactgtaatg caagtagcgt atgcgctcac gcaactggtc cagaaccttg
      541 accgaacgca gcggtggtaa cggcgcagtg gcggttttca tggcttgtta tgactgtttt
      601 tttgtacagt ctatgcctcg ggcatccaag cagcaagcgc gttacgccgt gggtcgatgt
      661 ttgatgttat ggagcagcaa cgatgttacg cagcagggca gtcgccctaa aacaaagtta
      721 ggcatcacaa agtacagcat catgacacag catagtcctt ggtaccgccg tccggtcaat
      781 ccctatctgt tgatgagcgt ggtcgcttta tttttgtcag cgacagcaaa cctaactttc
      841 tttgataaaa tcaccaatac ttatccgatg gcacaaaacg caggctttgt gatctcaacg
      901 gcgcttgtgc tatttggggc gatgctattg attactgtgc tgttatcgta tcgctatgtg
      961 cttaagcctg tgttgatttt gctgcttatc atgggtgcgg tgacgagcta ttttaccgat
     1021 acttatggca ccgtttatga caccaccatg ctccaaaatg ccttgcaaac tgaccaagcc
     1081 gagtctaagg acttgatgaa tatggcgttt tttgtgcgga ttatcgggct tggcgtgttg
     1141 ccaagtatct tggtggcgtg ggtcaaggtg gattatccga cattgggtaa gagtctgatt
     1201 cagcgtgcga tgacttgggg tgtggcagtg gtgatggcac ttgtgccgat tttggcattt
     1261 agtagtcact acgccagttt ctttcgtgaa cataagccac tgcgtagcta tgtcaatccc
     1321 gtgatgccga tttattcagt aggtaagctt gccagtattg agtacaaaaa agccaccgcg
     1381 ccaaaagaca ccatctatca tgccaaagat gctgtacaga cgacgacgcc tgccgagcgt
     1441 aagccacgac tcgtggtgtt cgtcgtcggt gagacggctc gagctgacca tgtgcagttt
     1501 aatggctata gtcgtgagac ttttccgcag cttgccaaga ttgacaacct agccaatttt
     1561 agccaagtga catcgtgtgg cacatcgacg gcgtactctg tgccgtgtat gttcagttat
     1621 ctgggtcaag atgactatga tgtcgatacc gccaaatacc aagaaaacgt gctggatacg
     1681 cttgaccgac tgggtgtggg tatcctgtgg cgggataata attcagactc aaaaggcgtg
     1741 atggataaac tgcctgcttc gcagtatttt gattataaat cagcgaccaa caacaccatc
     1801 tgtaacacca atccttacaa cgaatgtcgt gatgtcggta tgttggtggg gctagatgat
     1861 tatgtgagta ccaatcaagg caaagatatg ctcatcatgc tacaccaaat gggtaatcat
     1921 gggccggcgt acttcaagcg ttatgacgag caatttgcca aatacacccc tgtgtgcgaa
     1981 ggtaatgaac ttgccaagtg tgaacaccaa tcgctcatca acgcctatga taatgcactg
     2041 cttgcgaccg atgattttat cgccaaaagt atcgattggc taaaaacgca tcaggccaac
     2101 tatgatgttg ccatgctcta tgtcagcgac cacggcgaga gtctgggtga aaatggcgtc
     2161 tatctgcatg gtatgccaaa tgcctttgca ccaaaagaac agcgagcggt accggcattc
     2221 ttttggtcaa ataatccatc gttcacgcca actgccagcg acactgtgct gacacatgat
     2281 gcgattacgc cgactctact gaagctgttt gatgtcacag cggataaggt caaagaccgc
     2341 accgcattca tccgctgaat cggatcccgg gcccgtcgac tgcagaggcc tgcatgcaag
     2401 cttggcgtaa tcatggtcat agctgtttcc tgtgtgaaat tgttatccgc tcacaattcc
     2461 acacaacata cgagccggaa gcataaagtg taaagcctgg ggtgcctaat gagtgagcta
     2521 actcacatta attgcgttgc gctcactgcc cgctttccag tcgggaaacc tgtcgtgcca
     2581 gctgcattaa tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg ggcgctcttc
     2641 cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc
     2701 tcactcaaag gcggtaatac ggttatccac agaatcaggg gataacgcag gaaagaacat
     2761 gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt
     2821 ccataggctc cgcccccctg acgagcatca caaaaatcga cgctcaagtc agaggtggcg
     2881 aaacccgaca ggactataaa gataccaggc gtttccccct ggaagctccc tcgtgcgctc
     2941 tcctgttccg accctgccgc ttaccggata cctgtccgcc tttctccctt cgggaagcgt
     3001 ggcgctttct catagctcac gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa
     3061 gctgggctgt gtgcacgaac cccccgttca gcccgaccgc tgcgccttat ccggtaacta
     3121 tcgtcttgag tccaacccgg taagacacga cttatcgcca ctggcagcag ccactggtaa
     3181 caggattagc agagcgaggt atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa
     3241 ctacggctac actagaagaa cagtatttgg tatctgcgct ctgctgaagc cagttacctt
     3301 cggaaaaaga gttggtagct cttgatccgg caaacaaacc accgctggta gcggtggttt
     3361 ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag atcctttgat
     3421 cttttctacg gggtctgacg ctcagtggaa cgaaaactca cgttaaggga ttttggtcat
     3481 gagattatca aaaaggatct tcacctagat ccttttaaat taaaaatgaa gttttaaatc
     3541 aagcccaatc tgaataatgt tacaaccaat taaccaattc tgattagaaa aactcatcga
     3601 gcatcaaatg aaactgcaat ttattcatat caggattatc aataccatat ttttgaaaaa
     3661 gccgtttctg taatgaagga gaaaactcac cgaggcagtt ccataggatg gcaagatcct
     3721 ggtatcggtc tgcgattccg actcgtccaa catcaataca acctattaat ttcccctcgt
     3781 caaaaataag gttatcaagt gagaaatcac catgagtgac gactgaatcc ggtgagaatg
     3841 gcaaaagttt atgcatttct ttccagactt gttcaacagg ccagccatta cgctcgtcat
     3901 caaaatcact cgcatcaacc aaaccgttat tcattcgtga ttgcgcctga gcgagacgaa
     3961 atacgcgatc gctgttaaaa ggacaattac aaacaggaat cgaatgcaac cggcgcagga
     4021 acactgccag cgcatcaaca atattttcac ctgaatcagg atattcttct aatacctgga
     4081 atgctgtttt tccggggatc gcagtggtga gtaaccatgc atcatcagga gtacggataa
     4141 aatgcttgat ggtcggaaga ggcataaatt ccgtcagcca gtttagtctg accatctcat
     4201 ctgtaacatc attggcaacg ctacctttgc catgtttcag aaacaactct ggcgcatcgg
     4261 gcttcccata caagcgatag attgtcgcac ctgattgccc gacattatcg cgagcccatt
     4321 tatacccata taaatcagca tccatgttgg aatttaatcg cggcctcgac gtttcccgtt
     4381 gaatatggct cataacaccc cttgtattac tgtttatgta agcagacagt tttattgttc
     4441 atgatgatat atttttatct tgtgcaatgt aacatcagag attttgagac acgggccaga
     4501 gctgca
//
