LOCUS       62056_22160        4506 bp DNA     circular SYN 18-SEP-2019
DEFINITION  Expression vector pUC57-Kan-mcr-2 DNA, complete sequence.
ACCESSION   LC498722
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  JOURNAL   Unpublished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 (04-SEP-2019) 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            "Unpublished"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 
            (04-SEP-2019) 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-2"
                     /label=mcr-2
     CDS             742..2358
                     /codon_start=1
                     /transl_table=11
                     /gene="mcr-2"
                     /product="lipid A phosphoethanolamine transferase MCR-2"
                     /label=mcr-2
                     /protein_id="BBN60807.1"
                     /translation="MTSHHSWYRYSINPFVLMGLVALFLAATANLTFFEKAMAVYPVSD
                     NLGFIISMAVAVMGAMLLIVVLLSYRYVLKPVLILLLIMGAVTSYFTDTYGTVYDTTML
                     QNAMQTDQAESKDLMNLAFFVRIIGLGVLPSVLVAVAKVNYPTWGKGLIQRAMTWGVSL
                     VLLLVPIGLFSSQYASFFRVHKPVRFYINPITPIYSVGKLASIEYKKATAPTDTIYHAK
                     DAVQTTKPSERKPRLVVFVVGETARADHVQFNGYGRETFPQLAKVDGLANFSQVTSCGT
                     STAYSVPCMFSYLGQDDYDVDTAKYQENVLDTLDRLGVGILWRDNNSDSKGVMDKLPAT
                     QYFDYKSATNNTICNTNPYNECRDVGMLVGLDDYVSANNGKDMLIMLHQMGNHGPAYFK
                     RYDEQFAKFTPVCEGNELAKCEHQSLINAYDNALLATDDFIAKSIDWLKTHEANYDVAM
                     LYVSDHGESLGENGVYLHGMPNAFAPKEQRAVPAFFWSNNTTFKPTASDTVLTHDAITP
                     TLLKLFDVTAGKVKDRAAFIQ"
     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 catgacatca catcactctt ggtatcgcta ttctatcaat
      781 ccttttgtgc tgatgggttt ggtggcgtta tttttggcag cgacagcgaa cctgacattt
      841 tttgaaaaag cgatggcggt ctatcctgta tcggataact taggctttat catctcaatg
      901 gcggtggcgg tgatgggtgc tatgctactg attgtcgtgc tgttatccta tcgctatgtg
      961 ctaaagcctg tcctgatttt gctactgatt atgggtgcgg tgacgagcta ttttaccgat
     1021 acttatggca cggtctatga caccaccatg ctccaaaatg ccatgcaaac cgaccaagcc
     1081 gagtctaagg acttgatgaa tttggcgttt tttgtgcgaa ttatcgggct tggcgtgttg
     1141 ccaagtgtgt tggtcgcagt tgccaaagtc aattatccaa catggggcaa aggtctgatt
     1201 cagcgtgcga tgacatgggg tgtcagcctt gtgctgttgc ttgtgccgat tggactattt
     1261 agcagtcagt atgcgagttt ctttcgggtg cataagccag tgcgttttta tatcaacccg
     1321 attacgccga tttattcggt gggtaagctt gccagtatcg agtacaaaaa agccactgcg
     1381 ccaacagaca ccatctatca tgccaaagac gccgtgcaga ccaccaagcc gagcgagcgt
     1441 aagccacgcc tagtggtgtt cgtcgtcggt gagacggcgc gtgctgacca tgtgcagttc
     1501 aatggctatg gccgtgagac tttcccgcag cttgccaaag ttgatggctt ggcgaatttt
     1561 agccaagtga catcgtgtgg cacatcgacg gcgtattctg tgccgtgtat gttcagctat
     1621 ttgggtcaag atgactatga tgtcgatacc gccaaatacc aagaaaatgt gctagatacg
     1681 cttgaccgct tgggtgtggg tatcttgtgg cgtgataata attcagactc aaaaggcgtg
     1741 atggataagc tacctgccac gcagtatttt gattataaat cagcaaccaa caataccatc
     1801 tgtaacacca atccctataa cgaatgccgt gatgtcggta tgcttgtcgg gctagatgac
     1861 tatgtcagcg ccaataatgg caaagatatg ctcatcatgc tacaccaaat gggcaatcat
     1921 gggccggcgt actttaagcg ttatgatgag caatttgcca aattcacccc cgtgtgcgaa
     1981 ggcaacgagc ttgccaaatg cgaacaccaa tcactcatca atgcctatga caatgcgcta
     2041 cttgcgactg atgattttat cgccaaaagc atcgattggc taaaaacgca tgaagcgaac
     2101 tacgatgtcg ccatgctcta tgtcagtgac cacggcgaga gcttgggcga aaatggtgtc
     2161 tatctgcatg gtatgccaaa tgcctttgca ccaaaagaac agcgagctgt gcctgcgttt
     2221 ttttggtcaa ataatacgac attcaagcca actgccagcg atactgtgct gacgcatgat
     2281 gcgattacgc caacactgct taagctgttt gatgtcacag cgggcaaggt caaagaccgc
     2341 gcggcattta tccagtaaat 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
//
