LOCUS       62056_22190        4587 bp DNA     circular SYN 13-JUN-2020
DEFINITION  Expression vector pUC57-Kan-mcr-8 DNA, complete seuence.
ACCESSION   LC548752
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4587)
  AUTHORS   Suzuki M.
  TITLE     Bacterial ARG expression libraries
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 4587)
  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 4587)
  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            "Unpublished"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..4587
                     /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..2439
                     /gene="mcr-8"
                     /label=mcr-8
     CDS             742..2439
                     /codon_start=1
                     /transl_table=11
                     /gene="mcr-8"
                     /product="lipid A phosphoethanolamine transferase MCR-8"
                     /label=mcr-8
                     /protein_id="BCG50375.1"
                     /translation="MFKYLLSFKLNPVQRTWAAAFFFTTIGNIALWQTLWINVDVHNIH
                     NLLFFASLPIFLFCFLSILLTPVMVIPYLCRPLLVVLILISACCSYFMMKYNILIDRSM
                     VQNFFETNQAELTSYLSVPFLSTLFLLGIVPAIILALPSTDNKRGAFRIELWWLAHICI
                     AVVLLAMVTMVFYKDYASLIRNNMQIKDQALPFNFVRNTNGYLKRKYQASSTILQSVGE
                     DAVRPIYSNAPPKLVVVVVGETARAQNFQLNGYSRVTNPYLSRRHDVISFKNVSSCGTA
                     TAISLPCMFSRMSRNEYNEVRAASEENLLDILKRTGVEVLWRNNNNGGCKGICKRVPTD
                     DMPAMKVIGECVNKDGTCFDEVLLNQLSSRINAMQGDALIVLHQMGSHGPTYFERYPST
                     SKVFSPTCDSNLIEKCSNKELVNTYDNTLVYTDRMLSKTIELLQRYSGMRDVAMIYLSD
                     HGESLGESGIYLHGTPYIIAPNEQTHIPMFMWFSSSFAQHSKLNLECLTGNADKQYSHD
                     NFYHSILGLFNVKTSVYKPELDMFTLCRQSDHTPLSSAVVREKTDGNG"
     primer_bind     complement(2497..2513)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    complement(2521..2537)
                     /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(2545..2575)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    complement(2590..2611)
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     rep_origin      complement(2899..3487)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(3668..4474)
                     /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 catgttcaag tatcttttat ctttcaaact gaacccggta
      781 caacggacct gggctgcagc attttttttc actacaatcg gcaacatagc actttggcaa
      841 acactatgga ttaatgtaga tgttcataat atacataatc tacttttttt tgccagtctg
      901 ccaatatttc ttttctgctt tctaagtatc ttacttacac cagtcatggt tattccatat
      961 ttatgcaggc ctctacttgt agttcttatt ctaatcagtg cctgctgtag ttatttcatg
     1021 atgaaataca acatattaat tgaccgcagc atggtgcaaa acttttttga gactaatcag
     1081 gctgaattaa catcatactt atccgttcct tttctttcca ctctatttct acttggcatt
     1141 gtaccagcaa ttatcctggc gttgccttca acagacaata agcggggagc ttttagaatt
     1201 gaattgtggt ggttggcgca tatttgcata gctgtagtct tattagccat ggttaccatg
     1261 gtgttttata aggattacgc atctctcata cgaaacaata tgcagattaa agaccaggct
     1321 ttacctttta actttgtgcg taatacgaat ggttacctta aaagaaaata ccaggcatct
     1381 tcaacaattc tacaaagcgt gggggaggat gctgtacgtc caatatattc aaatgctcca
     1441 ccgaaactgg tggttgtcgt cgtgggcgaa accgccagag cacagaattt ccagctgaat
     1501 ggctattcgc gggtaaccaa cccctatctt tccagacgac atgatgttat cagtttcaaa
     1561 aatgtgtcgt catgcggaac ggctaccgca atatcactac cctgcatgtt ctcgcgaatg
     1621 tcacgtaacg aatacaatga agtccgtgcc gcatcagaag aaaacttgct ggatatcctt
     1681 aaacgtacag gtgttgaggt gctatggcgc aacaataaca atggtggttg taagggaatc
     1741 tgcaagcgag tacccacaga tgatatgccg gcaatgaaag taattgggga atgtgttaac
     1801 aaagatggta catgctttga tgaggtgtta ttaaatcaac tctcatcccg aattaatgca
     1861 atgcagggtg atgcgcttat tgttttacat caaatgggca gtcatggacc aacatatttt
     1921 gaacgttatc cgtctacaag taaagtcttt agcccaactt gcgacagcaa cctgatcgaa
     1981 aaatgctcaa ataaagaact ggtcaataca tacgacaata cgctagttta tactgatcgt
     2041 atgctgagca aaactattga actgttgcaa cgttattccg ggatgcgtga cgttgctatg
     2101 atatatcttt ctgatcatgg agaatcgctg ggggaaagcg gaatatatct tcatggcaca
     2161 ccatatatta ttgcccccaa tgaacaaaca cacatcccga tgtttatgtg gttttcgtct
     2221 tcattcgcgc agcattccaa attaaatcta gaatgcctga ccggtaatgc cgacaaacaa
     2281 tacagtcatg ataattttta tcattcaata cttggtctct tcaacgtaaa aaccagtgta
     2341 tataaaccgg agttagatat gtttactcta tgtcgacaat ctgaccacac accactgtct
     2401 tccgcagttg taagagagaa aacagatggg aatggttaga tcggatcccg ggcccgtcga
     2461 ctgcagaggc ctgcatgcaa gcttggcgta atcatggtca tagctgtttc ctgtgtgaaa
     2521 ttgttatccg ctcacaattc cacacaacat acgagccgga agcataaagt gtaaagcctg
     2581 gggtgcctaa tgagtgagct aactcacatt aattgcgttg cgctcactgc ccgctttcca
     2641 gtcgggaaac ctgtcgtgcc agctgcatta atgaatcggc caacgcgcgg ggagaggcgg
     2701 tttgcgtatt gggcgctctt ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg
     2761 gctgcggcga gcggtatcag ctcactcaaa ggcggtaata cggttatcca cagaatcagg
     2821 ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa
     2881 ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc acaaaaatcg
     2941 acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg cgtttccccc
     3001 tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat acctgtccgc
     3061 ctttctccct tcgggaagcg tggcgctttc tcatagctca cgctgtaggt atctcagttc
     3121 ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc agcccgaccg
     3181 ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg acttatcgcc
     3241 actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg gtgctacaga
     3301 gttcttgaag tggtggccta actacggcta cactagaaga acagtatttg gtatctgcgc
     3361 tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg gcaaacaaac
     3421 caccgctggt agcggtggtt tttttgtttg caagcagcag attacgcgca gaaaaaaagg
     3481 atctcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga acgaaaactc
     3541 acgttaaggg attttggtca tgagattatc aaaaaggatc ttcacctaga tccttttaaa
     3601 ttaaaaatga agttttaaat caagcccaat ctgaataatg ttacaaccaa ttaaccaatt
     3661 ctgattagaa aaactcatcg agcatcaaat gaaactgcaa tttattcata tcaggattat
     3721 caataccata tttttgaaaa agccgtttct gtaatgaagg agaaaactca ccgaggcagt
     3781 tccataggat ggcaagatcc tggtatcggt ctgcgattcc gactcgtcca acatcaatac
     3841 aacctattaa tttcccctcg tcaaaaataa ggttatcaag tgagaaatca ccatgagtga
     3901 cgactgaatc cggtgagaat ggcaaaagtt tatgcatttc tttccagact tgttcaacag
     3961 gccagccatt acgctcgtca tcaaaatcac tcgcatcaac caaaccgtta ttcattcgtg
     4021 attgcgcctg agcgagacga aatacgcgat cgctgttaaa aggacaatta caaacaggaa
     4081 tcgaatgcaa ccggcgcagg aacactgcca gcgcatcaac aatattttca cctgaatcag
     4141 gatattcttc taatacctgg aatgctgttt ttccggggat cgcagtggtg agtaaccatg
     4201 catcatcagg agtacggata aaatgcttga tggtcggaag aggcataaat tccgtcagcc
     4261 agtttagtct gaccatctca tctgtaacat cattggcaac gctacctttg ccatgtttca
     4321 gaaacaactc tggcgcatcg ggcttcccat acaagcgata gattgtcgca cctgattgcc
     4381 cgacattatc gcgagcccat ttatacccat ataaatcagc atccatgttg gaatttaatc
     4441 gcggcctcga cgtttcccgt tgaatatggc tcataacacc ccttgtatta ctgtttatgt
     4501 aagcagacag ttttattgtt catgatgata tatttttatc ttgtgcaatg taacatcaga
     4561 gattttgaga cacgggccag agctgca
//
