LOCUS       62056_22155        4509 bp DNA     circular SYN 13-JUN-2020
DEFINITION  Expression vector pUC57-Kan-mcr-10 DNA, complete seuence.
ACCESSION   LC548754
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4509)
  AUTHORS   Suzuki M.
  TITLE     Bacterial ARG expression libraries
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 4509)
  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 4509)
  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            "Unpublished"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..4509
                     /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..2361
                     /gene="mcr-10"
                     /label=mcr-10
     CDS             742..2361
                     /codon_start=1
                     /transl_table=11
                     /gene="mcr-10"
                     /product="lipid A phosphoethanolamine transferase MCR-10"
                     /label=mcr-10
                     /protein_id="BCG50379.1"
                     /translation="MPVLFRMRVIPLVLLLALVFAFLLNWPVLLHFYDILSRLEHVRAG
                     FVISIPFVLVAALNFVFMPFSVRYLLKPFFALLLVTGSVVSYATLKYKVMFDQSMIENI
                     LETNPQEAHAYLNGSLVLWLVFMGILPAILLFLIKIEYADKWYKGVAHRLLSMLASLIL
                     IAGVAALYYQDYASVGRNNPTLNKEIIPANYAYSTFHYVKDTYFTTKMPFRTLGDDARR
                     VTRNGKPTLMFLVIGETARSQNFSMNGYPRDTNAFTSKIDGVISFRNMRSCGTATAVSV
                     PCMFSDMNRTDYDGKKAAGSENVLDIVQKTGVSLLWKENDGGCKGVCSRIPTVEINPGI
                     SKKLCDGKTCYDDVMLENLDTEIGKMAGDKLIAFHMIGSHGPTYYQRYPAEHRHFMPEC
                     ARSDIENCTQEQLVNTYDNTIRHTDYVLAQMIEKLKQYSEQYNTVLLYVSDHGESLGES
                     GLYLHGTPYKLAPDQQTHIPMQLWMSPGFIAAKNINAACLQHNAVNRTYSHDNLFASVL
                     GLWDITTGAYLPESDLFRECRG"
     primer_bind     complement(2419..2435)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    complement(2443..2459)
                     /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(2467..2497)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    complement(2512..2533)
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     rep_origin      complement(2821..3409)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(3590..4396)
                     /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 catgcccgta cttttcagga tgagggtaat ccccttggtt
      781 ttacttctgg cactcgtttt tgcattctta cttaactggc cggtgttgct gcatttctac
      841 gatatcctga gccgtcttga acatgtgagg gcggggttcg tcatctccat tccgtttgtg
      901 ctggttgcag cgcttaactt tgtgtttatg cccttctcgg ttcgctacct gctcaaaccc
      961 ttctttgccc tgttgctggt caccggttcg gtggtgagtt acgccacact gaaatataaa
     1021 gtgatgtttg atcagtccat gatcgaaaat atactggaaa caaacccaca ggaagcgcat
     1081 gcctacctga atggctcact ggtgctgtgg ctggtcttca tgggcattct tccggctatc
     1141 ctgttgtttt tgattaaaat tgaatatgca gacaaatggt acaaaggggt tgcccaccgg
     1201 ctgctttcca tgctcgcttc gctgatcctg attgcaggtg ttgccgctct gtattaccag
     1261 gattatgctt ctgtcgggcg caataacccg acgctgaaca aagaaattat cccggcaaac
     1321 tatgcgtaca gcactttcca ttacgtgaag gatacctatt ttacgacgaa aatgcctttc
     1381 cggacgctgg gggatgatgc aaggcgcgtt acccggaatg gtaaacccac gctgatgttc
     1441 ctggtaattg gcgaaacggc acggagccag aatttctcca tgaacggcta cccgcgtgac
     1501 acaaatgcct ttaccagcaa aatcgatggc gttatttcgt tcaggaatat gcgttcctgt
     1561 ggcacggcga ccgcagtctc ggtgccctgt atgttctcgg atatgaaccg gacggattac
     1621 gatggtaaaa aggctgccgg cagtgaaaat gtcctcgaca tcgtgcagaa aacgggggtt
     1681 tcgctgttgt ggaaagaaaa cgatggcggg tgtaaaggcg tatgcagccg tatcccgact
     1741 gtcgaaatta atcccggtat cagtaaaaaa ctgtgtgacg gtaaaacctg ctatgacgat
     1801 gttatgctgg aaaacctgga taccgaaatc ggcaaaatgg ccggagacaa gctgatcgcc
     1861 ttccatatga ttggcagcca tggaccgacc tattaccagc gttatccggc agagcatcgt
     1921 cacttcatgc cggaatgtgc gcgcagcgat atcgaaaact gcacgcagga acagctggtt
     1981 aatacctacg acaataccat tcgccacacc gactatgtgt tagcgcagat gattgaaaag
     2041 cttaagcaat acagcgaaca gtacaacacc gtactgctgt atgtgtccga tcacggcgaa
     2101 tctctgggag agagcggact gtatctgcac ggtaccccct acaaactggc accggatcag
     2161 cagacgcaca tcccgatgca gctctggatg tcgccaggct tcattgctgc taaaaatatt
     2221 aacgccgcgt gtctgcagca taatgccgtt aacaggacat attcccacga taaccttttc
     2281 gcgtccgtac tggggctctg ggacatcacc accggggcct atcttccgga aagcgacctg
     2341 ttccgcgaat gtcgtggata gatcggatcc cgggcccgtc gactgcagag gcctgcatgc
     2401 aagcttggcg taatcatggt catagctgtt tcctgtgtga aattgttatc cgctcacaat
     2461 tccacacaac atacgagccg gaagcataaa gtgtaaagcc tggggtgcct aatgagtgag
     2521 ctaactcaca ttaattgcgt tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg
     2581 ccagctgcat taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta ttgggcgctc
     2641 ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc
     2701 agctcactca aaggcggtaa tacggttatc cacagaatca ggggataacg caggaaagaa
     2761 catgtgagca aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt
     2821 tttccatagg ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg
     2881 gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggaagct ccctcgtgcg
     2941 ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag
     3001 cgtggcgctt tctcatagct cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc
     3061 caagctgggc tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa
     3121 ctatcgtctt gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccactgg
     3181 taacaggatt agcagagcga ggtatgtagg cggtgctaca gagttcttga agtggtggcc
     3241 taactacggc tacactagaa gaacagtatt tggtatctgc gctctgctga agccagttac
     3301 cttcggaaaa agagttggta gctcttgatc cggcaaacaa accaccgctg gtagcggtgg
     3361 tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag aagatccttt
     3421 gatcttttct acggggtctg acgctcagtg gaacgaaaac tcacgttaag ggattttggt
     3481 catgagatta tcaaaaagga tcttcaccta gatcctttta aattaaaaat gaagttttaa
     3541 atcaagccca atctgaataa tgttacaacc aattaaccaa ttctgattag aaaaactcat
     3601 cgagcatcaa atgaaactgc aatttattca tatcaggatt atcaatacca tatttttgaa
     3661 aaagccgttt ctgtaatgaa ggagaaaact caccgaggca gttccatagg atggcaagat
     3721 cctggtatcg gtctgcgatt ccgactcgtc caacatcaat acaacctatt aatttcccct
     3781 cgtcaaaaat aaggttatca agtgagaaat caccatgagt gacgactgaa tccggtgaga
     3841 atggcaaaag tttatgcatt tctttccaga cttgttcaac aggccagcca ttacgctcgt
     3901 catcaaaatc actcgcatca accaaaccgt tattcattcg tgattgcgcc tgagcgagac
     3961 gaaatacgcg atcgctgtta aaaggacaat tacaaacagg aatcgaatgc aaccggcgca
     4021 ggaacactgc cagcgcatca acaatatttt cacctgaatc aggatattct tctaatacct
     4081 ggaatgctgt ttttccgggg atcgcagtgg tgagtaacca tgcatcatca ggagtacgga
     4141 taaaatgctt gatggtcgga agaggcataa attccgtcag ccagtttagt ctgaccatct
     4201 catctgtaac atcattggca acgctacctt tgccatgttt cagaaacaac tctggcgcat
     4261 cgggcttccc atacaagcga tagattgtcg cacctgattg cccgacatta tcgcgagccc
     4321 atttataccc atataaatca gcatccatgt tggaatttaa tcgcggcctc gacgtttccc
     4381 gttgaatatg gctcataaca ccccttgtat tactgtttat gtaagcagac agttttattg
     4441 ttcatgatga tatattttta tcttgtgcaa tgtaacatca gagattttga gacacgggcc
     4501 agagctgca
//
