LOCUS       62056_22195        4509 bp DNA     circular SYN 13-JUN-2020
DEFINITION  Expression vector pUC57-Kan-mcr-9 DNA, complete seuence.
ACCESSION   LC548753
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  JOURNAL   Unpublished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 AMR Research Center, 
            National Institute of Infectious Diseases; 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            "Unpublished"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 AMR Research Center, National 
            Institute of Infectious Diseases; 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-9"
                     /label=mcr-9
     CDS             742..2361
                     /codon_start=1
                     /transl_table=11
                     /gene="mcr-9"
                     /product="lipid A phosphoethanolamine transferase MCR-9"
                     /label=mcr-9
                     /protein_id="BCG50377.1"
                     /translation="MPVLFRVKVIPLVLLLAMIFAFLLNWPILLHFYEILSHLEHVKIG
                     FVISIPFVLVAALNVVFMPFSVRFLLKPFFALLFITGSLVSYSTLKYKVMFDQTMIQNI
                     IETNPQEAHSYLNGSIIIWFVFTGILPAILLFSIKIQYPEKWYKGIAYRLLSVLASLSL
                     IAGVAALYYQDYASVGRNNSTLNKEIIPANYAYSTFQYVKDTYFTTKVPFQTLGNDAKR
                     VVAHEKPTLMFLVIGETARSQNFSMNGYSRDTNAFTSKSGGVISFKNMHSCGTATAISV
                     PCMFSNMNRTEYDSKKASNSENFLDIVQKTGVSLLWKENDGGCKGVCSRIPTVEIKPSD
                     NPKLCDGKTCHDEVMLENLDDEIAKMPGDKLVAFHIIGSHGPTYYLRYPAEHRHFMPEC
                     ARSDIENCTQEQLVNTYDNTLRYTDYVLAEMIEKLKNYSDQYNTVLLYVSDHGESLGES
                     GLYLHGTPYKLAPDQQTHIPMQVWMSPGFIAGKHINMSCLENNAAKKSYSHDNLFSSIL
                     GLWDVSTSVYNPDRDLFRECRG"
     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 catgcctgta cttttcaggg tgaaagttat tccgctggtt
      781 ttacttctgg caatgatctt tgcgttttta cttaactggc caatattgct gcatttttac
      841 gagattttgt cgcatttaga gcatgtcaaa attggttttg tcatttctat tccctttgtt
      901 ctggttgcgg cgcttaacgt tgtttttatg cctttctcag ttcgttttct gctgaaacct
      961 ttctttgctt tactgtttat cactggctca ctggtcagtt attcgacact aaaatataaa
     1021 gtaatgtttg atcaaacgat gattcaaaac attattgaaa ctaaccccca ggaagcgcat
     1081 tcctatctta atggctcaat tattatatgg ttcgtcttta ccggtatcct tcctgccatc
     1141 ctcctttttt caataaaaat tcaatatcct gaaaaatggt ataaaggcat tgcttaccgt
     1201 ttgctctccg tgctggcatc gttgagtttg attgcaggtg ttgccgcact ttattatcag
     1261 gattatgcct ctgtcggccg caataactcg acattgaata aagagatcat cccggcgaac
     1321 tacgcttaca gcactttcca gtatgttaag gatacgtact ttacgactaa agtgcctttc
     1381 cagacgctgg ggaatgatgc taaacgcgtc gtcgctcacg aaaaacccac gctgatgttc
     1441 ctggtgattg gcgaaacggc acgcagccag aatttctcga tgaacggtta ttcgcgtgat
     1501 accaatgcct ttaccagcaa atccggcggc gttatttcgt ttaaaaatat gcattcctgc
     1561 ggtaccgcta ccgcaatatc cgttccgtgc atgttctcga atatgaatcg caccgagtac
     1621 gacagtaaaa aagcatctaa cagtgaaaat ttcctcgaca tcgtgcagaa aaccggtgtc
     1681 tcgctgttat ggaaagagaa cgatggcggt tgtaaaggcg tatgtagccg catcccgact
     1741 gtcgaaatta agcctagtga taacccgaaa ctgtgcgatg gcaaaacgtg ccatgacgag
     1801 gtgatgctgg aaaaccttga tgatgaaatc gccaaaatgc caggtgataa gcttgtcgcc
     1861 ttccatatca ttggcagcca tggaccgact tattacctgc gttatccggc tgagcatcgc
     1921 cacttcatgc ccgaatgtgc acgtagcgat atcgaaaact gtactcagga acaattggtc
     1981 aacacctacg acaacaccct tcgttataca gactatgtat tagctgagat gattgaaaag
     2041 ctaaaaaatt acagcgatca gtacaacacc gtgctgcttt atgtgtccga tcatggtgaa
     2101 tcattgggcg aaagcgggct atatctgcac ggcacgccgt acaaactggc accggatcag
     2161 cagacgcata ttccgatgca ggtctggatg tcaccgggct ttatcgccgg gaaacacatc
     2221 aacatgtctt gccttgaaaa taatgcggcg aaaaaatcat attcccacga caacctgttc
     2281 tcatcgattt tggggctgtg ggacgtaagc accagcgtct ataatcctga ccgcgatttg
     2341 ttccgcgaat gccgtggcta aatcggatcc 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
//
