LOCUS       62056_22170        4515 bp DNA     circular SYN 13-JUN-2020
DEFINITION  Expression vector pUC57-Kan-mcr-4 DNA, complete seuence.
ACCESSION   LC548748
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4515)
  AUTHORS   Suzuki M.
  TITLE     Bacterial ARG expression libraries
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 4515)
  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 4515)
  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            "Unpublished"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..4515
                     /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..2367
                     /gene="mcr-4"
                     /label=mcr-4
     CDS             742..2367
                     /codon_start=1
                     /transl_table=11
                     /gene="mcr-4"
                     /product="lipid A phosphoethanolamine transferase MCR-4"
                     /label=mcr-4
                     /protein_id="BCG50367.1"
                     /translation="MISRFKTLSVNQFTFITALFYVAIFNLPLFGIVRKGIEKQPEVDP
                     LFIASMPLFLTFALSFLFSIFTVKYLLKPFFIVLTLLSSSVFFAAYQYNVVFDYGMIEN
                     TFQTHPAEALMYVNLASITNLLLTGLLPSYLIYKADIHYQPFFKELLHKLAFMLLMFVG
                     IGIVAFFYYQDYAAFVRNNSELRRYIVPTYFVSSASKYLNEHYLQTPMEYQQLGLDAKN
                     ASRNPNTKPNLLVVVVGETARSMSYQYYGYNKPTNAHTQNQGLIAFNDTSSCGTATAVS
                     LPCMFSRMGRADYDPRRANAQDTVIDVLSHSGIKVQWFDNDSGCKGVCDQVENLTIDLK
                     SDPKLCSGQYCFDQVLLNKLDKILAVAPSQDTVIFLHIIGSHGPTYYLRYPPEHRKFIP
                     DCPRSDIQNCSQEELINTYDNTILYTDFILSEVVNKLKGKQDMFDTAMLYLSDHGESLG
                     EKGMYLHGAPYSIAPKEQTSVPMLAWVSNDFSQDNQLNMTCVAQRAEQGGFSHDNLFDS
                     LLGLMNVKTTVYQSQLDIFAPCRY"
     primer_bind     complement(2425..2441)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    complement(2449..2465)
                     /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(2473..2503)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    complement(2518..2539)
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     rep_origin      complement(2827..3415)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(3596..4402)
                     /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 catgatttct agatttaaga cgttatcggt taaccaattc
      781 actttcatca ctgcgttgtt ttatgttgcc attttcaatc taccgctctt tggtatagtg
      841 cgaaaaggaa ttgaaaaaca accagaagtt gatccccttt tcatcgcatc tatgccgcta
      901 tttttaacat ttgcgctgag ttttttgttt tcaattttta ccgtcaaata cctgctgaag
      961 ccctttttta tcgtattgac gttactttcc tcaagtgtat tttttgcagc ctatcaatac
     1021 aatgtcgtgt ttgactacgg catgatagaa aacacgtttc aaacacatcc tgctgaagca
     1081 ttgatgtatg taaatcttgc atcaattacc aatctactgc tgactgggct attaccgtca
     1141 tatcttattt ataaggccga tattcattat cagccctttt ttaaggagtt attgcataaa
     1201 ttagccttta tgctgctaat gttcgttggc attgggatag tcgccttttt ttactatcaa
     1261 gattatgctg catttgttcg aaacaacagt gagttaaggc gttacattgt ccctacctat
     1321 tttgtcagta gtgcatctaa atatctcaat gagcactatt tgcagacgcc catggaatac
     1381 caacaacttg gcctagatgc gaagaatgcc agtcgtaacc cgaacactaa acctaactta
     1441 ttagtggttg ttgtgggtga aactgcgcgc tcaatgagct atcaatatta tggatataac
     1501 aagccaacca atgctcatac ccaaaatcag gggctgattg cgtttaacga tactagctca
     1561 tgcggcacgg ccacggcggt gtctctaccc tgtatgtttt cacgaatggg gcgggcagac
     1621 tatgatcctc gccgtgctaa tgctcaagac acagtgattg atgtgttaag tcatagtggt
     1681 ataaaagtac agtggtttga taatgattct ggctgtaaag gtgtgtgtga tcaggttgaa
     1741 aatctcacga tagatttgaa gagtgatccg aagctgtgtt ctggccaata ttgttttgac
     1801 caagtattgc tcaacaaatt agataaaatt ctggcagtag caccaagtca agatacagta
     1861 atttttttgc atatcattgg tagtcatgga ccaacttatt atcttagata cccgccagag
     1921 catcgtaaat ttataccgga ttgtccgcgc agtgatattc aaaattgcag tcaagaagaa
     1981 ctgattaaca cctacgacaa cactattcta tatacggatt ttattctcag tgaagtggtg
     2041 aataaattaa aaggtaagca ggatatgttc gatactgcaa tgctgtatct ctctgaccat
     2101 ggtgagtctt tgggtgaaaa gggcatgtat ttacatggtg cgccctatag tattgcaccg
     2161 aaagaacaaa ctagcgtacc aatgctggct tgggtatcta atgactttag ccaagataat
     2221 cagttgaaca tgacttgtgt tgcacagcga gcagaacagg gcggcttttc ccacgacaat
     2281 ttgttcgaca gtttgctagg acttatgaat gtaaaaacca ccgtctatca gagccaactc
     2341 gatatttttg caccttgcag gtattagatc ggatcccggg cccgtcgact gcagaggcct
     2401 gcatgcaagc ttggcgtaat catggtcata gctgtttcct gtgtgaaatt gttatccgct
     2461 cacaattcca cacaacatac gagccggaag cataaagtgt aaagcctggg gtgcctaatg
     2521 agtgagctaa ctcacattaa ttgcgttgcg ctcactgccc gctttccagt cgggaaacct
     2581 gtcgtgccag ctgcattaat gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg
     2641 gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc
     2701 ggtatcagct cactcaaagg cggtaatacg gttatccaca gaatcagggg ataacgcagg
     2761 aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct
     2821 ggcgtttttc cataggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagtca
     2881 gaggtggcga aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct
     2941 cgtgcgctct cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc
     3001 gggaagcgtg gcgctttctc atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt
     3061 tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc
     3121 cggtaactat cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc
     3181 cactggtaac aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg
     3241 gtggcctaac tacggctaca ctagaagaac agtatttggt atctgcgctc tgctgaagcc
     3301 agttaccttc ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag
     3361 cggtggtttt tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga
     3421 tcctttgatc ttttctacgg ggtctgacgc tcagtggaac gaaaactcac gttaagggat
     3481 tttggtcatg agattatcaa aaaggatctt cacctagatc cttttaaatt aaaaatgaag
     3541 ttttaaatca agcccaatct gaataatgtt acaaccaatt aaccaattct gattagaaaa
     3601 actcatcgag catcaaatga aactgcaatt tattcatatc aggattatca ataccatatt
     3661 tttgaaaaag ccgtttctgt aatgaaggag aaaactcacc gaggcagttc cataggatgg
     3721 caagatcctg gtatcggtct gcgattccga ctcgtccaac atcaatacaa cctattaatt
     3781 tcccctcgtc aaaaataagg ttatcaagtg agaaatcacc atgagtgacg actgaatccg
     3841 gtgagaatgg caaaagttta tgcatttctt tccagacttg ttcaacaggc cagccattac
     3901 gctcgtcatc aaaatcactc gcatcaacca aaccgttatt cattcgtgat tgcgcctgag
     3961 cgagacgaaa tacgcgatcg ctgttaaaag gacaattaca aacaggaatc gaatgcaacc
     4021 ggcgcaggaa cactgccagc gcatcaacaa tattttcacc tgaatcagga tattcttcta
     4081 atacctggaa tgctgttttt ccggggatcg cagtggtgag taaccatgca tcatcaggag
     4141 tacggataaa atgcttgatg gtcggaagag gcataaattc cgtcagccag tttagtctga
     4201 ccatctcatc tgtaacatca ttggcaacgc tacctttgcc atgtttcaga aacaactctg
     4261 gcgcatcggg cttcccatac aagcgataga ttgtcgcacc tgattgcccg acattatcgc
     4321 gagcccattt atacccatat aaatcagcat ccatgttgga atttaatcgc ggcctcgacg
     4381 tttcccgttg aatatggctc ataacacccc ttgtattact gtttatgtaa gcagacagtt
     4441 ttattgttca tgatgatata tttttatctt gtgcaatgta acatcagaga ttttgagaca
     4501 cgggccagag ctgca
//
