LOCUS       62056_22070        3515 bp DNA     circular SYN 13-JUN-2020
DEFINITION  Expression vector pUC57-Amp-aac(3)-Ia DNA, complete seuence.
ACCESSION   LC548760
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3515)
  AUTHORS   Suzuki M.
  TITLE     Bacterial ARG expression libraries
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 3515)
  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 3515)
  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            "Unpublished"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..3515
                     /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        450..478
                     /label=Pc promoter
                     /note="class 1 integron promoter"
     CDS             726..1256
                     /codon_start=1
                     /label=GmR
                     /note="gentamycin acetyltransferase"
                     /translation="MLRSSNDVTQQGSRPKTKLGGSSMGIIRTCRLGPDQVKSMRAALD
                     LFGREFGDVATYSQHQPDSDYLGNLLRSKTFIALAAFDQEAVVGALAAYVLPRFEQPRS
                     EIYIYDLAVSGEHRRQGIATALINLLKHEANALGAYVIYVQADYGDDPAVALYTKLGIR
                     EEVMHFDIDPSTAT"
     primer_bind     complement(1294..1310)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    complement(1318..1334)
                     /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(1342..1372)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    complement(1387..1408)
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     rep_origin      complement(1696..2284)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(2458..3315)
                     /codon_start=1
                     /label=AmpR
                     /note="beta-lactamase"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     promoter        complement(3316..3420)
                     /label=AmpR promoter
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 gacgcgtatt gggatcacac
      421 cgtggaaacg gatgaaggca cgaacccagt tgacataagc ctgttcggtt cgtaaactgt
      481 aatgcaagta gcgtatgcgc tcacgcaact ggtccagaac cttgaccgaa cgcagcggtg
      541 gtaacggcgc agtggcggtt ttcatggctt gttatgactg tttttttgta cagtctatgc
      601 ctcgggcatc caagcagcaa gcgcgttacg ccgtgggtcg atgtttgatg ttatggagca
      661 gcaacgatgt tacgcagcag ggcagtcgcc ctaaaacaaa gttaggcatc acaaagtaca
      721 gcatcatgtt acgcagcagc aacgatgtta cgcagcaggg cagtcgccct aaaacaaagt
      781 taggtggctc aagtatgggc atcattcgca catgtaggct cggccctgac caagtcaaat
      841 ccatgcgggc tgctcttgat cttttcggtc gtgagttcgg agacgtagcc acctactccc
      901 aacatcagcc ggactccgat tacctcggga acttgctccg tagtaagaca ttcatcgcgc
      961 ttgctgcctt cgaccaagaa gcggttgttg gcgctctcgc ggcttacgtt ctgcccaggt
     1021 ttgagcagcc gcgtagtgag atctatatct atgatctcgc agtctccggc gagcaccgga
     1081 ggcagggcat tgccaccgcg ctcatcaatc tcctcaagca tgaggccaac gcgcttggtg
     1141 cttatgtgat ctacgtgcaa gcagattacg gtgacgatcc cgcagtggct ctctatacaa
     1201 agttgggcat acgggaagaa gtgatgcact ttgatatcga cccaagtacc gccacctaaa
     1261 tcccaatggc gcgccgagct tggcgtaatc atggtcatag ctgtttcctg tgtgaaattg
     1321 ttatccgctc acaattccac acaacatacg agccggaagc ataaagtgta aagcctgggg
     1381 tgcctaatga gtgagctaac tcacattaat tgcgttgcgc tcactgcccg ctttccagtc
     1441 gggaaacctg tcgtgccagc tgcattaatg aatcggccaa cgcgcgggga gaggcggttt
     1501 gcgtattggg cgctcttccg cttcctcgct cactgactcg ctgcgctcgg tcgttcggct
     1561 gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg ttatccacag aatcagggga
     1621 taacgcagga aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc
     1681 cgcgttgctg gcgtttttcc ataggctccg cccccctgac gagcatcaca aaaatcgacg
     1741 ctcaagtcag aggtggcgaa acccgacagg actataaaga taccaggcgt ttccccctgg
     1801 aagctccctc gtgcgctctc ctgttccgac cctgccgctt accggatacc tgtccgcctt
     1861 tctcccttcg ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc tcagttcggt
     1921 gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg
     1981 cgccttatcc ggtaactatc gtcttgagtc caacccggta agacacgact tatcgccact
     2041 ggcagcagcc actggtaaca ggattagcag agcgaggtat gtaggcggtg ctacagagtt
     2101 cttgaagtgg tggcctaact acggctacac tagaagaaca gtatttggta tctgcgctct
     2161 gctgaagcca gttaccttcg gaaaaagagt tggtagctct tgatccggca aacaaaccac
     2221 cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc
     2281 tcaagaagat cctttgatct tttctacggg gtctgacgct cagtggaacg aaaactcacg
     2341 ttaagggatt ttggtcatga gattatcaaa aaggatcttc acctagatcc ttttaaatta
     2401 aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa acttggtctg acagttacca
     2461 atgcttaatc agtgaggcac ctatctcagc gatctgtcta tttcgttcat ccatagttgc
     2521 ctgactcccc gtcgtgtaga taactacgat acgggagggc ttaccatctg gccccagtgc
     2581 tgcaatgata ccgcgagacc cacgctcacc ggctccagat ttatcagcaa taaaccagcc
     2641 agccggaagg gccgagcgca gaagtggtcc tgcaacttta tccgcctcca tccagtctat
     2701 taattgttgc cgggaagcta gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt
     2761 tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc
     2821 cggttcccaa cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa aagcggttag
     2881 ctccttcggt cctccgatcg ttgtcagaag taagttggcc gcagtgttat cactcatggt
     2941 tatggcagca ctgcataatt ctcttactgt catgccatcc gtaagatgct tttctgtgac
     3001 tggtgagtac tcaaccaagt cattctgaga atagtgtatg cggcgaccga gttgctcttg
     3061 cccggcgtca atacgggata ataccgcgcc acatagcaga actttaaaag tgctcatcat
     3121 tggaaaacgt tcttcggggc gaaaactctc aaggatctta ccgctgttga gatccagttc
     3181 gatgtaaccc actcgtgcac ccaactgatc ttcagcatct tttactttca ccagcgtttc
     3241 tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa
     3301 atgttgaata ctcatactct tcctttttca atattattga agcatttatc agggttattg
     3361 tctcatgagc ggatacatat ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg
     3421 cacatttccc cgaaaagtgc cacctgacgt ctaagaaacc attattatca tgacattaac
     3481 ctataaaaat aggcgtatca cgaggccctt tcgtc
//
