LOCUS       V016064                 7335 bp    DNA     circular SYN 06-MAY-2019
DEFINITION  Exported.
ACCESSION   V016064
VERSION     V016064
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
            .
REFERENCE   1  (bases 1 to 7335)
  AUTHORS   Hu XM, Yan XH, Yang B, Wang Q, Zheng L.
  TITLE     Full-length Real-time PCR for Rapid Detection of Seventeen
            Carbapenemase Genes in Gram-negative bacteria
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 7335)
  AUTHORS   Hu XM, Yan XH, Yang B, Wang Q, Zheng L.
  TITLE     Direct Submission
  JOURNAL   Submitted (25-APR-2019) Department of Laboratory Medicine, Nanfang
            Hospital, Southern Medical University, No. 1383, Guangzhou Dadao
            Bei, Guangzhou, Guangdong 510515, China
REFERENCE   3  (bases 1 to 7335)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     ##Assembly-Data-START##
            Sequencing Technology :: Sanger dideoxy sequencing
            ##Assembly-Data-END##
            SGRef: number: 1; type: "Journal Article"; journalName:
            "Unpublished"
            SGRef: number: 2; type: "Journal Article"; journalName: "Submitted
            (25-APR-2019) Department of Laboratory Medicine, Nanfang Hospital,
            Southern Medical University, No. 1383, Guangzhou Dadao Bei,
            Guangzhou, Guangdong 510515, China"
FEATURES             Location/Qualifiers
     source          1..7335
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      12..467
                     /label="f1 ori"
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     CDS             complement(563..1375)
                     /label="KanR"
                     /note="aminoglycoside phosphotransferase"
     rep_origin      1497..2085
                     /label="ori"
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
     misc_feature    complement(2271..2413)
                     /label="bom"
                     /note="basis of mobility region from pBR322"
     CDS             complement(2518..2706)
                     /label="rop"
                     /note="Rop protein, which maintains plasmids at low copy
                     number"
     protein_bind    complement(3481..3502)
                     /label="CAP binding site"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     CDS             complement(3518..4597)
                     /label="lacI"
                     /note="lac repressor"
     promoter        complement(4598..4675)
                     /label="lacI promoter"
     promoter        4984..5002
                     /label="T7 promoter"
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    5003..5027
                     /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)."
     RBS             5042..5064
                     /label="RBS"
                     /note="efficient ribosome binding site from bacteriophage
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             5083..5100
                     /label="6xHis"
                     /note="6xHis affinity tag"
     CDS             5110..5127
                     /label="thrombin site"
                     /note="thrombin recognition and cleavage site"
     CDS             5131..5163
                     /label="T7 tag (gene 10 leader)"
                     /note="leader peptide from bacteriophage T7 gene 10"
     CDS             complement(5190..6077)
                     /codon_start=1
                     /transl_table=11
                     /product="transcriptional regulator NMC-R"
                     /label="transcriptional regulator NMC-R"
                     /protein_id="QCH41444.1"
                     /translation="MRARLPLNALRAFEASARYLNFTKAGLELHVSQAAVSQQVRTLEQ
                     MLGVALFTRVPRGLQLTDEGMHLLPSITEALQMMSSAMDKFHEGKIKEVLTIAVVGTFA
                     IGWLLPRITAFLNENPWIDIRILTHNNVVNLAAEGIDASIRFGTGGWINTENILLFQAP
                     HTVLCSPETSKKLYIPSDLKKVCLLRSYRKEEWNNWFKAAGIDPWTITGPIFDSTRLMI
                     DAVKLGDYAALVPYHMFQKELNERSVAKPFEIYATLGGYWLTLQKSRVNHNSEALNVFK
                     EWIIEHSREFVLKS"
     CDS             6219..7094
                     /gene="nmcA"
                     /label="Imipenem-hydrolyzing beta-lactamase"
                     /note="Imipenem-hydrolyzing beta-lactamase from
                     Enterobacter cloacae. Accession#: P52663"
     CDS             7179..7196
                     /label="6xHis"
                     /note="6xHis affinity tag"
     terminator      7263..7310
                     /label="T7 terminator"
                     /note="transcription terminator for bacteriophage T7 RNA
                     polymerase"
ORIGIN
        1 tggcgaatgg gacgcgccct gtagcggcgc attaagcgcg gcgggtgtgg tggttacgcg
       61 cagcgtgacc gctacacttg ccagcgccct agcgcccgct cctttcgctt tcttcccttc
      121 ctttctcgcc acgttcgccg gctttccccg tcaagctcta aatcgggggc tccctttagg
      181 gttccgattt agtgctttac ggcacctcga ccccaaaaaa cttgattagg gtgatggttc
      241 acgtagtggg ccatcgccct gatagacggt ttttcgccct ttgacgttgg agtccacgtt
      301 ctttaatagt ggactcttgt tccaaactgg aacaacactc aaccctatct cggtctattc
      361 ttttgattta taagggattt tgccgatttc ggcctattgg ttaaaaaatg agctgattta
      421 acaaaaattt aacgcgaatt ttaacaaaat attaacgttt acaatttcag gtggcacttt
      481 tcggggaaat gtgcgcggaa cccctatttg tttatttttc taaatacatt caaatatgta
      541 tccgctcatg aattaattct tagaaaaact catcgagcat caaatgaaac tgcaatttat
      601 tcatatcagg attatcaata ccatattttt gaaaaagccg tttctgtaat gaaggagaaa
      661 actcaccgag gcagttccat aggatggcaa gatcctggta tcggtctgcg attccgactc
      721 gtccaacatc aatacaacct attaatttcc cctcgtcaaa aataaggtta tcaagtgaga
      781 aatcaccatg agtgacgact gaatccggtg agaatggcaa aagtttatgc atttctttcc
      841 agacttgttc aacaggccag ccattacgct cgtcatcaaa atcactcgca tcaaccaaac
      901 cgttattcat tcgtgattgc gcctgagcga gacgaaatac gcgatcgctg ttaaaaggac
      961 aattacaaac aggaatcgaa tgcaaccggc gcaggaacac tgccagcgca tcaacaatat
     1021 tttcacctga atcaggatat tcttctaata cctggaatgc tgttttcccg gggatcgcag
     1081 tggtgagtaa ccatgcatca tcaggagtac ggataaaatg cttgatggtc ggaagaggca
     1141 taaattccgt cagccagttt agtctgacca tctcatctgt aacatcattg gcaacgctac
     1201 ctttgccatg tttcagaaac aactctggcg catcgggctt cccatacaat cgatagattg
     1261 tcgcacctga ttgcccgaca ttatcgcgag cccatttata cccatataaa tcagcatcca
     1321 tgttggaatt taatcgcggc ctagagcaag acgtttcccg ttgaatatgg ctcataacac
     1381 cccttgtatt actgtttatg taagcagaca gttttattgt tcatgaccaa aatcccttaa
     1441 cgtgagtttt cgttccactg agcgtcagac cccgtagaaa agatcaaagg atcttcttga
     1501 gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg
     1561 gtggtttgtt tgccggatca agagctacca actctttttc cgaaggtaac tggcttcagc
     1621 agagcgcaga taccaaatac tgtccttcta gtgtagccgt agttaggcca ccacttcaag
     1681 aactctgtag caccgcctac atacctcgct ctgctaatcc tgttaccagt ggctgctgcc
     1741 agtggcgata agtcgtgtct taccgggttg gactcaagac gatagttacc ggataaggcg
     1801 cagcggtcgg gctgaacggg gggttcgtgc acacagccca gcttggagcg aacgacctac
     1861 accgaactga gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga
     1921 aaggcggaca ggtatccggt aagcggcagg gtcggaacag gagagcgcac gagggagctt
     1981 ccagggggaa acgcctggta tctttatagt cctgtcgggt ttcgccacct ctgacttgag
     2041 cgtcgatttt tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg
     2101 gcctttttac ggttcctggc cttttgctgg ccttttgctc acatgttctt tcctgcgtta
     2161 tcccctgatt ctgtggataa ccgtattacc gcctttgagt gagctgatac cgctcgccgc
     2221 agccgaacga ccgagcgcag cgagtcagtg agcgaggaag cggaagagcg cctgatgcgg
     2281 tattttctcc ttacgcatct gtgcggtatt tcacaccgca tatatggtgc actctcagta
     2341 caatctgctc tgatgccgca tagttaagcc agtatacact ccgctatcgc tacgtgactg
     2401 ggtcatggct gcgccccgac acccgccaac acccgctgac gcgccctgac gggcttgtct
     2461 gctcccggca tccgcttaca gacaagctgt gaccgtctcc gggagctgca tgtgtcagag
     2521 gttttcaccg tcatcaccga aacgcgcgag gcagctgcgg taaagctcat cagcgtggtc
     2581 gtgaagcgat tcacagatgt ctgcctgttc atccgcgtcc agctcgttga gtttctccag
     2641 aagcgttaat gtctggcttc tgataaagcg ggccatgtta agggcggttt tttcctgttt
     2701 ggtcactgat gcctccgtgt aagggggatt tctgttcatg ggggtaatga taccgatgaa
     2761 acgagagagg atgctcacga tacgggttac tgatgatgaa catgcccggt tactggaacg
     2821 ttgtgagggt aaacaactgg cggtatggat gcggcgggac cagagaaaaa tcactcaggg
     2881 tcaatgccag cgcttcgtta atacagatgt aggtgttcca cagggtagcc agcagcatcc
     2941 tgcgatgcag atccggaaca taatggtgca gggcgctgac ttccgcgttt ccagacttta
     3001 cgaaacacgg aaaccgaaga ccattcatgt tgttgctcag gtcgcagacg ttttgcagca
     3061 gcagtcgctt cacgttcgct cgcgtatcgg tgattcattc tgctaaccag taaggcaacc
     3121 ccgccagcct agccgggtcc tcaacgacag gagcacgatc atgcgcaccc gtggggccgc
     3181 catgccggcg ataatggcct gcttctcgcc gaaacgtttg gtggcgggac cagtgacgaa
     3241 ggcttgagcg agggcgtgca agattccgaa taccgcaagc gacaggccga tcatcgtcgc
     3301 gctccagcga aagcggtcct cgccgaaaat gacccagagc gctgccggca cctgtcctac
     3361 gagttgcatg ataaagaaga cagtcataag tgcggcgacg atagtcatgc cccgcgccca
     3421 ccggaaggag ctgactgggt tgaaggctct caagggcatc ggtcgagatc ccggtgccta
     3481 atgagtgagc taacttacat taattgcgtt gcgctcactg cccgctttcc agtcgggaaa
     3541 cctgtcgtgc cagctgcatt aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat
     3601 tgggcgccag ggtggttttt cttttcacca gtgagacggg caacagctga ttgcccttca
     3661 ccgcctggcc ctgagagagt tgcagcaagc ggtccacgct ggtttgcccc agcaggcgaa
     3721 aatcctgttt gatggtggtt aacggcggga tataacatga gctgtcttcg gtatcgtcgt
     3781 atcccactac cgagatatcc gcaccaacgc gcagcccgga ctcggtaatg gcgcgcattg
     3841 cgcccagcgc catctgatcg ttggcaacca gcatcgcagt gggaacgatg ccctcattca
     3901 gcatttgcat ggtttgttga aaaccggaca tggcactcca gtcgccttcc cgttccgcta
     3961 tcggctgaat ttgattgcga gtgagatatt tatgccagcc agccagacgc agacgcgccg
     4021 agacagaact taatgggccc gctaacagcg cgatttgctg gtgacccaat gcgaccagat
     4081 gctccacgcc cagtcgcgta ccgtcttcat gggagaaaat aatactgttg atgggtgtct
     4141 ggtcagagac atcaagaaat aacgccggaa cattagtgca ggcagcttcc acagcaatgg
     4201 catcctggtc atccagcgga tagttaatga tcagcccact gacgcgttgc gcgagaagat
     4261 tgtgcaccgc cgctttacag gcttcgacgc cgcttcgttc taccatcgac accaccacgc
     4321 tggcacccag ttgatcggcg cgagatttaa tcgccgcgac aatttgcgac ggcgcgtgca
     4381 gggccagact ggaggtggca acgccaatca gcaacgactg tttgcccgcc agttgttgtg
     4441 ccacgcggtt gggaatgtaa ttcagctccg ccatcgccgc ttccactttt tcccgcgttt
     4501 tcgcagaaac gtggctggcc tggttcacca cgcgggaaac ggtctgataa gagacaccgg
     4561 catactctgc gacatcgtat aacgttactg gtttcacatt caccaccctg aattgactct
     4621 cttccgggcg ctatcatgcc ataccgcgaa aggttttgcg ccattcgatg gtgtccggga
     4681 tctcgacgct ctcccttatg cgactcctgc attaggaagc agcccagtag taggttgagg
     4741 ccgttgagca ccgccgccgc aaggaatggt gcatgcaagg agatggcgcc caacagtccc
     4801 ccggccacgg ggcctgccac catacccacg ccgaaacaag cgctcatgag cccgaagtgg
     4861 cgagcccgat cttccccatc ggtgatgtcg gcgatatagg cgccagcaac cgcacctgtg
     4921 gcgccggtga tgccggccac gatgcgtccg gcgtagagga tcgagatctc gatcccgcga
     4981 aattaatacg actcactata ggggaattgt gagcggataa caattcccct ctagaaataa
     5041 ttttgtttaa ctttaagaag gagatatacc atgggcagca gccatcatca tcatcatcac
     5101 agcagcggcc tggtgccgcg cggcagccat atggctagca tgactggtgg acagcaaatg
     5161 ggtcgcggat cctatacctt tagcagagac taggatttta atacaaactc tctgctatgc
     5221 tcgataatcc attccttgaa aacattaagt gcttcactat tatgattaac ccgagatttc
     5281 tgcaaagtaa gccaataacc gccaagcgtg gcataaattt caaaaggttt tgccacggat
     5341 cgctcattta attctttttg aaacatatga taaggaacta gtgctgcata atcacctaac
     5401 tttacagcat ctatcataag tctggtagaa tcaaagatag gccctgtaat agtccaaggg
     5461 tcgatcccag cagctttaaa ccaattattc cattcctcct ttcgatagga acgcagtaag
     5521 caaacttttt ttaagtctga tggaatgtac aatttcttgg atgtttccgg ggaacacaat
     5581 actgtatgcg gcgcctgaaa gagtaaaata ttttccgtat taatccagcc gcctgttcca
     5641 aatctaatag atgcatcaat accttctgca gcaagattga caacgttatt atgcgttaaa
     5701 attctaatat caatccatgg attttcatta agaaacgcag taatccgagg aaggagccat
     5761 cctattgcaa aagtccctac gacagcaata gtaagcacct ctttaatctt accttcatgg
     5821 aatttatcca tcgcagaact catcatctgc aaagcttctg taatcgaagg caaaagatgc
     5881 atgccttcgt ctgtcaactg taacccacgc ggtactcttg taaaaagagc cacaccaagc
     5941 atttgttcaa gtgttctgac ctgctggctt acggcggcct ggcttacatg aagttctagt
     6001 ccggctttag taaaatttag atatcttgct gaggcttcaa aagctcgtaa tgcatttaaa
     6061 ggtaatctgg cacgcatggt aacccaaaga aaaagttatg tctgtataaa aattatattg
     6121 cttcaaagaa aaaataacaa ctgtataatc gctcgctgtt gaaagttaat acacgctatg
     6181 tcctagttaa tatagaaaat tgtatataag gtaaaaccat gtcacttaat gtaaagcaaa
     6241 gtagaatagc catcttgttt agctcttgtt taatttcaat atcatttttc tcacaggcca
     6301 atacgaaggg cattgatgag attaaaaacc ttgaaacaga tttcaatggc aggattggtg
     6361 tctacgcttt agacactggc tcgggtaaat cattttcgta cagagcaaat gaacgatttc
     6421 cattatgtag ttcttttaaa ggttttttag ctgctgctgt attaaaaggc tctcaagata
     6481 atcgacttaa tcttaatcag attgtgaatt ataatacaag aagtttagag ttccattcac
     6541 ccatcacaac taaatataaa gataatggaa tgtcattagg tgatatggct gctgctgctt
     6601 tacaatatag cgacaatggt gctactaata ttattcttga acgttatatc ggtggtccag
     6661 agggtatgac taaattcatg cggtcgattg gagatgaaga ttttagactc gatcgttggg
     6721 agttagatct aaacacagct attccaggcg atgagcgtga cacatctaca cctgcagcag
     6781 tagccaagag tctgaaaacc cttgctctgg gtaacatact tagtgaacat gaaaaggaaa
     6841 cctatcagac atggttaaag ggtaacacaa ccggtgcagc gcgtattcgt gctagcgtac
     6901 caagcgattg ggtagttggc gataaaactg gtagttgcgg agcatacggt acggcaaatg
     6961 attatgcggt agtctggcca aagaaccggg ctcctcttat aatttctgta tacacaacaa
     7021 aaaacgaaaa agaagccaag catgaggata aagtaatcgc agaagcttca agaattgcaa
     7081 ttgataacct taaataaacc taactgtagc catcatttca ttatgatggc ttaacttcat
     7141 ctaagagctc cgtcgacaag cttgcggccg cactcgagca ccaccaccac caccactgag
     7201 atccggctgc taacaaagcc cgaaaggaag ctgagttggc tgctgccacc gctgagcaat
     7261 aactagcata accccttggg gcctctaaac gggtcttgag gggttttttg ctgaaaggag
     7321 gaactatatc cggat
//
