LOCUS       62056_18190        5361 bp DNA     circular SYN 17-JUL-2019
DEFINITION  Cloning vector pNPTS138 plasmid pNPTS138, complete sequence.
ACCESSION   MK533795
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5361)
  AUTHORS   Ferro JA, Facincani AP, Tezza RD, Varani AM, Ferreira H.
  TITLE     The nucleotide sequence of the plasmid pNPTS138
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 5361)
  AUTHORS   Ferro JA, Facincani AP, Tezza RD, Varani AM, Ferreira H.
  TITLE     Direct Submission
  JOURNAL   Submitted (18-FEB-2019) Departamento de Tecnologia, Univ. Estadual 
            Paulista 'Julio de Mesquita Filho' - UNESP-Jaboticabal, Via de 
            Acesso Prof.Paulo Donato Castellane s/n, Jaboticabal, SP 14884-900, 
            Brazil
REFERENCE   3  (bases 1 to 5361)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: 
            "Unpublished"
COMMENT     SGRef: number: 2; type: "Journal Article"; journalName: "Submitted 
            (18-FEB-2019) Departamento de Tecnologia, Univ. Estadual Paulista 
            'Julio de Mesquita Filho' - UNESP-Jaboticabal, Via de Acesso 
            Prof.Paulo Donato Castellane s/n, Jaboticabal, SP 14884-900, Brazil"
COMMENT     ##Assembly-Data-START##
            Sequencing Technology :: Sanger dideoxy sequencing 
            ##Assembly-Data-END##
            The plasmid was a gift from Prof. Lucy Shapiro, Department of 
            Developmental Biology, Stanford University School of Medicine 
            Beckman Center 279 Campus Drive, B300 Stanford, CA 94305.
FEATURES             Location/Qualifiers
     source          1..5361
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(158..174)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    complement(182..198)
                     /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(206..236)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    complement(251..272)
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     rep_origin      complement(333..922)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     oriT            1140..1249
                     /label=oriT
                     /note="incP origin of transfer"
     CDS             1282..1650
                     /codon_start=1
                     /label=traJ
                     /note="oriT-recognizing protein"
                     /translation="MADETKPTRKGSPPIKVYCLPDERRAIEEKAAAAGMSLSAYLLAV
                     GQGYKITGVVDYEHVRELARINGDLGRLGGLLKLWLTDDPRTARFGDATILALLAKIEE
                     KQDELGKVMMGVVRPRAEP"
     rep_origin      1824..2275
                     /label=M13 ori
                     /note="M13 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     CDS             2371..3183
                     /codon_start=1
                     /label=KanR
                     /note="aminoglycoside phosphotransferase"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKFLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     promoter        3258..3685
                     /label=sacB promoter
                     /note="sacB promoter and control region"
     CDS             3686..5104
                     /codon_start=1
                     /label=SacB
                     /note="secreted levansucrase that renders bacterial growth 
                     sensitive to sucrose"
                     /translation="MNIKKFAKQATVLTFTTALLAGGATQAFAKETNQKPYKETYGISH
                     ITRHDMLQIPEQQKNEKYQVPEFDSSTIKNISSAKGLDVWDSWPLQNADGTVANYHGYH
                     IVFALAGDPKNADDTSIYMFYQKVGETSIDSWKNAGRVFKDSDKFDANDSILKDQTQEW
                     SGSATFTSDGKIRLFYTDFSGKHYGKQTLTTAQVNVSASDSSLNINGVEDYKSIFDGDG
                     KTYQNVQQFIDEGNYSSGDNHTLRDPHYVEDKGHKYLVFEANTGTEDGYQGEESLFNKA
                     YYGKSTSFFRQESQKLLQSDKKRTAELANGALGMIELNDDYTLKKVMKPLIASNTVTDE
                     IERANVFKMNGKWYLFTDSRGSKMTIDGITSNDIYMLGYVSNSLTGPYKPLNKTGLVLK
                     MDLDPNDVTFTYSHFAVPQAKGNNVVITSYMTNRGFYADKQSTFAPSFLLNIKGKKTSV
                     VKDSILEQGQLTVNK"
     primer_bind     5316..5332
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
ORIGIN
        1 tagagggtcg acgcatgcct gtacatccgg agacgcgtca cggccgaagc tagcgaattc
       61 gtggatccag atatcctgca gagaagcttg gcgccagccg gcttcaattg cacgggcccc
      121 actagtgagt cgtattacgt agcttggcgt aatcatggtc atagctgttt cctgtgtgaa
      181 attgttatcc gctcacaatt ccacacaaca tacgagccgg aagcataaag tgtaaagcct
      241 ggggtgccta atgagtgagc taactcacat tacatgtgag caaaaggcca gcaaaaggcc
      301 aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc ccctgacgag
      361 catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact ataaagatac
      421 caggcgtttc cccctggaag ctccctcgtg cgctcttcct gttccgaccc tgccgcttac
      481 cggatacctg tccgcctttc tcccttcggg aagcgtggcg ctttctcata gctcacgctg
      541 taggtatctc agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc
      601 cgttcagccc gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag
      661 acacgactta tcgccactgg cagcagccac tggtaacagg attagcagag cgaggtatgt
      721 aggcggtgct acagagttct tgaagtggtg gcctaactac ggctacacta gaagaacagt
      781 atttggtatc tgcgctctgc tgaagccagt taccttcgga aaaagagttg gtagctcttg
      841 atccggcaaa caaaccaccg ctggtagcgg tggttttttt gtttgcaagc agcagattac
      901 gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt tctacggggt ctgacgctca
      961 gtggaacgaa aactcacgtt aagggatttt ggtcatggac ggatcttttc cgctgcataa
     1021 ccctgcttcg gggtcattat agcgattttt tcggtatatc catccttttt cgcacgatat
     1081 acaggatttt gccaaagggt tcgtgtagac tttccttggt gtatccaacg gcgtcagccg
     1141 ggcaggatag gtgaagtagg cccacccgcg agcgggtgtt ccttcttcac tgtcccttat
     1201 tcgcacctgg cggtgctcaa cgggaatcct gctctgcgag gctggccggc taccgccggc
     1261 gtaacagatg agggcaagcg gatggctgat gaaaccaagc caaccaggaa gggcagccca
     1321 cctatcaagg tgtactgcct tccagacgaa cgaagagcga ttgaggaaaa ggcggcggcg
     1381 gccggcatga gcctgtcggc ctacctgctg gccgtcggcc agggctacaa aatcacgggc
     1441 gtcgtggact atgagcacgt ccgcgagctg gcccgcatca atggcgacct gggccgcctg
     1501 ggcggcctgc tgaaactctg gctcaccgac gacccgcgca cggcgcggtt cggtgatgcc
     1561 acgatcctcg ccctgctggc gaagatcgaa gagaagcagg acgagcttgg caaggtcatg
     1621 atgggcgtgg tccgcccgag ggcagagcca tgactttttt agccgctaaa acggccgggg
     1681 ggtgcgcgtg attgccaagc acgtccccat gcgctccatc aagaagagcg acttcgcgga
     1741 gctggtgaag tacatcaccg acgagcaagg caagaccgat cccccatgag attatcaaaa
     1801 aggatcttca cctagatcct tttacgcgcc ctgtagcggc gcattaagcg cggcgggtgt
     1861 ggtggttacg cgcagcgtga ccgctacact tgccagcgcc ctagcgcccg ctcctttcgc
     1921 tttcttccct tcctttctcg ccacgttcgc tttccccgtc aagctctaaa tcgggggctc
     1981 cctttagggt tccgatttag tgctttacgg cacctcgacc ccaaaaaact tgatttgggt
     2041 gatggttcac gtagtgggcc atcgccctga tagacggttt ttcgcccttt gacgttggag
     2101 tccacgttct ttaatagtgg actcttgttc caaactggaa caacactcaa ccctatctcg
     2161 ggctattctt ttgatttata agggattttg ccgatttcgg cctattggtt aaaaaatgag
     2221 ctgatttaac aaaaatttaa cgcgaatttt aacaaaatat taacgtttac aatttctaga
     2281 gcttggccgg gttacattgc acaagataaa aatatatcat catgaacaat aaaactgtct
     2341 gcttacataa acagtaatac aaggggtgtt atgagccata ttcaacggga aacgtcttgc
     2401 tcgaggccgc gattaaattc caacatggat gctgatttat atgggtataa atgggctcgc
     2461 gataatgtcg ggcaatcagg tgcgacaatc tatcgattgt atgggaagcc cgatgcgcca
     2521 gagttgtttc tgaaacatgg caaaggtagc gttgccaatg atgttacaga tgagatggtc
     2581 agactaaact ggctgacgga atttatgcct cttccgacca tcaagcattt tatccgtact
     2641 cctgatgatg catggttact caccactgcg atccccggta aaacagcatt ccaggtatta
     2701 gaagaatatc ctgattcagg tgaaaatatt gttgatgcgc tggcagtgtt cctgcgccgg
     2761 ttgcattcga ttcctgtttg taattgtcct tttaacagcg atcgcgtatt tcgtctcgct
     2821 caggcgcaat cacgaatgaa taacggtttg gttgatgcga gtgattttga tgacgagcgt
     2881 aatggttggc ctgttgaaca agtctggaaa gaaatgcata agtttttgcc attctcaccg
     2941 gattcagtcg tcactcatgg tgatttctca cttgataacc ttatttttga cgaggggaaa
     3001 ttaataggtt gtattgatgt tggacgagtc ggaatcgcag accgatacca ggatcttgcc
     3061 atcctatgga actgcctcgg tgagttttct ccttcattac agaaacggct ttttcaaaaa
     3121 tatggtattg ataatcctga tatgaataaa ttgcagtttc atttgatgct cgatgagttt
     3181 ttctaatcag aattggttaa ttggttgtaa cactggcaga gcattacgct gacttgacgg
     3241 gacggcgccg gccaagctct attatttagt gaaatgagat attatgatat tttctgaatt
     3301 gtgattaaaa aggcaacttt atgcccatgc aacagaaact ataaaaaata cagagaatga
     3361 aaagaaacag atagattttt tagttcttta ggcccgtagt ctgcaaatcc ttttatgatt
     3421 ttctatcaaa caaaagagga aaatagacca gttgcaatcc aaacgagagt ctaatagaat
     3481 gaggtcgaaa agtaaatcgc gcgggtttgt tactgataaa gcaggcaaga cctaaaatgt
     3541 gtaaagggca aagtgtatac tttggcgtca ccccttacat attttaggtc tttttttatt
     3601 gtgcgtaact aacttgccat cttcaaacag gagggctgga agaagcagac cgctaacaca
     3661 gtacataaaa aaggagacat gaacgatgaa catcaaaaag tttgcaaaac aagcaacagt
     3721 attaaccttt actaccgcac tgctggcagg aggcgcaact caagcgtttg cgaaagaaac
     3781 gaaccaaaag ccatataagg aaacatacgg catttcccat attacacgcc atgatatgct
     3841 gcaaatccct gaacagcaaa aaaatgaaaa atatcaagtt cctgagttcg attcgtccac
     3901 aattaaaaat atctcttctg caaaaggcct ggacgtttgg gacagctggc cattacaaaa
     3961 cgctgacggc actgtcgcaa actatcacgg ctaccacatc gtctttgcat tagccggaga
     4021 tcctaaaaat gcggatgaca catcgattta catgttctat caaaaagtcg gcgaaacttc
     4081 tattgacagc tggaaaaacg ctggccgcgt ctttaaagac agcgacaaat tcgatgcaaa
     4141 tgattctatc ctaaaagacc aaacacaaga atggtcaggt tcagccacat ttacatctga
     4201 cggaaaaatc cgtttattct acactgattt ctccggtaaa cattacggca aacaaacact
     4261 gacaactgca caagttaacg tatcagcatc agacagctct ttgaacatca acggtgtaga
     4321 ggattataaa tcaatctttg acggtgacgg aaaaacgtat caaaatgtac agcagttcat
     4381 cgatgaaggc aactacagct caggcgacaa ccatacgctg agagatcctc actacgtaga
     4441 agataaaggc cacaaatact tagtatttga agcaaacact ggaactgaag atggctacca
     4501 aggcgaagaa tctttattta acaaagcata ctatggcaaa agcacatcat tcttccgtca
     4561 agaaagtcaa aaacttctgc aaagcgataa aaaacgcacg gctgagttag caaacggcgc
     4621 tctcggtatg attgagctaa acgatgatta cacactgaaa aaagtgatga aaccgctgat
     4681 tgcatctaac acagtaacag atgaaattga acgcgcgaac gtctttaaaa tgaacggcaa
     4741 atggtatctg ttcactgact cccgcggatc aaaaatgacg attgacggca ttacgtctaa
     4801 cgatatttac atgcttggtt atgtttctaa ttctttaact ggcccataca agccgctgaa
     4861 caaaactggc cttgtgttaa aaatggatct tgatcctaac gatgtaacct ttacttactc
     4921 acacttcgct gtacctcaag cgaaaggaaa caatgtcgtg attacaagct atatgacaaa
     4981 cagaggattc tacgcagaca aacaatcaac gtttgcgccg agcttcctgc tgaacatcaa
     5041 aggcaagaaa acatctgttg tcaaagacag catccttgaa caaggacaat taacagttaa
     5101 caaataaaaa cgcaaaagaa aatgccgata acaaaaaaaa gcccgccgaa gcgggcttta
     5161 ttaccaagcg aagcgccatt cgccattcag gctgcgcaac tgttgggaag ggcgatcggt
     5221 gcgggcctct tcgctattac gccagctggc gaaaggggga tgtgctgcaa ggcgattaag
     5281 ttgggtaacg ccagggtttt cccagtcacg acgttgtaaa acgacggcca gtccgtaata
     5341 cgactcactt aaggccttga c
//
