LOCUS       40924_43668        3147 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Cloning vector pTn7-M, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3147)
  AUTHORS   Zobel S, Benedetti I, Eisenbach L, de Lorenzo V, Wierckx N, Blank 
            LM.
  TITLE     A Tn7-based device for calibrated heterologous gene expression in 
            Pseudomonas putida
  JOURNAL   ACS Synth Biol (2015) In press
  PUBMED    26133359
REFERENCE   2  (bases 1 to 3147)
  AUTHORS   Zobel S, Benedetti I, Eisenbach L, de Lorenzo V, Wierckx N, Blank 
            LM.
  TITLE     Direct Submission
  JOURNAL   Submitted (28-MAY-2015) System Biology, Centro Nacional de 
            Biotecnologia, C/Darwin 3, Madrid 28049, Spain
REFERENCE   3  (bases 1 to 3147)
  AUTHORS   Zobel S, Benedetti I, Eisenbach L, de Lorenzo V, Wierckx N, Blank 
            LM.
  TITLE     Direct Submission
  JOURNAL   Submitted (13-NOV-2015) System Biology, Centro Nacional de 
            Biotecnologia, C/Darwin 3, Madrid 28049, Spain
REFERENCE   4  (bases 1 to 3147)
  TITLE     Direct Submission
REFERENCE   5  (bases 1 to 3147)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "ACS Synth 
            Biol (2015) In press"
COMMENT     SGRef: number: 2; type: "Journal Article"; journalName: "Submitted 
            (28-MAY-2015) System Biology, Centro Nacional de Biotecnologia, 
            C/Darwin 3, Madrid 28049, Spain"
COMMENT     SGRef: number: 3; type: "Journal Article"; journalName: "Submitted 
            (13-NOV-2015) System Biology, Centro Nacional de Biotecnologia, 
            C/Darwin 3, Madrid 28049, Spain"
COMMENT     SGRef: number: 4; type: "Journal Article"
COMMENT     ##Assembly-Data-START##
            Sequencing Technology :: Sanger dideoxy sequencing 
            ##Assembly-Data-END##
            On Nov 13, 2015 this sequence version replaced KR920750.1.
FEATURES             Location/Qualifiers
     source          1..3147
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    56..112
                     /label=MCS
                     /note="pUC18/19 multiple cloning site"
     terminator      155..249
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     misc_feature    254..452
                     /label=Tn7R
                     /note="Tn7R"
     CDS             562..1374
                     /codon_start=1
                     /label=KanR
                     /note="aminoglycoside phosphotransferase"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     oriT            1533..1641
                     /label=oriT
                     /note="incP origin of transfer"
     rep_origin      1660..2048
                     /label=R6K gamma ori
                     /note="gamma replication origin from E. coli plasmid R6K;
                     requires the R6K initiator protein pi for replication"
     mobile_element  2060..2225
                     /label=Tn7L
                     /note="mini-Tn7 element (left end of the Tn7 transposon)"
     terminator      complement(2239..2325)
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB
                     gene"
     CDS             complement(2394..2924)
                     /codon_start=1
                     /label=GmR
                     /note="gentamycin acetyltransferase"
                     /translation="MLRSSNDVTQQGSRPKTKLGGSSMGIIRTCRLGPDQVKSMRAALD
                     LFGREFGDVATYSQHQPDSDYLGNLLRSKTFIALAAFDQEAVVGALAAYVLPKFEQPRS
                     EIYIYDLAVSGEHRRQGIATALINLLKHEANALGAYVIYVQADYGDDPAVALYTKLGIR
                     EEVMHFDIDPSTAT"
     promoter        complement(3113..3141)
                     /label=Pc promoter
                     /note="class 1 integron promoter"
ORIGIN
        1 ttaattaaag cggataacaa tttcacacag gaggccgcct aggccgcggc cgcgcgaatt
       61 cgagctcggt acccggggat cctctagagt cgacctgcag gcatgcaagc ttgcggccgc
      121 gtcgtgactg ggaaaaccct ggcgactagt cttggactcc tgttgataga tccagtaatg
      181 acctcagaac tccatctgga tttgttcaga acgctcggtt gccgccgggc gttttttatt
      241 ggtgagaatc cagccacgcc cctctttaat acgacgggca atttgcactt cagaaaatga
      301 agagtttgct ttagccataa caaaagtcca gtatgctttt tcacagcata actggactga
      361 tttcagttta caactattct gtctagttta agactttatt gtcatagttt agatctattt
      421 tgttcagttt aagactttat tgtccgccca caatttaaat ttgtgtctca aaatctctga
      481 tgttacattg cacaagataa aaatatatca tcatgaacaa taaaactgtc tgcttacata
      541 aacagtaata caaggggtgt tatgagccat attcagcgtg aaacgagctg tagccgtccg
      601 cgtctgaaca gcaacatgga tgcggatctg tatggctata aatgggcgcg tgataacgtg
      661 ggtcagagcg gcgcgaccat ttatcgtctg tatggcaaac cggatgcgcc ggaactgttt
      721 ctgaaacatg gcaaaggcag cgtggcgaac gatgtgaccg atgaaatggt gcgtctgaac
      781 tggctgaccg aatttatgcc gctgccgacc attaaacatt ttattcgcac cccggatgat
      841 gcgtggctgc tgaccaccgc gattccgggc aaaaccgcgt ttcaggtgct ggaagaatat
      901 ccggatagcg gcgaaaacat tgtggatgcg ctggccgtgt ttctgcgtcg tctgcatagc
      961 attccggtgt gcaactgccc gtttaacagc gatcgtgtgt ttcgtctggc ccaggcgcag
     1021 agccgtatga acaacggcct ggtggatgcg agcgattttg atgatgaacg taacggctgg
     1081 ccggtggaac aggtgtggaa agaaatgcat aaactgctgc cgtttagccc ggatagcgtg
     1141 gtgacccacg gcgattttag cctggataac ctgattttcg atgaaggcaa actgattggc
     1201 tgcattgatg tgggccgtgt gggcattgcg gatcgttatc aggatctggc cattctgtgg
     1261 aactgcctgg gcgaatttag cccgagcctg caaaaacgtc tgtttcagaa atatggcatt
     1321 gataatccgg atatgaacaa actgcaattt catctgatgc tggatgaatt tttctaataa
     1381 ttaattggac cgcggtcctt ttccgctgca taaccctgct tcggggtcat tatagcgatt
     1441 ttttcggtat atccatcctt tttcgcacga tatacaggat tttgccaaag ggttcgtgta
     1501 gactttcctt ggtgtatcca acggcgtcag ccgggcagga taggtgaagt aggcccaccc
     1561 gcgagcgggt gttccttctt cactgtccct tattcgcacc tggcggtgct caacgggaat
     1621 cctgctctgc gaggctggcc gtaggccggc cggcgcgccg atctgaagat cagcagttca
     1681 acctgttgat agtacgtact aagctctcat gtttcacgta ctaagctctc atgtttaacg
     1741 tactaagctc tcatgtttaa cgaactaaac cctcatggct aacgtactaa gctctcatgg
     1801 ctaacgtact aagctctcat gtttcacgta ctaagctctc atgtttgaac aataaaatta
     1861 atataaatca gcaacttaaa tagcctctaa ggttttaagt tttataagaa aaaaaagaat
     1921 atataaggct tttaaagcct ttaaggttta acggttgtgg acaacaagcc agggatgtaa
     1981 cgcactgaga agcccttaga gcctctcaaa gcaattttga gtgacacagg aacacttaac
     2041 ggctgacatg gggcgcgcct gtgggcggac aaaatagttg ggaactggga ggggtggaaa
     2101 tggagttttt aaggattatt tagggaagag tgacaaaata gatgggaact gggtgtagcg
     2161 tcgtaagcta atacgaaaat taaaaatgac aaaatagttt ggaactagat ttcacttatc
     2221 tggtttctag ggcggcggat ttgtcctact caggagagcg ttcaccgaca aacaacagat
     2281 aaaacgaaag gcccagtctt tcgactgagc ctttcgtttt atttgatgcc gacgtccaat
     2341 ttaccgaaca actccgcggc cgggaagccg atctcggctt gaacgaattg ttaggtggcg
     2401 gtacttgggt cgatatcaaa gtgcatcact tcttcccgta tgcccaactt tgtatagaga
     2461 gccactgcgg gatcgtcacc gtaatctgct tgcacgtaga tcacataagc accaagcgcg
     2521 ttggcctcat gcttgaggag attgatgagc gcggtggcaa tgccctgcct ccggtgctct
     2581 ccggagactg cgagatcata gatatagatc tcactacgcg gctgctcaaa cttgggcaga
     2641 acgtaagccg cgagagcgcc aacaaccgct tcttggtcga aggcagcaag cgcgatgaat
     2701 gtcttactac ggagcaagtt cccgaggtaa tcggagtccg gctgatgttg ggagtaggtg
     2761 gctacgtctc cgaactcacg accgaaaaga tcaagagcag cccgcatgga tttgacttgg
     2821 tcagggccga gcctacatgt gcgaatgatg cccatacttg agccacctaa ctttgtttta
     2881 gggcgactgc cctgctgcgt aacatcgttg ctgctgcgta acatcgttgc tgctccataa
     2941 catcaaacat cgacccacgg cgtaacgcgc ttgctgcttg gatgcccgag gcataggctg
     3001 tacaaaaaaa cagtcataac aagccatgaa aaccgccact gcgccgttac caccgctgcg
     3061 ttcggtcaag gttctggacc agttgcgtga gcgcatacgc tacttgcatt acagtttacg
     3121 aaccgaacag gcttatgtca agacgtc
//
