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 //