LOCUS SYNPE194V 3728 bp DNA circular SYN 26-JUL-1993 DEFINITION pE194 cloning vector. ACCESSION L08862 VERSION L08862.1 KEYWORDS . SOURCE synthetic construct ORGANISM synthetic construct other sequences; artificial sequences. REFERENCE 1 (bases 1 to 3728) AUTHORS Gilbert,W. TITLE Obtained from VecBase 3.0 JOURNAL Unpublished COMMENT These data and their annotation were supplied to GenBank by Will Gilbert under the auspices of the GenBank Currator Program. pE194 - Cloning Vector ENTRY PE194 #TYPE DNA CIRCULAR TITLE pE194 - Cloning Vector DATE 11-JUN-1987 ACCESSION VB0131 SOURCE Staphylococcus aureus COLLECTION ATCC 37128 REFERENCE #number 1 #authors Gryczan T.J., Grandi G., Hahn J., Grandi R., Dubnau D. #journal Nucl. Acids Res. (1980) 8: 6081-6097 #title Conformational alteration of mRNA structure and the posttranscriptional regulation of erythromycin-induced drug resistance #comment (bases 1722 to 3074; complementary strand) REFERENCE #number 2 #authors Horinouchi S., Weisblum B. #journal Proc. Nat. Acad. Sci. USA (1980) 77: 7079-7083 #title Posttranscriptional modification of mRNA conformation: Mechanism that regulates erythromycin-induced resistance #comment (bases 1940 to 3386; complementary strand) REFERENCE #number 3 #authors Horinouchi S., Weisblum B. #journal Mol. Gen. Genet. (1981) 182: 341-348 #title The control region for erythromycin resistance: Free energy changes related to induction and mutation of constitutive expression #comment (bases 2629 to 2927; complementary strand) REFERENCE #number 4 #authors Horinouchi S., Weisblum B. #journal J. Bacteriol. (1982) 150: 804-814 #title Nucleotide sequence and functional map of pE194, a plasmid that specifies inducible resistance to macrolide, lincosamide, and streptogramin type B antibiotics #comment (bases 1 to 3727) REFERENCE #number 5 #authors Hahn J., Grandi G., Gryczan T.J., Dubnau D. #journal Mol. Gen. Genet. (1982) 186: 204-216 #title Translational attenuation of ermC: A deletion analysis #comment (bases 2858 to 3335; complementary strand) REFERENCE #number 6 #authors Pouwels P.H., Enger-Valk B.E., Brammar W.J. #book Cloning Vectors, Elsvier 1985 and supplements #comment vector IV-A-a-i-2 COMMENT fetched from GenBank(48) COMMENT this natural isolate from Staphylococcus aureus has been used as cloning vector in Bacillus subtilis and other gram-positive species. COMMENT from GenBank(48) [4] found five open reading frames, three of which (E, 1174-857; D, 1244-1594; and C, 1506-1859) lacked putative promoter regions. [3] proposes that the control region of the erythromycin resistance gene, located at the 5' end of the messenger, is transcribed along with the coding sequence. This region comprises a series of inverted complementary repeats that can redistribute and assume alternate double-stranded conformations. Activation of the messenger depends on the degree to which ribosomes, stalled by erythromycin, disrupt the secondary structure in a part of the control region that simultaneously codes for a small peptide and overlaps two of the inverted repeats. The result of the inverted repeat redistribution is the unmasking of a ribosome binding site for synthesis of the protein that mediates the resistance phenotype. [5] constructed deletants missing the ermC promoter and parts of the leader region. The ermC transcription was dependent on fusion to an upstream promoter. As the deletions extended deeper into the leader region, successively masking and unmasking sequences required for translation of the methylase, an alternation of high and low level methylase expression was observed. FEATURES from to/span description pept 2738 2004 (c) adenine methylase (ermC) pept 2857 2798 (c) leader peptide SITES conflict 1871 3 agg in [4]; aagag in [1] ->pept 2004 1 (c) ermC cds end pept<- 2738 1 (c) ermC cds start ->pept 2798 1 (c) leader pept cds end [1] pept<- 2857 1 (c) leader pept cds start [1] mRNA<- 2877 1 (c) ermC mRNA start [1] KEYWORDS adenine methylase \ erythromycin resistance \ leader peptide \ lincosamide resistance \ macrolide resistance \ streptogramin resistance CROSSREFERENCE #identical GenBank(48):PE1 PARENT Features of pE194 (3728 bp) original isolate, no homology to other sequences FEATURE 2004-2738 735-1 (c) erythromycin resistance protein (ermC) SELECTION #resistance erythromycin SUMMARY pE194 #length 3728 #checksum 1540. FEATURES Location/Qualifiers source 1..3728 /organism="synthetic construct" /mol_type="genomic DNA" /db_xref="taxon:32630" BASE COUNT 1363 a 460 c 721 g 1184 t ORIGIN 1 gatcagtaca agaaagatac tgtatttcat aaacaggaac tgcaagaagt taaggatgag 61 ttacagaagg caaataagca gttacagagt ggaatagagc atatgaggtc tacgaaaccc 121 tttgattatg aaaatgagcg tacaggtttg ttctctggac gtgaagagac tggtagaaag 181 atattaactg ctgatgaatt tgaacgcctg caagaaacaa tctcttctgc agaacggatt 241 gttgatgatt acgaaaatat taagagcaca gactattaca cagaaaatca agaattaaaa 301 aaacgtagag agagtttgaa agaagtagtg aatacatgga aagaggggta tcacgaaaaa 361 agtaaagagg ttaataaatt aaagcgagag aatgatagtt tgaatgagca gttgaatgta 421 tcagagaaat ttcaagctag tacagtgact ttatatcgtg ctgcgagggc gaatttccct 481 gggtttgaga aagggtttaa taggcttaaa gagaaattct ttaatgattc caaatttgag 541 cgtgtgggac agtttatgga tgttgtacag gataatgtcc agaaggtcga tagaaagcgt 601 gagaaacagc gtacagacga tttagagatg tagaggtact tttatgccga gaaaactttt 661 tgcgtgtgac agtccttaaa atatacttag agcgtaagcg aaagtagtag cgacagctat 721 taactttcgg ttgcaaagct ctaggatttt taatggacgc agcgcatcac acgcaaaaag 781 gaaattggaa taaatgcgaa atttgagatg ttaattaaag acctttttga ggtctttttt 841 tcttagattt ttggggttat ttaggggaga aaacataggg gggtactacg acctcccccc 901 taggtgtcca ttgtccattg tccaaacaaa taaataaata ttgggttttt aatgttaaaa 961 ggttgttttt tatgttaaag tgaaaaaaac agatgttggg aggtacagtg atagttgtag 1021 atagaaaaga agagaaaaaa gttgctgtta ctttaagact tacaacagaa gaaaatgaga 1081 tattaaatag aatcaaagaa aaatataata ttagcaaatc agatgcaacc ggtattctaa 1141 taaaaaaata tgcaaaggag gaatacggtg cattttaaac aaaaaaagat agacagcact 1201 ggcatgctgc ctatctatga ctaaattttg ttaagtgtat tagcaccgtt attatatcat 1261 gagcgaaaat gtaataaaag aaactgaaaa caagaaaaat tcaagaggac gtaattggac 1321 atttgtttta tatccagaat cagcaaaagc cgagtggtta gagtatttaa aagagttaca 1381 cattcaattt gtagtgtctc cattacatga tagggatact gatacagaag gtaggatgaa 1441 aaaagagcat tatcatattc tagtgatgta tgagggtaat aaatcttatg aacagataaa 1501 aataattaac agaagaattg aatgcgacta ttccgcagat tgcaggaagt gtgaaaggtc 1561 ttgtgagata tatgcttcac atggacgatc ctaataaatt taaatatcaa aaagaagata 1621 tgatagttta tggcggtgta gatgttgatg aattattaaa gaaaacaaca acagatagat 1681 ataaattaat taaagaaatg attgagttta ttgatgaaca aggaatcgta gaatttaaga 1741 gtttaatgga ttatgcaatg aagtttaaat ttgatgattg gttcccgctt ttatgtgata 1801 actcggcgta tgttattcaa gaatatataa aatcaaatcg gtataaatct gaccgataga 1861 ttttgaattt aggtgtcaca agacactctt ttttcgcacc agcgaaaact ggtttaagcc 1921 gactgcgcaa aagacataat cgattcacaa aaaataggca cacgaaaaac aagttaaggg 1981 atgcagttta tgcatccctt aacttactta ttaaataatt tatagctatt gaaaagagat 2041 aagaattgtt caaagctaat attgtttaaa tcgtcaattc ctgcatgttt taaggaattg 2101 ttaaattgat tttttgtaaa tattttcttg tattctttgt taacccattt cataacgaaa 2161 taattatact tttgtttatc tttgtgtgat attcttgatt tttttctact taatctgata 2221 agtgagctat tcactttagg tttaggatga aaatattctc ttggaaccat acttaatata 2281 gaaatatcaa cttctgccat taaaagtaat gccaatgagc gttttgtatt taataatctt 2341 ttagcaaacc cgtattccac gattaaataa atctcattag ctatactatc aaaaacaatt 2401 ttgcgtatta tatccgtact tatgttataa ggtatattac catatatttt ataggattgg 2461 tttttaggaa atttaaactg caatatatcc ttgtttaaaa cttggaaatt atcgtgatca 2521 acaagtttat tttctgtagt tttgcataat ttatggtcta tttcaatggc agttacgaaa 2581 ttacacctct ttactaattc aagggtaaaa tggccttttc ctgagccgat ttcaaagata 2641 ttatcatgtt catttaatct tatatttgtc attattttat ctatattatg ttttgaagta 2701 ataaagtttt gactgtgttt tatatttttc tcgttcatta taaccctctt taatttggtt 2761 atatgaattt tgcttattaa cgattcatta taaccactta ttttttgttt ggttgataat 2821 gaactgtgct gattacaaaa atactaaaaa tgcccatatt ttttcctcct tataaaatta 2881 gtataattat agcacgagct ctgataaata tgaacatgat gagtgatcgt taaatttata 2941 ctgcaatcgg atgcgattat tgaataaaag atatgagaga tttatctaat ttcttttttc 3001 ttgtaaaaaa agaaagttct taaaggtttt atagttttgg tcgtagagca cacggtttaa 3061 cgacttaatt acgaagtaaa taagtctagt gtgttagact ttatgaaatc tatatacgtt 3121 tatatatatt tattatccgg aggtgtagca tgtctcattc aattttgagg gttgccagag 3181 ttaaaggatc aagtaataca aacgggatac aaagacataa tcaaagagag aataaaaact 3241 ataataataa agacataaat catgaggaaa catataaaaa ttatgatttg attaacgcac 3301 aaaatataaa gtataaagat aaaattgatg aaacgattga tgagaattat tcagggaaac 3361 gtaaaattcg gtcagatgca attcgacatg tggacggact ggttacaagt gataaagatt 3421 tctttgatga tttaagcgga gaagaaatag aacgattttt taaagatagc ttggagtttc 3481 tagaaaatga atacggtaag gaaaatatgc tgtatgcgac tgtccatctg gatgaaagag 3541 tcccacatat gcactttggt tttgtccctt taacagagga cgggagattg tctgcaaaag 3601 aacagttagg caacaagaaa gactttactc aattacaaga tagatttaat gagtatgtga 3661 atgagaaagg ttatgaactt gaaagaggca cgtccaaaga ggttacagaa cgagaacata 3721 aagcgatg //