LOCUS AJ606312 8798 bp DNA circular SYN 14-NOV-2006
DEFINITION Cloning vector pMR10 complete sequence.
ACCESSION AJ606312
VERSION AJ606312.1
KEYWORDS kanamaycin resistance; kanamycin phosphotransferase; lacZ gene;
parA gene; parB gene; parC gene; parD gene; parE gene; plasmid
replication initiation protein; plasmid stabilisation protein;
resolvase; trfA gene.
SOURCE Cloning vector pMR10
ORGANISM Cloning vector pMR10
other sequences; artificial sequences; vectors.
REFERENCE 1 (bases 1 to 8798)
AUTHORS Bourniquel A.A.
JOURNAL Submitted (11-NOV-2003) to the INSDC. Bourniquel A.A., Dept. of
Molecular Microbiology, Biozentrum der Universitaet Basel,
Klingelbergstr. 50-70, CH-4056 Basel, SWITZERLAND.
REFERENCE 2
AUTHORS Bourniquel A.A., Hamburger F., Mollet B., Bickle T.A.
TITLE Overexpression, purification and characterisation of type I
restriction enzymes.
JOURNAL Unpublished.
REFERENCE 3
AUTHORS Mohr C., Roberts R.
TITLE pMR10, a RK2-based low copy cloning vector for Escherichia coli.
JOURNAL Unpublished.
FEATURES Location/Qualifiers
source 1..8798
/organism="Cloning vector pMR10"
/focus
/lab_host="Escherichia coli"
/mol_type="other DNA"
/note="Gram-negative broad host range RK2-based low copy
number cloning vector"
/db_xref="taxon:255043"
source 251..4112
/organism="Plasmid RP4"
/plasmid="RP4"
/mol_type="other DNA"
/note="Birmingham IncP-alpha plasmid"
/db_xref="taxon:2503"
source 4113..5416
/organism="Cosmid vector pRR54"
/mol_type="other DNA"
/note="derived from Klebsiella aeruginosa gram negative
broad host range Birmingham IncP-alpha plasmid RK2"
/db_xref="taxon:232937"
source 5417
/organism="synthetic construct"
/mol_type="other DNA"
/db_xref="taxon:32630"
source 5418..5828
/organism="Cosmid vector pRR54"
/mol_type="other DNA"
/note="derived from Klebsiella aeruginosa gram negative
broad host range Birmingham IncP-alpha plasmid RK2"
/db_xref="taxon:232937"
source 5829..8575
/organism="Cosmid vector pRR54"
/mol_type="other DNA"
/note="derived from Klebsiella aeruginosa gram negative
broad host range Birmingham IncP-alpha plasmid RK2"
/db_xref="taxon:232937"
CDS 1..>21
/gene="lacZ"
/product="LacZ protein"
/note="truncated"
/protein_id="CAE54487.1"
/translation="MTMITPS"
misc_feature 17..134
/note="polylinker sequence"
/experiment="experimental evidence, no additional details
recorded"
regulatory 298..371
/note="T4 terminator"
/regulatory_class="terminator"
oriT 430..758
/note="IncP origin of transfer"
CDS 888..1703
/product="kanamycin phosphotransferase"
/function="kanamaycin resistance"
/experiment="experimental evidence, no additional details
recorded"
/protein_id="CAE54488.1"
/translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLY
GKPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIP
GKTAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGL
VDASDFDDERNGWPVEQVWKEMHKFLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVG
RVGIADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
CDS complement(2386..2697)
/gene="parE"
/product="plasmid stabilisation protein ParE"
/protein_id="CAE54489.1"
/translation="MTAYILTAEAEADLRGIIRYTRREWGAAQVRRYIAKLEQGIARL
AAGEGPFKDMSELFPALRMARCEHHYVFCLPRAGEPALVVAILHERMDLMTRLADRLK
G"
CDS complement(2694..2945)
/gene="parD"
/product="plasmid stabilisation protein ParD"
/protein_id="CAE54490.1"
/translation="MSRLTIDMTDQQHQSLKALAALQGKTIKQYALERLFPGDADADQ
AWQELKTMLGNRINDGLAGKVSTKSVGEILDEELSGDRA"
CDS 3096..3389
/gene="parC"
/product="plasmid stabilisation protein ParC"
/protein_id="CAE54491.1"
/translation="MGIPNLTKGDAMRAAKHYRQLLSIDFNIEALAFVPGPDGTRGRR
IHVLGREVRDRPGLVEYLSPAFGSRVALDGYCKANFDAVLHLAYPDHQQWGHA"
CDS 3386..3919
/gene="parB"
/product="plasmid stabilisation protein ParB"
/protein_id="CAE54492.1"
/translation="MKRRSYAMLRAAAALAVLVVASPAWAELRGEVVRIIDGDTIDVL
VDKQPVRVRLVDIDAPEKRQAFGERARQALAGMVFRRHVLVDEKDTDRYGRTLGTVWV
NMELASRPPQPRNVNAAMVHQGMAWAYRFHGRAADPEMLRLEQEARGKRVGLWSDPHA
VEPWKWRRESNNRRDEG"
CDS 3916..4677
/gene="parA"
/product="plasmid stabilisation protein ParA"
/function="resolvase"
/protein_id="CAE54493.1"
/translation="MKVARIYLRASTDEQNLERQESLVAATRAAGYYVAGIYREKASG
ARADRPELLRMIADLQPGEVVVESLVAATRAAGYYVAGIYREKASGARADRPELLRMI
ADLQPGEVVVAEKIDRISRLPLAEAERLVASIRAKGAKLAVPGVVDLSELAAEANGVA
KIVLESVQDMLLKLALQMARDDYEDRRERQRQGVQLAKAAGRYTGRKRDAGMHDRIIT
LRSGGSSIAKTAKLVGCSPSQVKRVWAAWNAQQQK"
rep_origin 6004..6715
/direction=RIGHT
/note="oriV, origin of vegetative replication"
regulatory 7229..7278
/gene="aph"
/note="neo/kan resistance promoter"
/regulatory_class="promoter"
CDS 7386..8534
/gene="trfA"
/product="plasmid replication initiation protein TrfA"
/protein_id="CAE54494.1"
/translation="MNRTFDRKAYRQELIDAGFSAEDAETIASRTVMRAPRETFQSVG
SMVQQATAKIERDSVQLAPPALPAPSAAVERSRRLEQEAAGLAKSMTIDTRGTMTTKK
RKTAGEDLAKQVSEAKQAALLKHTKQQIKEMQLSLFDIAPWPDTMRAMPNDTARSALF
TTRNKKIPREALQNKVIFHVNKDVKITYTGVELRADDDELVWQQVLEYAKRTPIGEPI
TFTFYELCQDLGWSINGRYYTKAEECLSRLQATAMGFTSDRVGHLESVSLLHRFRVLD
RGKKTSRCQVLIDEEIVVLFAGDHYTKFIWEKYRKLSPTARRMFDYFSSHREPYPLKL
ETFRLMCGSDSTRVKKWREQVGEACEELRGSGLVEHAWVNDDLVHCKR"
BASE COUNT 1839 a 2556 c 2647 g 1756 t
ORIGIN
1 atgaccatga ttacgccaag cttccatggg atatcgagat ctcctgcaga gctctagagt
61 cgagactagt ctcgacgggc ccggtacccc ctcgaggggg ccgcacttaa gttacgcgtg
121 gatccccggg aattcactgg ccgtcgtttt acaacgtcgt gactgggaaa accctggcgt
181 tacccaactt aatcgccttg cagcacatcc ccctttcgcc agctggcgta atagcgaaga
241 ggcccgcacc gatcgccctt cccaacagtt gcgcagcctg aatggcgaat ggcgatagat
301 ccggtggatg accttttgaa tgacctttaa tagattatat tactaattaa ttggggaccc
361 tagaggtccc cttttttatt ttaaaaattt tttcacaaaa cggtttacaa gcataaagct
421 cggacggatc ttttccgctg cataaccctg cttcggggtc attatagcga ttttttcggt
481 atatccatcc tttttcgcac gatatacagg attttgccaa agggttcgtg tagactttcc
541 ttggtgtatc caacggcgtc agccgggcag gataggtgaa gtaggcccac ccgcgagcgg
601 gtgttccttc ttcactgtcc cttattcgca cctggcggtg ctcaacggga atcctgctct
661 gcgaggctgg ccggctaccg ccggcgtaac agatgagggc aagcggatgg ctgatgaaac
721 caagccaacc aggaagggca gcccacctat caaggtgtcg atgcaggggg gggggggaaa
781 gccacgttgt gtctcaaaat ctctgatgtt acattgcaca agataaaaat atatcatcat
841 gaacaataaa actgtctgct tacataaaca gtaatacaag gggtgttatg agccatattc
901 aacgggaaac gtcttgctca aggccgcgat taaattccaa catggatgct gatttatatg
961 ggtataaatg ggctcgcgat aatgtcgggc aatcaggtgc gacaatctat cgattgtatg
1021 ggaagcccga tgcgccagag ttgtttctga aacatggcaa aggtagcgtt gccaatgatg
1081 ttacagatga gatggtcaga ctaaactggc tgacggaatt tatgcctctt ccgaccatca
1141 agcattttat ccgtactcct gatgatgcat ggttactcac cactgcgatc cccggaaaaa
1201 cagcattcca ggtattagaa gaatatcctg attcaggtga aaatattgtt gatgcgctgg
1261 cagtgttcct gcgccggttg cattcgattc ctgtttgtaa ttgtcctttt aacagcgatc
1321 gcgtatttcg tctcgctcag gcgcaatcac gaatgaataa cggtttggtt gatgcgagtg
1381 attttgatga cgagcgtaat ggctggcctg ttgaacaagt ctggaaagaa atgcataagt
1441 ttttgccatt ctcaccggat tcagtcgtca ctcatggtga tttctcactt gataacctta
1501 tttttgacga ggggaaatta ataggttgta ttgatgttgg acgagtcgga atcgcagacc
1561 gataccagga tcttgccatc ctatggaact gcctcggtga gttttctcct tcattacaga
1621 aacggctttt tcaaaaatat ggtattgata atcctgatat gaataaattg cagtttcatt
1681 tgatgctcga tgagtttttc taatcagaat tggttaattg gttgtaacac tggcagagca
1741 ttacgctgac ttgacgggac ggcggctttg ttgaataaat cgaacttttg ctgagttgaa
1801 ggatcagatc acgcatcttc ccgacaacgc agaccgttcc gtggcaaagc aaaagttcaa
1861 aatcaccaac tggtccacct acaacaaagc tctcatcaac cgtggctccc tcactttctg
1921 gctggatgat ggggcgattc aggcctggta tgagtcagca acaccttctt cacgaggcag
1981 acctcagacg gtatcggatc gatcccccga tgtgtagcag tggcggacca tataggcaga
2041 tcagaaggcg cggttctcct acatgttcgc cggtgaacgc gttgaggaag ccgggcagtg
2101 cctcggcaaa atccttgcgt gtagacaaga catctgcgta gcagttgtcc tcaacaacga
2161 tgtcgaaatc caaatcggag tgctcatcga gtcctccgtg aacgtaagag ccgccgatca
2221 gaagagcgcg gaagcgaaca tcggaagcga ccgcatcgcg gatgcggttc aagaaagttg
2281 catgagcttg tggaagtgtg ctgagcataa atgattctcc tagctgttct ttgggtaagt
2341 acgccatcag gacgttgtga gtggcgcgat ttttagcggc tgaaatcagc ccttgagcct
2401 gtcggcaagt cgcgtcatga ggtccatgcg ctcatgcagg atcgccacga ccaacgcggg
2461 ttcgcccgca cgcggcaggc aaaaaacgta gtggtgttcg cagcgggcca tccgcagcgc
2521 gggaaagagt tcgctcatgt ccttaaacgg gccttcgccg gcggcaagcc tggctatgcc
2581 ctgttccagc ttagcgatat agcggcgcac ctgcgccgcg ccccactccc ggcgcgtgta
2641 gcggatgatg ccgcgtagat cggcttcggc ctcagccgtg aggatgtagg ccgtcaagcg
2701 cgatccccgc tgagttcttc atcaagaatt tcgccgacgc tcttggtgga caccttgccg
2761 gcaagcccat cgttgatgcg gttccccagc atggttttca gttcctgcca tgcctgatcg
2821 gcatcagcgt caccggggaa cagacgttcg agggcgtatt gcttaatggt cttgccctgc
2881 aaggcggcca gggctttcag gctctggtgc tgctggtccg tcatgtcgat tgtcaggcgg
2941 ctcattggat aacctccata aaatacacgt aaccacatta gcacatatgt gggcgtgagg
3001 ctacagcgcg aggcgcatta aggtcgggaa aatgcgctag gcgcatttaa attgcgtatt
3061 gctgtaatgc gccatgccgg ctagactagg cccaaatggg tatacccaat ttgaccaagg
3121 gggacgcgat gagggcggcc aagcactacc gacaacttct atccatcgac ttcaacatcg
3181 aggcgctggc cttcgtgcct ggacccgacg gcacacgcgg ccggcgcatc cacgtcctgg
3241 ggcgcgaggt ccgcgaccgg cccggcctgg tcgagtacct ttcgccggcg ttcggctcgc
3301 gggtggcgct ggacggctac tgcaaggcca atttcgatgc agtgctgcac ctggcgtacc
3361 ccgatcatca gcaatggggc cacgcatgaa gcgccgaagc tacgccatgc tgcgcgccgc
3421 tgccgcgctg gccgtcctgg tcgttgcctc gccggcatgg gccgagctgc gcggcgaggt
3481 cgtgcgcatc atcgacggcg acaccatcga cgtgctggta gacaagcagc cggtgcgcgt
3541 gcgcctggtg gacattgacg cgccggaaaa gcggcaagcc ttcggcgaac gtgcgcgcca
3601 ggcgctggcc ggcatggtgt tccgccggca cgtcctggtc gacgagaagg acaccgaccg
3661 ttacggccgc acgctgggca ccgtgtgggt caacatggag ctggccagcc ggccgccgca
3721 gccgcgcaac gtcaacgccg cgatggttca ccagggcatg gcgtgggcct atcgcttcca
3781 cggccgcgcg gccgaccctg aaatgctgcg gctcgaacag gaggcgcgag gcaagcgcgt
3841 cggcctctgg tccgatccgc acgccgtcga gccgtggaaa tggcgacgcg agagcaacaa
3901 ccggagggac gaaggttgaa ggtcgcccgc atctacctgc gcgccagtac ggacgagcag
3961 aatcttgaac gccaggagag ccttgtagcg gccacgcggg ccgccgggta ctacgtcgcc
4021 ggcatctacc gcgagaaggc gtccggcgca cgcgccgacc ggcccgagct gctgcgcatg
4081 atcgcggacc tgcaacctgg tgaagtcgtc gtggagagcc ttgtagcggc cacgcgggcc
4141 gccgggtact acgtcgccgg catctaccgc gagaaggcgt ccggcgcacg cgccgaccgg
4201 cccgagctgc tgcgcatgat cgcggacctg caacctggtg aagtcgtcgt tgcggagaag
4261 atcgaccgca tcagccgctt gccgttggcc gaggccgagc gcctggttgc gtcgatccgg
4321 gccaaagggg ccaagctggc cgtgcctggc gtggtggacc tgtcggagct ggccgccgag
4381 gcgaacggag tggcgaaaat cgttctggaa tccgtccagg acatgctttt gaagctcgcc
4441 ttgcagatgg cccgcgacga ctacgaggat cggcgcgagc gtcaacgtca gggtgtccag
4501 ttggcgaagg ccgccggccg ctacaccggc cgcaaacgtg acgccggcat gcacgaccgc
4561 atcatcacgc ttcgctccgg cggatcgagc attgccaaga cggccaagct ggtcggatgc
4621 agcccgagcc aggtcaaacg agtgtgggcg gcctggaacg cgcagcagca aaaataaagc
4681 cgggcagtgc ccggcttttc tcaccttttc gcgtcccgca gggccgctgc gagcgcccta
4741 cctagatcct cgctttcccc ctcggtgtag tccggccagg gcacgaaggg cgcggatgcg
4801 aacctgttga gcaggtacgc cttcgggcag cggtagacca ccggcgagtt cgccttttca
4861 tcccaccggg ccaggatcac gtccgcatcg cagtgcatgt ccttcacctg gtcgcggaag
4921 aagccgaagg ccaccatgcc gctatgttcg ccgaggaacg ccagttgctt cgcgctggcg
4981 atcgcgccga cgccgccggc caaaaccgac gccatcaccc agccgacgaa ccagaagctg
5041 gcatgcttgc ggttgaccac cgcacgcgca gccgcgacca ggacaacggc caagctgccg
5101 accagggcca tgacgaccgt gatccggccg ttgtggaaag cgatgggctt gccagcgtcc
5161 gcttgcacgg cgtcgtaaat gctggacccg atgggcgcgc acatcagcac gacaggcagc
5221 agcaccagga acatcgtccg cgtccattgc gcgagtgcct tgcggcgttc gccggcggca
5281 agcgcctcca tcatcggcgt gaagcccaac agggccaccg cagccgccaa gccggcaacg
5341 atgccgcagg cgattacata catacatcct ccctaatgcg ccttgcgcac ggttgtagtc
5401 agagtccgcg gtggggcgat aagctcatga ccaaaatccc ttaacgtgag ttttcgttcc
5461 actgagcgtc agaccccgta gaaaagatca aaggatcttc ttgagatcct ttttttctgc
5521 gggggatcag gaccgctgcc ggagcgcaac ccactcacta cagcagagcc atgtagacaa
5581 catcccctcc ccctttccac cgcgtcagac gcccgtagca gcccgctacg ggctttttca
5641 tgccctgccc tagcgtccaa gcctcacggc cgcgctcggc ctctctggcg gccttctggc
5701 gctcctgctg cggcgtccgc tcgtgggccg tggcgcgggt ccgcgcgccg gcctcgtgcg
5761 cctggcgctc gcgggcgagg tccagggcgg ccgtcttcac gttctgcctt gcgcagatga
5821 gatagatcga tctagcgtgg actcaaggct ctcgcgaatg gctcgcgttg gaaactttca
5881 ttgacacttg aggggcaccg cagggaaatt ctcgtccttg cgagaaccgg ctatgtcgtg
5941 ctgcgcatcg agcctgcgcc cttggcttgt ctcgcccctc tccgcgtcgc tacggggctt
6001 ccagcgcctt tccgacgctc accgggctgg ttgccctcgc cgctgggctg gcggccgtct
6061 atggccctgc aaacgcgcca gaaacgccgt cgaagccgtg tgcgagacac cgcggccgcc
6121 ggcgttgtgg atacctcgcg gaaaacttgg ccctcactga cagatgaggg gcggacgttg
6181 acacttgagg ggccgactca cccggcgcgg cgttgacaga tgaggggcag gctcgatttc
6241 ggccggcgac gtggagctgg ccagcctcgc aaatcggcga aaacgcctga ttttacgcga
6301 gtttcccaca gatgatgtgg acaagcctgg ggataagtgc cctgcggtat tgacacttga
6361 ggggcgcgac tactgacaga tgaggggcgc gatccttgac acttgagggg cagagtgctg
6421 acagatgagg ggcgcaccta ttgacatttg aggggctgtc cacaggcaga aaatccagca
6481 tttgcaaggg tttccgcccg tttttcggcc accgctaacc tgtcttttaa cctgctttta
6541 aaccaatatt tataaacctt gtttttaacc agggctgcgc cctgtgcgcg tgaccgcgca
6601 cgccgaaggg gggtgccccc ccttctcgaa ccctcccggc ccgctaacgc gggcctccca
6661 tccccccagg ggctgcgccc ctcggccgcg aacggcctca ccccaaaaat ggcagcgctg
6721 gcagtccttg ccattgccgg gatcggggca gtaacgggat gggcgatcag cccgagcgcg
6781 acgcccggaa gcattgacgt gccgcaggtg ctggcatcga cattcagcga ccaggtgccg
6841 ggcagtgagg gcggcggcct gggtggcggc ctgcccttca cttcggccgt cggggcattc
6901 acggacttca tggcggggcc ggcaattttt accttgggca ttcttggcat agtggtcgcg
6961 ggtgccgtgc tcgtgttcgg gggtgaatta attccccgga tcgatccgtc agcttcacgc
7021 tgccgcaagc actcagggcg caagggctgc taaaggaagc ggaacacgta gaaagccagt
7081 ccgcagaaac ggtgctgacc ccggatgaat gtcagctact gggctatctg gacaagggaa
7141 aacgcaagcg caaagagaaa gcaggtagct tgcagtgggc ttacatggcg atagctagac
7201 tgggcggttt tatggacagc aagcgaaccg gaattgccag ctggggcgcc ctctggtaag
7261 gttgggaagc cctgcaaagt aaactggatg gctttcttgc cgccaaggat ctgatggcgc
7321 aggggatcaa gatcgacgga tcgatccggg gaattaattc cggggcaatc ccgcaaggag
7381 ggtgaatgaa tcggacgttt gaccggaagg catacaggca agaactgatc gacgcggggt
7441 tttccgccga ggatgccgaa accatcgcaa gccgcaccgt catgcgtgcg ccccgcgaaa
7501 ccttccagtc cgtcggctcg atggtccagc aagctacggc caagatcgag cgcgacagcg
7561 tgcaactggc tccccctgcc ctgcccgcgc catcggccgc cgtggagcgt tcgcgtcgtc
7621 tcgaacagga ggcggcaggt ttggcgaagt cgatgaccat cgacacgcga ggaactatga
7681 cgaccaagaa gcgaaaaacc gccggcgagg acctggcaaa acaggtcagc gaggccaagc
7741 aggccgcgtt gctgaaacac acgaagcagc agatcaagga aatgcagctt tccttgttcg
7801 atattgcgcc gtggccggac acgatgcgag cgatgccaaa cgacacggcc cgctctgccc
7861 tgttcaccac gcgcaacaag aaaatcccgc gcgaggcgct gcaaaacaag gtcattttcc
7921 acgtcaacaa ggacgtgaag atcacctaca ccggcgtcga gctgcgggcc gacgatgacg
7981 aactggtgtg gcagcaggtg ttggagtacg cgaagcgcac ccctatcggc gagccgatca
8041 ccttcacgtt ctacgagctt tgccaggacc tgggctggtc gatcaatggc cggtattaca
8101 cgaaggccga ggaatgcctg tcgcgcctac aggcgacggc gatgggcttc acgtccgacc
8161 gcgttgggca cctggaatcg gtgtcgctgc tgcaccgctt ccgcgtcctg gaccgtggca
8221 agaaaacgtc ccgttgccag gtcctgatcg acgaggaaat cgtcgtgctg tttgctggcg
8281 accactacac gaaattcata tgggagaagt accgcaagct gtcgccgacg gcccgacgga
8341 tgttcgacta tttcagctcg caccgggagc cgtacccgct caagctggaa accttccgcc
8401 tcatgtgcgg atcggattcc acccgcgtga agaagtggcg cgagcaggtc ggcgaagcct
8461 gcgaagagtt gcgaggcagc ggcctggtgg aacacgcctg ggtcaatgat gacctggtgc
8521 attgcaaacg ctagggcctt gtggggtcag ttccggctgg gggttcagca gccactcgat
8581 cgaggtccca atacgcaaac cgcctctccc cgcgcgttgg ccgattcatt aatgcagctg
8641 gcacgacagg tttcccgact ggaaagcggg cagtgagcgc aacgcaatta atgtgagtta
8701 gctcactcat taggcacccc aggctttaca ctttatgctt ccggctcgta tgttgtgtgg
8761 aattgtgagc ggataacaat ttcacacagg aaacagct
//