pIJ10257

From ActinoBase


Use

This vector (originally produced and published by the Buttner group 1) can be used to introduce a gene or set of genes into Streptomyces under the control of the constitutive ermE* promoter 2. This can be useful for over expression or complementation experiments.

Features

  • Hygromycin resistance gene
  • ermE* promoter2
  • ΦBT1 phage integration site
  • Origin of transfer oriT from RK2

History

Backbone vector pMS81 was modified by cloning in a 330 bp fragment containing ermE*, a ribosome binding site and a multicloning site isolated from pIJ8723, to create pIJ102571.

Map

PIJ10257 Map2.png

Sequence links

"LOCUS pIJ10257 6430 bp ds-DNA linear 27-MAR-2019 DEFINITION Ligation of Fragment 2 into Fragment 2 ACCESSION pIJ10257 KEYWORDS . SOURCE Unknown. 

 ORGANISM  Unknown Unclassified.

REFERENCE 1 (bases 1 to 6424) 

 AUTHORS   Self
 JOURNAL   Unpublished.

COMMENT SECID/File created by SciEd Central, Scientific & Educational 

           Software

COMMENT COMMENT ApEinfo:methylated:1 FEATURES Location/Qualifiers 

    CDS             1259..1285
                    /gene="aac3 (IV)'"
                    /label=aac3 (IV)'
                    /ApEinfo_fwdcolor=#e9d024
                    /ApEinfo_revcolor=#e9d024
                    /ApEinfo_graphicformat=arrow_data {{0 1 2 0 0 -1} {} 0}
                    width 5 offset 0
    CDS             complement(1572..2570)
                    /gene="hyg"
                    /label=hyg
                    /ApEinfo_fwdcolor=#e9d024
                    /ApEinfo_revcolor=#e9d024
                    /ApEinfo_graphicformat=arrow_data {{0 1 2 0 0 -1} {} 0}
                    width 5 offset 0
    CDS             complement(2955..3036)
                    /gene="ermE1'"
                    /product="ErmE1"
                    /label=ErmE1
                    /ApEinfo_fwdcolor=#e9d024
                    /ApEinfo_revcolor=#e9d024
                    /ApEinfo_graphicformat=arrow_data {{0 1 2 0 0 -1} {} 0}
                    width 5 offset 0
    CDS             3116..3121
                    /gene="-35p2"
                    /label=-35p2
                    /ApEinfo_fwdcolor=#e9d024
                    /ApEinfo_revcolor=#e9d024
                    /ApEinfo_graphicformat=arrow_data {{0 1 2 0 0 -1} {} 0}
                    width 5 offset 0
    CDS             3140..3145
                    /gene="-10p2"
                    /label=-10p2
                    /ApEinfo_fwdcolor=#e9d024
                    /ApEinfo_revcolor=#e9d024
                    /ApEinfo_graphicformat=arrow_data {{0 1 2 0 0 -1} {} 0}
                    width 5 offset 0
    CDS             3186..3190
                    /gene="-35p1"
                    /product="TGG deletion (252-254) results in ermE
                    over-expression"
                    /label=TGG deletion (252-254) results in ermE
                    over-expression
                    /ApEinfo_fwdcolor=#e9d024
                    /ApEinfo_revcolor=#e9d024
                    /ApEinfo_graphicformat=arrow_data {{0 1 2 0 0 -1} {} 0}
                    width 5 offset 0
    CDS             3208..3213
                    /gene="-10p1"
                    /label=-10p1
                    /ApEinfo_fwdcolor=#e9d024
                    /ApEinfo_revcolor=#e9d024
                    /ApEinfo_graphicformat=arrow_data {{0 1 2 0 0 -1} {} 0}
                    width 5 offset 0
    misc_feature    4134..4245
                    /gene="oriT"
                    /label=oriT
                    /ApEinfo_fwdcolor=#7eff74
                    /ApEinfo_revcolor=#7eff74
                    /ApEinfo_graphicformat=arrow_data {{0 1 2 0 0 -1} {} 0}
                    width 5 offset 0
    misc_feature    complement(4454..4462)
                    /gene="attP"
                    /label=attP
                    /ApEinfo_fwdcolor=#7eff74
                    /ApEinfo_revcolor=#7eff74
                    /ApEinfo_graphicformat=arrow_data {{0 1 2 0 0 -1} {} 0}
                    width 5 offset 0
    CDS             4536..5965
                    /gene="int"
                    /label=int
                    /ApEinfo_fwdcolor=#e9d024
                    /ApEinfo_revcolor=#e9d024
                    /ApEinfo_graphicformat=arrow_data {{0 1 2 0 0 -1} {} 0}
                    width 5 offset 0

ORIGIN 

       1 GATCCGCGGC CGCGCGCGAT ATCGAATTCG TAATCATGTC ATAGCTGTTT CCTGTGTGAA
      61 ATTGTTATCC GCTCACAATT CCACACAACA TACGAGCCGG AAGCATAAAG TGTAAAGCCT
     121 GGGGTGCCTA ATGAGTGAGC TAACTCACAT TAATTGCGTT GCGCTCACTG CCCGCTTTCC
     181 AGTCGGGAAA CCTGTCGTGC CAGCTGCATT AATGAATCGG CCAACGCGCG GGGAGAGGCG
     241 GTTTGCGTAT TGGGCGCTCT TCCGCTTCCT CGCTCACTGA CTCGCTGCGC TCGGTCGTTC
     301 GGCTGCGGCG AGCGGTATCA GCTCACTCAA AGGCGGTAAT ACGGTTATCC ACAGAATCAG
     361 GGGATAACGC AGGAAAGAAC ATGTGAGCAA AAGGCCAGCA AAAGGCCAGG AACCGTAAAA
     421 AGGCCGCGTT GCTGGCGTTT TTCCATAGGC TCCGCCCCCC TGACGAGCAT CACAAAAATC
     481 GACGCTCAAG TCAGAGGTGG CGAAACCCGA CAGGACTATA AAGATACCAG GCGTTTCCCC
     541 CTGGAAGCTC CCTCGTGCGC TCTCCTGTTC CGACCCTGCC GCTTACCGGA TACCTGTCCG
     601 CCTTTCTCCC TTCGGGAAGC GTGGCGCTTT CTCATAGCTC ACGCTGTAGG TATCTCAGTT
     661 CGGTGTAGGT CGTTCGCTCC AAGCTGGGCT GTGTGCACGA ACCCCCCGTT CAGCCCGACC
     721 GCTGCGCCTT ATCCGGTAAC TATCGTCTTG AGTCCAACCC GGTAAGACAC GACTTATCGC
     781 CACTGGCAGC AGCCACTGGT AACAGGATTA GCAGAGCGAG GTATGTAGGC GGTGCTACAG
     841 AGTTCTTGAA GTGGTGGCCT AACTACGGCT ACACTAGAAG AACAGTATTT GGTATCTGCG
     901 CTCTGCTGAA GCCAGTTACC TTCGGAAAAA GAGTTGGTAG CTCTTGATCC GGCAAACAAA
     961 CCACCGCTGG TAGCGGTGGT TTTTTTGTTT GCAAGCAGCA GATTACGCGC AGAAAAAAAG
    1021 GATCTCAAGA AGATCCTTTG ATCTTTTCTA CGGGGTCTGA CGCTCAGTGG AACGAAAACT
    1081 CACGTTAAGG GATTTTGGTC ATGAGATTAT CAAAAAGGAT CTTCACCTAG ATCCTTTTGG
    1141 TTCATGTGCA GCTCCATCAG CAAAAGGGGA TGATAAGTTT ATCACCACCG ACTATTTGCA
    1201 ACAGTGCCGT TGATCGTGCT ATGATCGACT GATGTCATCA GCGGTGGAGT GCAATGTCGT
    1261 GCAATACGAA TGGCGAAAAG CCGAGAACNN NNNNNNNNNN NNNNNNNNNN NNNNNNNNNN
    1321 NNNNNNNNNN NNNCCCGGGT TGCCGAGCTG GGATGGAAGC TCGGCCGACC ACCCTGGAGG
    1381 AGATGATCGA GGATGCCAGG GCCTTTCACG CCCGCCGCTG CTGAGCGTCC GCCGCCGGGC
    1441 CCGCACCGCC GTCGGCCGGC CCGCTCCGGG CTCGCAGCAG CGGGCTTCGG CGCGGGCCCG
    1501 GGGCTCCCGA GCGCGGGCGG GGCTCCGGGC GGCCGCCGGG GGCCGGGGGC GGCGCCGGGC
    1561 GGCCCGGGGC GTCAGGCGCC GGGGGCGGTG TCCGGCGGCC CCCAGAGGAA CTGCGCCAGT
    1621 TCCTCCGGAT CGGTGAAGCC GGAGAGATCC AGCGGGGTCT CCTCGAACAC CTCGAAGTCG
    1681 TGCAGGAAGG TGAAGGCGAG CAGTTCGCGG GCGAAGTCCT CGGTCCGCTT CCACTGCGCC
    1741 CCGTCGAGCA GCGCGGCCAG GATCTCGCGG TCGCCCCGGA AGGCGTTGAG ATGCAGTTGC
    1801 ACCAGGCTGT AGCGGGAGTC TCCCGCATAG ACGTCGGTGA AGTCGACGAT CCCGGTGACC
    1861 TCGGTCGCGG CCAGGTCCAC GAAGATGTTG GTCCCGTGCA GGTCGCCGTG GACGAACCGG
    1921 GGTTCGCGGC CGGCCAGCAG CGTGTCCACG TCCGGCAGCC AGTCCTCCAG GCGGTCCAGC
    1981 AGCCGGGGCG AGAGGTAGCC CCACCCGCGG TGGTCCTCGA CGGTCGCCGC GCGGCGTTCC
    2041 CGCAGCAGTT CCGGGAAGAC CTCGGAATGG GGGGTGAGCA CGGTGTTCCC GGTCAGCGGC
    2101 ACCCTGTGCA GCCGGCCGAG CACCCGGCCG AGTTCGCGGG CCAGGGCGAG CAGCGCGTTC
    2161 CGGTCGGTCG TGCCGTCCAT CGCGGACCGC CAGGTGGTGC CGGTCATCCG GCTCATCACC
    2221 AGGTAGGGCC ACGGCCAGGC TCCGGTGCCG GGCCGCAGCT CGCCGCGGCC GAGGAGGCGG
    2281 GGCACCGGCA CCGGGGCGTC CGCCAGGACC GCGTACGCCT CCGACTCCGA CGCGAGGCTC
    2341 TCCGGACCGC ACCAGTGCTC GCCGAACAGC TTGATCACCG GGTCGGGCTC GCCGACCAGT
    2401 ACGGGGTTGG TGCTCTCGCC GGGCACCCGC AGCACCGGCG GCACCGGCAG CCCGAGCTCC
    2461 TCCAGGGCTC GGCGGGCCAG CGGCTCCCAG AATTCCTGGT CGTTCCGCAG GCTCGCGTAG
    2521 GAATCATCCG AATCAATACG GTCGAGAAGT AACAGGGATT CTTGTGTCAC AGCGGACCTC
    2581 TATTCACAGG GTACGGGCCG GCTTAATTCC GCACGGCCGG TCGCGACACG GCCTGTCCGC
    2641 ACCGCGGTCA GGCGTTGACG ATGACGGGCT GGTCGGCCAC GTCGGGGACG TTCTCGGTGG
    2701 TGCTGCGGTC GGGATCGCCA ATCTCTACGG GCCGACCGAG GCGACGGTGT ACGCCACCGC
    2761 CTGGTTCTGC GACGGCGAGG CGCCGCCAGG CCCCGCNNNN NNNNNNNNNN NNNNNNNNNN
    2821 NNNNNNNNNN NNNNNNNNNN NNNNNNNNNN NNNNNNNNNN NNNNNNNNNN NNNNNNNNNN
    2881 NNNNNNNNNN NNNNNNNNNN NNNNNNNNNN NNNNNNNNNN NNNNNNNNNN NNNCCCTNAG
    2941 GTACCAGCCC GACCCGAGCA CGCGCCGGCA CGCCTGGTCG ATGTCGGACC GGAGTTCGAG
    3001 GTACGCGGCT TGCAGGTCCA GGAAGGGGAC GTCCATGCGA GTGTCCGTTC GAGTGGCGGC
    3061 TTGCGCCCGA TGCTAGTCGC GGTTGATCGG CGATCGCAGG TGCACGCGGT CGATCTTGAC
    3121 GGCTGGCGAG AGGTGCGGGG AGGATCTGAC CGACGCGGTC CACACGTGGC ACCGCGATGC
    3181 TGTTGTGGGC ACAATCGTGC CGGTTGGTAG GATCGTCTAG AACAGGAGGC CCCATATGAG
    3241 ATCTCGAGTG ATTAATTAAC TAGAGTCGAC CTGCATAGAT Ctaagcttgg atccTAGGTT
    3301 CTCATGAGCG GAGAACGAGA TGACGTTGGA GGGGCAAGGT CGCGCTGATT GCTGGGGCAA
    3361 CACGTGGAGC GGATCGGGGA TTGTCTTTCT TCAGCTCGCT GATGATATGC TGACGCTCAA
    3421 TGCCGTTTGG CCTCCGACTA ACGAAAATCC CGCATTTGGA CGGCTGATCC GATTGGCACG
    3481 GCGGACGGCG AATGGCGGAG CAGACGCTCG TCCGGGGGCA ATGAGATATG AAAAAGCCTG
    3541 AACTCACCGC GACGTATCGG GCCCTGGCCA GCTAGCTAGA GTCGACCTGC AGGTCCCCGG
    3601 GGATCGGTCT TGCCTTGCTC GTCGGTGATG TACTTCACCA GCTCCGCGAA GTCGCTCTTC
    3661 TTGATGGAGC GCATGGGGAC GTGCTTGGCA ATCACGCGCA CCCCCCGGCC GTTTTAGCGG
    3721 CTAAAAAAGT CATGGCTCTG CCCTCGGGCG GACCACGCCC ATCATGACCT TGCCAAGCTC
    3781 GTCCTGCTTC TCTTCGATCT TCGCCAGCAG GGCGAGGATC GTGGCATCAC CGAACCGCGC
    3841 CGTGCGCGGG TCGTCGGTGA GCCAGAGTTT CAGCAGGCCG CCCAGGCGGC CCAGGTCGCC
    3901 ATTGATGCGG GCCAGCTCGC GGACGTGCTC ATAGTCCACG ACGCCCGTGA TTTTGTAGCC
    3961 CTGGCCGACG GCCAGCAGGT AGGCCGACAG GCTCATGCCG GCCGCCGCCG CCTTTTCCTC
    4021 AATCGCTCTT CGTTCGTCTG GAAGGCAGTA CACCTTGATA GGTGGGCTGC CCTTCCTGGT
    4081 TGGCTTGGTT TCATCAGCCA TCCGCTTGCC CTCATCTGTT ACGCCGGCGG TAGCCGGCCA
    4141 GCCTCGCAGA GCAGGATTCC CGTTGAGCAC CGCCAGGTGC GAATAAGGGA CAGTGAAGAA
    4201 GGAACACCCG CTCGCGGGTG GGCCTACTTC ACCTATCCTG CCCGGCTGAC GCCGTTGGAT
    4261 ACACCAAGGA AAGTCTACAC GAACCCTTTG GCAAAATCCT GTATATCGTG CGAAAAAGGA
    4321 TGGATATACC GAAAAAATCG CTATAATGAC CCCGAAGCAG GGTTATGCAG CGGAAAAGAT
    4381 CCGTCGACCT GCAGGCATGC TGGCGCCGGA CGGGGCTTCA GACGTTTCGG GTGCTGGGTT
    4441 GTTGTCTCTG GACAGTGATC CATGGGAAAC TACTCAGCAC CACCAATGTT CCCAAAAGAA
    4501 AGCGCAGGTC AGCGCCCATG AGCCAAGATC TAGGCATGTC GCCCTTCATC GCTCCCGACG
    4561 TCCCTGAGCA CCTTCTAGAC ACTGTTCGCG TCTTCCTGTA CGCGCGTCAG TCTAAGGGCC
    4621 GGTCCGACGG CTCAGACGTG TCGACCGAAG CACAGCTAGC GGCCGGTCGT GCGTTGGTCG
    4681 CGTCTCGCAA CGCCCAGGGG GGTGCGCGCT GGGTCGTGGC AGGTGAGTTC GTGGACGTCG
    4741 GGCGCTCCGG CTGGGACCCG AACGTGACCC GTGCCGACTT CGAGCGCATG ATGGGCGAAG
    4801 TCCGCGCCGG CGAAGGTGAC GTTGTCGTTG TGAATGAGCT TTCCCGGCTC ACTCGCAAGG
    4861 GCGCCCATGA CGCGCTCGAA ATCGACAACG AATTGAAGAA GCACGGCGTG CGCTTCATGT
    4921 CGGTTCTTGA GCCGTTCCTT GACACGTCTA CCCCTATCGG CGTCGCCATT TTCGCGCTGA
    4981 TCGCTGCCCT TGCGAAACAG GACAGTGACC TGAAGGCGGA GCGCCTGAAG GGTGCGAAAG
    5041 ACGAGATTGC CGCGCTGGGT GGCGTTCACT CGTCTTCCGC CCCGTTCGGA ATGCGCGCCG
    5101 TGCGCAAGAA GGTCGATAAT CTCGTGATCT CCGTTCTTGA GCCGGACGAA GACAACCCGG
    5161 ATCACGTCGA GCTAGTTGAG CGCATGGCGA AAATGTCGTT CGAAGGCGTG TCCGACAACG
    5221 CCATTGCAAC GACCTTCGAG AAGGAAAAGA TCCCGTCGCC CGGAATGGCT GAGAGACGCG
    5281 CCACGGAAAA GCGTCTTGCG TCCATCAAGG CACGTCGCCT GAACGGCGCT GAAAAGCCGA
    5341 TCATGTGGCG CGCTCAAACG GTCCGATGGA TTCTCAACCA TCCCGCAATC GGCGGTTTCG
    5401 CATTCGAGCG TGTGAAGCAC GGTAAGGCGC ACATCAACGT CATACGGCGC GACCCCGGCG
    5461 GCAAGCCGCT AACGCCCCAC ACGGGCATTC TCAGCGGCTC GAAGTGGCTT GAGCTTCAAG
    5521 AGAAGCGTTC CGGGAAGAAT CTCAGCGACC GGAAGCCTGG GGCCGAAGTC GAACCGACGC
    5581 TTCTGAGCGG GTGGCGTTTC CTGGGGTGCC GAATCTGCGG CGGCTCAATG GGTCAGTCCC
    5641 AGGGTGGCCG TAAGCGCAAC GGCGACCTTG CCGAAGGCAA TTACATGTGC GCCAACCCGA
    5701 AGGGGCACGG CGGCTTGTCG GTCAAGCGCA GCGAACTGGA CGAATTCGTT GCTTCGAAGG
    5761 TGTGGGCACG GCTCCGCACA GCCGACATGG AAGATGAACA CGATCAGGCA TGGATTGCCG
    5821 CCGCTGCGGA GCGCTTCGCC CTTCAGCACG ACCTAGCGGG GGTGGCCGAT GAGCGGCGCG
    5881 AACAACAGGC GCACCTAGAC AACGTGCGGC GCTCCATCAA GGACCTTCAG GCGGACCGTA
    5941 AGGCCGGTCT GTACGTCGGG CGTGAAGAGC TGGAAACGTG GCGCTCAACG GTGCTGCAAT
    6001 ACCGGTCCTA CGAAGCGGAG TGCACGACCC GACTCGCTGA GCTTGACGAG AAGATGAACG
    6061 GCAGCACCCG CGTTCCGTCT GAGTGGTTCA GCGGCGAAGA CCCGACGGCC GAAGGGGGCA
    6121 TCTGGGCAAG CTGGGACGTG TACGAGCGTC GGGAGTTCCT GAGCTTCTTC CTTGACTCCG
    6181 TCATGGTCGA CCGGGGGCGC CACCCTGAGA CGAAGAAATA CATCCCCCTG AAGGACCGTG
    6241 TGACGCTCAA GTGGGCGGAG CTGCTGAAGG AGGAAGACGA AGCGAGCGAA GCCACTGAGC
    6301 GGGAGCTTGC GGCGCTGTAG CGCACAGCGG GAGGGGTCGA GCCGGCGGAC GGTTCGGCCC
    6361 CTTTTTTGGC CTTGAAATCG TTAGTTAGGC TAACTAGTAG TTCCTTCGTC ACCATAGCGG
    6421 GCAGGGAGCG

// "

References

  1. Hong, H., Hutchings, M. I., Hill, L. M., Buttner, M. J. (2005). The Role of the Novel Fem Protein VanK in Vancomycin Resistance in Streptomyces coelicolor. Journal of Biological Chemistry, 280, pp. 13055-13061. doi: 10.1074/jbc.M413801200
  2. Bibb, M. J., Janssen, G. R., and Ward, J. M. (1985). Cloning and analysis of the promoter region of the erythromycin resistance gene (ermE) of Streptomyces erythraeus. ‘’Gene’’, 38, pp. 215-226. doi: 10.1016/0378-1119(85)90220-3
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