IS21

  • Family IS21
  • Group
MGE type ISRelated element(s) :
Isoform Synonym(s) IS8
Accession numberTranspositionOriginHost
X14793 Y Pseudomonas aeruginosa
Pseudomonas aeruginosa PAO25 plasmid pR68.45
DNA section
IS Length : 2131 bp

Ends


IR Length : 30/42

IRL : TGTCAGCGCCAGTGATATAAGACGGTAATTCACCATTTGGATTGTCCGCT
IRR : TGTCAACGCCACGATGTTTGACCGTTATTTGCCATTTTCATGCCCGCTAA

Insertion site


Left flankDirect repeatRight flankDR Length
NNNNNNNTTCTCGGAGCNNNNNNN4
NNNNNNNTTCTCGGAGCNNNNNNN4
NNNNNNNGCGCCTATATNNNNNNN4
NNNNNNNGCGCCTATATNNNNNNN4
NNNNNNNGTCAGGCACCNNNNNNN4
NNNNNNNCCACTATCGANNNNNNN4
NNNNNNNTGTTGGGCGCNNNNNNN4
NNNNNNNCCATTCCGACNNNNNNN4
NNNNNNNCCGGTACTGCNNNNNNN4
NNNNNNNAGTCAGGCACCNNNNNNN5

DNA sequence

TGTCAGCGCCAGTGATATAAGACGGTAATTCACCATTTGGATTGTCCGCTCCACCCAACATGTTGTTTCCTTAAGGTTCTCACACCAGAAAGGACATCAA
CATGCTGAGCAGAGAGGACTTTTACATGATAAAGCAAATGCGCCAGCAGGGCGCGTACATTGTCGATATTGCGACTCAGATTGGTTGCTCTGAACGGACG
GTCAGACGCTACCTCAAATACCCTGAACCGCCAGCCAGAAAGACCCGCCACAAAATGGTTAAGCTGAAACCGTTTATGGATTACATCGACATGCGCCTGG
CAGAGAATGTCTGGAATAGTGAGGTTATCTTTGCGGAGATTAAGGCAATGGGTTATACGGGCGGACGTTCCATGCTGCGTTACTACATCCAGCCCAAACG
TAAAATGCGTCCGTCAAAAAGAACAGTTCGCTTCGAAACTCAGCCTAGATACCAGCTCCAGCACGACTGGGGCGAAGTTGAGGTGGAGGTTGCCGGGCAA
CGGTGCAAAGTTAACTTTGCGGTTAATACGCTGGGGTTCTCCCGCCGCTTCCATGTCTTCGCCGCACCAAAACAGGATGCTGAGCATACCTACGAATCAC
TGGTTCGCGCCTTCCGCTACTTCGGTGGTTGTGTGAAAACGGTGCTGGTTGATAACCAGAAGGCTGCGGTGCTGAAGAATAACAACGGGAAAGTCGTGTT
CAACTCCGGATTCCTGTTGCTGGCCGACCACTATAACTTCCTGCCACGGGCATGCCGTCCACGCAGGGCCAGAACAAAAGGTAAGGTTGAGCGGATGGTG
AAATACCTCAAGGAGAACTTCTTCGTTCGGTACCGCAGGTTCGACAGCTTCACTCATGTTAATCAACAACTGGAGCAATGGATAGCCGATGTGGCTGACA
AACGGGAACTTCGCCAGTTCAAAGAAACGCCGGAACAGCGCTTCGCGCGTGAGCAGGAACATCTGCAGCCGTTACCGGATACGGACTTCGATACCAGTTA
CTTCGACATCCGCCATGTGTCCTGGGACAGCTATATCGAGGTTGGTGGTAATCGTTACAGCGTTCCCGAAGCGCTGTGTGGTCAGCCGGTATCGATACGA
ATATCGCTGGATGACGAGTTGCGGATCTACAGTAATGAGAAACTGGTGGCCTCACATCGCCTCTGTTCGGCATCGTCTGGCTGGCAGACAGTGCCGGAGC
ATCACGCCCCGCTCTGGCAGCAGGTCAGTCAGGTGGAACATCGACCACTGAGTGCCTATGAGGAGCTGTTGTGATGCATGAGCTGGAAGTCCTGCTGAGT
CGCCTGAAAATGGAGCATCTGAGTTATCACGTTGAAAGCCTGCTGGAACAGGCAGCTAAAAAAGAGCTGAACTACCGGGAGTTCCTGTGCATGGCGCTAC
AGCAGGAATGGAACGGCAGGCATCAGCGCGGTATGGAGTCCAGGCTGAAGCAGGCTCGTCTGCCGTGGGTCAAAACGCTGGAGCAGTTCGACTTTACCTT
CCAGCCGGGCATCGACCGTAAGGTTGTCCGGGAACTGGCTGGTCTGGCGTTCGTGGAGCGCAGCGAAAACGTGATCCTGCTGGGACCTCCTGGTGTCGGA
AAAACTCATCTGGCCATAGCTCTTGGCGTGAAAGCGGTGGATGCGGGACATCGGGTACTGTTTATGCCACTGGACAGACTGATCGCGACACTGATGAAAG
CGAAAGAGGAAAACCGGCTGGAGCGTCAGCTGCAGCAACTGAGTTATGCCCGGGTGTTGATCCTGGATGAAATAGGCTATCTGCCCATGAACAGAGAGGA
AGCCAGCCTGTTCTTCCGGCTACTGAACCGTCGATATGAAAAAGCGAGCATCATACTGACGTCAAACAAAGGGTTCGCAGACTGGGGAGAAATGTTCGGA
GATCACGTGCTGGCAACAGCGATACTGGATCGGTTGCTACATCACTCAACCACGCTGAATATCAAAGGAGAGAGTTACCGGTTAAAAGAGAAACGTAAAG
CTGGAGTGCTGACCAAAAACACAACGCCAATCAGTGATGATGAAATGGTGAAAAGCGGACAGCATCAGTAACGAAAGTATCTTAGCGGGCATGAAAATGG
CAAATAACGGTCAAACATCGTGGCGTTGACA
Protein section
ORF number : 2

 

ORF 1
LengthBeginEndStrandFusion ORF
1173 bp390 aa1021274+No
ORF function : Transposase
Chemistry : DDE

ORF sequence :

MLSREDFYMIKQMRQQGAYIVDIATQIGCSERTVRRYLKYPEPPARKTRHKMVKLKPFMDYIDMRLAENVWNSEVIFAEIKAMGYTGGRSMLRYYIQPKR
KMRPSKRTVRFETQPRYQLQHDWGEVEVEVAGQRCKVNFAVNTLGFSRRFHVFAAPKQDAEHTYESLVRAFRYFGGCVKTVLVDNQKAAVLKNNNGKVVF
NSGFLLLADHYNFLPRACRPRRARTKGKVERMVKYLKENFFVRYRRFDSFTHVNQQLEQWIADVADKRELRQFKETPEQRFAREQEHLQPLPDTDFDTSY
FDIRHVSWDSYIEVGGNRYSVPEALCGQPVSIRISLDDELRIYSNEKLVASHRLCSASSGWQTVPEHHAPLWQQVSQVEHRPLSAYEELL

 

Blast result :
ORF 2
LengthBeginEndStrandFusion ORF
798 bp265 aa12742071+No
ORF function : Accessory Gene
AG : IS21 helper

ORF sequence :

MHELEVLLSRLKMEHLSYHVESLLEQAAKKELNYREFLCMALQQEWNGRHQRGMESRLKQARLPWVKTLEQFDFTFQPGIDRKVVRELAGLAFVERSENV
ILLGPPGVGKTHLAIALGVKAVDAGHRVLFMPLDRLIATLMKAKEENRLERQLQQLSYARVLILDEIGYLPMNREEASLFFRLLNRRYEKASIILTSNKG
FADWGEMFGDHVLATAILDRLLHHSTTLNIKGESYRLKEKRKAGVLTKNTTPISDDEMVKSGQHQ

 

Blast result :
Comments
IS21 was first observed in R68.45 as a direct duplication of a 2 kb fragment of the original R68 plasmid. It is involved in conduction of the bacterial chromosome and the pTi plasmid, via "cointegrate formation", but also promotes adjacent deletions. However, cointegrate formation proceeds first through an adjacent transposition of IS21 in the donor molecule, and then transposition of that structure (tandem direct repeat of IS21) in the target molecule thereby generating a typical cointegrate molecule which can be resolved in a recA+ strain. In the "intermediate" structure (IS21)2, a new promoter is created at the junction (-35 box in the right IR of the right copy and a -10 box at the left end of the left IS21 copy) and activates the transpositional activity of the left copy (Reimmann et al., 1989). IS21 has the tendency to insert within or very close to the ends of IS21 (Reimmann and Haas, 1987). All the above results suggest a conservative, "cut-and-paste" transposition mechanism for IS21. The istA gene is responsible for the 3'-end cleavages but istB is also required for cointegrate formation (Reimmann and Haas, 1990). IS21 is located upstream of the kanamycin resistance determinant of plasmid RP4 (= R18, R68, RK2, or RP1), and carries the -35 promoter box of the aphA gene. IS21 is similar to IS232 and IS640. In Escherichia coli, the first 106-bp of IS21 were found downstream of the nadE (NH3-dependent NAD synthetase) gene, at position 38.1 min (M15328).
References
1] Jacob, A.E., Cresswell, J.M., and Hedges, R.W. (1977) FEMS Lett. 1, 71-74.
2] Depicker, A., De Block, M., Inzé, D., Van Montagu, M., and Schell, J. (1980) Gene 10, 329-338.
3] Leemans, J., Villarroel, R., Silva, B., Van Montagu, M., and Schell, J. (1980) Gene 10, 319-328.
4] Riess, G., Holloway, B.W., and Pühler, A. (1980) Genet. Res. 36, 99-109.
5] Willetts, N.S., Crowther, C., and Holloway, B.W. (1981) Plasmid 6, 30-52.
6] Currier, T.C., and Morgan, M.K. (1982) J. Bacteriol.150, 251-259.
7] Haas, D., and Riess, G. (1983) Plasmid 9, 42-52.
8] Riess, G., Masepohl, B., and Puehler, A. (1983) Plasmid 10, 111-118.
9] Dobritsa, A.P. (1984) Mol. Gen. Genet. 194, 206-210.
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11] Reimmann, C., and Haas, D. (1986) Mol. Gen. Genet. 203, 511-519.
12] Schurter, W., and Holloway, B.W. (1986) Plasmid 15, 8-18.
13] Pansegrau, W., Miele, L., Lurz, R., and Lanka, E. (1987) Plasmid 18, 193-204.
14] Reimmann, C., and Haas, D. (1987) Genetics 115, 619-625.
15] Schurter, W., and Holloway, B.W. (1987) Plasmid 17, 61-64.
16] Reimmann, C., Rella, M., and Haas, D. (1988) J. Gen. Microbiol. 134, 1515-1523.
17] Reimmann, C., Moore, R., Little, S., Savioz, A., Willetts, N.S., and Haas, D. (1989) Mol. Gen. Genet. 215, 416-424.
18] Gerlitz, M., Hrabak, O., and Schwab, H. (1990) J. Bacteriol. 172, 6194-6203.
19] Menou, G., Mahillon, J., Lecadet, M.-M., and Lereclus, D. (1990) J. Bacteriol. 172, 6689-6696.
20] Reimmann, C., and Haas, D. (1990) EMBO J. 9, 4055-4063.