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ID   RK2        preliminary; circular DNA; SYN; 56400 BP.
AC   X59971; S37640; X14165; M33117; M25423; ATCC37125;
DT   01-OCT-1993 (Rel. 7, Created)
DT   01-APR-1995 (Rel. 11, Last updated, Version 1)
DE   Gram-negative bacteria/E.coli incP plasmid RK2 - incomplete.
KW   cloning vector; incompatibility group P alpha; Birmingham; incP.
OS   Gram-negative bacteria
OC   Prokaryotae.
RN   [1]
RC   lambda trpED10T from lambda & trp gene
RC   pVH103 from ColE1 & lambda trpED10T
RC   pRM2 from pVH103 & RK2
RA   Meyer R., Figurski D., Helinski D.R.;
RT   "Molecular vehicle properties of the broad host range plasmid RK2";
RL   Science 190:1226-1228(1975).
RN   [2]
RP   1-1320
RC   RK2
RA   Thomas C.M.;
RT   ;
RL   Submitted (05-JUN-1991) by:
RL   Thomas C.M., School of Biological Sciences,
RL   Univ. of Birmingham, P O Box 363, Birmingham B15 2TT, UNITED KINGDOM.
RN   [3]
RP   1-1320
RC   RK2 trfAts from RK2
RC   pCT662delta from pRK229
RC   pMMV103.1, pMMV112.1, pMMV100 series from pCT662delta & Tn1723
RC   pMMV200 series from pCT662delta & Tn1723
RC   pMMV400 from pCT662delta
RC   pMMV61 from pCT662delta
RC   pDB776 from RP4 & pT7-7
RC   pMS470delta from R751
RC   pMS54, pMS55 from pMS470delta & pDB776
RC   pGBT46 from pMMV103.1 & pCT673
RC   pGBT50 from pMMV112.1 & pCT673
RC   pGBT48 from pGBT46 & lacIqtacp
RC   pGBT47 from pGBT48
RC   pGBT49 from pGBT50
RC   pGBT51 from pGBT49 & lacIqtacp
RA   Motallebi-Veshareh M., Balzer D., Lanka E., Jagura-Burdzy G.,
RA   Thomas C.M.;
RT   "Conjugative transfer functions of broad-host-range plasmid RK2 are
RT   coregulated with vegetative replication";
RL   Mol. Microbiol. 6:907-920(1992).
RN   [4]
RC   pRK2501ts3 from pRK2501
RC   pCT86, pCT87 from pRK248 & pRK353
RC   Charon 3/86 from Charon 3 & pCT86
RC   [pCT153.1 from RK2 trfAts & pRK353]
RC   RP1, RP4, RK2, RP8, R1822, R68, R68.45 from Pseudomonas incP1 plasmids
RC   R91a, R751, R906, R26, R527, R772 from Pseudomonas incP1 plasmids
RC   pAV1 from Pseudomonas incP1 plasmids
RA   Thomas C.M.;
RT   "Molecular genetics of broad host range plasmid RK2";
RL   Plasmid 5:10-19(1981).
RN   [5]
RC   pRR41-8R from pRR10-ts97 & RK2
RC   pRR41-3.2 from pRR10-ts97 & RK2, par
RC   pRR41-0.8 from pRR10-ts97 & RK2, par
RC   pRR41-0.8D from pRR41-0.8 & linker
RC   pRR91 from pRR10 & RK2, par
RC   pRR93 from pRR10-ts97 & RK2, par
RC   pRR97 from pRR61 & RK2, par
RC   pRR110 from pBluescript II SK+ & ColE1, cer site
RC   pRR116 series from pRR series & ColE1, cer site
RC   pRR120 from pBluescript II SK+ & RK2, par
RC   pRR121 from pBluescript II SK+ & parD gene & parE gene
RC   pRR136 from pBluescript II SK+ & parD gene
RC   pRR137 from pBluescript II SK+ & parE gene
RC   pRR138 from pRR121 & linker
RC   pRR146 series from pRR120 & lacZ
RC   pRR series from pRR120
RA   Roberts R.C., Helinski D.R.;
RT   "Definition of a minimal plasmid stabilization system from the
RT   broad-host-range plasmid RK2";
RL   J. Bacteriol. 174:8119-8132(1992).
RN   [6]
RC   pRR91 from pRR10 & parD region
RC   pRR105-1 from pDK6 & parD gene
RC   pTD3 from pBluescript II SK+ & terminator
RC   pTD4 from pBluescript II SK+ & parCBA genes
RC   pTD5 from pBluescript II SK+ & parA gene
RC   pTD10 from pAL4000 & parCBA promoter & luc gene
RC   pTD11 from pAL4000 & parD promoter & luc gene
RC   pTD30 from pTD3 & parCBA promoter
RC   pTD31 from pTD3 & parD promoter
RA   Davis T.D., Helinski D.R., Roberts R.C.;
RT   "Transcription and autoregulation of the stabilizing functions of
RT   broad host range plasmid RK2 in Escherichia coli, Agrobacterium
RT   tumefaciens and Pseudomonas aeruginosa";
RL   Mol. Microbiol. 6:1981-1994(1992).
RN   [7]
RC   pMM40 from pKK223-3 & pMJR1560
RC   pMM63 from pKK223-3 & pMJR1560
RC   plasmid from pMM63
RC   pDK5 from plasmid & pBGS18
RC   pDK6 from pDK5 & pBGS18
RC   pDK7 from pDK5 & pACYC184
RA   Kleiner D., Paul W., Merrick M.J.;
RT   "Construction of multicopy expression vectors for regulated
RT   overproduction of proteins in Klebsiella pneumoniae and other
RT   enteric bacteria";
RL   J. Gen. Microbiol. 134:1779-1784(1988).
RN   [8]
RC   pKS45, pKS451 from pUC9 & ColE1, cer site
RC   pKS490 from pUC8 & ColE1, cer site
RC   pKS494 from pUC8 & ColE1, cer site
RA   Summers D.K., Sherratt D.J.;
RT   "Resolution of ColE1 dimers requires a DNA sequence implicated
RT   in the three-dimensional organization of the cer site";
RL   EMBO J. 7:851-858(1988).
RN   [9]
RC   pQV11 from pUC19
RC   pRR10 from pFF1 & pQV11
RC   pRR10-ts97 from pRR10 & trfA ts97 gene
RC   pRR12 from pSV16 & pUC19
RC   pRR13 from pUC19 & RK2 & pUC4K
RC   pRR14 from pUC19 & RK2 & pUC4K
RC   pRR15 from pUC19 & pRK419, R6K ori
RC   plasmid from RK2 & pUC19
RC   plasmid2 from plasmid & pUC4K
RC   plasmid3 from plasmid2 & RK2
RC   pRB7 from plasmid3 & pUC9Cm
RC   pRR38 from pRR15 & RK2, par
RC   pRR41 from pRR10-ts97 & RK2, par
RC   pRR53 from pRR10 & pBK280
RC   pRR54 from pRR10 & RK2, par
RC   plasmid4 from pBK280 & linker
RC   pRR61 from plasmid4 & pUC19
RC   pRR62 from pRR61 & RK2, par
RA   Roberts R.C., Burioni R., Helinski D.R.;
RT   "Genetic characterization of the stabilizing functions of a region
RT   of braod-host-range plasmid RK2";
RL   J. Bacteriol. 172:6204-6216(1990).
RN   [10]
RC   pRK212.1 from RK2
RC   pRK229 from RK2
RC   pRK233 from RK2
RC   pRK248 from pRK229
RC   pRK259 from pCR1 & pRK229
RC   pDG2 from pRK215.1 & pBR322
RC   pCT1, pCT2 from pRK248 & pMK20
RC   pCT3 from pCT1
RC   pCT4, pCT5 from pCT1
RC   pCT4.3 from pCT4
RC   pCT6, pCT7, pCT8, pCT17, pCT27 from pCT4.3
RC   pCT14, pCT15, pCT16, pCT32, pCT33 from pCT2
RC   pRK261 from RK2
RC   pRK24 from RK2 & lambda & trp gene
RC   pRK2045 from pSC101 & pRK212.2
RC   pRK2067 from RK2
RA   Thomas C.M., Meyer R., Helinski D.R.;
RT   "Regions of the broad-host-range plasmid RK2 which are essential
RT   for replication and mainenance";
RL   J. Bacteriol. 141:213-222(1980).
RN   [11]
RC   from pCT87 & Tn1723
RC   pVI3 from pKO-6 & pCT294.5
RC   pCT409 from pCT407 & pCT319.1
RC   pCT410 from pRK259.2 & pBR322
RC   pCT411 from pCT410 & pHV60; [similar to pHV32]
RC   pCT415 from pCT407 & pCT411
RA   Shingler V., Thomas C.M.;
RT   "Transcription in the trfA region of broad host range plasmid RK2
RT   is regulated by trfB and korB";
RL   Mol. Gen. Genet. 195:523-529(1984).
RN   [12]
RC   pDS3 from p15A & cat gene
RC   pRK2501ts3R0, pRK2501ts3R14, pRK2501ts3R15 from pRK2501ts3
RC   pCT110 from kan gene
RC   pCT113 from pBR322 & pRK2501ts3
RC   pCT138 from pBR322 & pRK2501ts3R0
RC   pCT139 from pCT138
RC   pCT140 from pBR322 & pRK2501
RC   pCT143 from pCT140
RC   pCT495 from pRK2501 & pDS3
RC   pCT502 from pCT495
RC   pCT503 from pRK2501ts3R0 & pDS3
RC   pCT505 from pRK2501ts3R14 & pDS3
RC   pCT506 from pRK2501ts3R15 & pDS3
RC   [pCT536 from pCT series]
RC   pCT549 from pRK2501 & korB gene
RC   pCT625 from pCT139 & pCT110
RC   pCT626 from pCT625
RC   pCT627 from pCT625 & pCT140
RC   pCT628 from pCT627
RC   pCT629 from pCT628 & pBR322
RC   pCT640 from pCT139 & pCT143
RC   pCT648 from pCT628 & pCT536
RC   pCT670 from pBR322 & galK gene
RC   pCT671 from pCT670 & pCT143, trfB gene
RC   pCT673 from pBR322 & galK gene & lambda, tR1
RC   pCT674 from pCT673 & trfB gene
RC   pCT675 from pRK2501ts3R0 & pCT673
RC   pCT676 from pCT673 & pCT143, trfB gene
RA   Thomas C.M.;
RT   "Evidence for the involvement of the incC locus of
RT   braod-host-range plasmid RK2 in plasmid maintenance";
RL   Plasmid 16:15-29(1986).
RN   [13]
RC   pGBT28 from pCT673 & pGBT19
RC   pGBT29 from pCT673 & pGBT19
RC   pGBT30 from pCT673 & pGBT19
RC   pGBT40 from pCT673 & pGBT25
RC   pGBT41 from pCT673 & pGBT25
RA   Jagura-Burdzy G., Ibbotson J.P., Thomas C.M.;
RT   "The korF region of broad-host-range plasmid RK2 encodes two
RT   polypeptides with transcriptional repressor activity";
RL   J. Bacteriol. 173:826-833(1991).
RN   [14]
RC   pJL23 from pACYC184 & lambda, cI857
RA   Lodge J., Roberts T.;
RT   ;
RL   Unpublished (1985).
RN   [15]
RC   pKB280-cI857 from pKB280 & lambda, cI857
RA   Hecht M.H., Nelson H.C., Sauer R.T.;
RT   "Mutations in lambda repressor's amino-terminal domain:
RT   implications for protein stability and DNA binding";
RL   Proc. Natl. Acad. Sci. U.S.A. 80:2676-2680(1983).
RN   [16]
RC   RK2 from E.coli P inc group
RA   Ingram L., Richmond M.H., Sykes R.B.;
RT   "Molecular characterization of the R factors implicated in the
RT   carbanicillin resistance of a sequence of Pseudomonas aeruginosa
RT   strains isolated from burns";
RL   Antimicrob. Agents Chemother. 3:279-288(1973).
RN   [17]
RC   pCT570 from pSC101 & RK2
RA   Hasnain S., Thomas C.M.;
RT   "Construction of a novel gene bank of B. subtilis using a low copy
RT   number vector in E. coli";
RL   J. Gen. Microbiol. 132:1863-1874(1986).
RN   [18]
RC   pCT720 from RK2 & korA gene & xylE gene
RA   Thomas C.M., Ibbotson J.P., Wang N., Smith C.A., Tipping R.,
RA   Loader N.M.;
RT   "Gene regulation on braod host range plasmid RK2: identification of
RT   three novel operons whose transcription is repressed by both KorA and
RT   KorC";
RL   Nucleic Acids Res. 16:5345-5359(1988).
RN   [19]
RC   pCT730 from pBR322 & kfrA gene & galK gene
RC   pCT731 from pCT730
RC   pGBT19 from pBR322 & korF gene
RC   pGBT25 from pBR322 & korF gene & tac promoter & xylE gene
RA   Thomas C.M., Theophilus B.D., Johnston L., Jagura-Burdzy G.,
RA   Schilf W., Lurz R., Lanka E.;
RT   "Identification of a seventh operon on plasmid RK2 regulated by the
RT   korA gene product";
RL   Gene 89:29-35(1990).
RN   [20]
RC   pFF1 from RK2
RC   pSV16 from RK2
RA   Durland R.H., Toukdarian A., Fang F., Helinski D.R.;
RT   "Mutations in the trfA replication gene of the broad-host-range
RT   plasmid RK2 result in elevated plasmid copy numbers";
RL   J. Bacteriol. 172:3859-3867(1990).
RN   [21]
RC   pBK280 from F factor, ccd gene
RA   Hogan J.E., Kline B.C., Levy S.B.;
RT   "Regions on the F plasmid which affect plasmid maintenance and the
RT   ability to segregate into Escherichia coli minicells";
RL   Plasmid 8:36-44(1982).
RN   [22]
RC   pRD110-cop267L from pBR322 & cop 267L gene
RA   Durland R.;
RT   ;
RL   Unpublished (1990).
RN   [23]
RC   pCT407 from R300B & pCT16
RC   pCT408 from R300B & pCT14
RA   Thomas C.M., Hussain A.A., Smith C.A.;
RT   "Maintenance of broad host range plasmid RK2 replicons in
RT   Pseudomonas aeruginosa";
RL   Nature 298:674-676(1982).
RN   [24]
RC   pVI100.3 from pVI3 & tet gene
RC   pVI104.3 from pVI3 & tet gene
RC   pVI106.3 from pVI3 & tet gene
RC   pVI107.3 from pVI3 & tet gene
RC   pVI108.2 from pVI3
RC   pVI109.2 from pVI3
RC   pVI113.2 from pVI3
RC   pVI114.2 from pVI3
RC   pVI115.2 from pVI3
RC   pVI123.2 from pVI3
RC   [pCT409 from pCT407]
RA   Shingler V., Thomas C.M.;
RT   "Analysis of the trfA region of broad host range plasmid RK2
RT   by transposon mutagenesis and identification of polypeptide products";
RL   J. Mol. Biol. 175:229-249(1984).
RN   [25]
RC   pCT294.5 from pCT series
RC   pCT319.1 from pCT series
RC   pRK259.2 from pRK series
RA   Smith C.A., Thomas C.M.;
RT   "Deletion mapping of kil and kor functions in the trfA and trfB
RT   regions of braod host range plasmid RK2";
RL   Mol. Gen. Genet. 190:245-254(1983).
RN   [26]
RC   pCT549 from pRK2501 & korB gene & kilB gene
RA   Thomas C.M., Hussain A.A.;
RT   "The korB gene of broad host range plasmid RK2 is a major copy number
RT   control element which may act together with trfB by limiting trfA
RT   expression";
RL   EMBO J. 3:1513-1519(1984).
RN   [27]
RC   pAv1 from A.calcoaceticus
RA   Hinchchliffe E., Vivian A.;
RT   "Naturally occurring plasmids in Acinetobacter calcoaceticus: A P
RT   class R factor of restricted host range";
RL   J. Gen. Microbiol. 116:75-86(1980).
RN   [28]
RC   pAS8 from RP4 & ColE1
RC   pAS9 from pAS8
RA   Urlapova S.V., Myakinin V.B., Stepanov A.L.;
RT   "Temperature-sensitive mutant of the plasmid RP1";
RL   Genetika 15:433-443(1979).
RN   [29]
RC   pSDL2 from Salmonella
RA   Krause M., Guiney D.G.;
RT   "Identification of a multimer resolution system involved in
RT   stabilization of the Salmonella dublin virulence plasmid pSDL2";
RL   J. Bacteriol. 173:5754-5762(1991).
CC   IncP plasmid
CC   ATCC depositor: Helinski D.R.
CC   Naturally occuring Pseudomonas plasmid containing relaxation complex
CC   site (rlx). (ATCC staff)
CC   RP4 may be identical to RK2.
CC   Medium is 1227 LB plus ampicillin.
CC   NCBI gi: 48928
CC   NM (RK2)
CC   CM (no)
CC   NA (ds-DNA)
CC   TP (circular)
CC   ST ()
CC   TY (plasmid)
CC   HO (E.coli C600)(E.coli K802)(broad host range)(Pseudomonas)(E.coli)
CC   HO (Gram-negative bacteria)
CC   CP ()
CC   FN (transfer)
CC   SE ()
CC   PA (RK2)(IncP alpha plasmid)
CC   BR (RP4)(RP1)(R18)(R68)
CC   OF ()
CC   OR ()
FH   Key             Location/Qualifiers
FT   misc_feature    0..0
FT                   /note="1. Pseudomonas
FT                   -> RK2"
FT   rep_origin      0..0
FT                   /note="ORI E. coli promiscuous IncP plasmid oriT"
FT   misc_binding    0..0
FT                   /note="SIT EcoRI"
FT   misc_binding    0..0
FT                   /note="SIT BglII"
FT   misc_binding    0..0
FT                   /note="SIT BamHI"
FT   misc_binding    0..0
FT                   /note="SIT HpaI"
FT   misc_binding    0..0
FT                   /note="SIT HindIII"
FT   CDS             0..0
FT                   /note="ANT E. coli beta-lactamase gene (bla)
FT                   ampicillin resistance gene (apr/amp)"
FT   CDS             0..0
FT                   /note="ANT E. coli kanamycin resistance gene (kan)"
FT   CDS             0..0
FT                   /note="ANT E. coli tetracycline resistance gene (tet)"
FT   promoter        49..61
FT                   /note="PRO KorB operator"
FT   CDS             290..1249
FT                   /note="GEN E. coli kilB gene; NCBI gi: 48929"
SQ   Sequence 570 BP; 133 A; 148 C; 182 G; 107 T; 0 other;
     ctggttggct tggtttcatc agccatccgc ttgccctcat ctgttacgcc ggcggtagcc
     ggccagcctc gcagagcagg attcccgttg agcaccgcca ggtgcgaata agggacagtg
     aagaaggaac acccgctcgc gggtgggcct acttcaccta tcctgcccgg ctgacgccgt
     tggatacacc aaggaaagtc tacgcgaacc ctttggcaaa atcctgtata tcgtgcgaaa
     aaggatggat ataccgaaaa aatcgctata atgaccccga agcagggtta tgcagcggaa
     aagcgctgct tccctgctgt tttgtggaat atctaccgac tggaaacagg caaatgcagg
     aaattactga actgagggga caggcgagag acgatgccaa agagctacac cgacgagctg
     gccgagtggg ttgaatcccg cgcggccaag aagcgccggc gtgatgaggc tgcggttgcg
     ttcctggcgg tgagggcgga tgtcgaggcg gcgttagcgt ccggctatgc gctcgtcacc
     atttgggagc acatgcggga aacggggagg