Return to this vector's summary.
ID PMMB66EH preliminary; circular DNA; PRO; 8807 BP.
XX
AC X15234; M28829; V00294; X15194; ATCC37620;
XX
DT 20-DEC-1989 (Rel. 4, Created)
DT 01-JUL-1995 (Rel. 12, Last updated, Version 1)
XX
DE Broad host range/E.coli plasmid vector pMMB66EH - complete.
XX
KW cloning vector.
XX
OS Pseudomonas
OC Prokaryota; Bacteria; Gracilicutes; Scotobacteria;
OC Facultatively anaerobic rods; Enterobacteriaceae.
XX
RN [1]
RP 1-8807
RC pMMB66EH
RA Morales V.M.;
RT ;
RL Submitted (08-MAY-1989) on tape to the EMBL Data Library by:
RL Morales V.M., Michigan Biotechnology Institute, 3900 Collins Road,
RL Lansing MI 48909, USA.
XX
RN [2]
RP 1-8807
RC RP4, R751 from E. coli IncQ plasmids
RA Morales V.M., Baeckman A., Bagdasarian M.;
RT "Promiscuous plasmids of the IncQ group. Mode of replication and
RT use for gene cloning in Gram-negative bacteria.";
RL Pseudomonas. Biotransformations, Pathogenesis and ... 0:1-1(1990).
RL Silver S., Chakrabarty A.M., Gunsalus R.P., Iglewski B.;
RL (eds.), Academic Press, New York.
XX
RN [3]
RC pJF118EH, pJF118HE from pMMB22 & pKK223-3
RC pMMB66EH from RSF1010 & pJF118EH
RC pMMB66HE from pMMB66EH & pJF118HE
RC pJF119EH from pJF118EH & M13mp18
RC pJF119HE from pJF118HE & M13mp18
RC pMMB67EH from pMMB66EH & M13mp18
RC pMMB67HE from pMMB66HE & M13mp18
RC pMS324a from R300B & RP4
RC pFG101 series from pJF118EH & pMS324a
RC pFG102 series from pJF118HE & pMS324a
RC pWP101 series from pMMB66EH & pMS324a
RC pWP102 series from pMMB66HE & pMS324a
RA Furste J.P., Pansegrau W., Frank R., Blocker H., Scholz P.,
RA Bagdasarian M., Lanka E.;
RT "Molecular cloning of the plasmid RP4 primase
RT region in a multi-host-range tacP expression vector";
RL Gene 48:119-131(1986).
XX
RN [4]
RC pMMB66EH from RSF1010 & pJF118EH
RA Morales V.M., Backman A., Bagdasarian M.;
RT "Sequence of junctions of expression vector pMMB66EH";
RL Nucleic Acids Res. 17:0-0(1989).
XX
CC This sequence overlaps M28829 and V00294.
CC Restriction digests of the clone give the following sizes (kb):
CC AccI--3.9, 3.0, 1.9; BamHI--8.8; HindIII/PvuII--7.6, 1.3. (ATCC staff)
CC Contains the following sites separated by (bp): EcoRI-764 -AhaIII-97
CC -ScaI-112 -PvuI-2101 -AccI-1848 -ScaI-55 -AccI-2039 -BstEII-372
CC -PvuII-93 -PvuII-94 -HpaI-320 -BstEII-182 -MluI-736 -EcoRI. [3]
CC The sequence of RSF1010 is given in this reference. [4]
CC A 1246 bp lacIQ HindIII fragment was inserted at XmaIII/BamHI of
CC pKK223-3. The 2995 bp NruI/AhaIII fragment of the resulting plasmid
CC was cloned between PstI (bp 7768) and PvuII (bp 1951) of RSF1010. [3]
CC In the pMMB66 (ATCC 37620, 37621) and pMMB67 (ATCC 37622, 37623)
CC vector series,the EH and HE in the name designate the orientation of
CC the multiple cloning site to the tac promoter.
CC (personal communication)
CC An autoregulated high-level expression vector utilizing the tac
CC promoter, rrnB terminators and lacIq for lac repression.
CC Concentration of induced gene product may be regulated by varying the
CC lac inducer concentration. [3]
CC Can be mobilized by conjugative IncP helper plasmids [e.g. pRK2013
CC (ATCC 37159), pUB307] into Klebsiella aerogenes, Proteus mirabilis,
CC Pseudomonas aeruginosa, Serratia marcescens, and other gram-negative
CC bacteria. [3]
CC Medium is 1227 LB plus ampicillin.
CC NM (pMMB66EH)
CC CM (yes)
CC NA (ds-DNA)
CC TP (circular)
CC ST ()
CC TY (plasmid)
CC SP (ATCC)
CC HO (Gram-negative bacteria)(E.coli K-12)(E.coli HB101)(E.coli)
CC HO (broad host range)
CC CP (low)
CC FN (cloning)
CC SE (color blue/white)
CC PA ()
CC BR (pMMB190)(pMMB206)
CC OF ()
CC OR ()
XX
FH Key Location/Qualifiers
FH
FT misc_feature 0..0
FT /note="1. pMMB22 HindIII-HindIII 1247bp, lacIq gene
FT 2. pKK223-3 remove BamHI-XmaIII 564bp 257..821
FT \ 4020bp [547bp]
FT -> pJF118EH 5267bp
FT 1. RSF1010 remove PstI-PvuII 2754bp 7910..8684..1980,
FT \ 5930bp
FT 2. pJF118EH NruI-AhaIII 2996bp, pKK223-3 NruI-? 856..?
FT -> pMMB66EH 8807bp"
FT promoter 482..509
FT /note="PRO E. coli bla gene"
FT CDS 552..1409
FT /note="ANT E. coli beta-lactamase gene (bla)
FT ampicillin resistance gene (apr/amp) (AA 1-286)"
FT rep_origin 1850..2245
FT /note="ORI E. coli IncQ plasmid origin of vegetative
FT replication oriV"
FT CDS complement(2273..2554)
FT /note="GEN E. coli mobC gene
FT (conjugational DNA transfer) (AA 1-94)"
FT promoter 2546..2576
FT /note="PRO E. coli promoter P1"
FT promoter 2565..2595
FT /note="PRO E. coli promoter P3"
FT rep_origin 2585..2672
FT /note="ORI E. coli IncQ plasmid origin of conjugative
FT DNA transfer oriT"
FT promoter complement(2587..2617)
FT /note="PRO E. coli promoter"
FT CDS 2753..4879
FT /note="GEN E. coli repB/mobA gene (primase)
FT (AA 1-709)"
FT CDS 3501..3911
FT /note="GEN E. coli mobB gene
FT (conjugational DNA transfer) (AA 1-137)"
FT CDS 3911..4879
FT /note="GEN E. coli repB' gene (primase) (AA 1-323)"
FT promoter 4884..4915
FT /note="PRO E. coli promoter P4"
FT CDS 4943..5152
FT /note="GEN E. coli E gene (function unknown)
FT (AA 1-70)"
FT CDS 5157..5360
FT /note="GEN E. coli cac gene
FT (regulates repA and repC gene expression) (AA 1-68)"
FT CDS 5393..6229
FT /note="GEN E. coli repA gene (DNA helicase)
FT (AA 1-279)"
FT CDS 6219..7067
FT /note="GEN E. coli repC gene (replication initiator
FT protein) (AA 1-283)"
FT promoter 7231..7261
FT /note="PRO E. coli promoter P5"
FT promoter complement(8463..8492)
FT /note="PRO E. coli lacIq repressor gene"
FT promoter 8736..8764
FT /note="PRO E. coli tac (trp and lac)"
FT misc_binding 0..0
FT /note="MCS unique EcoRI-SmaI-BamHI-SalI-PstI-HindIII"
FT rep_origin 0..0
FT /note="ORI E. coli RSF1010"
FT CDS 0..0
FT /note="REP E. coli lacIq repressor gene"
XX
SQ Sequence 8807 BP; 1904 A; 2577 C; 2631 G; 1695 T; 0 other;
ttcccgggga tccgtcgacc tgcagccaag cttggctgtt ttggcggatg agagaagatt
ttcagcctga tacagattaa atcagaacgc agaagcggtc tgataaaaca gaatttgcct
ggcggcagta gcgcggtggt cccacctgac cccatgccga actcagaagt gaaacgccgt
agcgccgatg gtagtgtggg gtctccccat gcgagagtag ggaactgcca ggcatcaaat
aaaacgaaag gctcagtcga aagactgggc ctttcgtttt atctgttgtt tgtcggtgaa
cgctctcctg agtaggacaa atccgccggg agcggatttg aacgttgcga agcaacggcc
cggagggtgg cgggcaggac gcccgccata aactgccagg catcaaatta agcagaaggc
catcctgacg gatggccttt ttgcgtttct acaaactctt ttgtttattt ttctaaatac
attcaaatat gtatccgctc atgagacaat aaccctgata aatgcttcaa taatattgaa
aaaggaagag tatgagtatt caacatttcc gtgtcgccct tattcccttt tttgcggcat
tttgccttcc tgtttttgct cacccagaaa cgctggtgaa agtaaaagat gctgaagatc
agttgggtgc acgagtgggt tacatcgaac tggatctcaa cagcggtaag atccttgaga
gttttcgccc cgaagaacgt tttccaatga tgagcacttt taaagttctg ctatgtggcg
cggtattatc ccgtgttgac gccgggcaag agcaactcgg tcgccgcata cactattctc
agaatgactt ggttgagtac tcaccagtca cagaaaagca tcttacggat ggcatgacag
taagagaatt atgcagtgct gccataacca tgagtgataa cactgcggcc aacttacttc
tgacaacgat cggaggaccg aaggagctaa ccgctttttt gcacaacatg ggggatcatg
taactcgcct tgatcgttgg gaaccggagc tgaatgaagc cataccaaac gacgagcgtg
acaccacgat gcctgtagca atggcaacaa cgttgcgcaa actattaact ggcgaactac
ttactctagc ttcccggcaa caattaatag actggatgga ggcggataaa gttgcaggac
cacttctgcg ctcggccctt ccggctggct ggtttattgc tgataaatct ggagccggtg
agcgtgggtc tcgcggtatc attgcagcac tggggccaga tggtaagccc tcccgtatcg
tagttatcta cacgacgggg agtcaggcaa ctatggatga acgaaataga cagatcgctg
agataggtgc ctcactgatt aagcattggt aactgtcaga ccaagtttac tcatatatac
tttagattga tttctgaaag cgaccaggtg ctcggcgtgg caagactcgc agcgaacccg
tagaaagcca tgctccagcc gcccgcattg gagaaattct tcaaattccc gttgcacata
gcccggcaat tcctttccct gctctgccat aagcgcagcg aatgccgggt aatactcgtc
aacgatctga tagagaaggg tttgctcggg tcggtggctc tggtaacgac cagtatcccg
atcccggctg gccgtcctgg ccgccacatg aggcatgttc cgcgtccttg caatactgtg
tttacataca gtctatcgct tagcggaaag ttcttttacc ctcagccgaa atgcctgccg
ttgctagaca ttgccagcca gtgcccgtca ctcccgtact aactgtcacg aacccctgca
ataactgtca cgcccccctg caataactgt cacgaacccc tgcaataact gtcacgcccc
caaacctgca aacccagcag gggcgggggc tggcggggtg ttggaaaaat ccatccatga
ttatctaaga ataatccact aggcgcggtt atcagcgccc ttgtggggcg ctgctgccct
tgcccaatat gcccggccag aggccggata gctggtctat tcgctgcgct aggctacaca
ccgccccacc gctgcgcggc agggggaaag gcgggcaaag cccgctaaac cccacaccaa
accccgcaga aatacgctgg agcgctttta gccgctttag cggcctttcc ccctacccga
agggtggggg cgcgtgtgca gccccgcagg gcctgtctcg gtcgatcatt cagcccggct
catccttctg gcgtggcggc agaccgaaca aggcgcggtc gtggtcgcgt tcaaggtacg
catccattgc cgccatgagc cgatcctccg gccactcgct gctgttcacc ttggccaaaa
tcatggcccc caccagcacc ttgcgccttg tttcgttctt gcgctcttgc tgctgttccc
ttgcccgctc ccgctgaatt tcggcattga ttcgcgctcg ttgttcttcg agcttggcca
gccgatccgc cgccttgttg ctccccttaa ccatcttgac accccattgt taatgtgctg
tctcgtaggc tatcatggag gcacagcggc ggcaatcccg accctacttt gtaggggagg
gcgcacttac cggtttctct tcgagaaact ggcctaacgg ccacccttcg ggcggtgcgc
tctccgaggg ccattgcatg gagccgaaaa gcaaaagcaa cagcgaggca gcatggcgat
ttatcacctt acggcgaaaa ccggcagcag gtcgggcggc caatcggcca gggccaaggc
cgactacatc cagcgcgaag gcaagtatgc ccgcgacatg gatgaagtct tgcacgccga
atccgggcac atgccggagt tcgtcgagcg gcccgccgac tactgggatg ctgccgacct
gtatgaacgc gccaatgggc ggctgttcaa ggaggtcgaa tttgccctgc cggtcgagct
gaccctcgac cagcagaagg cgctggcgtc cgagttcgcc cagcacctga ccggtgccga
gcgcctgccg tatacgctgg ccatccatgc cggtggcggc gagaacccgc actgccacct
gatgatctcc gagcggatca atgacggcat cgagcggccc gccgctcagt ggttcaagcg
gtacaacggc aagaccccgg agaagggcgg ggcacagaag accgaagcgc tcaagcccaa
ggcatggctt gagcagaccc gcgaggcatg ggccgaccat gccaaccggg cattagagcg
ggctggccac gacgcccgca ttgaccacag aacacttgag gcgcagggca tcgagcgcct
gcccggtgtt cacctggggc cgaacgtggt ggagatggaa ggccggggca tccgcaccga
ccgggcagac gtggccctga acatcgacac cgccaacgcc cagatcatcg acttacagga
ataccgggag gcaatagacc atgaacgcaa tcgacagagt gaagaaatcc agaggcatca
acgagttagc ggagcagatc gaaccgctgg cccagagcat ggcgacactg gccgacgaag
cccggcaggt catgagccag accaagcagg ccagcgaggc gcaggcggcg gagtggctga
aagcccagcg ccagacaggg gcggcatggg tggagctggc caaagagttg cgggaggtag
ccgccgaggt gagcagcgcc gcgcagagcg cccggagcgc gtcgcggggg tggcactgga
agctatggct aaccgtgatg ctggcttcca tgatgcctac ggtggtgctg ctgatcgcat
cgttgctctt gctcgacctg acgccactga caaccgagga cggctcgatc tggctgcgct
tggtggcccg atgaagaacg acaggacttt gcaggccata ggccgacagc tcaaggccat
gggctgtgag cgcttcgata tcggcgtcag ggacgcaccc accggccaga tgatgaaccg
ggaatggtca gccgccgaag tgctccagaa cacgccatgg ctcaagcgga tgaatgccca
gggcaatgac gtgtatatca ggcccgccga gcaggagcgg catggtctgg tgctggtgga
cgacctcagc gagtttgacc tggatgacat gaaagccgag ggccgggagc ctgccctggt
agtggaaacc agcccgaaga actatcaggc atgggtcaag gtggccgacg ccgcaggcgg
tgaacttcgg gggcagattg cccggacgct ggccagcgag tacgacgccg acccggccag
cgccgacagc cgccactatg gccgcttggc gggcttcacc aaccgcaagg acaagcacac
cacccgcgcc ggttatcagc cgtgggtgct gctgcgtgaa tccaagggca agaccgccac
cgctggcccg gcgctggtgc agcaggctgg ccagcagatc gagcaggccc agcggcagca
ggagaaggcc cgcaggctgg ccagcctcga actgcccgag cggcagctta gccgccaccg
gcgcacggcg ctggacgagt accgcagcga gatggccggg ctggtcaagc gcttcggtca
tgacctcagc aagtgcgact ttatcgccgc gcagaagctg gccagccggg gccgcagtgc
cgaggaaatc ggcaaggcca tggccgaggc cagcccagcg ctggcagagc gcaagcccgg
ccacgaagcg gattacatcg agcgcaccgt cagcaaggtc atgggtctgc ccagcgtcca
gcttgcgcgg gccgagctgg cacgggcacc ggcaccccgc cagcgaggca tggacagggg
cgggccagat ttcagcatgt agtgcttgcg ttggtactca cgcctgttat actatgagta
ctcacgcaca gaagggggtt ttatggaata cgaaaaaagc gcttcagggt cggtctacct
gatcaaaagt gacaagggct attggttgcc cggtggcttt ggttatacgt caaacaaggc
cgaggctggc cgcttttcag tcgctgatat ggccagcctt aaccttgacg gctgcacctt
gtccttgttc cgcgaagaca agcctttcgg ccccggcaag tttctcggtg actgatatga
aagaccaaaa ggacaagcag accggcgacc tgctggccag ccctgacgct gtacgccaag
cgcgatatgc cgagcgcatg aaggccaaag ggatgcgtca gcgcaagttc tggctgaccg
acgacgaata cgaggcgctg cgcgagtgcc tggaagaact cagagcggcg cagggcgggg
gtagtgaccc cgccagcgcc taaccaccaa ctgcctgcaa aggaggcaat caatggctac
ccataagcct atcaatattc tggaggcgtt cgcagcagcg ccgccaccgc tggactacgt
tttgcccaac atggtggccg gtacggtcgg ggcgctggtg tcgcccggtg gtgccggtaa
atccatgctg gccctgcaac tggccgcaca gattgcaggc gggccggatc tgctggaggt
gggcgaactg cccaccggcc cggtgatcta cctgcccgcc gaagacccgc ccaccgccat
tcatcaccgc ctgcacgccc ttggggcgca cctcagcgcc gaggaacggc aagccgtggc
tgacggcctg ctgatccagc cgctgatcgg cagcctgccc aacatcatgg ccccggagtg
gttcgacggc ctcaagcgcg ccgccgaggg ccgccgcctg atggtgctgg acacgctgcg
ccggttccac atcgaggaag aaaacgccag cggccccatg gcccaggtca tcggtcgcat
ggaggccatc gccgccgata ccgggtgctc tatcgtgttc ctgcaccatg ccagcaaggg
cgcggccatg atgggcgcag gcgaccagca gcaggccagc cggggcagct cggtactggt
cgataacatc cgctggcagt cctacctgtc gagcatgacc agcgccgagg ccgaggaatg
gggtgtggac gacgaccagc gccggttctt cgtccgcttc ggtgtgagca aggccaacta
tggcgcaccg ttcgctgatc ggtggttcag gcggcatgac ggcggggtgc tcaagcccgc
cgtgctggag aggcagcgca agagcaaggg ggtgccccgt ggtgaagcct aagaacaagc
acagcctcag ccacgtccgg cacgacccgg cgcactgtct ggcccccggc ctgttccgtg
ccctcaagcg gggcgagcgc aagcgcagca agctggacgt gacgtatgac tacggcgacg
gcaagcggat cgagttcagc ggcccggagc cgctgggcgc tgatgatctg cgcatcctgc
aagggctggt ggccatggct gggcctaatg gcctagtgct tggcccggaa cccaagaccg
aaggcggacg gcagctccgg ctgttcctgg aacccaagtg ggaggccgtc accgctgaat
gccatgtggt caaaggtagc tatcgggcgc tggcaaagga aatcggggca gaggtcgata
gtggtggggc gctcaagcac atacaggact gcatcgagcg cctttggaag gtatccatca
tcgcccagaa tggccgcaag cggcaggggt ttcggctgct gtcggagtac gccagcgacg
aggcggacgg gcgcctgtac gtggccctga accccttgat cgcgcaggcc gtcatgggtg
gcggccagca tgtgcgcatc agcatggacg aggtgcgggc gctggacagc gaaaccgccc
gcctgctgca ccagcggctg tgtggctgga tcgaccccgg caaaaccggc aaggcttcca
tagatacctt gtgcggctat gtctggccgt cagaggccag tggttcgacc atgcgcaagc
gccgcaagcg ggtgcgcgag gcgttgccgg agctggtcgc gctgggctgg acggtaaccg
agttcgcggc gggcaagtac gacatcaccc ggcccaaggc ggcaggctga ccccccccac
tctattgtaa acaagacatt tttatctttt atattcaatg gcttattttc ctgctaattg
gtaataccat gaaaaatacc atgctcagaa aaggcttaac aatattttga aaaattgcct
actgagcgct gccgcacagc tccataggcc gctttcctgg ctttgcttcc agatgtatgc
tcttctgctc ccgaacgcca gcaagacgta gcccagcgcg tcggccagct tgcaattcgc
gctaacttac attaattgcg ttgcgctcac tgcccgcttt ccagtcggga aacctgtcgt
gccagctgca ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt attgggcgcc
agggtggttt ttcttttcac cagtgagacg ggcaacagct gattgccctt caccgcctgg
ccctgagaga gttgcagcaa gcggtccacg tggtttgccc cagcaggcga aaatcctgtt
tgatggtggt taacggcggg atataacatg agctgtcttc ggtatcgtcg tatcccacta
ccgagatatc cgcaccaacg cgcagcccgg actcggtaat ggcgcgcatt gcgcccagcg
ccatctgatc gttggcaacc agcatcgcag tgggaacgat gccctcattc agcatttgca
tggtttgttg aaaaccggac atggcactcc agtcgccttc ccgttccgct atcggctgaa
tttgattgcg agtgagatat ttatgccagc cagccagacg cagacgcgcc gagacagaac
ttaatgggcc cgctaacagc gcgatttgct ggtgacccaa tgcgaccaga tgctccacgc
ccagtcgcgt accgtcttca tgggagaaaa taatactgtt gatgggtgtc tggtcagaga
catcaagaaa taacgccgga acattagtgc aggcagcttc cacagcaatg gcatcctggt
catccagcgg atagttaatg atcagcccac tgacgcgttg cgcgagaaga ttgtgcaccg
ccgctttaca ggcttcgacg ccgcttcgtt ctaccatcga caccaccacg ctggcaccca
gttgatcggc gcgagattta atcgccgcga caatttgcga cggcgcgtgc agggccagac
tggaggtggc aacgccaatc agcaacgact gtttgcccgc cagttgttgt gccacgcggt
tgggaatgta attcagctcc gccatcgccg cttccacttt ttcccgcgtt ttcgcagaaa
cgtggctggc ctggttcacc acgcgggaaa cggtctgata agagacaccg gcatactctg
cgacatcgta taacgttact ggtttcacat tcaccaccct gaattgactc tcttccgggc
gctatcatgc cataccgcga aaggttttgc accattcgat ggtgtcaacg taaatgccgc
ttcgccttcg cgcgcgaatt gcaagctgat ccgggcttat cgactgcacg gtgcaccaat
gcttctggcg tcaggcagcc atcggaagct gtggtatggc tgtgcaggtc gtaaatcact
gcataattcg tgtcgctcaa ggcgcactcc cgttctggat aatgtttttt gcgccgacat
cataacggtt ctggcaaata ttctgaaatg agctgttgac aattaatcat cggctcgtat
aatgtgtgga attgtgagcg gataacaatt tcacacagga aacagaa
//