Return to this vector's summary.
ID PMMB66HE preliminary; circular DNA; SYN; 8821 BP.
XX
AC M28829; L23118; ATCC37621;
XX
DT 01-JUL-1993 (Rel. 7, Created)
DT 01-JUL-1995 (Rel. 12, Last updated, Version 1)
XX
DE Broad host range/E.coli plasmid vector pMMB66HE - complete.
XX
KW cloning vector.
XX
OS Cloning vector
OC Artificial sequences; Cloning vehicles.
XX
RN [1]
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 region in a
RT multi-host-range tacP expression vector";
RL Gene 48:119-131(1986).
XX
RN [2]
RC pMG from pMMB66HE & pCH58, phoA gene
RA Blanco D.R., Giladi M., Champion C.I., Haake D.A., Chikami G.K.,
RA Miller J.N., Lovett M.A.;
RT "Identification of Treponema pallidum subspecies pallidum genes
RT encoding signal peptides and membrane-spanning sequences using a
RT novel alkaline phosphatase expression vector";
RL Mol. Microbiol. 5:2405-2415(1991).
XX
CC Created by Moore, July 1995, under contract with NCBI.
CC Restriction digests of the clone give the following sizes (kb):
CC AccI--3.8, 3.1, 1.9; BamHI--8.6; HindIII/PvuII--7.4, 1.3. (ATCC staff)
CC Contains the following sites separated by (bp): EcoRI - 739 - AhaIII -
CC 97 - ScaI - 112 - PvuI - 2101 - AccI - 1848 - ScaI - 55 - AccI - 2039
CC - BstEII - 372 - PvuII - 93 - PvuII - 94 - HpaI - 320 - BstEII - 182
CC - MluI - 771 - EcoRI. (personal communication)
CC The sequence of RSF1010 is given in this reference. [2]
CC A 1246 bp lacIQ HindIII fragment inserted at XmaIII/BamHI of pKK223-3.
CC A M13mp9 polylinker replaced the M13mp8 polylinker of pMMB66EH
CC (ATCC 37620).
CC THe 3005 bp NruI/AhaIII fragment was cloned between PstI (7768) and
CC PvuII (1951) of RSF1010. [1]
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. (personal
CC communication)
CC An autoregulated high-level expression vector utilizing the tac
CC promoter, rrnB terminators and lacIq for lac repression. Concentration
CC of induced gene product may be regulated by varying the lac inducer
CC concentration. [1]
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. [1]
CC Medium is 1227 LB plus ampicillin.
CC NM (pMMB66HE)
CC CM (yes)
CC NA (ds-DNA)
CC TP (circular)
CC ST ()
CC TY (plasmid)
CC SP (ATCC)
CC HO (E.coli HB101)(broad host range)(E.coli)
CC CP ()
CC FN (cloning)
CC SE ()
CC PA (pMMB66EH)(pJF118HE)
CC BR ()
CC OF ()
CC OR ()
XX
FH Key Location/Qualifiers
FH
FT misc_feature 0..0
FT /note="1. pMMB22 remove, 12700bp-
FT 2. pKK223-3 4584bp
FT -> pJF118EH
FT 1. pJF118EH remove EcoRI-HindIII, MCS
FT fill in:fill in
FT 2. oligo HindIII-PstI-SalI-BamHI-SmaI-EcoRI 40bp
FT \ aattaagcttggctgcaggtcgacggatccccgggaattc
FT -> pJF118HE
FT 1. pMMB66EH remove MluI-PvuI 1704bp 8074..8807..971,
FT \ 7103bp
FT 2. pJF118HE MluI-PvuI 1719bp
FT \ pKK223-3 MluI 3617
FT -> pMMB66HE 8820bp"
FT - 1..40
FT /note="aattaagcttggctgcaggtcgacggatccccgggaattc 40bp
FT \ ...gaattc
FT HindIII = A^AGCTT"
FT - 41..8821
FT /note="pMMB66EH 29..8807..2 8781bp
FT EcoRI = G^AATT C
FT \ aatta..."
FT misc_binding 0..0
FT /note="MCS HindIII-PstI-SalI-BamHI-SmaI-EcoRI"
FT misc_binding 0..0
FT /note="SIT unique HindIII-PstI-SalI-BamHI-SmaI-EcoRI"
FT rep_origin 0..0
FT /note="ORI E. coli IncQ plasmid RSF1010 oriT and oriV"
FT promoter 0..0
FT /note="PRO E. coli tac"
FT CDS 0..0
FT /note="REP E. coli lacIq repressor gene"
FT CDS 0..0
FT /note="ANT E. coli beta-lactamase gene (bla)
FT ampicillin resistance gene (apr/amp)"
XX
SQ Sequence 8821 BP; 1909 A; 2576 C; 2635 G; 1701 T; 0 other;
aattaagctt ggctgcaggt cgacggatcc ccgggaattc agcttggctg ttttggcgga
tgagagaaga ttttcagcct gatacagatt aaatcagaac gcagaagcgg tctgataaaa
cagaatttgc ctggcggcag tagcgcggtg gtcccacctg accccatgcc gaactcagaa
gtgaaacgcc gtagcgccga tggtagtgtg gggtctcccc atgcgagagt agggaactgc
caggcatcaa ataaaacgaa aggctcagtc gaaagactgg gcctttcgtt ttatctgttg
tttgtcggtg aacgctctcc tgagtaggac aaatccgccg ggagcggatt tgaacgttgc
gaagcaacgg cccggagggt ggcgggcagg acgcccgcca taaactgcca ggcatcaaat
taagcagaag gccatcctga cggatggcct ttttgcgttt ctacaaactc ttttgtttat
ttttctaaat acattcaaat atgtatccgc tcatgagaca ataaccctga taaatgcttc
aataatattg aaaaaggaag agtatgagta ttcaacattt ccgtgtcgcc cttattccct
tttttgcggc attttgcctt cctgtttttg ctcacccaga aacgctggtg aaagtaaaag
atgctgaaga tcagttgggt gcacgagtgg gttacatcga actggatctc aacagcggta
agatccttga gagttttcgc cccgaagaac gttttccaat gatgagcact tttaaagttc
tgctatgtgg cgcggtatta tcccgtgttg acgccgggca agagcaactc ggtcgccgca
tacactattc tcagaatgac ttggttgagt actcaccagt cacagaaaag catcttacgg
atggcatgac agtaagagaa ttatgcagtg ctgccataac catgagtgat aacactgcgg
ccaacttact tctgacaacg atcggaggac cgaaggagct aaccgctttt ttgcacaaca
tgggggatca tgtaactcgc cttgatcgtt gggaaccgga gctgaatgaa gccataccaa
acgacgagcg tgacaccacg atgcctgtag caatggcaac aacgttgcgc aaactattaa
ctggcgaact acttactcta gcttcccggc aacaattaat agactggatg gaggcggata
aagttgcagg accacttctg cgctcggccc ttccggctgg ctggtttatt gctgataaat
ctggagccgg tgagcgtggg tctcgcggta tcattgcagc actggggcca gatggtaagc
cctcccgtat cgtagttatc tacacgacgg ggagtcaggc aactatggat gaacgaaata
gacagatcgc tgagataggt gcctcactga ttaagcattg gtaactgtca gaccaagttt
actcatatat actttagatt gatttctgaa agcgaccagg tgctcggcgt ggcaagactc
gcagcgaacc cgtagaaagc catgctccag ccgcccgcat tggagaaatt cttcaaattc
ccgttgcaca tagcccggca attcctttcc ctgctctgcc ataagcgcag cgaatgccgg
gtaatactcg tcaacgatct gatagagaag ggtttgctcg ggtcggtggc tctggtaacg
accagtatcc cgatcccggc tggccgtcct ggccgccaca tgaggcatgt tccgcgtcct
tgcaatactg tgtttacata cagtctatcg cttagcggaa agttctttta ccctcagccg
aaatgcctgc cgttgctaga cattgccagc cagtgcccgt cactcccgta ctaactgtca
cgaacccctg caataactgt cacgcccccc tgcaataact gtcacgaacc cctgcaataa
ctgtcacgcc cccaaacctg caaacccagc aggggcgggg gctggcgggg tgttggaaaa
atccatccat gattatctaa gaataatcca ctaggcgcgg ttatcagcgc ccttgtgggg
cgctgctgcc cttgcccaat atgcccggcc agaggccgga tagctggtct attcgctgcg
ctaggctaca caccgcccca ccgctgcgcg gcagggggaa aggcgggcaa agcccgctaa
accccacacc aaaccccgca gaaatacgct ggagcgcttt tagccgcttt agcggccttt
ccccctaccc gaagggtggg ggcgcgtgtg cagccccgca gggcctgtct cggtcgatca
ttcagcccgg ctcatccttc tggcgtggcg gcagaccgaa caaggcgcgg tcgtggtcgc
gttcaaggta cgcatccatt gccgccatga gccgatcctc cggccactcg ctgctgttca
ccttggccaa aatcatggcc cccaccagca ccttgcgcct tgtttcgttc ttgcgctctt
gctgctgttc ccttgcccgc tcccgctgaa tttcggcatt gattcgcgct cgttgttctt
cgagcttggc cagccgatcc gccgccttgt tgctcccctt aaccatcttg acaccccatt
gttaatgtgc tgtctcgtag gctatcatgg aggcacagcg gcggcaatcc cgaccctact
ttgtagggga gggcgcactt accggtttct cttcgagaaa ctggcctaac ggccaccctt
cgggcggtgc gctctccgag ggccattgca tggagccgaa aagcaaaagc aacagcgagg
cagcatggcg atttatcacc ttacggcgaa aaccggcagc aggtcgggcg gccaatcggc
cagggccaag gccgactaca tccagcgcga aggcaagtat gcccgcgaca tggatgaagt
cttgcacgcc gaatccgggc acatgccgga gttcgtcgag cggcccgccg actactggga
tgctgccgac ctgtatgaac gcgccaatgg gcggctgttc aaggaggtcg aatttgccct
gccggtcgag ctgaccctcg accagcagaa ggcgctggcg tccgagttcg cccagcacct
gaccggtgcc gagcgcctgc cgtatacgct ggccatccat gccggtggcg gcgagaaccc
gcactgccac ctgatgatct ccgagcggat caatgacggc atcgagcggc ccgccgctca
gtggttcaag cggtacaacg gcaagacccc ggagaagggc ggggcacaga agaccgaagc
gctcaagccc aaggcatggc ttgagcagac ccgcgaggca tgggccgacc atgccaaccg
ggcattagag cgggctggcc acgacgcccg cattgaccac agaacacttg aggcgcaggg
catcgagcgc ctgcccggtg ttcacctggg gccgaacgtg gtggagatgg aaggccgggg
catccgcacc gaccgggcag acgtggccct gaacatcgac accgccaacg cccagatcat
cgacttacag gaataccggg aggcaataga ccatgaacgc aatcgacaga gtgaagaaat
ccagaggcat caacgagtta gcggagcaga tcgaaccgct ggcccagagc atggcgacac
tggccgacga agcccggcag gtcatgagcc agaccaagca ggccagcgag gcgcaggcgg
cggagtggct gaaagcccag cgccagacag gggcggcatg ggtggagctg gccaaagagt
tgcgggaggt agccgccgag gtgagcagcg ccgcgcagag cgcccggagc gcgtcgcggg
ggtggcactg gaagctatgg ctaaccgtga tgctggcttc catgatgcct acggtggtgc
tgctgatcgc atcgttgctc ttgctcgacc tgacgccact gacaaccgag gacggctcga
tctggctgcg cttggtggcc cgatgaagaa cgacaggact ttgcaggcca taggccgaca
gctcaaggcc atgggctgtg agcgcttcga tatcggcgtc agggacgcac ccaccggcca
gatgatgaac cgggaatggt cagccgccga agtgctccag aacacgccat ggctcaagcg
gatgaatgcc cagggcaatg acgtgtatat caggcccgcc gagcaggagc ggcatggtct
ggtgctggtg gacgacctca gcgagtttga cctggatgac atgaaagccg agggccggga
gcctgccctg gtagtggaaa ccagcccgaa gaactatcag gcatgggtca aggtggccga
cgccgcaggc ggtgaacttc gggggcagat tgcccggacg ctggccagcg agtacgacgc
cgacccggcc agcgccgaca gccgccacta tggccgcttg gcgggcttca ccaaccgcaa
ggacaagcac accacccgcg ccggttatca gccgtgggtg ctgctgcgtg aatccaaggg
caagaccgcc accgctggcc cggcgctggt gcagcaggct ggccagcaga tcgagcaggc
ccagcggcag caggagaagg cccgcaggct ggccagcctc gaactgcccg agcggcagct
tagccgccac cggcgcacgg cgctggacga gtaccgcagc gagatggccg ggctggtcaa
gcgcttcggt catgacctca gcaagtgcga ctttatcgcc gcgcagaagc tggccagccg
gggccgcagt gccgaggaaa tcggcaaggc catggccgag gccagcccag cgctggcaga
gcgcaagccc ggccacgaag cggattacat cgagcgcacc gtcagcaagg tcatgggtct
gcccagcgtc cagcttgcgc gggccgagct ggcacgggca ccggcacccc gccagcgagg
catggacagg ggcgggccag atttcagcat gtagtgcttg cgttggtact cacgcctgtt
atactatgag tactcacgca cagaaggggg ttttatggaa tacgaaaaaa gcgcttcagg
gtcggtctac ctgatcaaaa gtgacaaggg ctattggttg cccggtggct ttggttatac
gtcaaacaag gccgaggctg gccgcttttc agtcgctgat atggccagcc ttaaccttga
cggctgcacc ttgtccttgt tccgcgaaga caagcctttc ggccccggca agtttctcgg
tgactgatat gaaagaccaa aaggacaagc agaccggcga cctgctggcc agccctgacg
ctgtacgcca agcgcgatat gccgagcgca tgaaggccaa agggatgcgt cagcgcaagt
tctggctgac cgacgacgaa tacgaggcgc tgcgcgagtg cctggaagaa ctcagagcgg
cgcagggcgg gggtagtgac cccgccagcg cctaaccacc aactgcctgc aaaggaggca
atcaatggct acccataagc ctatcaatat tctggaggcg ttcgcagcag cgccgccacc
gctggactac gttttgccca acatggtggc cggtacggtc ggggcgctgg tgtcgcccgg
tggtgccggt aaatccatgc tggccctgca actggccgca cagattgcag gcgggccgga
tctgctggag gtgggcgaac tgcccaccgg cccggtgatc tacctgcccg ccgaagaccc
gcccaccgcc attcatcacc gcctgcacgc ccttggggcg cacctcagcg ccgaggaacg
gcaagccgtg gctgacggcc tgctgatcca gccgctgatc ggcagcctgc ccaacatcat
ggccccggag tggttcgacg gcctcaagcg cgccgccgag ggccgccgcc tgatggtgct
ggacacgctg cgccggttcc acatcgagga agaaaacgcc agcggcccca tggcccaggt
catcggtcgc atggaggcca tcgccgccga taccgggtgc tctatcgtgt tcctgcacca
tgccagcaag ggcgcggcca tgatgggcgc aggcgaccag cagcaggcca gccggggcag
ctcggtactg gtcgataaca tccgctggca gtcctacctg tcgagcatga ccagcgccga
ggccgaggaa tggggtgtgg acgacgacca gcgccggttc ttcgtccgct tcggtgtgag
caaggccaac tatggcgcac cgttcgctga tcggtggttc aggcggcatg acggcggggt
gctcaagccc gccgtgctgg agaggcagcg caagagcaag ggggtgcccc gtggtgaagc
ctaagaacaa gcacagcctc agccacgtcc ggcacgaccc ggcgcactgt ctggcccccg
gcctgttccg tgccctcaag cggggcgagc gcaagcgcag caagctggac gtgacgtatg
actacggcga cggcaagcgg atcgagttca gcggcccgga gccgctgggc gctgatgatc
tgcgcatcct gcaagggctg gtggccatgg ctgggcctaa tggcctagtg cttggcccgg
aacccaagac cgaaggcgga cggcagctcc ggctgttcct ggaacccaag tgggaggccg
tcaccgctga atgccatgtg gtcaaaggta gctatcgggc gctggcaaag gaaatcgggg
cagaggtcga tagtggtggg gcgctcaagc acatacagga ctgcatcgag cgcctttgga
aggtatccat catcgcccag aatggccgca agcggcaggg gtttcggctg ctgtcggagt
acgccagcga cgaggcggac gggcgcctgt acgtggccct gaaccccttg atcgcgcagg
ccgtcatggg tggcggccag catgtgcgca tcagcatgga cgaggtgcgg gcgctggaca
gcgaaaccgc ccgcctgctg caccagcggc tgtgtggctg gatcgacccc ggcaaaaccg
gcaaggcttc catagatacc ttgtgcggct atgtctggcc gtcagaggcc agtggttcga
ccatgcgcaa gcgccgcaag cgggtgcgcg aggcgttgcc ggagctggtc gcgctgggct
ggacggtaac cgagttcgcg gcgggcaagt acgacatcac ccggcccaag gcggcaggct
gacccccccc actctattgt aaacaagaca tttttatctt ttatattcaa tggcttattt
tcctgctaat tggtaatacc atgaaaaata ccatgctcag aaaaggctta acaatatttt
gaaaaattgc ctactgagcg ctgccgcaca gctccatagg ccgctttcct ggctttgctt
ccagatgtat gctcttctgc tcccgaacgc cagcaagacg tagcccagcg cgtcggccag
cttgcaattc gcgctaactt acattaattg cgttgcgctc actgcccgct ttccagtcgg
gaaacctgtc gtgccagctg cattaatgaa tcggccaacg cgcggggaga ggcggtttgc
gtattgggcg ccagggtggt ttttcttttc accagtgaga cgggcaacag ctgattgccc
ttcaccgcct ggccctgaga gagttgcagc aagcggtcca cgtggtttgc cccagcaggc
gaaaatcctg tttgatggtg gttaacggcg ggatataaca tgagctgtct tcggtatcgt
cgtatcccac taccgagata tccgcaccaa cgcgcagccc ggactcggta atggcgcgca
ttgcgcccag cgccatctga tcgttggcaa ccagcatcgc agtgggaacg atgccctcat
tcagcatttg catggtttgt tgaaaaccgg acatggcact ccagtcgcct tcccgttccg
ctatcggctg aatttgattg cgagtgagat atttatgcca gccagccaga cgcagacgcg
ccgagacaga acttaatggg cccgctaaca gcgcgatttg ctggtgaccc aatgcgacca
gatgctccac gcccagtcgc gtaccgtctt catgggagaa aataatactg ttgatgggtg
tctggtcaga gacatcaaga aataacgccg gaacattagt gcaggcagct tccacagcaa
tggcatcctg gtcatccagc ggatagttaa tgatcagccc actgacgcgt tgcgcgagaa
gattgtgcac cgccgcttta caggcttcga cgccgcttcg ttctaccatc gacaccacca
cgctggcacc cagttgatcg gcgcgagatt taatcgccgc gacaatttgc gacggcgcgt
gcagggccag actggaggtg gcaacgccaa tcagcaacga ctgtttgccc gccagttgtt
gtgccacgcg gttgggaatg taattcagct ccgccatcgc cgcttccact ttttcccgcg
ttttcgcaga aacgtggctg gcctggttca ccacgcggga aacggtctga taagagacac
cggcatactc tgcgacatcg tataacgtta ctggtttcac attcaccacc ctgaattgac
tctcttccgg gcgctatcat gccataccgc gaaaggtttt gcaccattcg atggtgtcaa
cgtaaatgcc gcttcgcctt cgcgcgcgaa ttgcaagctg atccgggctt atcgactgca
cggtgcacca atgcttctgg cgtcaggcag ccatcggaag ctgtggtatg gctgtgcagg
tcgtaaatca ctgcataatt cgtgtcgctc aaggcgcact cccgttctgg ataatgtttt
ttgcgccgac atcataacgg ttctggcaaa tattctgaaa tgagctgttg acaattaatc
atcggctcgt ataatgtgtg gaattgtgag cggataacaa tttcacacag gaaacagaat
t
//