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ID PMGU preliminary; circular DNA; SYN; 4185 BP.
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AC S46756; IG1361; ATCC77367;
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DT 01-OCT-1993 (Rel. 7, Created)
DT 01-JUL-1995 (Rel. 12, Last updated, Version 1)
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DE E. coli phagemid vector pMGU - incomplete.
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KW cloning vector.
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OS Cloning vector
OC Artificial sequences; Cloning vehicles.
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RN [1]
RC [lambda 2690 from lambda]
RC lambda MGU2 from lambda 2690 & modified gam gene
RC pKG1 from pKSS & modified gam gene
RC pUM13 from BlueScribe M13+
RC pKG2 from pKG1 & pUM13
RC pLP1 from pRH536 & oligo
RC pMGU from pKG2 & pLP1
RC lambda MGU1 from pMGU & lambda 2690
RC lambda MGU2 from lambda MGU1
RC pCRE1 from pRH147 & pBR322
RA Maruyama I.N., Brenner S.;
RT "A selective lambda phage cloning vector with automatic excision
RT of the insert in a plasmid";
RL Gene 120:135-141(1992).
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CC The following is for the lambda phage vector ATCC77367:
CC lambda phage vector is 41700 bp.
CC Deposited by: Ichiro N. Maruyama
CC Restriction digests of the clone give the following sizes (kb):
CC HindIII--24.1, 17.6; EcoRI--22.0, 20.0; BamHI--24.8, 16.9;
CC XbaI--32.7,9.0; NotI--41.7. (ATCC staff)
CC Vector useful for constructing cDNA libraries. Permits positive
CC selection for inserts using the Spi- phenotype, and excision of
CC phagemid by lox/cre site-specific recombination. [1]
CC To prepare phagemid from lambdaMGU2, grow recombinants on a RecA- host
CC expressing the Cre protein (E. coli 1046[pCRE1], ATCC 77368) and
CC select for ampicillin resistance. The pMGU product is 4.185 kb. [1]
CC The order of the major features in the cloning region of the lambda
CC vector is: lambda J - SmaI - SalI - loxP - EcoRI - M13 ori - ampR -
CC pMB1 ori - HindIII - 3'gam/BamHI/5'gam - XhoI - loxP - SalI -
CC lambda N. [1]
CC Inserts can be amplified using the following primers flanking the
CC BamHI cloning site: upstream 5'-AAGAGGCAGAACTGGCAG-3' and downstream
CC 5'-ATCGATGCATAGCGATTC-3'. [1]
CC Efficiency of phagemid recovery is approximately 20%. Plasmid pCRE1
CC may be a low level contaminant, but is easily distinguished from pMGU
CC DNA. [1]
CC To enable the positive selection of inserts, the library should be
CC plated on a P2 lysogen such as Escherichia coli Q359 (ATCC 47019). [1]
CC Medium is 1592 SM buffer.
CC
CC EcoRI-HindIII fragment is 1039 bp.
CC This sequence comes from Fig. 3. [1]
CC NCBI gi: 258428
CC NM (pMGU)
CC CM (no)
CC NA (ds-DNA)
CC TP (circular)
CC ST ()
CC TY (phagemid)
CC SP (ATCC)
CC HO (E.coli)(E.coli Q358)(bacteria-free lysate)
CC CP ()
CC FN (cloning cDNA)
CC SE ()
CC PA (pKG1 from pKSS)(pLP1 from pRH536)(pUM13 from BlueScribe M13+)
CC PA (pCRE1 from pBR322 & pRH147)(lambda 2690)
CC BR ()
CC OF (lambda MGU1)(lambda MGU2)
CC OR ()
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FH Key Location/Qualifiers
FH
FT misc_feature 0..0
FT /note="1. lambda SalI-SalI 499bp 32746..33245,gam gene
FT 2. M13mp8 SalI 7229bp 6248..6248
FT -> phage 7728bp
FT 1. phage 7728bp
FT 2. oligo 18bp gaaaaagggatcccccag
FT mutagenesis
FT -> phage2 7746bp
FT 1. lambda remove SalI-SalI 499bp, gam gene/48003bp
FT -> lambda 2690 48003bp
FT 1. lambda 2690 48003bp
FT 2. phage2 SalI-ScaI 500bp, modified gam gene
FT -> lambda MGU 48500bp
FT 1. pKSS XhoI-SmaI 4091bp 59..4133..17, neo gene
FT 2. phage2 SalI-ScaI 500bp, modified gam gene
FT -> pKG1 4600bp
FT 1. pBlueScript M13+ 3204bp, in MCS
FT 2. oligo XhoI-NsiI
FT -> pUM13 3200bp
FT 1. pKG1 PstI-PstI 1170bp, modified gam gene
FT oligo PstI-SacI-KpnI-SphI-XhoI
FT 2. pUM13 XhoI 3200bp, oligo
FT -> pKG2 4400bp
FT 1. phage P1
FT -> pRH536
FT 1. pRH536 remove BamHI-HindIII, neo gene
FT 2. oligo BamHI-HindIII 10bp agctgtcgac
FT -> pLP1
FT 1. pLP1 EcoRI-XhoI 162bp, loxP sites
FT 2. pKG2 XhoI-HindIII 871bp, modified gam gene
FT -> fragment 1033bp
FT 1. fragment EcoRI-HindIII 1033bp, modified gam/loxP
FT 2. pUM13 remove EcoRI-HindIII, MCS/3200bp
FT -> pMGU 4185bp"
FT rep_origin complement(239..694)
FT /note="ORI bacteriophage M13 intergenic region"
FT misc_binding 881..881
FT /note="SIT EcoRI"
FT misc_feature 911..944
FT /note="SIT bacteriophage lambda loxP"
FT misc_binding 964..964
FT /note="SIT SalI"
FT misc_feature 1002..1035
FT /note="SIT bacteriophage lambda loxP"
FT misc_binding 1033..1033
FT /note="SIT XhoI"
FT promoter 1096..1101
FT /note="PRO E. coli kanamycin gene -35 region,
FT from Tn903"
FT promoter 1119..1124
FT /note="PRO E. coli kanamycin gene -10 region,
FT from Tn903"
FT misc_feature 1364..1656
FT /note="bacteriophage lambda gam"
FT misc_binding 1905..1905
FT /note="SIT HindIII"
FT rep_origin complement(0..0)
FT /note="ORI E. coli pMB1 (ColE1 and pBR322)"
FT CDS complement(0..0)
FT /note="ANT E. coli beta-lactamase gene (bla)
FT ampicillin resistance gene (apr/amp)"
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SQ Sequence 919 BP; 267 A; 194 C; 216 G; 242 T; 0 other;
gaattccgat catattcaat aacccttaat ataacttcgt ataatgtatg ctatacgaag
ttattaggtc tgaagaggag tttacgtcga gccaagctgt cgacgatccg gaacccttaa
tataacttcg tataatgtat gctatacgaa gttattaggt ccctcgaggg taccgcatgc
gagctcgacc tgcagggggg ggggggaaag ccacgttgtg tctcaaaatc tctgatgtta
cattgcacaa gataaaaata tatcatcatg aacaataaaa ctgtctgctt acataaacag
taatacaagg ggtgttatga gccatattca acgggaaacg tcttgctcga cgcttataaa
aaatggatat taatactgaa actgagatca agcaaaagca ttcactaacc ccctttcctg
ttttcctaat cagcccggca tttcgcgggc gatattttca cagctatttc aggagttcag
ccatgaacgc ttattacatt caggatcgtc ttgaggctca gagctgggcg cgtcactacc
agcagctcgc ccgtgaagag aaagaggcag aactggcaga cgacatggaa aaagggatcc
cccagcacct gtttgaatcg ctatgcatcg atcatttgca acgccacggg gccagcaaaa
acgaatgagt gggaaaacag cattccaggt attagaagaa tatcctgatt caggtgaaaa
tattgttgat gcgctggcag tgttcctgcg ccggttgcat tcgattcctg tttgtaattg
tccttttaac agcgatcgcg tatttcgtct cgctcaggcg caatcacgaa tgaataacgg
tttggttgat gcgagtgatt ttgatgacga gcgtaatggc tggcctgttg aacaagtctg
gaaagaaatg cataagctt
//