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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
//