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ID   LAMMGU2    preliminary; circular DNA; SYN; 41700 BP.
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AC   S46756; ATCC77367;
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DT   03-FEB-1994 (Rel. 8, Created)
DT   01-JUL-1995 (Rel. 12, Last updated, Version 1)
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DE   E. coli phage vector lambda MGU2 - incomplete, cloning sites/loxP sit.
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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   pUM113 from BlueScribe M13+
RC   pKG2 from pKG1 & pUM113
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   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;
CC   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   EcoRI-HindIII fragment is 1039 bp.
CC   This sequence comes from Fig. 3. [1]
CC   NCBI gi: 258428
CC   NM (lambda MGU2)
CC   CM (no)
CC   NA (ds-DNA)
CC   TP (circular)
CC   ST ()
CC   TY (phage)
CC   SP (ATCC)
CC   HO (E.coli)(E.coli Q358)(bacteria-free lysate)
CC   CP ()
CC   FN (cloning)
CC   SE ()
CC   PA ()
CC   BR ()
CC   OF ()
CC   OR ()
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FH   Key             Location/Qualifiers
FH
FT   misc_feature    0..0
FT                   /note="1. lambda MGU1 remove SmaI-SmaI 3200bp,41700bp
FT                   -> lambda MGU2 41700bp"
FT   misc_binding    0..0
FT                   /note="SIT unique BamHI"
FT   rep_origin      0..0
FT                   /note="ORI bacteriophage lambda"
FT   rep_origin      0..0
FT                   /note="ORI E. coli pMB1 (ColE1 and pBR322)"
FT   rep_origin      0..0
FT                   /note="ORI bacteriophage M13"
FT   CDS             0..0
FT                   /note="GEN E. coli Spi+ reporter gene"
FT   CDS             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
//