ID   LAMDL10    preliminary; circular DNA; SYN; 39000 BP.
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AC   ATCC37489;
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DT   01-JUL-1993 (Rel. 7, Created)
DT   01-JUL-1995 (Rel. 12, Last updated, Version 1)
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DE   E. coli phage vector lambda DL10 - 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 D69 from lambda]
RC   lambda DL9 from lambda D69 & pUC9
RC   lambda DL9-neo from lambda DL9 & neo gene
RC   lambda DL10 from M13mp10 & lambda DL9-neo
RC   lambda DL11 from M13mp11 & lambda DL9-neo
RC   pPR410 from lambda DL10 & pCQV2
RC   pPR110 from pPR410
RC   pPR411 from lambda DL11 & pCQV2
RC   pPR111 from pPR411
RA   Windle B.E.;
RT   "Phage lambda and plasmid expression vectors with multiple cloning
RT   sites and lacZ alpha-complementation";
RL   Gene 45:95-99(1986).
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CC   A BamHI digest removes a 930 bp fragment containing the lacZalpha
CC   coding frame. Thus use of BamHI as a cloning site and selection of
CC   clear plaques on Xgal media may not indicate the presence of a
CC   recombinant insert.
CC   lambdaDL10 (ATCC 37489) and lambdaDL11 (ATCC 37490) have opposite
CC   orientations of the restriction sites within lacZ.
CC   A 435 bp HaeII fragment of pUC9, encoding beta-galactosidase, was
CC   inserted into the HindIII site of lambdaD69 to produce lambdaDL9.
CC   The multiple cloning sites of M13mp10 were crossed into lambdaDL9 to
CC   produce lambdaDL10.
CC   E.coli JM109 is not a good host for this vector. (personal
CC   communication)
CC   If HindIII, SacI, XbaI, XhoI, or BamHI are used as cloning sites, the
CC   vector can accept fragments of 0 - 12 kb. Inserts in HindIII, SacI,
CC   XbaI, BamHI, and SalI inactivate lacZ.
CC   If SalI is used as a cloning site, the vector can accept fragments of
CC   9 - 20 kb. Replacement of the SalI fragment with a DNA insert results
CC   in a lambdacIII- phage that forms virtually clear plaques.
CC   Medium is 1227 LB plus ampicillin.
CC   NM (lambda DL10)
CC   CM (no)
CC   NA (ds-DNA)
CC   TP (circular)
CC   ST ()
CC   TY (phage)
CC   SP (ATCC)
CC   HO (bacteria-free lysate)(E.coli C600)(E.coli JM101)
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
FT                   -> lambda D69
FT                   1. lambda D69
FT                   T4 DNA polymerase, [to connect cos sites]
FT                   HindIII, 5' to imm21 gene
FT                   \ lambda 27480 or 36896
FT                   T4 DNA polymerase
FT                   2. pUC19 HaeII-HaeII 445bp 240..685, lacZ alpha
FT                   T4 DNA polymerase:T4 DNA polymerase
FT                   -> lambda DL9
FT                   1. lambda DL9 HindIII
FT                   2. Tn5 HindIII-HindIII 3424bp 1196..4620, neo/kan gene
FT                   -> lambda DL9-neo
FT                   1. M13mp10 EcoRI-HindIII 45bp 6232..6277, MCS
FT                   2. lambda DL9-neo remove EcoRI-HindIII 51bp,
FT                   \ MCS/pUC19 397..448
FT                   -> lambda DL10 39000bp"
FT   misc_binding    0..0
FT                   /note="MCS EcoRI-SacI-SmaI-BamHI-XbaI-SalI-PstI-
FT                   HindIII"
FT   misc_binding    0..0
FT                   /note="SIT unique HindIII-SacI-XbaI-XhoI-BamHI-SalI"
FT   rep_origin      0..0
FT                   /note="ORI bacteriophage lambda"
FT   promoter        0..0
FT                   /note="PRO E. coli lac gene"
FT   CDS             0..0
FT                   /note="GEN E. coli beta-galactosidase gene (lacZ);
FT                   reporter gene"
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SQ   Sequence 1 BP; 0 A; 0 C; 0 G; 0 T; 1 other;
     n
//