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ID   EMBL3COS   preliminary; circular DNA; SYN; 48200 BP.
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AC   ATCC37602;
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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 EMBL3cos - 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   mpAC5 from pWP14 & M13mp8
RC   [lambda NM1265, lambda NM1266 from lambda]
RC   lambda NM1274 from lambda NM1265 & mpAC5 & lambda NM1266
RC   lambda NM1275 from lambda NM1274 & EMBL3
RC   EMBL3cos from lambda NM1275
RC   lambda NM1277 from lambda & EMBL301
RC   lambda NM1278 from EMBL3cos & lambda NM1277
RC   EMBL3cos-Not from EMBL3cos & lambda NM1278
RA   Whittaker P.A., Campbell A.J., Southern E.M., Murray N.E.;
RT   "Enhanced recovery and restriction mapping of DNA fragments cloned
RT   in a new lambda vector";
RL   Nucleic Acids Res. 16:6725-6736(1988).
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RN   [2]
RC   pWP14 from pUC9 & lambda, cos
RC   pWP32 from pUC9 & lambda, cos 154
RA   Shinder G., Gold M.;
RT   "The Nul subunit of bacteriophage lambda terminase binds to
RT   specific sites in cos DNA";
RL   J. Virol. 62:387-392(1988).
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RN   [3]
RC   ptk4 from pBR322 & HSV TK gene
RA   Brown C.;
RT   ;
RL   Unpublished (1988).
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CC   Restriction digests of the clone give the following sizes (kb):
CC   EcoRI--33.5, 14.5; BamHI--33.5, 14.5. (ATCC staff)
CC   By labelling the left cohesive end, which is 200 bp from cloning
CC   sites, reliable restriction maps of clones can be made from partial
CC   digests. (personal communication)
CC   For library construction, a double digest of the vector with BamHI +
CC   EcoRI and removal of the linker fragment reduces the number of
CC   background phage.
CC   Phosphatase treatment of digested vector eliminates background.
CC   (personal communication)
CC   A modified EMBL vector, with all phage coding sequences to the right
CC   of the cloning sites to facilitate restriction mapping of clones. Can
CC   be used for genomic library construction. (personal communication)
CC   Medium is 1592 SM buffer.
CC   NM (EMBL3cos)
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 MC1061)(E.coli K802)
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. pUC9 HincII 2665bp 250..250
FT                   2. lambda EcoRI-EcoRI 393bp, cos
FT                   \ 21227 26105 31748 39169 44973
FT                   HincII-HincII 403bp 48299..48502..200, lambda cos
FT                   -> pWP14 3068bp
FT                   1. pWP14 HincII-HincII 403bp 48299..48502..200,
FT                   \ lambda cos
FT                   2. M13mp8 HindIII 7229bp 6262..6262
FT                   Klenow:Klenow
FT                   -> M13mpAC5 7632bp
FT                   1. lambda
FT                   -> lambda NM1265
FT                   1. lambda
FT                   -> lambda NM1266
FT                   1. lambda NM1265 left end-NaeI 20040bp 1..20040,
FT                   \ left arm
FT                   2. M13mpAC5 EcoRI-FspI 578bp 6232..cos..6407, MCS/cos
FT                   3. lambda NM1266 EcoRI-right end 27276bp 21226..48502,
FT                   \ right arm
FT                   -> lambda NM1274 47894bp
FT                   1. lambda NM1274 left end-EcoRI 20618bp 1..20618,
FT                   \ left arm/cos
FT                   2. EMBL3 EcoRI-right end 9170bp 20067..29237,
FT                   \ right arm
FT                   -> lambda NM1275 29788bp [2 cos]
FT                   1. lambda NM1275 29788bp
FT                   S1 nuclease, remove ends
FT                   -> EMBL3cos 48200bp"
FT   misc_binding    0..0
FT                   /note="MCS PstI-SalI-BamHI-SmaI-EcoRI; EcoRI-BamHI-
FT                   SalI"
FT   misc_binding    0..0
FT                   /note="SIT double BamHI-EcoRI-SalI"
FT   rep_origin      0..0
FT                   /note="ORI bacteriophage lambda"
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SQ   Sequence 1 BP; 0 A; 0 C; 0 G; 0 T; 1 other;
     n
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