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ID   PJJS314    preliminary; circular DNA; SYN; 232 BP.
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AC   M10304; M10307;
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DT   15-MAR-1988 (Rel. 3, Created)
DT   01-JUL-1995 (Rel. 12, Last updated, Version 1)
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DE   E. coli plasmid vector pJJS314 - incomplete, lac promoter in cat gene.
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KW   cloning vector; cat gene; chloramphenicol acetyltransferase;
KW   promoter region.
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OS   Cloning vector
OC   Artificial sequences; Cloning vehicles.
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RN   [1]
RP   1-67, first
RP   1-52, second
RC   pJJS300 from pBR322 & lacZ gene promoter/SD
RC   pJJS310, pJJS301 from pJJS300 & pACYC184
RC   pJJS530 from pBR322 & pACYC184
RC   pJLS1000 from pBR322 & pJJS310
RC   pJLS1004 from pJLS1000 & pJJS310
RC   pJLS1021 from pJJS310 & pJLS1004
RC   pJJS312 from pJLS1021 & pJJS300
RC   pJLS1022, pJLS1023, pJLS313, pJJS313 from pJLS1021 & pJJS312
RC   pJLS314, pJJS314 from pJLS1021 & pJJS312
RC   pJLS1024, pJLS315 from pJLS1023 & pJJS314
RC   pJJS512 from pJLS1021 & pJJS500
RC   pJJS513 from pJJS512
RC   pJLS1030 from pJJS500 & pJLS1021
RC   pJJS501 from pJJS500
RC   pJJS1031 from pJJS501 & pACYC184
RC   pJJS712 from pJLS1021 & pJJS700
RC   pJJS713 from pJJS712
RC   pJLS1032 from pJJS700 & pJLS1021
RC   pJJS701 from pJJS700
RC   pJJS1033 from pJJS701 & pACYC184
RC   pJLS513 from pJJS513
RC   pJLS713 from pJJS713
RA   Schottel J.L., Sninsky J.J., Cohen S.N.;
RT   "Effects of alterations in the translation control region on
RT   bacterial gene expression: Use of cat gene constructs transcribed
RT   from the lac promoter as a model system";
RL   Gene 28:177-193(1984).
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CC   Clone pJJS314 was produced by filling in the BamHI site of pJJS312
CC   (#M10305) and inserting a ClaI site.
CC
CC   The region controlling translation of the cat gene has been varied
CC   structurally so that the gene is under control of the lac promoter.
CC   Altering the potential for secondary structure formation within the
CC   translation control region causes a ten-fold variation in the
CC   synthesis of the CAT enzyme, whereas varying the distance between
CC   the ribosome binding site and the start codon from 7 to 13 bases
CC   does not significantly affect the yield of CAT enzyme.  If the
CC   Shine Dalgarno sequence is situated in a region of mRNA capable of
CC   base pairing, the efficiency of translation is decreased.  The
CC   start codon can initiate translation efficiently even when located
CC   in a segment capable of duplex formation.  If translation is
CC   initiated upstream of the cat start codon, then out of frame
CC   overlapping translation reduces CAT production by 90% and in frame
CC   overlapping translation prevents detectable initiation of protein
CC   synthesis at the cat gene start codon and yields only fusion
CC   proteins.
CC   NCBI gi: 208707 & 208723
CC   NM (pJJS314)
CC   CM (no)
CC   NA (ds-DNA)
CC   TP (circular)
CC   ST ()
CC   TY (plasmid)
CC   SP ()
CC   HO (E.coli)
CC   CP ()
CC   FN (cloning)
CC   SE ()
CC   PA (lacZ)(pJJS312)
CC   BR ()
CC   OF ()
CC   OR ()
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FH   Key             Location/Qualifiers
FH
FT   misc_feature    0..0
FT                   /note="1. pJJS312 BamHI
FT                   fill in
FT                   ClaI linker 6bp atcgat
FT                   -> pJJS314"
FT   CDS             39..>66
FT                   /note="ANT E. coli chloramphenicol acetyltransferase
FT                   gene (cat); chloramphenicol resistance gene (cmr/cml);
FT                   NCBI gi: 208708; first"
FT   CDS             50..>52
FT                   /note="ANT E. coli chloramphenicol acetyltransferase
FT                   gene (cat); chloramphenicol resistance gene (cmr/cml);
FT                   NCBI gi: 208724; second"
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SQ   Sequence 232 BP; 43 A; 23 C; 26 G; 27 T; 113 other;
     aattgtgagc ggataacaat ttcacacagg aaacagctat gaccatgatt acggattcac
     tggccgtnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn
     nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn
     aattgtgagc ggataacaat ttcacacagg aaacaggatc gatcctaaaa tg
//