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ID   PCZ105     preliminary; circular DNA; SYN; 98 BP.
AC   K02755;
DT   11-NOV-1985 (Rel. 1, Created)
DT   01-APR-1995 (Rel. 11, Last updated, Version 1)
DE   E. coli plasmid vector pCZ105 - incomplete, trp promote/bovine growth.
KW   cloning vector; growth hormone; hormone.
OS   Cloning vector
OC   Artificial sequences; Cloning vehicles.
RN   [1]
RP   1-98
RC   pCZ101, pCZ108, pCZ104, pCZ100 from pI-MIA
RC   pCZ105, pCZ110, pCZ112, pCZ115 from pI-MIA & pHI7
RC   pCZ114 from pCZ101
RA   Schoner B.E., Hsiung H.M., Belagaje R.M., Mayne N.G.,
RA   Schoner R.G.;
RT   "Role of mRNA translational efficiency in bovine growth hormone
RT   expression in Escherichia coli";
RL   Proc. Natl. Acad. Sci. U.S.A. 81:5403-5407(1984).
CC   E.coli trp promoter DNA from pHI7.
CC   bovine cDNA provided by J.D.Baxter, grown in E.coli K-12 (RV308
CC   cells), clone pCZ105.
CC   [1] constructed expression plasmids to get more efficient production
CC   of growth hormone (bGH).  Besides an efficient promoter (trp)
CC   additional codons in the 5' end of the growth hormone coding
CC   sequence were required to raise the production level.  An
CC   artificial sequence (bp 22-97) was inserted, between the TaqI and
CC   HgiAI sites of which the first eight codons after the bGH start
CC   codon were foreign to the natural gene.  When the number of
CC   artificial codons just 3' of the 'atg' codon was decreased, the
CC   amount of growth hormone produced also decreased dramatically.
CC   Bases 30-53 are missing in pCZ110, and replaced by 'gat' and
CC   'gatgataag' in pCZ115 and pCZ112, respectively.  Variations, as
CC   annotated in the Sites Table, also caused different levels of bGH
CC   production.
CC   NCBI gi: 208981
CC   NM (pCZ105)
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 (pHI7)
CC   BR (pCZ110)(pCZ115)(pCZ112)
CC   OF ()
CC   OR ()
FH   Key             Location/Qualifiers
FT   misc_feature    0..0
FT                   /note="1. pKN402 remove small EcoRI-XbaI
FT                   2. E. coli EcoRI-XbaI, lpp prom/RBS/3' lpp gene/term
FT                   -> pI-MIA [kan gene]
FT                   1. pI-MIA remove small EcoRI-TaqI
FT                   2. pHI7 EcoRI-TaqI, trp promoter
FT                   -> plasmid
FT                   1. plasmid
FT                   2. bovine bGH cDNA
FT                   -> pCZ105 10800bp"
FT   misc_recomb     21..22
FT                   /note="E. coli DNA end/artificial DNA start"
FT   variation       56..59
FT                   /note="CCCA in pCZ105; TCCG in pCZ110, pCZ115, pCZ112"
FT   variation       62..62
FT                   /note="C in pCZ105; T in pCZ110, pCZ115, pCZ112"
FT   variation       68..69
FT                   /note="CT in pCZ105; TC in pCZ110, pCZ115, pCZ112"
FT   misc_recomb     97..98
FT                   /note="artificial DNA end/bovine DNA start"
FT   CDS             27..>98
FT                   /note="GEN artificial bovine growth hormone;
FT                   NCBI gi: 208982"
FT   misc_binding    0..0
FT                   /note="SIT TaqI"
FT   misc_binding    0..0
FT                   /note="SIT HgiAI"
SQ   Sequence 98 BP; 23 A; 24 C; 23 G; 28 T; 0 other;
     aagttcacgt aaaaagggta tcgacaatgt tcccattgga tgatgatgat aagttcccag
     ccatgtcctt gtccggcctg tttgccaacg ctgtgctc