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ID   PCZ101     preliminary; circular DNA; SYN; 110 BP.
AC   K02754;
DT   11-NOV-1985 (Rel. 1, Created)
DT   01-APR-1995 (Rel. 11, Last updated, Version 1)
DE   E. coli plasmid vector pCZ101 - incomplete, lpp promote/bovine growth.
KW   cloning vector; growth hormone; hormone.
OS   Cloning vector
OC   Artificial sequences; Cloning vehicles.
RN   [1]
RP   1-110
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).
RN   [2]
RC   pKN402A from pKN402 & R1drd-19, amp gene
RC   pKN402K from pKN402 & R1drd-19, kan gene
RC   pKN410K from pKN410 & R1drd-19, kan gene
RA   Uhlin B.E., Molin B.J., Gustafsson P., Nordstrom K.;
RT   "Plasmids with temperature-dependent copy number for amplification
RT   of cloned genes and their products";
RL   Gene 6:91-106(1979).
RN   [3]
RC   pPR1 from pIA7delta4delta1 & lambda cI857 Sam7
RC   pPR3, pPR4 from pIB7delta4delta1 & lambda cI857 Sam7
RC   pPR12 from pPR3 & pBR322
RC   pPR17 from pPR12 & pIA7delta4delta1
RC   pPR18 from pPR12 & pIB7delta4delta1
RC   pPR19 from pPR12 & pHI7delta4delta1
RA   Rosteck P.R., Hershberger C.L.;
RT   "Selective retention of recombinant plasmids coding for human
RT   insulin";
RL   Gene 25:29-38(1983).
RN   [4]
RC   pIA7 from trp promoter & human insulin A gene
RC   pIB7 from trp promoter & human insulin B gene
RC   pHI7 from trp promoter & human insulin genes
RC   pIA7delta4delta1 from pIA7, trp promoter
RC   pIB7delta4delta1 from pIB7, trp promoter
RC   pHI7delta4delta1 from pHI7, trp promoter
RA   Kleid D., Yansura D.G., Heyneker H.L., Miozzari G.F.;
RT   "A method of producing a polypeptide product and a plasmidic
RT   expression vehicle therefore, a method of creating an expression
RT   plasmid, a method of cleaving double stranded DNA, and specific
RT   plasmids";
RL   Unpublished (1981).
RL   European Patent Application 81301227.5.
RN   [5]
RC   pKN400, pKN401 from R1drd-19
RC   pKN402 from pKN400
RC   pKN410 from pKN401
RA   Uhlin B.E., Nordstrom K.;
RT   "A runaway-replication mutant of plasmid R1drd-19:
RT   temperature-dependent loss of copy number control";
RL   Mol. Gen. Genet. 165:167-179(1978).
RN   [6]
RC   ColE1-ampA from ColE1
RA   Edlund T.;
RT   ;
RL   Unpublished (1979).
CC   lpp promoter DNA from pIMIA.
CC   bovine cDNA provided by J.D.Baxter, grown in E.coli K-12 (RV308
CC   cells), clone pCZ101.
CC   [1] constructed expression plasmids to get more efficient
CC   production of growth hormone (bGH).  Besides an efficient promoter
CC   (lpp) 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 23-109) was inserted between the XbaI and
CC   HgiAI sites, of which the first eight codons after the bGH start
CC   codon are foreign to the natural gene.  When the number of
CC   artificial codons directly 3' to the 'atg' codon was decreased, the
CC   amount of growth hormone produced also decreased dramatically.
CC   Bases 42-65 are missing in pCZ108 and pCZ104 and are replaced by
CC   'gct' in pCZ100.  Variations, as annotated in the Sites Table also
CC   caused different levels of bGH production.
CC   NCBI gi: 208979
CC   NM (pCZ101)
CC   CM (no)
CC   NA (ds-DNA)
CC   TP (circular)
CC   ST ()
CC   TY (plasmid)
CC   SP ()
CC   HO (E.coli K-12)(E.coli RV308)
CC   CP ()
CC   FN (cloning)(expression eukaryote)
CC   SE ()
CC   PA (pIMIA)(bovine bGH cDNA)
CC   BR (pCZ108)(pCZ104)(pCZ100)
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
FT                   2. bovine bGH cDNA
FT                   -> pCZ101 10800bp"
FT   misc_recomb     22..23
FT                   /note="E. coli DNA end/artificial DNA start"
FT   variation       68..71
FT                   /note="CCCA in pCZ101, pCZ108; TCCG in pCZ104, pCZ100"
FT   variation       74..74
FT                   /note="C in pCZ101, pCZ108; T in pCZ104, pCZ100"
FT   variation       80..80
FT                   /note="C in pCZ101, pCZ108; T in pCZ104, pCZ100"
FT   misc_recomb     109..110
FT                   /note="artificial DNA end/bovine DNA start"
FT   CDS             39..>110
FT                   /note="GEN artificial bovine growth hormone;
FT                   NCBI gi: 208980"
FT   misc_binding    0..0
FT                   /note="SIT XbaI"
FT   misc_binding    0..0
FT                   /note="SIT HgiAI"
SQ   Sequence 110 BP; 26 A; 25 C; 25 G; 34 T; 0 other;
     gctacatgga gattaactca atctagaggg tattaataat gttcccattg gatgatgatg
     ataagttccc agccatgtcc ttgtccggcc tgtttgccaa cgctgtgctc