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ID   PER33      preliminary; circular DNA; SYN; 131 BP.
AC   J02550;
DT   22-FEB-1985 (Rel. 1, Created)
DT   01-APR-1995 (Rel. 11, Last updated, Version 1)
DE   E. coli K12 plasmid vector pER33 - incomplete, trp/RBS/IFN.
KW   cloning vector; tryptophan operon; RBS; interferon;
KW   interferon-alpha A.
OS   Cloning vector
OC   Artificial sequences; Cloning vehicles.
RN   [1]
RP   1-131
RC   [pBP101 or pB101 from Serratia marcescens]
RC   pER103, pER20 from pBR322 & pBP101 or pB101
RC   pER33, pER363 from pER103 & alpha-interferon A
RC   pER78 from pER103 & alpha-interferon A
RA   Dworkin-Rastl E., Swetly P., Dworkin M.B.;
RT   "Construction of expression plasmids producing high levels of human
RT   leukocyte-type interferon in Escherichia coli";
RL   Gene 21:237-248(1983).
RN   [2]
RC   pGM14 from pBR322 & Serratia marcescens, trp operon
RC   pGM63 from pGM14
RA   Miozzari G.F., Yanofsky C.;
RT   "The regulatory region of the trp operon of Serratia marcescens";
RL   Nature 276:684-689(1978).
CC   Serratia marcescens clone pB101.
CC   pER33 is a recombinant expression plasmid that directs the
CC   synthesis of type A preinterferon, type A mature interferon and
CC   type C mature interferon in E.coli K-12.
CC   NCBI gi: 208583
CC   NM (pER33)
CC   CM (no)
CC   NA (ds-DNA)
CC   TP (circular)
CC   ST ()
CC   TY (plasmid)
CC   SP ()
CC   HO (E.coli K-12)(Serratia marcescens)(human)
CC   CP ()
CC   FN (cloning)(expression eukaryote)
CC   SE ()
CC   PA (pBR322)(trp)(human IFN-alpha A)(pB101)
CC   PA (human lymphoblastoid Namalwa cell cDNA library, clone 1F7)
CC   BR ()
CC   OF ()
CC   OR ()
FH   Key             Location/Qualifiers
FT   misc_feature    0..0
FT                   /note="1. Serratia marcescens EcoRI-SalI, trp operon
FT                   2. pBR322 remove small EcoRI-SalI
FT                   -> pBP101
FT                   1. pBR322 remove EcoRI-HindIII 29bp
FT                   2. pBP101 EcoRI-EcoRI 200bp, trp promoter/operator
FT                   EcoRI-HaeII 90bp, trp promoter/operator
FT                   :RBS linker
FT                   EcoRI-HindIII 100bp
FT                   -> pER103 [unique HindIII]
FT                   1. pER103 HindIII
FT                   2. human leukocyte PstI-PstI, alpha-interferon A
FT                   BAL31 nuclease
FT                   Klenow, fill in:Klenow, fill in
FT                   HindIII linker:HindIII linker
FT                   HindIII-HindIII
FT                   -> pER33"
FT   misc_recomb     4..5
FT                   /note="EcoRI linker end/S.marcescens trp operon
FT                   promoter/operator start"
FT   misc_recomb     91..92
FT                   /note="S.marcescens trp operon promoter/operator
FT                   end/artificial ribosome binding site start"
FT   RBS             92..107
FT                   /note="RBS"
FT   misc_recomb     107..108
FT                   /note="artificial ribosome binding site end/HindIII
FT                   linker start"
FT   misc_recomb     115..116
FT                   /note="HindIII linker end/human IFN-alpha A start"
FT   CDS             113..>131
FT                   /note="GEN human interferon-alpha A gene;
FT                   NCBI gi: 208584"
SQ   Sequence 131 BP; 42 A; 26 C; 30 G; 33 T; 0 other;
     aattcacgct gatcgctaaa acattgtgca aaaagagggt tgactttgcc ttcgcgaacc
     agttaactag tacacaagtt cacggcaacg gtaaggaggt ttaagcttaa agatgtgtga
     tctgcctcaa a