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ID   PRX3       preliminary; circular DNA; SYN; 2853 BP.
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AC   M28835; M28836; M28837; ATCC37679;
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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 plasmid vector pRX-3 - complete.
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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   pRX-1, pRX-2, pRX-3 from pATH1 & linkers
RA   Rimm D.L., Pollard T.D.;
RT   "New plasmid vectors for high level synthesis of eukaryotic fusion
RT   proteins in Escherichia coli";
RL   Gene 75:323-327(1989).
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CC   Created by Moore, July 1995, under contract with NCBI.
CC   Restriction digests of the clone give the following sizes (kb):
CC   EcoRI--2.85; ClaI--2.85. (ATCC staff)
CC   When E.coli CAG-456 (a protease deficient host) is used this
CC   vector yields large amounts (100 mg/liter or 45% of the soluble
CC   protein) of the fusion protein.
CC   This vector contains the trpE operon and 51 bp of the trpE gene,
CC   followed by a 12-mer EcoRI linker and the M13mp13 multiple cloning
CC   site (MCS).
CC   There is a TGA stop codon in frame from the GAA of the EcoRI site
CC   33 bp beyond the MSC ClaI site.
CC   One of a series of plasmid expression vectors (ATCC 37677 to 37679)
CC   representing multiple reading frames for expression from trpE
CC   promoter.
CC   The fusion proteins produced contain less than 25 amino acids of the
CC   trpE protein.
CC   Use E.coli SG935 (ATCC 39623), SG936 (ATCC 39624) or WM6 (ATCC
CC   47020) as host for high level production of recombinant fusion
CC   protein.
CC   (ATCC staff)
CC   Medium is 1227 LB plus ampicillin.
CC   NM (pRX-3)
CC   CM (yes)
CC   NA (ds-DNA)
CC   TP (circular)
CC   ST ()
CC   TY (plasmid)
CC   SP (ATCC)
CC   HO (E.coli HB101)(E.coli CAG-456)(E.coli DH1)
CC   CP ()
CC   FN (cloning)
CC   SE ()
CC   PA (pATH1)
CC   BR ()
CC   OF ()
CC   OR ()
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FH   Key             Location/Qualifiers
FH
FT   misc_feature    0..0
FT                   /note="1. pATH1 NruI 3779bp 476..476,
FT                   \ 51 bp 3' to trpE init
FT                   2. EcoRI linker 12bp nnngaattcnnn
FT                   -> plasmid 3791bp
FT                   1. plasmid remove EcoRI-EcoRI 930bp 476..1406,
FT                   \ trpE gene/2857bp
FT                   -> pRX-3 2878bp"
FT   -               1..475
FT                   /note="pATH1 1..475 475bp
FT                   NruI = TCG^CGA
FT                   \          nnng"
FT   -               476..479
FT                   /note="nnng 4bp
FT                   \    nnng
FT                   EcoRI = G^AATTC"
FT   -               480..2853
FT                   /note="pATH1 1406..3779 2374bp"
FT   misc_binding    0..0
FT                   /note="MCS EcoRI-SacI-SmaI-BamHI-XbaI-SalI-HindIII-
FT                   ClaI"
FT   misc_binding    0..0
FT                   /note="SIT unique EcoRI-SacI-SmaI-BamHI-XbaI-SalI-
FT                   HindIII-ClaI"
FT   rep_origin      0..0
FT                   /note="ORI E. coli pMB1 (ColE1 and pBR322)"
FT   promoter        0..0
FT                   /note="PRO E. coli trpE gene"
FT   CDS             0..0
FT                   /note="ANT E. coli beta-lactamase gene (bla)
FT                   ampicillin resistance gene (apr/amp)"
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SQ   Sequence 2853 BP; 718 A; 695 C; 716 G; 721 T; 3 other;
     cagctgtggt gtcatggtcg gtgatcgcta gggtgccgag cgcatctcga ctgcacggtg
     caccaatgct tctggcgtca ggtagttatt ggaaagctgt ggtatggctg tgcaggtcgt
     aaatcactgc ataactcgct gctgcctaag gcgcactccc gttctggata atgttttttg
     cgccgacatc ataacggttc tggcaaatat tctgaaatga gctgttgaca attaatcatc
     gaactagtta actagtacgc aagttcacgt aaaaagggta tcgacaatga aagcaatttt
     cgtactgaaa ggttggtggc gcacttcctg aaacgggcag tgtattcacc atgcgtaaag
     caatcagata cccagcccgc ctaatgagcg ggcttttttt tgaacaaaat tagagaataa
     caatgcaaac acaaaaaccg actctcgaac tgctaacctg cgaaggcgct tatcgnnnga
     attcgagctc gcccggggat cctctagagt cgacctgcag cccaagctta tcgatgataa
     gctgtcaaac atgagaatta attcttgaag acgaaagggc ctcgtgatac gcctattttt
     ataggttaat gtcatgataa taatggtttc ttagacgtca ggtggcactt ttcggggaaa
     tgtgcgcgga acccctattt gtttattttt ctaaatacat tcaaatatgt atccgctcat
     gagacaataa ccctgataaa tgcttcaata atattgaaaa aggaagagta tgagtattca
     acatttccgt gtcgccctta ttcccttttt tgcggcattt tgccttcctg tttttgctca
     cccagaaacg ctggtgaaag taaaagatgc tgaagatcag ttgggtgcac gagtgggtta
     catcgaactg gatctcaaca gcggtaagat ccttgagagt tttcgccccg aagaacgttt
     tccaatgatg agcactttta aagttctgct atgtggcgcg gtattatccc gtgttgacgc
     cgggcaagag caactcggtc gccgcataca ctattctcag aatgacttgg ttgagtactc
     accagtcaca gaaaagcatc ttacggatgg catgacagta agagaattat gcagtgctgc
     cataaccatg agtgataaca ctgcggccaa cttacttctg acaacgatcg gaggaccgaa
     ggagctaacc gcttttttgc acaacatggg ggatcatgta actcgccttg atcgttggga
     accggagctg aatgaagcca taccaaacga cgagcgtgac accacgatgc ctgcagcaat
     ggcaacaacg ttgcgcaaac tattaactgg cgaactactt actctagctt cccggcaaca
     attaatagac tggatggagg cggataaagt tgcaggacca cttctgcgct cggcccttcc
     ggctggctgg tttattgctg ataaatctgg agccggtgag cgtgggtctc gcggtatcat
     tgcagcactg gggccagatg gtaagccctc ccgtatcgta gttatctaca cgacggggag
     tcaggcaact atggatgaac gaaatagaca gatcgctgag ataggtgcct cactgattaa
     gcattggtaa ctgtcagacc aagtttactc atatatactt tagattgatt taaaacttca
     tttttaattt aaaaggatct aggtgaagat cctttttgat aatctcatga ccaaaatccc
     ttaacgtgag ttttcgttcc actgagcgtc agaccccgta gaaaagatca aaggatcttc
     ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc
     agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt
     cagcagagcg cagataccaa atactgtcct tctagtgtag ccgtagttag gccaccactt
     caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc
     tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa
     ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac
     ctacaccgaa ctgagatacc tacagcgtga gcattgagaa agcgccacgc ttcccgaagg
     gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga
     gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact
     tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa
     cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgt tctttcctgc
     gttatcccct gattctgtgg ataaccgtat taccgccttt gagtgagctg ataccgctcg
     ccgcagccga acgaccgagc gcagcgagtc agtgagcgag gaagcggaag agcgcctgat
     gcggtatttt ctccttacgc atctgtgcgg tatttcacac cgcatatggt gcactctcag
     tacaatctgc tctgatgccg catagttaag ccagtataca ctccgctatc gctacgtgac
     tgggtcatgg ctgcgccccg acacccgcca acacccgctg acgcgccctg acgggcttgt
     ctgctcccgg catccgctta cagacaagct gtgaccgtct ccgggagctg catgtgtcag
     aggttttcac cgtcatcacc gaaacgcgcg agg
//