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ID   PRX2       preliminary; circular DNA; SYN; 2852 BP.
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AC   M28835; M28836; M28837; ATCC37678;
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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-2 - 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   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   This vector contains the trpE operon and 51 bp of the trpE gene,
CC   followed by a 10-mer EcoRI linker and the M13mp13 multiple cloning
CC   site (MCS).
CC   There are stop codons after th MCS XbaI and ClaI sites, in frame from
CC   the GAA of the EcoRI site.
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   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-2)
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 10bp nngaattcnn
FT                   -> plasmid 3789bp
FT                   1. plasmid remove EcoRI-EcoRI 930bp 476..1406,
FT                   \ trpE gene/2859bp
FT                   -> pRX-2 2877bp"
FT   -               1..475
FT                   /note="pATH1 1..475 475bp
FT                   NruI = TCG^CGA
FT                   \          nng"
FT   -               476..478
FT                   /note="nng 3bp
FT                   \     nng
FT                   EcoRI = G^AATTC"
FT   -               479..2852
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 2852 BP; 718 A; 695 C; 716 G; 721 T; 2 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 tatcgnngaa
     ttcgagctcg cccggggatc ctctagagtc gacctgcagc ccaagcttat cgatgataag
     ctgtcaaaca tgagaattaa ttcttgaaga cgaaagggcc tcgtgatacg cctattttta
     taggttaatg tcatgataat aatggtttct tagacgtcag gtggcacttt tcggggaaat
     gtgcgcggaa cccctatttg tttatttttc taaatacatt caaatatgta tccgctcatg
     agacaataac cctgataaat gcttcaataa tattgaaaaa ggaagagtat gagtattcaa
     catttccgtg tcgcccttat tccctttttt gcggcatttt gccttcctgt ttttgctcac
     ccagaaacgc tggtgaaagt aaaagatgct gaagatcagt tgggtgcacg agtgggttac
     atcgaactgg atctcaacag cggtaagatc cttgagagtt ttcgccccga agaacgtttt
     ccaatgatga gcacttttaa agttctgcta tgtggcgcgg tattatcccg tgttgacgcc
     gggcaagagc aactcggtcg ccgcatacac tattctcaga atgacttggt tgagtactca
     ccagtcacag aaaagcatct tacggatggc atgacagtaa gagaattatg cagtgctgcc
     ataaccatga gtgataacac tgcggccaac ttacttctga caacgatcgg aggaccgaag
     gagctaaccg cttttttgca caacatgggg gatcatgtaa ctcgccttga tcgttgggaa
     ccggagctga atgaagccat accaaacgac gagcgtgaca ccacgatgcc tgcagcaatg
     gcaacaacgt tgcgcaaact attaactggc gaactactta ctctagcttc ccggcaacaa
     ttaatagact ggatggaggc ggataaagtt gcaggaccac ttctgcgctc ggcccttccg
     gctggctggt ttattgctga taaatctgga gccggtgagc gtgggtctcg cggtatcatt
     gcagcactgg ggccagatgg taagccctcc cgtatcgtag ttatctacac gacggggagt
     caggcaacta tggatgaacg aaatagacag atcgctgaga taggtgcctc actgattaag
     cattggtaac tgtcagacca agtttactca tatatacttt agattgattt aaaacttcat
     ttttaattta aaaggatcta ggtgaagatc ctttttgata atctcatgac caaaatccct
     taacgtgagt tttcgttcca ctgagcgtca gaccccgtag aaaagatcaa aggatcttct
     tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca
     gcggtggttt gtttgccgga tcaagagcta ccaactcttt ttccgaaggt aactggcttc
     agcagagcgc agataccaaa tactgtcctt ctagtgtagc cgtagttagg ccaccacttc
     aagaactctg tagcaccgcc tacatacctc gctctgctaa tcctgttacc agtggctgct
     gccagtggcg ataagtcgtg tcttaccggg ttggactcaa gacgatagtt accggataag
     gcgcagcggt cgggctgaac ggggggttcg tgcacacagc ccagcttgga gcgaacgacc
     tacaccgaac tgagatacct acagcgtgag cattgagaaa gcgccacgct tcccgaaggg
     agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa caggagagcg cacgagggag
     cttccagggg gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca cctctgactt
     gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac
     gcggcctttt tacggttcct ggccttttgc tggccttttg ctcacatgtt ctttcctgcg
     ttatcccctg attctgtgga taaccgtatt accgcctttg agtgagctga taccgctcgc
     cgcagccgaa cgaccgagcg cagcgagtca gtgagcgagg aagcggaaga gcgcctgatg
     cggtattttc tccttacgca tctgtgcggt atttcacacc gcatatggtg cactctcagt
     acaatctgct ctgatgccgc atagttaagc cagtatacac tccgctatcg ctacgtgact
     gggtcatggc tgcgccccga cacccgccaa cacccgctga cgcgccctga cgggcttgtc
     tgctcccggc atccgcttac agacaagctg tgaccgtctc cgggagctgc atgtgtcaga
     ggttttcacc gtcatcaccg aaacgcgcga gg
//