ID   PNF3R      preliminary; circular DNA; SYN; 9400 BP.
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AC   M29400; M29401; M29402; ATCC37616;
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DT   01-JUL-1993 (Rel. 7, Created)
DT   01-APR-1995 (Rel. 11, Last updated, Version 1)
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DE   Rhodobacter/E.coli plasmid vector pNF3 - incomplete.
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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   pCB532 from Rhodobacter capsulatus nifHDK
RC   pNBR from pBR322
RC   M13::nif from M13mp19 & R.capsulans nifHDK operon
RC   pNF1 from M13::nif & pNBR
RC   pNF2 from pNF1 & pHP45omega
RC   pNF3 from pNF2 & kan gene
RC   pNF:Q-Zomega from pNF1 & pCB532
RA   Pollock D., Bauer C.E., Scolnik P.A.;
RT   "Transcription of the Rhodobacter capsulatus nifHDK operon is
RT   modulated by the nitrogen source. Construction of plasmid
RT   expression vectors based on the nifHDK promoter";
RL   Gene 65:269-275(1988).
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CC   The spcR marker is expressed in R. capsulatus.
CC   A wide range of transcription levels can be obtained by varying the
CC   nitrogen source in the medium. Transcription can be immediately
CC   stopped by the introduction of oxygen and ammonia.
CC   A Rhodobacter capsulatus expression vector with transcription of the
CC   insert under the control of the nifH promoter.
CC   Restriction digests of the clone give the following sizes (kb):
CC   BamHI--8.2, 1.0; HindIII--8.2, 1.0. (ATCC staff)
CC   Medium is 1227 LB plus ampicillin.
CC   NM (pNF3)
CC   CM (no)
CC   NA (ds-DNA)
CC   TP (circular)
CC   ST ()
CC   TY (plasmid)
CC   SP (ATCC)
CC   HO (E.coli)(Rhodobacter)
CC   CP ()
CC   FN (cloning)
CC   SE ()
CC   PA ()
CC   BR ()
CC   OF ()
CC   OR ()
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FH   Key             Location/Qualifiers
FH
FT   misc_feature    0..0
FT                   /note="1. pNF2 remove small SalI-SalI
FT                   T4 DNA polymerase fill in:T4 DNA polymerase fill in
FT                   2. pUC4-KIXX EcoRI-EcoRI, MCS-kan gene-MCS
FT                   T4 DNA polymerase fill in:T4 DNA polymerase fill in
FT                   -> pNF3"
FT   misc_binding    0..0
FT                   /note="MCS BamHI-SacI-SmaI-HindIII-XhoI-BamHI-SacI-
FT                   SmaI-HindIII-XhoI"
FT   misc_binding    0..0
FT                   /note="SIT unique NdeI-BamHI-SacI-SmaI-HindIII-XhoI"
FT   rep_origin      0..0
FT                   /note="ORI E. coli pMB1 (ColE1 and pBR322)"
FT   CDS             0..0
FT                   /note="ANT E. coli beta-lactamase gene (bla)
FT                   ampicillin resistance gene (apr/amp)"
FT   CDS             0..0
FT                   /note="ANT E. coli kanamycin resistance gene (kan)"
FT   CDS             0..0
FT                   /note="ANT E. coli spectinomycin resistance gene
FT                   (spc)"
FT   CDS             0..0
FT                   /note="ANT E. coli streptomycin resistance gene (str)"
FT   promoter        0..0
FT                   /note="PRO E. coli nifH gene"
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SQ   Sequence 381 BP; 88 A; 119 C; 98 G; 76 T; 0 other;
     agttcatccc gcgcgatgaa gacgtgatgg tgcatttcac cggcaccccc ggcggctcgg
     tctgaaaccg ccgcaaggac cggaaaacgc cgcgctggcg gggtttttca tttcctcccg
     acagagggcc cgacaaaacc ccgcccgaca ccgcaggccc tacaatttgt cggccttgtt
     ccatttcgaa caaaaccttc aacaccatga tttcgcgtca ttatttgcga aaattccggt
     tggcacgatg gctgctgtag aagctgtgag cccggttagg aaccgtctcg atattcgtga
     agcaccaacc ccaaggagcc accacatggg caaactccgt cagatcgcct tctacggcaa
     aggtggtatc ggcaagtcga c
//