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ID   PFD666     preliminary; circular DNA; SYN; 5250 BP.
AC   ATCC77286;
DT   01-JUL-1993 (Rel. 7, Created)
DT   01-JUL-1995 (Rel. 12, Last updated, Version 1)
DE   Actinomycetes/E.coli cosmid vector pFD666 - incomplete.
KW   cloning vector.
OS   Cloning vector
OC   Artificial sequences; Cloning vehicles.
RN   [1]
RC   pFD18 from pIBI76 & pNU81, Tn5 neo
RC   plasmid from Lorist6 & pUC18
RC   plasmid2 from plasmid & pIBI76
RC   pFD204 from plasmid2 & pFD18
RC   pFD300 from pFD204 & pUC118, M13 ori
RC   pFD304 from pFD300 & pcos2EMBL, lambda cos
RC   pFD666 from pFD304 & pWOR126
RA   Denis F., Brzezinski R.;
RT   "A versatile shuttle cosmid vector for use in Escherichia coli and
RT   actinomycetes";
RL   Gene 111:115-118(1992).
RN   [2]
RC   pFDNEO from pUC18 & pNU81
RC   pFDNEO-S from pFDNEO & oligo
RA   Denis F., Brzezinski R.;
RT   "An improved aminoglycoside resistance gene cassette for use in
RT   gram-negative bacteria and Streptomyces";
RL   FEMS Microbiol. Lett. 81:261-264(1991).
RN   [3]
RC   from pKN402 & pNU5
RA   Jaurin B., Grundstrom T.;
RT   "ampC cephalosporinase of Escherichia coli K-12 has a different
RT   evolutionary origin from that of beta-lactamases of the
RT   penicillinase type";
RL   Proc. Natl. Acad. Sci. U.S.A. 78:4897-4901(1981).
RN   [4]
RC   pNU79 from pBR322 & E. coli ampC gene
RC   pNU80 from pNU79 & pBY06, kan gene
RC   pNU81 from pNU80 & linker
RC   pNU100 series from pNU81
RA   Jaurin B., Grundstrom T., Normark D.A.;
RT   "Sequence elements determining ampC promoter strength in E. coli";
RL   EMBO J. 1:875-881(1982).
RN   [5]
RC   pWOR101, pWOR103, pWOR191 from pIJ702 & pJV1; [compatible with pIJ101]
RC   pWOR109 from pIJ702 & pJV1
RC   pWOR120 from pWOR109
RC   pWOR121, pWOR122 from pWOR120 & pAT153
RC   pWOR123, pWOR124, pWOR125, pWOR126 from pWOR120 & pIJ702
RC   from pWOR109 & pSKS101
RA   Bailey C.R., Bruton C.J., Butler M.J., Chater K.F., Harris J.E.,
RA   Hopwood D.A.;
RT   "Properties of in vitro recombinant derivatives of pJV1, a
RT   multy-copy plasmid from Streptomyces phaeochromogenes";
RL   J. Gen. Microbiol. 132:2071-2078(1986).
RN   [6]
RC   pNU5, pNU6 from pBR322 & E. coli amp gene
RA   Edlund T., Grundstrom T., Normark D.A.;
RT   "Isolation and characterization of DNA repetitions carrying the
RT   chromosomal beta-lactamase gene of Escherichia coli K-12";
RL   Mol. Gen. Genet. 173:115-125(1979).
RN   [7]
RC   pBY06 from kan gene
RA   Bystrom A.;
RT   ;
RL   Unpublished (1982).
RN   [8]
RC   pBI136 from Bacteroides ovatus ery gene
RC   pFD214 from pBI136
RA   Smith C.J.;
RT   "Development and use of cloning systems for Bacteroides fragilis:
RT   cloning of a plasmid-encoded clindamycin resistance determinant";
RL   J. Bacteriol. 164:294-301(1985).
RN   [9]
RC   pJV1 from Streptomyces phaeochromogenes
RA   Doull J.L., Vining L.C., Stuttard C.;
RT   "A cryptic plasmid in the chloramphenicol-producing actinomycete,
RT   Streptomyces phaeochromogenes";
RL   FEMS Microbiol. Lett. 16:349-352(1983).
CC   The order of the major features in this plasmid is:rrnC terminator -
CC   SP6 - SphI/MCS/HindIII - T7 - trpA terminator - neo - synthetic
CC   promoter -pMB1 ori - cos - M13 ori - pJV1 ori. [1]
CC   Cosmid shuttle vector permitting production of ssDNA and containing
CC   promoters for in vitro RNA synthesis.  Terminators flanking the
CC   polylinker protect the vector from transcripts originating in cloned
CC   inserts. [1]
CC   The ori of pJV1 from Streptomyces phaeochromogenes is compatible with
CC   plasmids derived from pIJ101.  This vector can be used for
CC   complementation studies using multiple cloned genes in one host. [1]
CC   M13K07 helper phage improves the yield of ssDNA in E.coli with this
CC   vector.  M13mp18 may also be used. [1]
CC   The neo promoter was replaced by a synthetic promoter recognized by
CC   both hosts. [1]
CC   Restriction digests of the clone give the following sizes (kb):
CC   BamHI--5.4; PstI--5.4; PvuII--3.7, 1.7; SacI--4.5, 0.9. (ATCC staff)
CC   Medium is 1236 LB plus kanamycin.
CC   NM (pFD666)
CC   CM (no)
CC   NA (ds-DNA)
CC   TP (circular)
CC   ST ()
CC   TY (cosmid)
CC   HO (E.coli)(Actinomycetes)
CC   HO (Streptomyces lividans)(Saccharopolyspora erythraea)
CC   CP ()
CC   FN (cloning)
CC   SE ()
CC   PA (pFDNEO)(pJV1)
CC   BR ()
CC   OF ()
CC   OR ()
FH   Key             Location/Qualifiers
FT   misc_feature    0..0
FT                   /note="1. Tn5, neo/kan gene
FT                   -> pBY06
FT                   1. pBR322 PstI 4361bp 3614..3614, amp gene
FT                   2. E. coli DNA PstI-PstI, ampC gene
FT                   -> pNU79
FT                   1. pNU79 PstI 3614..3614
FT                   2. pBY06 PstI-PstI, kan/neo gene
FT                   -> pNU80
FT                   1. pNU80 RsaI 3849..3849, -84 to -81 of ampC gene
FT                   BamHI linker 10bp ccggatccgg
FT                   remove small BamHI-BamHI 3500bp 376..3849, tet gene
FT                   -> pNU81 [unique BamHI]
FT                   1. pIBI76 remove HindIII-SmaI, MCS/pBR322 HindIII 30
FT                   Klenow:Klenow
FT                   2. pNU81 HindIII-BstBI, Tn5 neo gene
FT                   Klenow:Klenow
FT                   -> plasmid [NheI between HindIII's]
FT                   1. plasmid XbaI-DraI
FT                   Klenow:Klenow
FT                   remove HaeII-EcoRV, lac operator/promoter
FT                   T4 DNA polymerase
FT                   2. oligo, E. coli/actinomyces vegatative promoters
FT                   -> pFD18
FT                   1. Lorist6 EcoRI-EcoRI 1862bp,  cloning cassette
FT                   \ LoristB 531..2393
FT                   2. pUC18 EcoRI 2686bp 451..451
FT                   -> plasmid2 4548bp
FT                   1. plasmid2 remove SacI-BglI 312bp, LoristB 541..853
FT                   \ 4236bp
FT                   T4 DNA polymerase
FT                   -> plasmid3 4236bp
FT                   1. plasmid3 BamHI 4236bp, LoristB 902..902
FT                   \ pUC18 430..430
FT                   T4 DNA polymerase
FT                   2. pIBI76 ClaI-HindIII 695bp, pBR322 25..-..30
FT                   T4 DNA polymerase:T4 DNA polymerase
FT                   -> plasmid4 4931bp [BamHI at ClaI]
FT                   1. plasmid4 BamHI-BamHI 650bp
FT                   -> plasmid5
FT                   1. plasmid5 SalI, MCS
FT                   T4 DNA polymerase, to make NruI
FT                   EcoRI-EcoRI, cloning cassette
FT                   T4 DNA polymerase
FT                   2. pFD18 ClaI, pBR322 25..25
FT                   T4 DNA polymerase
FT                   -> pFD204 [NruI in MCS, no SalI in MCS]
FT                   1. pFD204 remove small NruI-EcoRI, MCS/2100bp
FT                   T4 DNA polymerase
FT                   2. pUC118 ApaLI-RsaI 465bp 178..643, M13 ori [463bp]
FT                   T4 DNA polymerase
FT                   -> pFD300 2600bp
FT                   1. pFD300 BspHI 2600bp
FT                   T4 DNA polymerase
FT                   2. pcos2EMBL EcoRI-PvuII 650bp, lambda cos
FT                   HaeIII-HaeII 223bp, lambda cos
FT                   -> pFD304 2800bp
FT                   1. Streptomyces phaeochromogene
FT                   -> pJV1 10800bp
FT                   1. pIJ702 BclI-BclI 1000bp, tsr gene
FT                   bacterial alkaline phosphatase
FT                   2. pJV1 MboI-MboI 5000bp
FT                   3. pJV1 MboI-MboI 2000bp
FT                   -> pWOR109 8000bp
FT                   1. pWOR109 remove KpnI-KpnI 2400bp, 5600bp
FT                   -> pWOR120 5600bp
FT                   1. pWOR120 remove small BamHI-BglII
FT                   2. pIJ702 BclI-BclI 1550bp, mel gene
FT                   -> pWOR126
FT                   1. pFD304 SspI 2800bp
FT                   2. pWOR126 NruI-KpnI 2420bp, pJV1 ori
FT                   T4 DNA polymerase:T4 DNA polymerase
FT                   -> pFD666 5250bp"
FT   misc_binding    0..0
FT                   /note="MCS SphI-MluI-XhoI-ApaI-PstI-NruI-XbaI-BamHI-
FT                   ScaI-NotI-HindIII"
FT   misc_binding    0..0
FT                   /note="SIT unique SphI-XhoI-PstI-NruI-XbaI-BamHI-
FT                   ScaI-NotI-HindIII"
FT   rep_origin      0..0
FT                   /note="ORI E. coli pMB1 (ColE1 and pBR322)"
FT   rep_origin      0..0
FT                   /note="ORI bacteriophage M13"
FT   rep_origin      0..0
FT                   /note="ORI Actinomycetes pJV1"
FT   promoter        0..0
FT                   /note="PRO bacteriophage SP6"
FT   promoter        0..0
FT                   /note="PRO bacteriophage T7"
FT   CDS             0..0
FT                   /note="ANT E. coli kanamycin resistance gene (kan)"
SQ   Sequence 1 BP; 0 A; 0 C; 0 G; 0 T; 1 other;