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ID   PHPS9      preliminary; circular DNA; SYN; 5700 BP.
AC   ATCC37816; ATCC37817;
DT   01-JUL-1993 (Rel. 7, Created)
DT   01-JUL-1995 (Rel. 12, Last updated, Version 1)
DE   Bacillus/E.coli plasmid vector pHPS9 - incomplete.
KW   cloning vector.
OS   Cloning vector
OC   Artificial sequences; Cloning vehicles.
RN   [1]
RC   pHP13 from pUC9 & pTA1060 & pE194 & pC194
RC   plasmid from pMLB1034 & pEB112
RC   pSG2 from plasmid & pPJ1
RC   pSG4 from pSG2 & pHP13
RC   pGK15 from pGK12
RC   pGKS1 from pSG4 & pGK15
RC   pUC9deltaM15 from pUC9 & pSG4
RC   pMG29 from pWV01
RC   pMGS2 from pUC9deltaM15 & pMG29
RC   pGHS1 from pMGS2 & pGKV223
RC   pLGS1 from pGKS1 & pLG339
RC   pGKH1 from pUC9 & pGKV259
RC   pHP13-2 from pHP13
RC   pHPS9 from pGKH1 & pHP13-2
RA   Haima P., Van Sinderen D., Schotting H., Bron S., Venema G.;
RT   "Development of a beta-galactosidase alpha-complementation system
RT   for molecular cloning in Bacillus subtilis";
RL   Gene 86:63-69(1990).
RN   [2]
RC   pHP9 from pHP3
RC   pHP9R from pHP9
RC   pHP9D from pHP9
RC   pHP9D0 from pHP9
RC   pHP9R1 from pHP9D0
RC   pHP9D1100 from pHP9D0 & E.coli DNA
RC   pHP9D1600 from pHP9D0 & E.coli DNA
RC   pHP9D3000 from pHP9D0 & E.coli DNA
RC   pHP9D4000 from pHP9D0 & E.coli DNA
RC   pHP9R2 from pHP9R1
RC   pHP9D1 from pHP9D0
RC   pHPS9R from pHP13-2
RA   Haima P., Bron S., Venema G.;
RT   "Novel plasmid marker rescue transformation system for molecular
RT   cloning in Bacillus subtilis enabling direct selection of
RT   recombinants";
RL   Mol. Gen. Genet. 223:185-191(1990).
RN   [3]
RC   pHP3 from pBR322 & pTA1060 & pE194 & pC194
RC   pHP3Ff from pHP3 & phage f1
RC   pHP3::Tn5 from pHP3 & Tn5
RC   pPJ1 from pUC7 & pJH1, kan gene
RC   pHP35B, pHP35H, pHP35F, pHP35P, pHP35X, pHP35I from pHP3::Tn5
RC   from pHP35B & pUR222
RC   from pHP35H & pUR222
RC   pHP300 series from pHP3Ff
RA   Peeters B.P., De Boer J., Bron S., Venema G.;
RT   "Structural plasmid instability in Bacillus subtilis: efect of
RT   direct and inverted repeats";
RL   Mol. Gen. Genet. 212:450-458(1988).
RN   [4]
RC   pHP13 from pUC9 & pTA1060 & pE194 & pC194
RC   pC19, pC23, pC37 from pHP13
RC   pG1, pG2, pG6, pG26, pG11, pG18 from pHP13
RC   pC5, pC14 from pHP13
RC   pC17, pC19, pC33 from pHP13
RA   Haima P., Bron S., Venema G.;
RT   "The effect of restriction on shotgun cloning and plasmid
RT   stability in Bacillus subtilis";
RL   Mol. Gen. Genet. 209:335-342(1987).
RN   [5]
RC   pbeta-C1 from pC1i & pHV1301
RC   pbeta-C2 from pC2i & pHV1301
RC   p52-C1 from pC1i & pTB53
RC   p52-C2 from pC2i & pTB53
RC   pHV1431 from pHV1301 & pHV60
RC   pHV1432 from pHV1431
RC   pHV1435 from pHV1301 & pHV60
RC   pHV1436 from pHV60 & pTB19
RA   Janniere L., Bruand C., Ehrlich S.D.;
RT   "Structurally stable DNA cloning vectors";
RL   Gene 87:53-61(1990).
RN   [6]
RC   pTB11, pTB12, pTB16 from B.stearothermophilus, ery gene
RC   pTB13, pTB17 from B.stearothermophilus, kan gene
RC   pTB14 from B.stearothermophilus, strep gene
RC   pTB18, pTB19 from B.stearothermophilus, kan gene/tet gene
RC   pTB15, pTB20 from B.stearothermophilus, tet gene
RC   pTB51, pTB52, pTB53 from pTB19
RA   Imanaka T., Fujii M., Aiba S.;
RT   "Isolation and Characterization of antibiotic resistance plasmids
RT   from thermophilic bacilli and construction of deletion plasmids";
RL   J. Bacteriol. 146:1091-1097(1981).
RN   [7]
RC   pRAT1 from pTB19 & pUB110
RC   pRAT11 from pRAT1
RA   Imanaka T., Ishikawa H., Aiba S.;
RT   "Complete nucleotide sequence of the low copy number plasmid pRAT11
RT   and replication control by the RepA protein in Bacillus subtilis";
RL   Mol. Gen. Genet. 205:90-96(1986).
RN   [8]
RC   pIL252 from pHV1301
RA   Simon D., Chopin A.;
RT   "Construction of a vector plasmid family and its use for molecular
RT   cloning in Streptococcus lactis";
RL   Biochimie 76:559-566(1988).
RN   [9]
RC   pHV1301 from pAMbeta1
RA   Noirot P.;
RT   ;
RL   Unpublished (1990).
RN   [10]
RC   pC1i, pC2i from pBR322 & pE194 & pC194 & Tn5
RA   Janniere L., Ehrlich S.D.;
RT   "Recombination between short repeated sequences is more frequent
RT   in plasmids than in the chromosome of Bacillus subtilis";
RL   Mol. Gen. Genet. 210:116-121(1987).
CC   Cloning into the NdeI, NheI, BamHI, SmaI or EcoRI sites inactivates
CC   lacZalpha.  The NcoI site interrupts the chloramphenicol resistance
CC   sequence. [1]
CC   Permits alpha-complementation to identify recombinants when used with
CC   Bacillus subtilis 6GM15, and alpha-complementation with plasmid marker
CC   rescue when used with Bacillis subtilis 6GM15[pHPS9R] (ATCC 37818).[3]
CC   The cat86::lacZalpha fusion is in-frame.  Expression is controlled by
CC   the P59 promoter from Lactococcus lactis subsp. cremoris Wg2. [1]
CC   This is the preferred strain for isolating the plasmid, because the
CC   copy number is higher in E.coli than in B. subtilis. (personal
CC   communication)
CC   Restriction digests of the clone give the following sizes (kb):
CC   BamHI--5.7, EcoRI--5.7, PstI--4.9, 0.7. (ATCC staff)
CC   Medium is -1 1065 plus erythromycin (200 ug/ml).
CC   Plasmid was not verified by restriction analysis.  Strain was checked
CC   for the following phenotypes:showed growth on LB + chloramphenicol
CC   (5 ug/ml, but not 10 ug/ml); blue colonies on IPTG + Xgal (80 ug/ml);
CC   (ATCC staff)
CC   blue colonies on IPTG + Xgal (80 ug/ml) + erythromycin (150 ug/ml).
CC   (ATCC staff)
CC   Medium is -1 1065 plus chloramphenicol (5 ug/ml).
CC   NM (pHPS9)
CC   CM (no)
CC   NA (ds-DNA)
CC   TP (circular)
CC   ST ()
CC   TY (plasmid)
CC   HO (E.coli NM522)(Bacillus subtilis)(E.coli)
CC   HO (Bacillus subtilis 6GM15 [ATCC 47051])
CC   CP ()
CC   FN (cloning)
CC   SE ()
CC   PA ()
CC   BR ()
CC   OF ()
CC   OR ()
FH   Key             Location/Qualifiers
FT   misc_feature    0..0
FT                   /note="1. pUC9 DraI/AhaIII-AatII 1613bp
FT                   \ 1785..2665..733, ori/lacZ gene
FT                   2. pTA1060 1300bp, ori
FT                   3. pE194 SacI-ClaI 960bp 1940..2900, ery gene
FT                   4. pC194 HpaII-MboI 1030bp 975..2005, cat gene
FT                   -> pHP13 4900bp
FT                   1. pUC19 AsuI/Sau96I-HindIII 161bp 287..448,lacZ alpha
FT                   fill in:fill in
FT                   2. pGKV259 HindIII 5000bp
FT                   fill in:fill in
FT                   -> pGKH1 5100bp [cat-86/lacZalpha in frame fusion]
FT                   1. pHP13 remove 233bp, lacZpo/pUC9 447..680/4700bp
FT                   remove 41bp, lacZalpha/pUC9 235..276/4600bp
FT                   -> pHP13-2 4600bp
FT                   1. pGKH1 ScaI-EcoRI 1052bp, promoter/cat-86/lacZalpha
FT                   2. pHP13-2 remove HindIII-EcoRI 30bp,
FT                   \ MCS/pUC9 234..264/4600bp
FT                   fill in:
FT                   -> pHPS9 5700bp"
FT   misc_binding    0..0
FT                   /note="MCS PstI-SalI-BamHI-SmaI-EcoRI"
FT   misc_binding    0..0
FT                   /note="SIT unique NdeI-NheI-BamHI-SmaI-EcoRI-NcoI"
FT   rep_origin      0..0
FT                   /note="ORI E. coli pMB1 (ColE1 and pBR322)"
FT   rep_origin      0..0
FT                   /note="ORI Bacillus pTA1060"
FT   promoter        0..0
FT                   /note="PRO Lactococcus lactis subsp. cremoris Wg2
FT                   P59 gene"
FT   CDS             0..0
FT                   /note="GEN E. coli beta-galactosidase gene (lacZ')"
FT   CDS             0..0
FT                   /note="ANT E. coli chloramphenicol acetyltransferase
FT                   gene (cat); chloramphenicol resistance gene (cmr/cml)"
FT   CDS             0..0
FT                   /note="ANT E. coli erythromycin resistance gene (ery)"
SQ   Sequence 1 BP; 0 A; 0 C; 0 G; 0 T; 1 other;