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ID PKSS preliminary; circular DNA; SYN; 4133 BP.
XX
AC U01668;
XX
DT 29-SEP-1993 (Rel. 7, Created)
DT 01-JUL-1995 (Rel. 12, Last updated, Version 1)
XX
DE E. coli phagemid vector pKSS - complete.
XX
KW cloning vector.
XX
OS Cloning vector
OC Artificial sequences; Cloning vehicles.
XX
RN [1]
RP 4091-4133; 1-61; 1237-1336
RC pKSS from pKSB1-M4G
RA Kast P.;
RT "pKSS - A second-generation general purpose cloning vector for
RT efficient positive selection of recombinant clones";
RL Gene 138:109-114(1994).
XX
RN [2]
RP 365-366; 1026-1026
RC pKSC-A, pKSC-K from pUC19 & E.coli pheS mutant chromosome
RC pKSC-W from pUC19 & pHW0
RC [pKSC-M4G from pUC19]
RC pKSB1-A, pKSB2-A from pKSC-A & pBluescript KS(+) or pBLS
RC pKSB1-K, pKSB2-K from pKSC-K & pBluescript KS(+) or pBLS
RC pKSB1-W, pKSB2-W from pKSC-W & pBluescript KS(+) or pBLS
RC pKSB1-M4G, pKSB2-M4G from pKSC-M4G & pBluescript KS(+) or pBLS
RA Kast P., Hennecke H.;
RT "Amino acid substrate specificity of Escherichia coli
RT phenylalanyl-tRNA synthetase altered by distinct mutations";
RL J. Mol. Biol. 222:99-124(1991).
XX
RN [3]
RP 59-1239
RC pBH10d from pB5-10
RC pBH57d from pB5-57
RC pXH57 or pB9-1 from pB5-57
RA Fayat G., Mayaux J.F., Sacerdot C., Fromant M., Springer M.,
RA Grunberg-Manago M., Blanquet S.;
RT "Escherichia coli phenylalanyl-tRNA synthetase operon region.
RT Evidence for an attenuation mechanism. Identification of the gene
RT for the ribosomal protein L20";
RL J. Mol. Biol. 171:239-261(1983).
XX
RN [4]
RP 1-4133
RC pKSS
RA Kast P.;
RT ;
RL Submitted (10-SEP-1993) by:
RL Kast P., Dept. of Chemistry and Molecular Biology,
RL Scripps Research Inst., 10666 N. Torrey Pines Rd.,
RL La Jolla, CA 92037 USA.
XX
RN [5]
RC lambda p2delta1 from lambda p2
RC lambda IBPC B1 from lambda p2delta1 & lambda NM540 or lambda 540
RC pA1 from pACYC184 & lambda p2
RC pB1 or pHW0 from pBR322 & lambda p2
RC pB5-10, pB5-57, pB8-7, pB8-9, pB9-1 from pBR322 & pB1
RC pHWO from pBR322 & lambda p2, pheS gene
RA Plumbridge J.A., Springer M., Graffe M., Goursot R.,
RA Grunberg-Manago M.;
RT "Physical localisation and cloning of the structural gene for E.
RT coli initiation factor IF3 from a group of genes concerned with
RT translation";
RL Gene 11:33-42(1980).
XX
RN [6]
RC from lambda p2
RC pHW0 or pB1 from pBR322 & E.coli chromosome
RA Elhardt D., Wirth R., Bock A.;
RT "Regulation of formation of threonyl-tRNA synthetase,
RT phenylalanyl-tRNA synthetase and protein synthesis initiation
RT factor 3 from Escherichia coli in vivo and in vitro";
RL Eur. J. Biochem. 123:477-482(1982).
XX
RN [7]
RC lambda p2 from lambda
RA Springer M., Graffe M., Hennecke H.;
RT "Specialized transducing phage for the initiation factor IF3 gene in
RT E. coli";
RL Proc. Natl. Acad. Sci. U.S.A. 74:3970-3974(1977).
XX
RN [8]
RC lambda 512 from lambda
RC lambda + or lambda papa from lambda
RA Kourilsky P., Perricaudet M., Gros D., Garapin A., Gottesman M.,
RA Fritch A., Tiollais P.;
RT "Description and properties of bacteriophage lambda vectors useful
RT for the cloning of EcoRI fragments";
RL Biochimie 60:183-187(1978).
XX
CC NM (pKSS)
CC CM (yes)
CC NA (ds-DNA)
CC TP (circular)
CC ST ()
CC TY (plasmid)
CC SP ()
CC HO (E.coli)
CC CP ()
CC FN (cloning)
CC SE ()
CC PA ()
CC BR ()
CC OF ()
CC OR ()
XX
FH Key Location/Qualifiers
FH
FT misc_feature 0..0
FT /note="1. pUC19 remove SmaI-HindII 33bp 415..448,
FT \ MCS/2653bp
FT 2. E. coli SmaI-HindII 1181bp, mutant pheS
FT -> pKSC-M4G 3834bp
FT 1. pKSC-M4G SmaI-HindII 1181bp, mutant pheS gene
FT 2. pBluescript KS(+) SmaI 2507bp
FT -> pKSB1-M4G 4138bp
FT 1. pKSB1-M4G PstI 4138bp, MCS
FT T4 DNA polymerase
FT -> pKSS 4133bp [unique PstI in pheS gene]"
FT CDS 4079..4093
FT /note="GEN E. coli beta-galactosidase gene (lacZ);
FT partial [1] [3] [4];
FT N-terminal 5 residues of beta-galactosidase"
FT misc_feature join(1..55,1240..4133)
FT /note="from GenBank accession X52331"
FT misc_feature 1..61
FT /note="MCS polylinker A:
FT KpnI-Eco0109I-ApaI-AvaI-XhoI-SalI-AccI-HincII-ClaI-
FT HindIII-EcoRV-EcoRI-AvaI-SmaI [1]"
FT misc_feature 1243..1284
FT /note="MCS polylinker B [1]:
FT BamHI-SpeI-XbaI-EagI-NotI-DsaI-SacII(KspI)-SacI(SstI)"
FT promoter complement(1292..1311)
FT /note="PRO bacteriophage T7 promoter [1] [3] [4]"
FT primer_bind complement(1318..1334)
FT /note="PRI bacteriophage M13 universal (-20)
FT sequencing primer [1] [3]"
FT rep_origin 1476..1931
FT /note="ORI bacteriophage f1; origin for single strand
FT DNA replication [3] [4]; direction right;
FT nicking site of gene II protein is between
FT 1768 and 1769"
FT sig_peptide 2062..2130
FT /note="E. coli beta-lactamase gene signal peptide [6]"
FT mat_peptide 2131..2919
FT /note="E. coli beta-lactamase gene mature peptide;
FT EC number 3.5.2.6 [6]"
FT rep_origin 3082..3696
FT /note="ORI E. coli pMB1 (ColE1 and pBR322)
FT [6] [8]; direction right;
FT the DNA region indicated is sufficient for a
FT functional replication origin [8]; DNA replication
FT initiates at 3681 to 3683 [6]"
FT promoter 3957..4061
FT /note="PRO E. coli lac promoter-operator (wild type)
FT [3] [4]; for details and references to the lac
FT promoter-operator region see Miller J.H.,
FT Reznikoff W.S., (ed.), The Operon, 2nd ed. (1980),
FT Cold Spring Harbor, pp. 177-243."
FT protein_bind 3957..3994
FT /note="SIT catabolite activator protein (CAP)
FT CAP binding site (wild-type sequence) [3], [4]"
FT -35_signal 4005..4010
FT /note="S35 E. coli lac gene"
FT -10_signal 4029..4034
FT /note="S10 E. coli lac gene"
FT protein_bind 4041..4061
FT /note="SIT E. coli lac repressor protein;
FT lacOlac operator"
FT promoter 4104..4123
FT /note="PRO bacteriophage T3 promoter [1], [3], [4]"
FT CDS 146..1129
FT /note="GEN E. coli pheS gene;
FT EC_number 6.1.1.20; Gly294 mutant;
FT confers p-Cl-phenylalanine sensitivity [2] [5]
FT phenylalanyl-tRNA synthetase alpha subunit
FT (Gly294 variant)"
FT CDS 1314..1474
FT /note="GEN E. coli beta-galactosidase gene (lacZ);
FT EC_number 3.2.1.23; partial [1] [3] [4]
FT N-truncated alpha-fragment of beta-galactosidase;
FT blue/white screening with the lacZ system is
FT irrelevant"
FT CDS 2062..2922
FT /note="ANT E. coli beta-lactamase gene (bla)
FT ampicillin resistance gene (apr/amp)
FT EC_number 3.5.2.6; [4] [6] [7]; precursor"
FT CDS 3864..3956
FT /note="REP E. coli lacI repressor gene;
FT partial [4] [7];
FT vestigal C-terminal portion of lac repressor;
FT this 3' portion of lacI is not functional"
XX
SQ Sequence 4133 BP; 1010 A; 1052 C; 1053 G; 1018 T; 0 other;
ggtaccgggc cccccctcga ggtcgacggt atcgataagc ttgatatcga attcccccgg
gaccaaaatg gcaagtaaaa tagcctgatg ggataggctc taagtccaac gaaccagtgt
caccactgac acaatgagga aaaccatgtc acatctcgca gaactggttg ccagtgcgaa
ggcggccatt agccaggcgt cagatgttgc cgcgttagat aatgtgcgcg tcgaatattt
gggtaaaaaa gggcacttaa cccttcagat gacgaccctg cgtgagctgc cgccagaaga
gcgtccggca gctggtgcgg ttatcaacga agcgaaagag caggttcagc aggcgctgaa
tgcgcgtaaa gcggaactgg aaagcgctgc actgaatgcg cgtctggcgg cggaaacgat
tgatgtctct ctgccaggtc gtcgcattga aaacggcggt ctgcatccgg ttacccgtac
catcgaccgt atcgaaagtt tcttcggtga gcttggcttt accgtggcaa ccgggccgga
aatcgaagac gattatcata acttcgatgc tctgaacatt cctggtcacc acccggcgcg
cgctgaccac gacactttct ggtttgacac tacccgcctg ctgcgtaccc agacctctgg
cgtacagatc cgcaccatga aagcccagca gccaccgatt cgtatcatcg cgcctggccg
tgtttatcgt aacgactacg accagactca cacgccgatg ttccatcaga tggaaggtct
gattgttgat accaacatca gctttaccaa cctgaaaggc acgctgcacg acttcctgcg
taacttcttt gaggaagatt tgcagattcg cttccgtcct tcctacttcc cgtttaccga
accttctgca gaagtggacg tcatgggtaa aaacggtaaa tggctggaag tgctgggctg
cgggatggtg catccgaacg tgttgcgtaa cgttggcatc gacccggaag tttactctgg
tttcggcttc gggatgggga tggagcgtct gactatgttg cgttacggcg tcaccgacct
gcgttcattc ttcgaaaacg atctgcgttt cctcaaacag tttaaataag gcaggaatag
attatgaaat tcagtgaact gtggttacgc gaatgggtga acccggcgat tgatagcgat
gcgctggcaa atcaaatcac tatggcgggc ctggaagttg ggggatccac tagttctaga
gcggccgcca ccgcggtgga gctccaattc gccctatagt gagtcgtatt acaattcact
ggccgtcgtt ttacaacgtc gtgactggga aaaccctggc gttacccaac ttaatcgcct
tgcagcacat ccccctttcg ccagctggcg taatagcgaa gaggcccgca ccgatcgccc
ttcccaacag ttgcgcagcc tgaatggcga atgggacgcg ccctgtagcg gcgcattaag
cgcggcgggt gtggtggtta cgcgcagcgt gaccgctaca cttgccagcg ccctagcgcc
cgctcctttc gctttcttcc cttcctttct cgccacgttc gccggctttc cccgtcaagc
tctaaatcgg gggctccctt tagggttccg atttagtgct ttacggcacc tcgaccccaa
aaaacttgat tagggtgatg gttcacgtag tgggccatcg ccctgataga cggtttttcg
ccctttgacg ttggagtcca cgttctttaa tagtggactc ttgttccaaa ctggaacaac
actcaaccct atctcggtct attcttttga tttataaggg attttgccga tttcggccta
ttggttaaaa aatgagctga tttaacaaaa atttaacgcg aattttaaca aaatattaac
gcttacaatt taggtggcac ttttcgggga aatgtgcgcg gaacccctat ttgtttattt
ttctaaatac attcaaatat gtatccgctc atgagacaat aaccctgata aatgcttcaa
taatattgaa aaaggaagag tatgagtatt caacatttcc gtgtcgccct tattcccttt
tttgcggcat tttgccttcc tgtttttgct cacccagaaa cgctggtgaa agtaaaagat
gctgaagatc agttgggtgc acgagtgggt tacatcgaac tggatctcaa cagcggtaag
atccttgaga gttttcgccc cgaagaacgt tttccaatga tgagcacttt taaagttctg
ctatgtggcg cggtattatc ccgtattgac gccgggcaag agcaactcgg tcgccgcata
cactattctc agaatgactt ggttgagtac tcaccagtca cagaaaagca tcttacggat
ggcatgacag taagagaatt atgcagtgct gccataacca tgagtgataa cactgcggcc
aacttacttc tgacaacgat cggaggaccg aaggagctaa ccgctttttt gcacaacatg
ggggatcatg taactcgcct tgatcgttgg gaaccggagc tgaatgaagc cataccaaac
gacgagcgtg acaccacgat gcctgtagca atggcaacaa cgttgcgcaa actattaact
ggcgaactac ttactctagc ttcccggcaa caattaatag actggatgga ggcggataaa
gttgcaggac cacttctgcg ctcggccctt ccggctggct ggtttattgc tgataaatct
ggagccggtg agcgtgggtc tcgcggtatc attgcagcac tggggccaga tggtaagccc
tcccgtatcg tagttatcta cacgacgggg agtcaggcaa ctatggatga acgaaataga
cagatcgctg agataggtgc ctcactgatt aagcattggt aactgtcaga ccaagtttac
tcatatatac tttagattga tttaaaactt catttttaat ttaaaaggat ctaggtgaag
atcctttttg ataatctcat gaccaaaatc ccttaacgtg agttttcgtt ccactgagcg
tcagaccccg tagaaaagat caaaggatct tcttgagatc ctttttttct gcgcgtaatc
tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc ggatcaagag
ctaccaactc tttttccgaa ggtaactggc ttcagcagag cgcagatacc aaatactgtt
cttctagtgt agccgtagtt aggccaccac ttcaagaact ctgtagcacc gcctacatac
ctcgctctgc taatcctgtt accagtggct gctgccagtg gcgataagtc gtgtcttacc
gggttggact caagacgata gttaccggat aaggcgcagc ggtcgggctg aacggggggt
tcgtgcacac agcccagctt ggagcgaacg acctacaccg aactgagata cctacagcgt
gagctatgag aaagcgccac gcttcccgaa gggagaaagg cggacaggta tccggtaagc
ggcagggtcg gaacaggaga gcgcacgagg gagcttccag ggggaaacgc ctggtatctt
tatagtcctg tcgggtttcg ccacctctga cttgagcgtc gatttttgtg atgctcgtca
ggggggcgga gcctatggaa aaacgccagc aacgcggcct ttttacggtt cctggccttt
tgctggcctt ttgctcacat gttctttcct gcgttatccc ctgattctgt ggataaccgt
attaccgcct ttgagtgagc tgataccgct cgccgcagcc gaacgaccga gcgcagcgag
tcagtgagcg aggaagcgga agagcgccca atacgcaaac cgcctctccc cgcgcgttgg
ccgattcatt aatgcagctg gcacgacagg tttcccgact ggaaagcggg cagtgagcgc
aacgcaatta atgtgagtta gctcactcat taggcacccc aggctttaca ctttatgctt
ccggctcgta tgttgtgtgg aattgtgagc ggataacaat ttcacacagg aaacagctat
gaccatgatt acgccaagct cgaaattaac cctcactaaa gggaacaaaa gct
//