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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
//