LOCUS       V005182                 4133 bp    DNA     circular SYN 18-DEC-2018
DEFINITION  Exported.
ACCESSION   V005182
VERSION     V005182
KEYWORDS    direct-selection cloning vehicle; positive-selection; pheS
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
            .
REFERENCE   1  (bases 1 to 4133)
  AUTHORS   Selzer G, Som T, Itoh T, Tomizawa J.
  TITLE     The origin of replication of plasmid p15A and comparative studies on
            the nucleotide sequences around the origin of related plasmids
  JOURNAL   Cell 32 (1), 119-129 (1983)
   PUBMED   6186390
REFERENCE   2  (bases 1 to 4133)
  AUTHORS   Fayat G, Mayaux JF, Sacerdot C, Fromant M, Springer M,
            Grunberg-Manago M, Blanquet S.
  TITLE     Escherichia coli phenylalanyl-tRNA synthetase operon region.
            Evidence for an attenuation mechanism. Identification of the gene
            for the ribosomal protein L20
  JOURNAL   J. Mol. Biol. 171 (3), 239-261 (1983)
   PUBMED   6317865
REFERENCE   3  (bases 3864 to 3956)
  AUTHORS   Yanisch-Perron C, Vieira J, Messing J.
  TITLE     Improved M13 phage cloning vectors and host strains: nucleotide
            sequences of the M13mp18 and pUC19 vectors
  JOURNAL   Gene 33 (1), 103-119 (1985)
   PUBMED   2985470
REFERENCE   4  (bases 1 to 4133)
  AUTHORS   Balbas P, Soberon X, Merino E, Zurita M, Lomeli H, Valle F, Flores
            N, Bolivar F.
  TITLE     Plasmid vector pBR322 and its special-purpose derivatives--a review
  JOURNAL   Gene 50 (1-3), 3-40 (1986)
   PUBMED   3034735
REFERENCE   5  (bases 4133 to 4133)
  AUTHORS   Short JM, Fernandez JM, Sorge JA, Huse WD.
  TITLE     Lambda ZAP: a bacteriophage lambda expression vector with in vivo
            excision properties
  JOURNAL   Nucleic Acids Res. 16 (15), 7583-7600 (1988)
   PUBMED   2970625
REFERENCE   6  (bases 1 to 4133)
  AUTHORS   Kast P, Hennecke H.
  TITLE     Amino acid substrate specificity of Escherichia coli
            phenylalanyl-tRNA synthetase altered by distinct mutations
  JOURNAL   J. Mol. Biol. 222 (1), 99-124 (1991)
   PUBMED   1942071
REFERENCE   7  (bases 1 to 4133)
  AUTHORS   Alting-Mees MA, Sorge JA, Short JM.
  TITLE     pBluescriptII: multifunctional cloning and mapping vectors
  JOURNAL   Meth. Enzymol. 216, 483-495 (1992)
   PUBMED   1362236
REFERENCE   8  (bases 1 to 4133)
  AUTHORS   Lin-Chao S, Chen WT, Wong TT.
  TITLE     High copy number of the pUC plasmid results from a
            Rom/Rop-suppressible point mutation in RNA II
  JOURNAL   Mol. Microbiol. 6 (22), 3385-3393 (1992)
   PUBMED   1283002
REFERENCE   9  (bases 4133 to 4133)
  AUTHORS   Kast P.
  TITLE     pKSS--a second-generation general purpose cloning vector for
            efficient positive selection of recombinant clones
  JOURNAL   Gene 138 (1-2), 109-114 (1994)
   PUBMED   8125286
REFERENCE   10  (bases 1 to 4133)
  AUTHORS   Kast P.
  TITLE     Direct Submission
  JOURNAL   Submitted (10-SEP-1993) Dept. of Chemistry and Molecular Biology,
            The Scripps Research Institute, 10550 North Torrey Pines Road, La
            Jolla, CA 92037, USA
REFERENCE   11  (bases 1 to 4133)
  TITLE     Direct Submission
REFERENCE   12  (bases 1 to 4133)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Cell";
            date: "1983"; volume: "32"; issue: "1"; pages: "119-129"
            SGRef: number: 2; type: "Journal Article"; journalName: "J. Mol.
            Biol."; date: "1983"; volume: "171"; issue: "3"; pages: "239-261"
            SGRef: number: 3; type: "Journal Article"; journalName: "Gene";
            date: "1985"; volume: "33"; issue: "1"; pages: "103-119"
            SGRef: number: 4; type: "Journal Article"; journalName: "Gene 50
            (1-3), 3-40 (1986)"
            SGRef: number: 5; type: "Journal Article"; journalName: "Nucleic
            Acids Res."; date: "1988"; volume: "16"; issue: "15"; pages:
            "7583-7600"
            SGRef: number: 6; type: "Journal Article"; journalName: "J. Mol.
            Biol."; date: "1991"; volume: "222"; issue: "1"; pages: "99-124"
            SGRef: number: 7; type: "Journal Article"; journalName: "Meth.
            Enzymol. 216, 483-495 (1992)"
            SGRef: number: 8; type: "Journal Article"; journalName: "Mol.
            Microbiol."; date: "1992"; volume: "6"; issue: "22"; pages:
            "3385-3393"
            SGRef: number: 9; type: "Journal Article"; journalName: "Gene 138
            (1-2), 109-114 (1994)"
            SGRef: number: 10; type: "Journal Article"; journalName: "Submitted
            (10-SEP-1993) Dept. of Chemistry and Molecular Biology, The Scripps
            Research Institute, 10550 North Torrey Pines Road, La Jolla, CA
            92037, USA"
            SGRef: number: 11; type: "Journal Article"
            On Oct 6, 1993 this sequence version replaced gi:403935.
FEATURES             Location/Qualifiers
     source          1..4133
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     18..34
                     /label="KS primer"
                     /note="common sequencing primer, one of multiple similar
                     variants"
     CDS             146..1126
                     /gene="pheS"
                     /label="Phenylalanine--tRNA ligase alpha subunit"
                     /note="Phenylalanine--tRNA ligase alpha subunit from
                     Escherichia coli O139:H28 (strain E24377A / ETEC).
                     Accession#: A7ZMI2"
     primer_bind     complement(1244..1260)
                     /label="SK primer"
                     /note="common sequencing primer, one of multiple similar
                     variants"
     promoter        complement(1293..1311)
                     /label="T7 promoter"
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1318..1334)
                     /label="M13 fwd"
                     /note="common sequencing primer, one of multiple similar
                     variants"
     rep_origin      1476..1931
                     /label="f1 ori"
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     promoter        1957..2061
                     /label="AmpR promoter"
     CDS             2062..2919
                     /label="AmpR"
                     /note="beta-lactamase"
     rep_origin      3093..3681
                     /label="ori"
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
     CDS             3864..3956
                     /codon_start=1
                     /gene="'lacI"
                     /product="lac repressor fragment"
                     /label="'lacI"
                     /note="vestigal C-terminal portion of lac repressor; this
                     3' portion of lacI is not functional"
                     /citation=[3]
                     /citation=[7]
                     /protein_id="AAB61949.1"
                     /translation="APNTQTASPRALADSLMQLARQVSRLESGQ"
     gene            3864..3956
                     /gene="'lacI"
                     /label="'lacI"
     protein_bind    3969..3990
                     /label="CAP binding site"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     promoter        4005..4035
                     /label="lac promoter"
                     /note="promoter for the E. coli lac operon"
     protein_bind    4043..4059
                     /label="lac operator"
                     /note="The lac repressor binds to the lac operator to
                     inhibit transcription in E. coli. This inhibition can be
                     relieved by adding lactose or
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     primer_bind     4067..4083
                     /label="M13 rev"
                     /note="common sequencing primer, one of multiple similar
                     variants"
     promoter        4104..4122
                     /label="T3 promoter"
                     /note="promoter for bacteriophage T3 RNA polymerase"
     old_sequence    4133
                     /replace="tg"
                     /note="all pBluescript I KS (+/-) vectors (and pKSS)
                     exhibit the deletion of the G nucleotide 5' to the KpnI
                     site [8,9]"
                     /citation=[5]
                     /citation=[7]
                     /citation=[9]
ORIGIN
        1 ggtaccgggc cccccctcga ggtcgacggt atcgataagc ttgatatcga attcccccgg
       61 gaccaaaatg gcaagtaaaa tagcctgatg ggataggctc taagtccaac gaaccagtgt
      121 caccactgac acaatgagga aaaccatgtc acatctcgca gaactggttg ccagtgcgaa
      181 ggcggccatt agccaggcgt cagatgttgc cgcgttagat aatgtgcgcg tcgaatattt
      241 gggtaaaaaa gggcacttaa cccttcagat gacgaccctg cgtgagctgc cgccagaaga
      301 gcgtccggca gctggtgcgg ttatcaacga agcgaaagag caggttcagc aggcgctgaa
      361 tgcgcgtaaa gcggaactgg aaagcgctgc actgaatgcg cgtctggcgg cggaaacgat
      421 tgatgtctct ctgccaggtc gtcgcattga aaacggcggt ctgcatccgg ttacccgtac
      481 catcgaccgt atcgaaagtt tcttcggtga gcttggcttt accgtggcaa ccgggccgga
      541 aatcgaagac gattatcata acttcgatgc tctgaacatt cctggtcacc acccggcgcg
      601 cgctgaccac gacactttct ggtttgacac tacccgcctg ctgcgtaccc agacctctgg
      661 cgtacagatc cgcaccatga aagcccagca gccaccgatt cgtatcatcg cgcctggccg
      721 tgtttatcgt aacgactacg accagactca cacgccgatg ttccatcaga tggaaggtct
      781 gattgttgat accaacatca gctttaccaa cctgaaaggc acgctgcacg acttcctgcg
      841 taacttcttt gaggaagatt tgcagattcg cttccgtcct tcctacttcc cgtttaccga
      901 accttctgca gaagtggacg tcatgggtaa aaacggtaaa tggctggaag tgctgggctg
      961 cgggatggtg catccgaacg tgttgcgtaa cgttggcatc gacccggaag tttactctgg
     1021 tttcggcttc gggatgggga tggagcgtct gactatgttg cgttacggcg tcaccgacct
     1081 gcgttcattc ttcgaaaacg atctgcgttt cctcaaacag tttaaataag gcaggaatag
     1141 attatgaaat tcagtgaact gtggttacgc gaatgggtga acccggcgat tgatagcgat
     1201 gcgctggcaa atcaaatcac tatggcgggc ctggaagttg ggggatccac tagttctaga
     1261 gcggccgcca ccgcggtgga gctccaattc gccctatagt gagtcgtatt acaattcact
     1321 ggccgtcgtt ttacaacgtc gtgactggga aaaccctggc gttacccaac ttaatcgcct
     1381 tgcagcacat ccccctttcg ccagctggcg taatagcgaa gaggcccgca ccgatcgccc
     1441 ttcccaacag ttgcgcagcc tgaatggcga atgggacgcg ccctgtagcg gcgcattaag
     1501 cgcggcgggt gtggtggtta cgcgcagcgt gaccgctaca cttgccagcg ccctagcgcc
     1561 cgctcctttc gctttcttcc cttcctttct cgccacgttc gccggctttc cccgtcaagc
     1621 tctaaatcgg gggctccctt tagggttccg atttagtgct ttacggcacc tcgaccccaa
     1681 aaaacttgat tagggtgatg gttcacgtag tgggccatcg ccctgataga cggtttttcg
     1741 ccctttgacg ttggagtcca cgttctttaa tagtggactc ttgttccaaa ctggaacaac
     1801 actcaaccct atctcggtct attcttttga tttataaggg attttgccga tttcggccta
     1861 ttggttaaaa aatgagctga tttaacaaaa atttaacgcg aattttaaca aaatattaac
     1921 gcttacaatt taggtggcac ttttcgggga aatgtgcgcg gaacccctat ttgtttattt
     1981 ttctaaatac attcaaatat gtatccgctc atgagacaat aaccctgata aatgcttcaa
     2041 taatattgaa aaaggaagag tatgagtatt caacatttcc gtgtcgccct tattcccttt
     2101 tttgcggcat tttgccttcc tgtttttgct cacccagaaa cgctggtgaa agtaaaagat
     2161 gctgaagatc agttgggtgc acgagtgggt tacatcgaac tggatctcaa cagcggtaag
     2221 atccttgaga gttttcgccc cgaagaacgt tttccaatga tgagcacttt taaagttctg
     2281 ctatgtggcg cggtattatc ccgtattgac gccgggcaag agcaactcgg tcgccgcata
     2341 cactattctc agaatgactt ggttgagtac tcaccagtca cagaaaagca tcttacggat
     2401 ggcatgacag taagagaatt atgcagtgct gccataacca tgagtgataa cactgcggcc
     2461 aacttacttc tgacaacgat cggaggaccg aaggagctaa ccgctttttt gcacaacatg
     2521 ggggatcatg taactcgcct tgatcgttgg gaaccggagc tgaatgaagc cataccaaac
     2581 gacgagcgtg acaccacgat gcctgtagca atggcaacaa cgttgcgcaa actattaact
     2641 ggcgaactac ttactctagc ttcccggcaa caattaatag actggatgga ggcggataaa
     2701 gttgcaggac cacttctgcg ctcggccctt ccggctggct ggtttattgc tgataaatct
     2761 ggagccggtg agcgtgggtc tcgcggtatc attgcagcac tggggccaga tggtaagccc
     2821 tcccgtatcg tagttatcta cacgacgggg agtcaggcaa ctatggatga acgaaataga
     2881 cagatcgctg agataggtgc ctcactgatt aagcattggt aactgtcaga ccaagtttac
     2941 tcatatatac tttagattga tttaaaactt catttttaat ttaaaaggat ctaggtgaag
     3001 atcctttttg ataatctcat gaccaaaatc ccttaacgtg agttttcgtt ccactgagcg
     3061 tcagaccccg tagaaaagat caaaggatct tcttgagatc ctttttttct gcgcgtaatc
     3121 tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc ggatcaagag
     3181 ctaccaactc tttttccgaa ggtaactggc ttcagcagag cgcagatacc aaatactgtt
     3241 cttctagtgt agccgtagtt aggccaccac ttcaagaact ctgtagcacc gcctacatac
     3301 ctcgctctgc taatcctgtt accagtggct gctgccagtg gcgataagtc gtgtcttacc
     3361 gggttggact caagacgata gttaccggat aaggcgcagc ggtcgggctg aacggggggt
     3421 tcgtgcacac agcccagctt ggagcgaacg acctacaccg aactgagata cctacagcgt
     3481 gagctatgag aaagcgccac gcttcccgaa gggagaaagg cggacaggta tccggtaagc
     3541 ggcagggtcg gaacaggaga gcgcacgagg gagcttccag ggggaaacgc ctggtatctt
     3601 tatagtcctg tcgggtttcg ccacctctga cttgagcgtc gatttttgtg atgctcgtca
     3661 ggggggcgga gcctatggaa aaacgccagc aacgcggcct ttttacggtt cctggccttt
     3721 tgctggcctt ttgctcacat gttctttcct gcgttatccc ctgattctgt ggataaccgt
     3781 attaccgcct ttgagtgagc tgataccgct cgccgcagcc gaacgaccga gcgcagcgag
     3841 tcagtgagcg aggaagcgga agagcgccca atacgcaaac cgcctctccc cgcgcgttgg
     3901 ccgattcatt aatgcagctg gcacgacagg tttcccgact ggaaagcggg cagtgagcgc
     3961 aacgcaatta atgtgagtta gctcactcat taggcacccc aggctttaca ctttatgctt
     4021 ccggctcgta tgttgtgtgg aattgtgagc ggataacaat ttcacacagg aaacagctat
     4081 gaccatgatt acgccaagct cgaaattaac cctcactaaa gggaacaaaa gct
//
