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ID   PGSR15184D preliminary; circular DNA; SYN; 1890 BP.
XX
AC   D13798; D00365;
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DT   05-DEC-1992 (Rel. 7, Created)
DT   01-APR-1995 (Rel. 11, Last updated, Version 1)
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DE   Saccharomyces/E.coli plasmid vector pGSR1518-4d - incomplete, prom.
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KW   cloning vector; GSH-I; gamma-glutamylcysteine synthetase.
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OS   Cloning vector
OC   Artificial sequences; Cloning vehicles.
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RN   [1]
RP   1-1890
RC   pGSR1 from pMC1585, URA3/yeast 2 micron ori/amp gene
RC   pGSR150 from pGSR100 & pGSR1
RC   pGSR151 from pGSR150
RC   pGSR1518 from pGSR151 & yeast P8 promoter
RC   pGSR2518-4, pGSR2518-7 from pGSR1518
RC   pGSR15184D from pGSR2518-4
RA   Ohtake Y., Watanabe K., Tezuka H., Ogata T., Yabuuchi S., Murata K.,
RA   Kimura A.;
RT   "The expression of the gamma-glutamylcysteine synthetase gene of
RT   Escherichia coli B in Saccharomyces cerevisiae";
RL   Agric. Biol. Chem. 52:2753-2762(1988).
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CC   gshI = gamma-glutamylcysteine synthetase
CC   NCBI gi: 220957
CC   NM (pGSR1518-4d)
CC   CM (no)
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 (E.coli gshI gene)(2 micron plasmid)(pGSR1518)
CC   BR ()
CC   OF ()
CC   OR (no)
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FH   Key             Location/Qualifiers
FH
FT   misc_feature    0..0
FT                   /note="1. pMC1585
FT                   -> pGSR1 5900bp
FT                   1. pMC1585 BamHI, 5' lacZ gene
FT                   2. Saccharomyces cerevisiae YNN27 BamHI-BamHI 3800bp
FT                   -> P8
FT                   1. pGS100
FT                   -> pGSR100 8000bp
FT                   1. pGSR100 ScaI-HpaI 2500bp
FT                   BamHI linker:BamHI linker
FT                   2. pGSR1 BamHI
FT                   -> pGSR150 8200bp
FT                   1. pGSR150 BamHI, 3' gshI gene
FT                   T4, fill in
FT                   -> pGSR151 8200bp
FT                   1. pGSR151 BamHI, 5' gshI gene
FT                   2. P8 BamHI-BamHI 3800bp
FT                   -> pGSR1518 12000bp
FT                   1. pGSR1518 BamHI, 5' P8
FT                   T4, fill in
FT                   -> pGSR2518 12000bp
FT                   1. pGSR2518 BamHI, 5' P8
FT                   BAL31
FT                   XhoI linker
FT                   -> pGSR2518-4 11200bp
FT                   1. pGSR2518-4 small SmaI-SacI
FT                   2. pGSR1518 remove SmaI-SacI, with BamHI
FT                   -> pGSR1518-4
FT                   1. pGSR1518-4 remove small BamHI-NruI
FT                   T4, fill in
FT                   -> pGSR1518-4d 9700bp"
FT   CAAT_signal     113..116
FT                   /note="PRO E. coli gshI gene"
FT   CAAT_signal     143..147
FT                   /note="PRO E. coli gshI gene"
FT   TATA_signal     226..232
FT                   /note="PRO E. coli gshI gene"
FT   TATA_signal     366..370
FT                   /note="PRO E. coli gshI gene"
FT   misc_feature    374..374
FT                   /note="fused gshI mRNA start site (alt.)"
FT   misc_feature    384..384
FT                   /note="fused gshI mRNA start site (alt.)"
FT   misc_feature    444..444
FT                   /note="fused gshI mRNA start site (alt.)"
FT   misc_feature    1851..1858
FT                   /note="SIT unique XhoI; linker"
FT   CDS             391..>1890
FT                   /note="GEN E. coli fused GSH-I;
FT                   NCBI gi: 220958"
XX
SQ   Sequence 1890 BP; 575 A; 400 C; 374 G; 541 T; 0 other;
     ggatccgaac gtgcaaaagg ccaatatttt tcgctagagt catcgcagtc atggcagctc
     tttcgctcta tctcccggtc gcaaaactgt ggtagtcata gctcgttctg ctcaattgag
     aactgtgaat gtgaatatgg aacaaatgcg atagatgcac taatttaagg gaagctagct
     agttttccca actgcgaaag aaaaaaagga aagaaaaaaa aattctatat aagtgataga
     tatttccatc tttactagca ttagtttctc ttttacgtat tcaatatttt tgttaaactc
     ttcctttatc ataaaaaagc aagcatctaa gagcattgac aacactctaa gaaacaaaat
     accaatataa tttcaaagta catatcaaaa atggttcaat ctgctgtctt agggttccca
     agaatcggtc caaacagaga attaaagaag gccactgaag gttactggaa cggtaaaatc
     actgtcgatg aattattcaa ggtcggtaag gatttgagaa ctcaaaactg gaagttgcaa
     aaggaggctg gtgttgatat catcccatcc aatgacttct ccttttacga ccaagttttg
     gattttgtct ttgttgttca atgtcattca gaccgttaca ctaagtacga tctatctcca
     atcgacactt tgtttgctat gggtagaggt ttacaaagaa aggccactga aactgaaaag
     gctgtcgacg tcactgcttt ggaaatggtt aaatggttcg actctaacta ccattacgtt
     agaccaactt tctccaagac cactcaattt aagttgaacg gccaaaagcc agttgacgaa
     tttttggaag ccaaggagtt aggtattcat actagacctg tcttgttggg tccagtttct
     tacttattct tgggtaaggc tgacaaggat tctctagatt tggaaccatt gtccctattg
     gaacaattgt tgcctctata cactgaaatc ctatctaaat tggcttctgc tggtgccact
     gaagttcaaa ttgacgaacc tgtcttagtt ttggacttgc ctgccaacgc ccaagccgcc
     attaagaagg cttacactta cttcggtgaa caaagcaatc taccaaagat tactttggct
     acttacttcg gtaccgttgt ccctaactta gacgccatca agggcttgcc agttgctgcc
     ttacacgttg actttgttag agctccagaa caatttgatg aagtcgttgc cgccattggt
     aacaaacaaa ccttgtccgt tggtattgtt gatggtagaa acatttggaa gaatgatttc
     aagaagtctt ccgctatcgt taacaaggct attgaaaagt tgggtgctga cagagtcgtt
     gttgccactt cttcttctct attgcacaca ccagttgatt tgaacaacga aaccaagttg
     gacgctgaaa tcaagggctt tttctctttc gccactcaaa aattggatga agttgttgtg
     atcaccaaga acgtttccgg tcaagacgtt gctgctgccc tagaagctaa cgctaaatct
     gttgaatcca gaggtaaatc caagtttatc cacgatgctg ccgttaaggc cagagttgcc
     tctatcgacg aaaaaatgtc tactagagca gctccatttg aacaaagatt gcctgaacaa
     caaaaagtct tcaacttgcc attgttccca acaacaacta ttggttcctt ccctcaaacc
     aaggacatca gaattaacag aaacaaattc aacaagggca ccatctctgc tgaagaatat
     gaaaaattca tcaattctga aattgaaaag gtcatcagat tccaagaaga cctcgaggtg
     atcccggacg tatcacaggc gctggcctgg
//