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ID   PLR60      preliminary; circular DNA; SYN; 3694 BP.
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AC   K01486; ATCC77335;
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DT   03-FEB-1994 (Rel. 8, Created)
DT   01-JUL-1995 (Rel. 12, Last updated, Version 1)
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DE   E. coli plasmid vector pLR60 - complete.
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KW   cloning vector.
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OS   Cloning vector
OC   Artificial sequences; Cloning vehicles.
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RN   [1]
RC   pLR310 from pBluescript SK- & pLK8, GAL4
RC   pLR60 from pLR310
RA   Raycroft L., Lozano G.;
RT   "A convenient cloning vector containing the GAL4 DNA-binding
RT   domain";
RL   Gene 118:143-144(1992).
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RN   [2]
RP   1-3694
RC   from pG525
RA   Laughon A.S., Gesteland R.F.;
RT   "Primary structure of the Saccharomyces cerevisiae GAL4 gene";
RL   Mol. Cell. Biol. 4:260-267(1984).
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RN   [3]
RC   pLK8 from GAL4 gene
RA   Silver P.;
RT   ;
RL   Unpublished (1992).
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RN   [4]
RC   YEpG202, YEpG501 from YEp13 & GAL4 gene
RC   YEpG213, YEpG215 from YEp13 & YEpG202
RC   YEpG515, YEpG525, YEpG528 from YEp13 & YEpG501
RC   pG525 from YEpG525
RC   pG4dh from pG525
RC   YEpG4dP from YEpG525
RC   YEpG4dH from pG4dh & YEp13
RA   Laughon A.S., Gesteland R.F.;
RT   "Isolation and preliminary characterization of the GAL4 gene, a
RT   positive regulator of transcription in yeast";
RL   Proc. Natl. Acad. Sci. U.S.A. 79:6827-6831(1982).
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CC   ATCC length is 3350 bp.
CC   Deposited by: Guillermina Lozano
CC   Convenient cloning vector allowing generation of a fusion protein with
CC   the DNA-binding domain of GAL4 (aa 3-147). [1]
CC   Multiple cloning sites present at both ends of the GAL4 segment permit
CC   in-frame fusions with the DNA-binding domain in the middle or at the
CC   C-terminus of the fusion product. [1]
CC   The order of the major features of this plasmid is:  pMB1 ori - lacI
CC   (non-functional) - lacZ'/ - T3 promoter - SacI/MCS/SpeI - GAL4 -
CC   ClaI/MCS/KpnI - T7 promoter - /lacZ' - f1 ori - ampR. [1]
CC   Transcription from the T3 promoter gives the sense strand. [1]
CC   pBR322 was modified by deletion of bp 75-2352, destruction (by filling
CC   in) of the HindIII site, deletion of a 100 bp AatII/ClaI fragment
CC   (nt 4285 and 22 of pBR322), and inserting an XbaI linker into the
CC   latter deletion junction. [2]
CC   Restriction digests of the clone give the following sizes (kb):
CC   SalI--3.35; SacI/KpnI--2.8, 0.5; XhoI--3.1, 0.25. (ATCC staff)
CC   Medium is 1227 LB plus ampicillin.
CC
CC   GAL4, which is expressed at low levels, encodes a positive
CC   regulator of the galactose inducible genes GAL1, GAL2, GAL7, GAL10
CC   and MEL1. GAL4 and the genes it regulates are not linked.
CC   Regulation takes place at the transcriptional level.
CC   [1] presents data suggesting GAL4 may encode two overlapping
CC   proteins. However, the second smaller protein has not yet been
CC   identified in vivo.
CC   NCBI gi: 171557
CC   NM (pLR60)
CC   CM (yes)
CC   NA (ds-DNA)
CC   TP (circular)
CC   ST ()
CC   TY (plasmid)
CC   SP (ATCC)
CC   HO (E.coli)(E.coli HB101)
CC   CP ()
CC   FN (cloning)
CC   SE ()
CC   PA (pG525)
CC   BR ()
CC   OF ()
CC   OR (DpnI recognition site)
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FH   Key             Location/Qualifiers
FH
FT   misc_feature    0..0
FT                   /note="1. yeast, GAL4 gene
FT                   -> pLK8
FT                   1. pBluescript SK- remove ClaI-SmaI 31bp 685..716,
FT                   \ MCS/2933bp
FT                   2. pLK8 AccI-ClaI 761bp, 55bp 5' to GAL4 gene start
FT                   \ codon to AA 147 of GAL4 gene
FT                   BAL31 nuclease:
FT                   Klenow:
FT                   -> pLR310 3694bp [GAL4 N-terminal]
FT                   1. pLR310 BamHI 3694bp 720..720
FT                   Klenow
FT                   -> pLR60 3694bp"
FT   misc_binding    0..0
FT                   /note="MCS SacI-SacII-NotI-XbaI-SpeI-...-
FT                   ClaI-HincII-AccI-SalI-XhoI-ApaI-DraII-KpnI"
FT   misc_binding    0..0
FT                   /note="SIT unique SacI-SacII-NotI-XbaI-SpeI-...-
FT                   ClaI-HincII-AccI-SalI-ApaI-DraII-KpnI"
FT   rep_origin      0..0
FT                   /note="ORI E. coli pMB1 (ColE1 and pBR322)"
FT   promoter        0..0
FT                   /note="PRO bacteriophage T3"
FT   promoter        0..0
FT                   /note="PRO bacteriophage T7"
FT   CDS             0..0
FT                   /note="ANT E. coli beta-lactamase gene (bla)
FT                   ampicillin resistance gene (apr/amp)"
FT   CDS             443..3088
FT                   /note="ANT yeast GAL4 protein gene; NCBI gi: 171558"
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SQ   Sequence 3694 BP; 1166 A; 796 C; 677 G; 1055 T; 0 other;
     gatcccttaa gtttaaacaa caacagcaag caggtgtgca agacactaga gactcctaac
     atgatgtatg ccaataaaac acaagagata aacaacattg catggaggcc ccagaggggc
     gattggtttg ggtgcgtgag cggcaagaag tttcaaaacg tccgcgtcct ttgagacagc
     attcgcccag tatttttttt attctacaaa ccttctataa tttcaaagta tttacataat
     tctgtatcag tttaatcacc ataatatcgt tttctttgtt tagtgcaatt aatttttcct
     attgttactt cgggcctttt tctgttttat gagctatttt ttccgtcatc cttccccaga
     ttttcagctt catctccaga ttgtgtctac gtaatgcacg ccatcatttt aagagaggac
     agagaagcaa gcctcctgaa agatgaagct actgtcttct atcgaacaag catgcgatat
     ttgccgactt aaaaagctca agtgctccaa agaaaaaccg aagtgcgcca agtgtctgaa
     gaacaactgg gagtgtcgct actctcccaa aaccaaaagg tctccgctga ctagggcaca
     tctgacagaa gtggaatcaa ggctagaaag actggaacag ctatttctac tgatttttcc
     tcgagaagac cttgacatga ttttgaaaat ggattcttta caggatataa aagcattgtt
     aacaggatta tttgtacaag ataatgtgaa taaagatgcc gtcacagata gattggcttc
     agtggagact gatatgcctc taacattgag acagcataga ataagtgcga catcatcatc
     ggaagagagt agtaacaaag gtcaaagaca gttgactgta tcgattgact cggcagctca
     tcatgataac tccacaattc cgttggattt tatgcccagg gatgctcttc atggatttga
     ttggtctgaa gaggatgaca tgtcggatgg cttgcccttc ctgaaaacgg accccaacaa
     taatgggttc tttggcgacg gttctctctt atgtattctt cgatctattg gctttaaacc
     ggaaaattac acgaactcta acgttaacag gctcccgacc atgattacgg atagatacac
     gttggcttct agatccacaa catcccgttt acttcaaagt tatctcaata attttcaccc
     ctactgccct atcgtgcact caccgacgct aatgatgttg tataataacc agattgaaat
     cgcgtcgaag gatcaatggc aaatcctttt taactgcata ttagccattg gagcctggtg
     tatagagggg gaatctactg atatagatgt tttttactat caaaatgcta aatctcattt
     gacgagcaag gtcttcgagt caggttccat aattttggtg acagccctac atcttctgtc
     gcgatataca cagtggaggc agaaaacaaa tactagctat aattttcaca gcttttccat
     aagaatggcc atatcattgg gcttgaatag ggacctcccc tcgtccttca gtgatagcag
     cattctggaa caaagacgcc gaatttggtg gtctgtctac tcttgggaga tccaattgtc
     cctgctttat ggtcgatcca tccagctttc tcagaataca atctccttcc cttcttctgt
     cgacgatgtg cagcgtacca caacaggtcc caccatatat catggcatca ttgaaacagc
     aaggctctta caagttttca caaaaatcta tgaactagac aaaacagtaa ctgcagaaaa
     aagtcctata tgtgcaaaaa aatgcttgat gatttgtaat gagattgagg aggtttcgag
     acaggcacca aagtttttac aaatggatat ttccaccacc gctctaacca atttgttgaa
     ggaacaccct tggctatcct ttacaagatt cgaactgaag tggaaacagt tgtctcttat
     catttatgta ttaagagatt ttttcactaa ttttacccag aaaaagtcac aactagaaca
     ggatcaaaat gatcatcaaa gttatgaagt taaacgatgc tccatcatgt taagcgatgc
     agcacaaaga actgttatgt ctgtaagtag ctatatggac aatcataatg tcaccccata
     ttttgcctgg aattgttctt attacttgtt caatgcagtc ctagtaccca taaagactct
     actctcaaac tcaaaatcga atgctgagaa taacgagacc gcacaattat tacaacaaat
     taacactgtt ctgatgctat taaaaaaact ggccactttt aaaatccaga cttgtgaaaa
     atacattcaa gtactggaag aggtatgtgc gccgtttctg ttatcacagt gtgcaatccc
     attaccgcat atcagttata acaatagtaa tggtagcgcc attaaaaata ttgtcggttc
     tgcaactatc gcccaatacc ctactcttcc ggaggaaaat gtcaacaata tcagtgttaa
     atatgtttct cctggctcag tagggccttc acctgtgcca ttgaaatcag gagcaagttt
     cagtgatcta gtcaagctgt tatctaaccg tccaccctct cgtaactctc cagtgacaat
     accaagaagc acaccttcgc atcgctcagt cacgcctttt ctagggcaac agcaacagct
     gcaatcatta gtgccactga ccccgtctgc tttgtttggt ggcgccaatt ttaatcaaag
     tgggaatatt gctgatagct cattgtcctt cactttcact aacagtagca acggtccgaa
     cctcataaca actcaaacaa attctcaagc gctttcacaa ccaattgcct cctctaacgt
     tcatgataac ttcatgaata atgaaatcac ggctagtaaa attgatgatg gtaataattc
     aaaaccactg tcacctggtt ggacggacca aactgcgtat aacgcgtttg gaatcactac
     agggatgttt aataccacta caatggatga tgtatataac tatctattcg atgatgaaga
     taccccacca aacccaaaaa aagagtaaaa tgaatcgtag atactgaaaa accccgcaag
     ttcacttcaa ctgtgcatcg tgcaccatct caatttcttt catttataca tcgttttgcc
     ttcttttatg taactatact cctctaagtt tcaatcttgg ccatgtaacc tctgatctat
     agaatttttt aaatgactag aattaatgcc catctttttt ttggacctaa attcttcatg
     aaaatatatt acgagggctt attcagaagc ttcgctcata taacgaaaaa aaagggtttg
     gatcgaacgt aattgagatt gattagttaa tactcaaaat aaaacagctc ctaccaccag
     tgtaaagtag aacgttaata gagcaatgtc ttcagacaaa tctattgaga aaaatacaga
     tacgatcgcc tctgaagttc acgaaggtga taatcattcg aataatttgg gttcaatgga
     ggaagagata aaatcaacgc catcagacca atatgaagag atagctataa ttccaactga
     gcccctccat tcggacaaag aactaaatga caagcaacaa agtttaggcc atgaagcacc
     cacaaatgta tcaagagaag aacctattgg gatc
//