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ID   PUC18CMR   preliminary; circular DNA; SYN; 4276 BP.
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AC   ATCC37718;
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DT   01-JUL-1993 (Rel. 7, Created)
DT   01-JUL-1995 (Rel. 12, Last updated, Version 1)
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DE   E. coli plasmid vector pUC18CMR - complete, MCS.
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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   pUC4CM from pUC4K & pBR325, E. coli cat gene
RC   M13mp18CML, M13mp18CMR from M13mp18 & pUC4CM, E. coli cat gene
RC   pUC18CML from pUC18 & M13mp18CML, E. coli cat gene
RC   pUC18CMR from pUC18 & M13mp18CMR, E. coli cat gene
RA   Schweizer H.P.;
RT   "The pUC18CM plasmids: a chloramphenicol resistance gene cassette
RT   for site-directed insertion and deletion mutagenesis in
RT   Escherichia coli";
RL   Biotechniques 8:612-616(1990).
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CC   Created by Moore, July 1995, under contract with NCBI.
CC   cmlR sequences (pBR325 cut with BclI/blunt ended with Klenow/cut with
CC   HincII) were ligated to a 2.6 kb HincII fragment of pUC4K. The 1.6 kb
CC   BamHI fragment was cloned into M13mp18;a 1.6 kb KpnI/HindIII fragment
CC   was cloned into the BamHI site of pUC18.
CC   pUC18CML (ATCC 37717) and pUC18CMR (ATCC 37718) differ only in the
CC   orientation of the cml cassette. In pUC18CMR, the lac promoter and the
CC   cml gene are in the same orientation.
CC   The cml cassette can be isolated by digestion with one enzyme (BamHI),
CC   by double digestion yielding blunt ends (HincII/SmaI), or by double
CC   digestion yielding overhangs for directional cloning.
CC   A plasmid containing the gene determining chloramphenicol resistance
CC   which is flanked by polylinker sites for ease in manipulation.
CC   Restriction digests of the clone give the following sizes (kb):
CC   EcoRI--3.4, 0.89; PstI--4.27; EcoRI/HindIII--2.45, 0.89, 0.75. (ATCC
CC   staff)
CC   Medium is 1235 LB plus tetracycline plus ampicillin.
CC   NM (pUC18CMR)
CC   CM (yes)
CC   NA (ds-DNA)
CC   TP (circular)
CC   ST ()
CC   TY (plasmid)
CC   SP (ATCC)
CC   HO (E.coli XL-1)(E.coli)
CC   CP ()
CC   FN (cloning)
CC   SE ()
CC   PA (pBR325)(pUC18)(pUC4K)(M13mp18
CC   BR (pUC18CML)
CC   OF ()
CC   OR ()
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FH   Key             Location/Qualifiers
FH
FT   misc_feature    0..0
FT                   /note="1. pUC4K HincII-HincII 2662bp 1666..3914..414
FT                   2. pBR325 HincII-BclI 1574bp 3910..5484, E. coli cat
FT                   :Klenow
FT                   -> pUC4CM 4236bp [no HincII sites]
FT                   1. M13mp18 BamHI 7249bp 6252..6252
FT                   2. pUC4CM BamHI-BamHI 1586bp 406..414..-..1666..1670,
FT                   \ E. coli cat gene
FT                   -> M13mp18CMR 8835bp [cat gene]
FT                   1. pUC18 BamHI 2686bp 430..430
FT                   2. M13mp18CMR BamHI-BamHI 1586bp, E. coli cat gene
FT                   -> pUC18CMR 4273bp [cat gene]"
FT   -               1..429
FT                   /note="pUC18 1..429 429bp
FT                   BamHI = G^GATCC"
FT   -               430..437
FT                   /note="pUC4K 406..413 8bp
FT                   HindII = HincII = GTY^RAC"
FT   -               438..2015
FT                   /note="pBR325 3910..5487 1578bp
FT                   BclI =         T^GATC A
FT                   HindII = HincII = GTY^RAC"
FT   -               2016..2019
FT                   /note="pUC4K 1666..1669 4bp
FT                   BamHI = G^GATCC"
FT   -               2020..4276
FT                   /note="pUC18 430..2686 2257bp"
FT   misc_binding    0..0
FT                   /note="MCS unique EcoRI-SstI-KpnI-SmaI-BamHI"
FT   CDS             0..0
FT                   /note="ANT E. coli chloramphenicol acetyltransferase
FT                   gene (cat); chloramphenicol resistance gene (cmr/cml)"
FT   misc_binding    0..0
FT                   /note="SIT EcoRI"
FT   misc_binding    0..0
FT                   /note="MCS unique BamHI-XbaI-SalI-PstI-SphI-HindIII"
FT   rep_origin      0..0
FT                   /note="ORI E. coli pMB1 (ColE1 and pBR322)"
FT   CDS             0..0
FT                   /note="ANT E. coli beta-lactamase gene (bla)
FT                   ampicillin resistance gene (apr/amp)"
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SQ   Sequence 4276 BP; 1116 A; 1019 C; 1044 G; 1097 T; 0 other;
     tcgcgcgttt cggtgatgac ggtgaaaacc tctgacacat gcagctcccg gagacggtca
     cagcttgtct gtaagcggat gccgggagca gacaagcccg tcagggcgcg tcagcgggtg
     ttggcgggtg tcggggctgg cttaactatg cggcatcaga gcagattgta ctgagagtgc
     accatatgcg gtgtgaaata ccgcacagat gcgtaaggag aaaataccgc atcaggcgcc
     attcgccatt caggctgcgc aactgttggg aagggcgatc ggtgcgggcc tcttcgctat
     tacgccagct ggcgaaaggg ggatgtgctg caaggcgatt aagttgggta acgccagggt
     tttcccagtc acgacgttgt aaaacgacgg ccagtgccaa gcttgcatgc ctgcaggtcg
     actctagagg atccgtcaac acgggataat accgcgccac atagcagaac tttaaaagtg
     ctcatcattg gaaaacgttc ttcggggcga aaactctcaa ggatcttacc gctgttgaga
     tccagttcga tgtaacccac tcgtgcaccc aactgatctt cagcatcttt tactttcacc
     agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg aataagggcg
     acacggaaat gttgaatact catactcttc ctttttcaat attattgaag catttatcag
     ggttattgtc tcatgagcgg atacatattt gaatgtattt agaaaaataa acaaataggg
     gttccgcgca catttccccg aaaagtgcca cctgacgtct aagaaaccat tattatcatg
     acattaacct ataaaaatag gcgtatcacg aggccctttc gtcttcgaat aaatacctgt
     gacggaagat cacttcgcag aataaataaa tcctggtgtc cctgttgata ccgggaagcc
     ctgggccaac ttttggcgaa aatgagacgt tgatcggcac gtaagaggtt ccaactttca
     ccataatgaa ataagatcac taccgggcgt attttttgag ttatcgagat tttcaggagc
     taaggaagct aaaatggaga aaaaaatcac tggatatacc accgttgata tatcccaatg
     gcatcgtaaa gaacattttg aggcatttca gtcagttgct caatgtacct ataaccagac
     cgttcagctg gatattacgg cctttttaaa gaccgtaaag aaaaataagc acaagtttta
     tccggccttt attcacattc ttgcccgcct gatgaatgct catccggaat tccgtatggc
     aatgaaagac ggtgagctgg tgatatggga tagtgttcac ccttgttaca ccgttttcca
     tgagcaaact gaaacgtttt catcgctctg gagtgaatac cacgacgatt tccggcagtt
     tctacacata tattcgcaag atgtggcgtg ttacggtgaa aacctggcct atttccctaa
     agggtttatt gagaatatgt ttttcgtctc agccaatccc tgggtgagtt tcaccagttt
     tgatttaaac gtggccaata tggacaactt cttcgccccc gttttcacca tgggcaaata
     ttatacgcaa ggcgacaagg tgctgatgcc gctggcgatt caggttcatc atgccgtttg
     tgatggcttc catgtcggca gaatgcttaa tgaattacaa cagtactgcg atgagtggca
     gggcggggcg taattttttt aaggcagtta ttggtgccct taaacgcctg gtgctacgcc
     tgaataagtg ataataagcg gatgaatggc agaaattcga aagcaaattc gacccggtcg
     tcggttcagg gcagggtcgt taaatagccg cttatgtcta ttgctggttt accggtttat
     tgactaccgg aagcagtgtg accgtgtgct tctcaaatgc ctgaggccag tttgctcagg
     ctctccccgt ggaggtaata attgacgata tgatcgacgg atccccgggt accgagctcg
     aattcgtaat catggtcata gctgtttcct gtgtgaaatt gttatccgct cacaattcca
     cacaacatac gagccggaag cataaagtgt aaagcctggg gtgcctaatg agtgagctaa
     ctcacattaa ttgcgttgcg ctcactgccc gctttccagt cgggaaacct gtcgtgccag
     ctgcattaat gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg gcgctcttcc
     gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct
     cactcaaagg cggtaatacg gttatccaca gaatcagggg ataacgcagg aaagaacatg
     tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc
     cataggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagtca gaggtggcga
     aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct cgtgcgctct
     cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc gggaagcgtg
     gcgctttctc aaagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag
     ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc cggtaactat
     cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc cactggtaac
     aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg gtggcctaac
     tacggctaca ctagaagaac agtatttggt atctgcgctc tgctgaagcc agttaccttc
     ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag cggtggtttt
     tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc
     ttttctacgg ggtctgacgc tcagtggaac gaaaactcac gttaagggat tttggtcatg
     agattatcaa aaaggatctt cacctagatc cttttaaatt aaaaatgaag ttttaaatca
     atctaaagta tatatgagta aacttggtct gacagttacc aatgcttaat cagtgaggca
     cctatctcag cgatctgtct atttcgttca tccatagttg cctgactccc cgtcgtgtag
     ataactacga tacgggaggg cttaccatct ggccccagtg ctgcaatgat accgcgagac
     ccacgctcac cggctccaga tttatcagca ataaaccagc cagccggaag ggccgagcgc
     agaagtggtc ctgcaacttt atccgcctcc atccagtcta ttaattgttg ccgggaagct
     agagtaagta gttcgccagt taatagtttg cgcaacgttg ttgccattgc tacaggcatc
     gtggtgtcac gctcgtcgtt tggtatggct tcattcagct ccggttccca acgatcaagg
     cgagttacat gatcccccat gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc
     gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg ttatggcagc actgcataat
     tctcttactg tcatgccatc cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag
     tcattctgag aatagtgtat gcggcgaccg agttgctctt gcccggcgtc aatacgggat
     aataccgcgc cacatagcag aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg
     cgaaaactct caaggatctt accgctgttg agatccagtt cgatgtaacc cactcgtgca
     cccaactgat cttcagcatc ttttactttc accagcgttt ctgggtgagc aaaaacagga
     aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat actcatactc
     ttcctttttc aatattattg aagcatttat cagggttatt gtctcatgag cggatacata
     tttgaatgta tttagaaaaa taaacaaata ggggttccgc gcacatttcc ccgaaaagtg
     ccacctgacg tctaagaaac cattattatc atgacattaa cctataaaaa taggcgtatc
     acgaggccct ttcgtc
//