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ID PK18 preliminary; circular DNA; SYN; 2661 BP.
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AC M17626;
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DT 15-MAR-1989 (Rel. 4, Created)
DT 01-JUL-1995 (Rel. 12, Last updated, Version 1)
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DE E. coli plasmid vector pK18 - 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]
RP 1-2661
RC pK18 from pUC18 & pBR-neo
RC pK19 from pUC19 & pBR-neo
RA Pridmore R.D.;
RT "New and versatile cloning vectors with kanamycin-resistance marker";
RL Gene 56:309-312(1987).
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CC NM (pK18)
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 (pBRNeo)(pUC18)
CC BR ()
CC OF ()
CC OR ()
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FH Key Location/Qualifiers
FH
FT misc_feature 0..0
FT /note="1. pBR322 4361bp
FT transposition
FT 2. Tn5 5818bp
FT -> pRZ102 10179bp
FT 1. pRZ102 remove pBR322 sequences
FT -> pRZ112
FT 1. pRZ112 HincII-HincII 2499bp 188..2687, neo gene
FT BamHI linker 10bp nnggatccnn:BamHI linker 10bp
FT \ nnggatccnn
FT HindIII-BamHI 1493bp 1196..2689, neo/kan gene
FT 2. pBR322 remove HindIII-BamHI 346bp 30..376, 4015bp
FT -> pBR-neo 5508bp
FT 1. pUC18 NarI-DraI/AhaIII 1329bp 237..1566,
FT \ ori/lacZ
FT 2. pBR-neo ClaI-SmaI 1324bp 25..30/1774..3093, neo/kan
FT \ pBR322 ClaI 25/pSV2-neo SmaI 1774
FT -> plasmid 2614bp
FT 1. plasmid HindIII 2614bp, kan gene
FT Klenow, [makes NheI]
FT PstI 2614bp, kan gene
FT mutagenesis 27bp ccctgaatgaactccaagacgaggcag
FT SphI 2641bp, kan gene
FT mutagenesis 20bp caaggcgcggatgccccgac
FT -> pK18 2661bp"
FT misc_binding 0..0
FT /note="SIT NheI"
FT misc_binding 0..0
FT /note="SIT BglII"
FT CDS 365..1159
FT /note="ANT E. coli neomycin phosphotransferase gene
FT (NPT), neomycin resistance gene (neo),
FT Tn5 kanamycin resistance gene (kan)"
FT misc_binding 0..0
FT /note="SIT NcoI"
FT misc_binding 0..0
FT /note="SIT AsuII"
FT misc_recomb 1332..1333
FT /note="pBRNeo DNA end/pUC18 DNA start"
FT rep_origin 0..0
FT /note="ORI E. coli pMB1 (ColE1 and pBR322)"
FT CDS 2400..2661
FT /note="GEN E. coli lacZ gene"
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SQ Sequence 2661 BP; 584 A; 724 C; 769 G; 584 T; 0 other;
cgataagcta gcttcacgct gccgcaagca ctcagggcgc aagggctgct aaaggaagcg
gaacacgtag aaagccagtc cgcagaaacg gtgctgaccc cggatgaatg tcagctactg
ggctatctgg acaagggaaa acgcaagcgc aaagagaaag caggtagctt gcagtgggct
tacatggcga tagctagact gggcggtttt atggacagca agcgaaccgg aattgccagc
tggggcgccc tctggtaagg ttgggaagcc ctgcaaagta aactggatgg ctttcttgcc
gccaaggatc tgatggcgca ggggatcaag atctgatcaa gagacaggat gaggatcgtt
tcgcatgatt gaacaagatg gattgcacgc aggttctccg gccgcttggg tggagaggct
attcggctat gactgggcac aacagacaat cggctgctct gatgccgccg tgttccggct
gtcagcgcag gggcgcccgg ttctttttgt caagaccgac ctgtccggtg ccctgaatga
actccaagac gaggcagcgc ggctatcgtg gctggccacg acgggcgttc cttgcgcagc
tgtgctcgac gttgtcactg aagcgggaag ggactggctg ctattgggcg aagtgccggg
gcaggatctc ctgtcatctc accttgctcc tgccgagaaa gtatccatca tggctgatgc
aatgcggcgg ctgcatacgc ttgatccggc tacctgccca ttcgaccacc aagcgaaaca
tcgcatcgag cgagcacgta ctcggatgga agccggtctt gtcgatcagg atgatctgga
cgaagagcat caggggctcg cgccagccga actgttcgcc aggctcaagg cgcggatgcc
cgacggcgag gatctcgtcg tgacccatgg cgatgcctgc ttgccgaata tcatggtgga
aaatggccgc ttttctggat tcatcgactg tggccggctg ggtgtggcgg accgctatca
ggacatagcg ttggctaccc gtgatattgc tgaagagctt ggcggcgaat gggctgaccg
cttcctcgtg ctttacggta tcgccgctcc cgattcgcag cgcatcgcct tctatcgcct
tcttgacgag ttcttctgag cgggactctg gggttcgaaa tgaccgacca agcgacgccc
aacctgccat cacgagattt cgattccacc gccgccttct atgaaaggtt gggcttcgga
atcgttttcc gggacgccgg ctggatgatc ctccagcgcg gggatctcat gctggagttc
ttcgcccacc ccaaaaggat ctaggtgaag atcctttttg ataatctcat gaccaaaatc
ccttaacgtg agttttcgtt ccactgagcg tcagaccccg tagaaaagat caaaggatct
tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta
ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc tttttccgaa ggtaactggc
ttcagcagag cgcagatacc aaatactgtc cttctagtgt agccgtagtt aggccaccac
ttcaagaact ctgtagcacc gcctacatac ctcgctctgc taatcctgtt accagtggct
gctgccagtg gcgataagtc gtgtcttacc gggttggact caagacgata gttaccggat
aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac agcccagctt ggagcgaacg
acctacaccg aactgagata cctacagcgt gagcattgag 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 acgaattcga gctcggtacc
cggggatcct ctagagtcga cctgcaggca tgcaagcttg gcactggccg tcgttttaca
acgtcgtgac tgggaaaacc ctggcgttac ccaacttaat cgccttgcag cacatccccc
tttcgccagc tggcgtaata gcgaagaggc ccgcaccgat cgcccttccc aacagttgcg
cagcctgaat ggcgaatggc g
//