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ID PK194 preliminary; circular DNA; SYN; 2435 BP.
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AC U00800; ATCC37767;
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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 pK194 - 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 pK184 from pK18 & pACYC184
RC pK194 from pK19 & pACYC184
RA Jobling M.G., Holmes R.K.;
RT "Construction of vectors with the p15a replicon, kanamycin
RT resistance, inducible lacZ alpha and pUC18 or pUC19 multiple
RT cloning sites";
RL Nucleic Acids Res. 18:5315-5315(1990).
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CC Created by Moore, July 1995, under contract with NCBI.
CC pK184 (ATCC 37766) and pK194 (ATCC 37767) have the multiple cloning
CC sites in opposite orientations, interrupting the lacZalpha coding
CC sequence.
CC A general purpose plasmid vector for use in complementation analysis
CC with plasmids having a ColE1 or pMB1 origin of replication.
CC Constructed by M. G. Jobling. (personal communication)
CC Restriction digests of the clone give the following sizes (kb):
CC EcoRI--2.4; HindIII--2.4; PstI--2.4; BamHI--2.4; XbaI--2.4. (ATCC
CC staff)
CC Medium is 1236 LB plus kanamycin.
CC NM (pK194)
CC CM (yes)
CC NA (ds-DNA)
CC TP (circular)
CC ST ()
CC TY (plasmid)
CC SP (ATCC)
CC HO (E.coli XL-1-Blue)(E.coli)
CC CP ()
CC FN (cloning)
CC SE ()
CC PA (pK18)(pUC19)(pACYC184)
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, Tn5 neo gene
FT BamHI linker 10bp nnggatccnn:BamHI linker 10bp
FT \ nnggatccnn
FT HindIII-BamHI 1493bp 1196..2689, Tn5 neo/kan gene
FT 2. pBR322 remove HindIII-BamHI 346bp 30..376, 4015bp
FT -> pBR-neo 5508bp
FT 1. pUC19 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 -> pK19 2661bp
FT 1. pK19 remove BstBI/AsuII-NspI/NspHI 916bp
FT \ 1176..2092, pMB1 ori to kan gene/1745bp
FT \ pK18 NspI/NspHI 2092 2493
FT \ pUC18 NspI = 42 410 811
FT \ pUC19 NspI = 42 446 811
FT T4 DNA polymerase:T4 DNA polymerase
FT 2. pACYC184 SacII-ClaI 684bp 836..1520, p15A ori
FT T4 DNA polymerase:T4 DNA polymerase
FT -> pK194 2432bp [like pK184, except pK19]"
FT - 1..1177
FT /note="pK18 1..1177 1177bp
FT AsuII = BstBI = TT^CG AA
FT SacII = CC GC^GG"
FT - 1178..1865
FT /note="pACYC184 834..1521 688bp
FT ClaI = AT^CG AT
FT NspI = R CATG^Y"
FT - 1866..2216
FT /note="pK18 2092..2442 351bp"
FT - 2217..2275
FT /note="pUC19 394..452 59bp complement"
FT - 2276..2435
FT /note="pK18 2502..2661 160bp"
FT misc_binding 0..0
FT /note="MCS HindIII-SphI-PstI-SfeI-HincII-AccI-SalI-
FT XbaI-BamHI-SmaI-AvaI-KpnI-SstI-EcoRI"
FT misc_binding 0..0
FT /note="SIT unique HindIII-SphI-PstI-HincII-SalI-
FT AccI-BamHI-XmaI-SmaI-KpnI-SacI-EcoRI"
FT rep_origin 0..0
FT /note="ORI E. coli miniplasmid p15A"
FT promoter 0..0
FT /note="PRO E. coli lac gene"
FT CDS 0..0
FT /note="GEN E. coli beta-galactosidase gene (lacZ');
FT reporter gene"
FT CDS 0..0
FT /note="ANT E. coli kanamycin resistance gene (kan)"
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SQ Sequence 2435 BP; 545 A; 668 C; 675 G; 547 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 gggttcggcg gcaaagccgt ttttccatag
gctccgcccc cctgacaagc atcacgaaat ctgacgctca aatcagtggt ggcgaaaccc
gacaggacta taaagatacc aggcgtttcc ccctggcggc tccctcgtgc gctctcctgt
tcctgccttt cggtttaccg gtgtcattcc gctgttatgg ccgcgtttgt ctcattccac
gcctgacact cagttccggg taggcagttc gctccaagct ggactgtatg cacgaacccc
ccgttcagtc cgaccgctgc gccttatccg gtaactatcg tcttgagtcc aacccggaaa
gacatgcaaa agcaccactg gcagcagcca ctggtaattg atttagagga gttagtcttg
aagtcatgcg ccggttaagg ctaaactgaa aggacaagtt ttggtgactg cgctcctcca
agccagttac ctcggttcaa agagttggta gctcagagaa ccttcgaaaa accgccctgc
aaggcggttt tttcgttttc agagcaagag attacgcgca gaccaaaacg atctcaagaa
gatcatctta ttaatcagat aaaatatttc tagatttcag tgcaatttat ctcttcaaat
gtagcacctg aagtcagccc catacgatat aagttgtaat tctcatgttt gacagcttat
catcgttctt tcctgcgtta tcccctgatt ctgtggataa ccgtattacc gcctttgagt
gagctgatac cgctcgccgc agccgaacga ccgagcgcag cgagtcagtg agcgaggaag
cggaagagcg cccaatacgc aaaccgcctc tccccgcgcg ttggccgatt cattaatgca
gctggcacga caggtttccc gactggaaag cgggcagtga gcgcaacgca attaatgtga
gttagctcac tcattaggca ccccaggctt tacactttat gcttccggct cgtatgttgt
gtggaattgt gagcggataa caatttcaca caggaaacag ctatgaccat gattacaagc
ttgcatgcct gcaggtcgac tctagaggat ccccgggtac cgagctcgaa ttcaccactg
gccgtcgttt tacaacgtcg tgactgggaa aaccctggcg ttacccaact taatcgcctt
gcagcacatc cccctttcgc cagctggcgt aatagcgaag aggcccgcac cgatcgccct
tcccaacagt tgcgcagcct gaatggcgaa tggcg
//