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ID   PKM1       preliminary; circular DNA; SYN; 4378 BP.
XX
AC   L08921; VB0124; ATCC37285;
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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 pKM1 - 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   pKM-1 from pKO-1
RC   pPCV from pBR322 & linker
RC   pBR322tacI from pHGH207-1 & pHGH107-11 & pBR322
RC   pHGH807tacI from pBR322tacI & pHGH207-1
RC   pHGH907tacII from pBR322tacI & pHGH207-1
RC   pKM-lac from pKM-1 & pHGH107-11
RC   pKM-trp from pKM-1 & pHGH207-1
RC   pKM-tacI from pKM-1 & pHGH807tacI
RC   pKM-tacII from pKM-1 & pHGH907tacII
RA   De Boer H.A., Comstock L.J., Vasser M.;
RT   "The tac promoter: a functional hybrid derived from the trp and lac
RT   promoters";
RL   Proc. Natl. Acad. Sci. U.S.A. 80:21-25(1983).
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RN   [2]
RP   1-4378
RC   pKM-1
RA   Gilbert W.;
RT   "Obtained from VecBase 3.0";
RL   Unpublished (1991).
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CC   Assembled from pKO1 and phage lambda by F. Pfeiffer.
CC   This is a promoter probe vector. The promoterless galK gene is
CC   used as selectable marker. Translationsal stop codons in all reading
CC   frames are present on a 11-bp oligonucleotide before the galKgene.
CC   Also, a polylinker was cloned before this gene. In addition, the
CC   lambda tr1 transcription terminator was inserted BEFORE the galK gene
CC   to allow measurement of very strong promoters due to read-through
CC   transcription.
CC
CC   Promoter-cloning vector for isolating and characterizing very strong
CC   promoters. [1]
CC   This vector was constructed by inserting a rho-dependent terminator
CC   (lambdaTRI) into the SmaI site of pKO1 (ATCC 37126).  This reduces the
CC   number of transcripts reaching the galK coding sequences to 30% or
CC   less & is thus useful for strong promoters. [2]
CC   Medium is 1227 LB plus ampicillin.
CC   NM (pKM1)
CC   CM (yes)
CC   NA (ds-DNA)
CC   TP (circular)
CC   ST ()
CC   TY (plasmid)
CC   SP (ATCC)
CC   HO (E.coli N100)(E.coli)
CC   CP ()
CC   FN (promoter analysis)
CC   SE ()
CC   PA (EcoGalK)(pBR322)(pKO-1)(lambda)
CC   BR ()
CC   OF (pKM2)
CC   OR ()
XX
FH   Key             Location/Qualifiers
FH
FT   misc_feature    0..0
FT                   /note="1. pKO-1 SmaI 3980bp 312..312
FT                   2. lambda SmaI-SmaI 400bp, tR1 rho-dependent term
FT                   -> pKM-1 4378bp"
FT   rep_origin      0..0
FT                   /note="ORI E. coli pMB1 (ColE1 and pBR322)"
FT   misc_feature    1..294
FT                   /note="from lambda"
FT   misc_feature    312..709
FT                   /note="from lambda"
FT   CDS             721..2083
FT                   /note="GEN E. coli galactokinase gene (galK)"
FT   misc_feature    2084..4378
FT                   /note="from pBR322"
FT   CDS             3313..4101
FT                   /note="ANT E. coli beta-lactamase gene (bla)
FT                   ampicillin resistance gene (apr/amp)"
XX
SQ   Sequence 4378 BP; 1122 A; 1111 C; 1120 G; 1025 T; 0 other;
     ttctggcgaa tcctctgacc agccagaaaa cgacctttct gtggtgaaac cggatgctgc
     aattcagagc gccagcaagt gggggacagc agaagacctg accgccgcag agtggatgtt
     tgacatggtg aagactatcg caccatcagc cagaaaaccg aattttgctg ggtgggctaa
     cgatatccgc ctgatgcgtg aacgtgacgg acgtaaccac cgcgacatgt gtgtgctgtt
     ccgctgggca tgccaggaca acttctggtc cggtaacgtg ctgagcccgg ccaagcttac
     tccccatccc cccgaaagat ttttttaact ataaacgctg atggaagcgt ttatgcggaa
     gaggtaaagc ccttcccgag taacaaaaaa acaacagcat aaataacccc gctcttacac
     attccagccc tgaaaaaggg catcaaatta aaccacacct atggtgtatg catttatttg
     catacattca atcaattgtt atctaaggaa atacttacat atggttcgtg caaacaaacg
     caacgaggct ctacgaatcg agagtgcgtt gcttaacaaa atcgcaatgc ttggaactga
     gaagacagcg gaagctgtgg gcgttgataa gtcgcagatc agcaggtgga agagggactg
     gattccaaag ttctcaatgc tgcttgctgt tcttgaatgg ggggtcgttg ggcaataagg
     gctgcacgcg cacttttatc cgcctctgct gcgctccgcc accgtacgta aatttatggt
     tggttatgaa atgctggcag agacccagcg agacctgacc gcagaacagg cagcagagcg
     tttgcgcgca gtcagcgata tccattttcg cgaatccgga gtgtaagaaa tgagtctgaa
     agaaaaaaca caatctctgt ttgccaacgc atttggctac cctgccactc acaccattca
     ggcgcctggc cgcgtgaatt tgattggtga acacaccgac tacaacgacg gtttcgttct
     gccctgcgcg attgattatc aaaccgtgat cagttgtgca ccacgcgatg accgtaaagt
     tcgcgtgatg gcagccgatt atgaaaatca gctcgacgag ttttccctcg atgcgcccat
     tgtcgcacat gaaaactatc aatgggctaa ctacgttcgt ggcgtggtga aacatctgca
     actgcgtaac aacagcttcg gcggcgtgga catggtgatc agcggcaatg tgccgacggg
     tgccgggtta agttcttccg cttcactgga agtcgcggtc ggaaccgtat tgcagcagct
     ttatcatctg ccgctggacg gcgcacaaat cgcgcttaac ggtcaggaag cagaaaacca
     gtttgtaggc tgtaactgcg ggatcatgga tcagctaatt tccgcgctcg gcaagaaaga
     tcatgccttg ctgatcgatt gccgctcact ggggaccaaa gcagtttcca tgcccaaagg
     tgtggctgtc gtcatcatca acagtaactt caaacgtacc ctggttggca gcgaatacaa
     cacccgtcgt gaacagtgcg aaaccggtgc gcgtttcttc cagcagccag ccctgcgtga
     tgtcaccatt gaagagttca acgctgttgc gcatgaactg gacccgatcg tggcaaaacg
     cgtgcgtcat atactgactg aaaacgcccg caccgttgaa gctgccagcg cgctggagca
     aggcgacctg aaacgtatgg gcgagttgat ggcggagtct catgcctcta tgcgcgatga
     tttcgaaatc accgtgccgc aaattgacac tctggtagaa atcgtcaaag ctgtgattgg
     cgacaaaggt ggcgtacgca tgaccggcgg cggatttggc ggctgtatcg tcgcgctgat
     cccggaagag ctggtgcctg ccgcacagca agctgtcgct gaacaatatg aagcaaaaac
     aggtattaaa gagacttttt acgtttgtaa accatcacaa ggagcaggac agtgctgaac
     gaaactcccg cactggcacc cgatggcagc cgtaccgact gttctgcctc gcgcgtttcg
     gtgatgacgg tgaaaacctc tgacacatgc agctcccgga gacggtcaca gcttgtctgt
     aagcggatgc cgggagcaga caagcccgtc agggcgcgtc agcgggtgtt ggcgggtgtc
     ggggcgcagc catgacccag tcacgtagcg atagcggagt gtatactggc ttaactatgc
     ggcatcagag cagattgtac tgagagtgca ccatatgcgg tgtgaaatac cgcacagatg
     cgtaaggaga aaataccgca tcaggcgctc ttccgcttcc tcgctcactg actcgctgcg
     ctcggtcgtt cggctgcggc gagcggtatc agctcactca aaggcggtaa tacggttatc
     cacagaatca ggggataacg caggaaagaa catgtgagca aaaggccagc aaaaggccag
     gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg ctccgccccc ctgacgagca
     tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg acaggactat aaagatacca
     ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg
     atacctgtcc gcctttctcc cttcgggaag cgtggcgctt tctcatagct cacgctgtag
     gtatctcagt tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt
     tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggtaagaca
     cgacttatcg ccactggcag cagccactgg taacaggatt agcagagcga ggtatgtagg
     cggtgctaca gagttcttga agtggtggcc taactacggc tacactagaa ggacagtatt
     tggtatctgc gctctgctga agccagttac cttcggaaaa agagttggta gctcttgatc
     cggcaaacaa accaccgctg gtagcggtgg tttttttgtt tgcaagcagc agattacgcg
     cagaaaaaaa ggatctcaag aagatccttt gatcttttct acggggtctg acgctcagtg
     gaacgaaaac tcacgttaag ggattttggt catgagatta tcaaaaagga tcttcaccta
     gatcctttta aattaaaaat gaagttttaa atcaatctaa agtatatatg agtaaacttg
     gtctgacagt taccaatgct taatcagtga ggcacctatc tcagcgatct gtctatttcg
     ttcatccata gttgcctgac tccccgtcgt gtagataact acgatacggg agggcttacc
     atctggcccc agtgctgcaa tgataccgcg agacccacgc tcaccggctc cagatttatc
     agcaataaac cagccagccg gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc
     ctccatccag tctattaatt gttgccggga agctagagta agtagttcgc cagttaatag
     tttgcgcaac gttgttgcca ttgctgcagg catcgtggtg tcacgctcgt cgtttggtat
     ggcttcattc agctccggtt cccaacgatc aaggcgagtt acatgatccc ccatgttgtg
     caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt
     gttatcactc atggttatgg cagcactgca taattctctt actgtcatgc catccgtaag
     atgcttttct gtgactggtg agtactcaac caagtcattc tgagaatagt gtatgcggcg
     accgagttgc tcttgcccgg cgtcaacacg ggataatacc gcgccacata gcagaacttt
     aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct
     gttgagatcc agttcgatgt aacccactcg tgcacccaac tgatcttcag catcttttac
     tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat
     aagggcgaca cggaaatgtt gaatactcat actcttcctt tttcaatatt attgaagcat
     ttatcagggt tattgtctca tgagcggata catatttgaa tgtatttaga aaaataaaca
     aataggggtt ccgcgcacat ttccccgaaa agtgccacct gacgtctaag aaaccattat
     tatcatgaca ttaacctata aaaataggcg tatcacgagg ccctttcgtc ttcaagaa
//