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ID PDO19 preliminary; circular DNA; SYN; 11581 BP.
XX
AC U03461; ATCC87041;
XX
DT 03-FEB-1994 (Rel. 8, Created)
DT 01-JUL-1995 (Rel. 12, Last updated, Version 1)
XX
DE E. coli cosmid vector pDO19 - complete.
XX
KW cloning vector.
XX
OS Cloning vector
OC Artificial sequences; Cloning vehicles.
XX
RN [1]
RP 1-11581
RC pDO2 from pUC19 & lambda::Tn5 & pNEB193
RC pDO6 from pDO2
RC pDO17 from pDO2 & pBR322
RC pDO19 from pDO17
RC pDO184 from pK184 & pBR322
RA Ahmed A.;
RT "Double origin vectors for isolating bidirectional deletions for DNA
RT sequence analysis";
RL Gene 141:71-73(1994).
XX
RN [2]
RP 1-11581
RC pDO19
RA Ahmed A.;
RT ;
RL Submitted (12-NOV-1993) by:
RL Ahmed A., Genetics, Biological Sciences Bldg Rm G502,
RL University of Alberta, Edmonton, Alberta, T6G 2E1, CANADA.
XX
RN [3]
RC pDO19 from pBR322 & lambda & kan gene
RA Ahmed A.L.;
RT "Isolation of nested, bidirectional deletions by the use of
RT exonuclease III on double-origin vectors";
RL Gene 151:97-101(1994).
XX
CC Restriction digests of the clone give the following sizes (kb):
CC EcoRI--11.5; NdeI--5.5, 4.2, 1.8; PvuII--5.8, 3.15, 0.78 (doublet),
CC 0.66, 0.28; PstI--8.4, 2.1, 0.96. (ATCC staff)
CC Plasmid contains the following restriction sites (approximate kb from
CC nt 1): BamHI--6.86; BglI--6.49, 7.42, 7.65, 9.97, 11.18; EcoRI-3.77;
CC HindIII--2.25; NdeI--2.96, 4.74, 8.79; PstI--0.65, 1.72, 10.10;
CC PvuI--5.50, 10.22; SalI--7.14; SmaI--0.93, 4.50. (GenBank/EMBL/DDBJ)
CC A series of overlapping deletion segments can be generated by
CC partial digestion with a restriction enzyme not found in the vector,
CC followed by complete digestion with lambda terminase.
CC The order of the major features in the plasmid is: ren - P - O -
CC ori lambda - cII - cro - kanR - cosR/L - ampR - oriP - tetR. [1]
CC Cosmid vector containing two compatible origins of replication, for
CC generating bidirectional deletions of large inserts to facilitate
CC sequencing. Resulting fragments are blunted and circularized by
CC ligation, and transformants are selected for either ampicillin or
CC kanamycin resistance. AmpR transformants can be sequenced in the
CC clockwise direction using the cosL primer 5'-TCATAAATAGCGAAAACC-3'.
CC KanR transformants can be sequenced in the counterclockwise direction
CC using the cosR primer 5'-ACTTTACGGGTCCTTTCCG-3'. [2]
CC Growth: 1065 plus ampicillin (50 ug/ml) plus kanamycin (20 ug/ml) 30C
CC Deposited by: Ahmed A.L.
CC NCBI gi: 424109
CC NM (pDO19)
CC CM (yes)
CC NA (ds-DNA)
CC TP (circular)
CC ST ()
CC TY (cosmid)
CC SP (ATCC)
CC HO (E.coli)(E.coli AA102)
CC CP ()
CC FN (cloning)(sequencing)
CC SE (color)
CC PA (pBR322)
CC BR (pDO2)(pDO6)(pDO184)(pDO17)
CC OF ()
CC OR ()
XX
FH Key Location/Qualifiers
FH
FT misc_feature 0..0
FT /note="1. pDO17 remove, 12035bp-
FT -> pDO19 11581bp"
FT rep_origin 0..0
FT /note="ORI E. coli pMB1 (ColE1 and pBR322)"
FT rep_origin 0..0
FT /note="ORI E. coli oriP"
FT rep_origin 0..0
FT /note="ORI E. coli phage lambda cos"
FT CDS 0..0
FT /note="ANT E. coli amp gene"
FT CDS 0..0
FT /note="ANT E. coli kan gene"
FT CDS 0..0
FT /note="ANT E. coli tet gene"
FT misc_binding 0..0
FT /note="MCS AatII-ClaI-NheI-BamHI-SgrAI-SalI"
XX
SQ Sequence 11581 BP; 2863 A; 3081 C; 2985 G; 2652 T; 0 other;
aggtcgccgc cccgtaacct gtcggatcac cggaaaggac ccgtaaagtg ataatgatta
tcatctacat atcacaacgt gcgtggaggc catcaaacca cgtcaaataa tcaattatga
cgcaggtatc gtattaattg atctgcatca acttaacgta aaaacaactt cagacaatac
aaatcagcga cactgaatac ggggcaacct catgtcaacg aagaacagaa cccgcagaac
aacaacccgc aacatccgct ttcctaacca aatgattgaa caaattaaca tcgctcttga
gcaaaaaggg tccgggaatt tctcagcctg ggtcattgaa gcctgccgtc ggagactaac
gtcagaaaag agagcatata catcaattaa aagtgatgaa gaatgaacat cccgcgttct
tccctccgaa caggacgata ttgtaaattc acttaattac gagggcattg cagtaattgg
gtgatgctgc caacttactg atttagtgta tgatggtgtt tttgaggtgc tccagtggct
tctgtttcta tcagctgtcc ctcctgttca gctactgacg gggtggtgcg taacggcaaa
agcaccgccg gacatcagcg ctatctctgc tctcactgcc gtaaaacatg gcaactgcag
ttcacttaca ccgcttctca acccggtacg caccagaaaa tcattgatat ggccatgaat
ggcgttggat gccgggcaac agcccgcatt atgggcgttg gcctcaacac gattttacgt
cacttaaaaa actcaggccg cagtcggtaa ccggcctctt catcgggaat gcgcgcgacc
ttcagcatcg ccggcatgtc cccctggcgg acgggaagta tccagctcga tcaggcagca
gctgaaccaa ctcgcgaggg gatcgagccc ggggtgggcg aagaactcca gcatgagatc
cccgcgctgg aggatcatcc agccggcgtc ccggaaaacg attccgaagc ccaacctttc
atagaaggcg gcggtggaat cgaaatctcg tgatggcagg ttgggcgtcg cttggtcggt
catttcgaac cccagagtcc cgctcagaag aactcgtcaa gaaggcgata gaaggcgatg
cgctgcgaat cgggagcggc gataccgtaa agcacgagga agcggtcagc ccattcgccg
ccaagctctt cagcaatatc acgggtagcc aacgctatgt cctgatagcg gtccgccaca
cccagccggc cacagtcgat gaatccagaa aagcggccat tttccaccat gatattcggc
aagcaggcat cgccatgggt cacgacgaga tcctcgccgt cgggcatgcg cgccttgagc
ctggcgaaca gttcggctgg cgcgagcccc tgatgctctt cgtccagatc atcctgatcg
acaagaccgg cttccatccg agtacgtgct cgctcgatgc gatgtttcgc ttggtggtcg
aatgggcagg tagccggatc aagcgtatgc agccgccgca ttgcatcagc catgatggat
actttctcgg caggagcaag gtgagatgac aggagatcct gccccggcac ttcgcccaat
agcagccagt cccttcccgc ttcagtgaca acgtcgagca cagctgcgca aggaacgccc
gtcgtggcca gccacgatag ccgcgctgcc tcgtcctgca gttcattcag ggcaccggac
aggtcggtct tgacaaaaag aaccgggcgc ccctgcgctg acagccggaa cacggcggca
tcagagcagc cgattgtctg ttgtgcccag tcatagccga atagcctctc cacccaagcg
gccggagaac ctgcgtgcaa tccatcttgt tcaatcatgc gaaacgatcc tcatcctgtc
tcttgatcag atcttgatcc cctgcgccat cagatccttg gcggcaagaa agccatccag
tttactttgc agggcttccc aaccttacca gagggcgccc cagctggcaa ttccggttcg
cttgctgtcc ataaaaccgc ccagtctagc tatcgccatg taagcccact gcaagctacc
tgctttctct ttgcgcttgc gttttccctt gtccagatag cccagtagct gacattcatc
cggggtcagc accgtttctg cggactggct ttctacgtgt tccgcttcct ttagcagccc
ttgcgccctg agtgcttgcg gcagcgtgaa gcttcaggtg agaacatccc tgcctgaaca
tgagaaaaaa cagggtactc atactcactt ctaagtgacg gctgcatact aaccgcttca
tacatctcgt agatttctct ggcgattgaa gggctaaatt cttcaacgct aactttgaga
atttttgcaa gcaatgcggc gttataagca tttaatgcat tgatgccatt aaataaagca
ccaacgcctg actgccccat ccccatcttg tctgcgacag attcctggga taagccaagt
tcatttttct ttttttcata aattgcttta aggcgacgtg cgtcctcaag ctgctcttgt
gttaatggtt tcttttttgt gctcatacgt taaatctatc accgcaaggg ataaatatct
aacaccgtgc gtgttgacta ttttacctct ggcggtgata atggttgcat gtactaagga
ggttgtatgg aacaacgcat aaccctgaaa gattatgcaa tgcgctttgg gcaaaccaag
acagctaaag atctcggcgt atatcaaagc gcgatcaaca aggccattca tgcaggccga
aagatttttt taactataaa cgctgatgga agcgtttatg cggaagaggt aaagcccttc
ccgagtaaca aaaaaacaac agcataaata accccgctct tacacattcc agccctgaaa
attgttatct aaggaaatac ttacatatgg ttcgtgcaaa caaacgcaac gaggctctac
gaatcgagag tgcgttgctt aacaaaatcg caatgcttgg aactgagaag acagcggaag
ctgtgggcgt tgataagtcg cagatcagca ggtggaagag ggactggatt ccaaagttct
caatgctgct tgctgttctt gaatgggggg tcgttgacga cgacatggct cgattggcgc
gacaagttgc tgcgattctc accaataaaa aacgcccggc ggcaaccgag cgttctgaac
aaatccagat ggagttctga ggtcattact ggatctatca acaggagtca ttatgacaaa
tacagcaaaa atactcaact tcggcagagg taactttgcc ggacaggagc gtaatgtggc
agatctcgat gatggttacg ccagactatc aaatatgctg cttgaggctt attcgggcgc
agatctgacc aagcgacagt ttaaagtgct gcttgccatt ctgcgtaaaa cctatgggtg
gaataaacca atggacagaa tcaccgattc tcaacttagc gagattacaa agttacctgt
caaacggtgc aatgaagcca agttagaact cgtcagaatg aatattatca agcagcaagg
cggcatgttt ggaccaaata aaaacatctc agaatggtgc atccctcaaa acgagggaaa
atcccctaaa acgagggata aaacatccct caaattgggg gattgctatc cctcaaaaca
gggggacaca aaagacacta ttacaaaaga aaaaagaaaa gattattcgt cagagaattc
tggcgaatcc tctgaccagc cagaaaacga cctttctgtg gtgaaaccgg atgctgcaat
tcagagcggc agcaagtggg ggacagcaga agacctgacc gccgcagagt ggatgtttga
catggtgaag actatcgcac catcagccag aaaaccgaat tttgctgggt gggctaacga
tatccgcctg atgcgtgaac gtgacggacg taaccaccgc gacatgtgtg tgctgttccg
ctgggcatgc caggacaact tctggtccgg taacgtgctg agcccggcca aactccgcga
taagtggacc caactcgaaa tcaaccgtaa caagcaacag gcaggcgtga cagccagcaa
accaaaactc gacctgacaa acacagactg gatttacggg gtggatctat gaaaaacatc
gccgcacaga tggttaactt tgaccgtgag cagatgcgtc ggatcgccaa caacatgccg
gaacagtacg acgaaaagcc gcaggtacag caggtagcgc agatcatcaa cggtgtgttc
agccagttac tggcaacttt cccggcgagc ctggctaacc gtgaccagaa cgaagtgaac
gaaatccgtc gccagtgggt tctggctttt cgggaaaacg ggatcaccac gatggaacag
gttaacgcag gaatgcgcgt agcccgtcgg cagaatcgac catttctgcc atcacccggg
cagtttgttg catggtgccg ggaagaagca tccgttaccg ccggactgcc aaacgtcagc
gagctggttg atatggttta cgagtattgc cggaagcgag gcctgtatcc ggatgcggag
tcttatccgt ggaaatcaaa cgcgcactac tggctggtta ccaacctgta tcagaacatg
cgggccaatg cgcttactga tgcggaatta cgccgtaagg ccgcagatga gcttgtccat
atgactgcga gaattaaccg tggtgaggcg atccctgaac cagtaaaaca acttcctgtc
atgggcggta gacctctaaa tcgtgcacag gctctggcga agatcgcaga aatcaaagct
aagttcggac tgaaaggagc aagtgtatga cgggcaaaga ggcaattatt cattacctgg
ggacgcataa tagcttctgt gcgccggacg ttgccgcgct aacaggcgca acagtaacca
gcataaatca ggccgcggct aaaatggcac gggcaggtct tctggttatc gaaggtaagg
tctggcgaac ggtgtattac cggtttgcta ccagggaaga acgggaagga aagatgagca
cgaacctggt ttttaaggag tgtcgccaga gtgccgcgat gaaacgggta ttggcggtat
atggagttaa aagatgacca tctacattac tgagctaata acaggcctgc tggtaatcgc
aggccttttt atttggggga gagggaagtc atgaaaaaac taacctttga aattcgatct
ccagcacatc agcaaaacgc tattcacgca gtacagcaaa tccttccaga cccaaccaaa
ccaatcgtag taaccattca ggaacgcaac cgcagcttag accaaaacag gaagctatgg
gcctgcttag gtgacgtcat cgtctgcgcg gaaatggacg aacagtgggg ctatgtcggg
gctaaatcgc gccagcgctg gctgttttac gcgttttgcc acgatcgggt ccagttcatg
cgcaacagcg ttgaactctt caatggtgac atcacgcagg gctggctgct ggaagaaacg
cgcaccggtt tcgcactgtt cacgacgggt gttgtattcg ctgccaacca gggtacgttt
gaagttactg ttgatgatga cgacagccac acctttgggc atggaaactg ctttggtccc
cagtgagcgg caatcgatca gcaaggcatg atctttcttg ccgagcgcgg aaattagctg
atccatgatc ccgcagttac agcctacaaa ctggttttct gcttcctgac cgttaagcgc
gatttgtgcg ccgtccagcg gcagatgata aagctgctgc aatacggttc cgaccgcgac
ttccagtgaa gcggaagaac ttaacccggc accctgcggc acattgccgt gatcaaccat
gtccacgccg ccgaagctgt tgttacgcag ttgcagatgt ttcaccacgc cacgaacgta
gttagcccat tgatagtttt catgtgcgac aatgggcgca tcgagggaaa actcgtcgag
ctgattttca taatcggctg ccatcacgcg aactttacgg tcatcgcgtg gtgcacaact
gatcacggtt tgataatcaa tcgcgcaggg cagaacgaaa ccgtcgttgt agtcggtgtg
ttcaccaatc aaattcacgc ggccaggcgc ctgaatggtg tgagtggcag ggtagccaaa
tgcgttggca aacagagatt gtgttttttc tttcagactc atttcttaca ctccggattc
gcgacggcct acagcaacct gggcgatgta tggcatcagg ttattcggaa tgccttgcgg
atcttcgccc atatcgcccg acggatgcgc gccaaccggg ttgaagtagc gcacgagggc
aatgctccag tccggctggg ctttttgcag atcggtgagg atctgttcca ccatcagctt
gcttttgccg taagggcttt gcggtgtgcc ggtcgggaag ctataatgcg gtagtttatc
acagttaaat tgctaacgca gtcaggcacc gtgtatgaaa tctaacaatg cgctcatcgt
catcctcggc accgtcaccc tggatgctgt aggcataggc ttggttatgc cggtactgcc
gggcctcttg cgggatatcg tccattccga cagcatcgcc agtcactatg gcgtgctgct
agcgctatat gcgttgatgc aatttctatg cgcacccgtt ctcggagcac tgtccgaccg
ctttggccgc cgcccagtcc tgctcgcttc gctacttgga gccactatcg actacgcgat
catggcgacc acacccgtcc tgtggatcct ctacgccgga cgcatcgtgg ccggcatcac
cggcgccaca ggtgcggttg ctggcgccta tatcgccgac atcaccgatg gggaagatcg
ggctcgccac ttcgggctca tgagcgcttg tttcggcgtg ggtatggtgg caggccccgt
ggccggggga ctgttgggcg ccatctcctt gcatgcacca ttccttgcgg cggcggtgct
caacggcctc aacctactac tgggctgctt cctaatgcag gagtcgcata agggagagcg
tcgaccgatg cccttgagag ccttcaaccc agtcagctcc ttccggtggg cgcggggcat
gactatcgtc gccgcactta tgactgtctt ctttatcatg caactcgtag gacaggtgcc
ggcagcgctc tgggtcattt tcggcgagga ccgctttcgc tggagcgcga cgatgatcgg
cctgtcgctt gcggtattcg gaatcttgca cgccctcgct caagccttcg tcactggtcc
cgccaccaaa cgtttcggcg agaagcaggc cattatcgcc ggcatggcgg ccgacgcgct
gggctacgtc ttgctggcgt tcgcgacgcg aggctggatg gccttcccca ttatgattct
tctcgcttcc ggcggcatcg ggatgcccgc gttgcaggcc atgctgtcca ggcaggtaga
tgacgaccat cagggacagc ttcaaggatc gctcgcggct cttaccagcc taacttcgat
cattggaccg ctgatcgtca cggcgattta tgccgcctcg gcgagcacat ggaacgggtt
ggcatggatt gtaggcgccg ccctatacct tgtctgcctc cccgcgttgc gtcgcggtgc
atggagccgg gccacctcga cctgaatgga agccggcggc acctcgctaa cggattcacc
actccaagaa ttggagccaa tcaattcttg cggagaactg tgaatgcgca aaccaaccct
tggcagaaca tatccatcgc gtccgccatc tccagcagcc gcacgcggcg catctcgggc
agcgttgggt cctggccacg ggtgcgcatg atcgtgctcc tgtcgttgag gacccggcta
ggctggcggg gttgccttac tggttagcag aatgaatcac cgatacgcga gcgaacgtga
agcgactgct gctgcaaaac gtctgcgacc tgagcaacaa catgaatggt cttcggtttc
cgtgtttcgt aaagtctgga aacgcggaag tcagcgccct gcaccattat gttccggatc
tgcatcgcag gatgctgctg gctaccctgt ggaacaccta catctgtatt aacgaagcgc
tggcattgac cctgagtgat ttttctctgg tcccgccgca tccataccgc cagttgttta
ccctcacaac gttccagtaa ccgggcatgt tcatcatcag taacccgtat cgtgagcatc
ctctctcgtt tcatcggtat cattaccccc atgaacagaa attccccctt acacggaggc
atcaagtgac caaacaggaa aaaaccgccc ttaacatggc ccgctttatc agaagccaga
cattaacgct tctggagaaa ctcaacgagc tggacgcgga tgaacaggca gacatctgtg
aatcgcttca cgaccacgct gatgagcttt accgcagctg cctcgcgcgt ttcggtgatg
acggtgaaaa cctctgacac atgcagctcc cggagacggt cacagcttgt ctgtaagcgg
atgccgggag cagacaagcc cgtcagggcg cgtcagcggg tgttggcggg tgtcggggcg
cagccatgac ccagtcacgt agcgatagcg gagtgtatac tggcttaact atgcggcatc
agagcagatt gtactgagag tgcaccatat gcggtgtgaa ataccgcaca gatgcgtaag
gagaaaatac cgcatcaggc gctcttccgc ttcctcgctc actgactcgc tgcgctcggt
cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt tatccacaga
atcaggggat aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg
taaaaaggcc gcgttgctgg cgtttttcca taggctccgc ccccctgacg agcatcacaa
aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga ctataaagat accaggcgtt
tccccctgga agctccctcg tgcgctctcc tgttccgacc ctgccgctta ccggatacct
gtccgccttt ctcccttcgg gaagcgtggc gctttctcaa tgctcacgct gtaggtatct
cagttcggtg taggtcgttc gctccaagct gggctgtgtg cacgaacccc ccgttcagcc
cgaccgctgc gccttatccg gtaactatcg tcttgagtcc aacccggtaa gacacgactt
atcgccactg gcagcagcca ctggtaacag gattagcaga gcgaggtatg taggcggtgc
tacagagttc ttgaagtggt ggcctaacta cggctacact agaaggacag tatttggtat
ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa
acaaaccacc gctggtagcg gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa
aaaaggatct caagaagatc ctttgatctt ttctacgggg tctgacgctc agtggaacga
aaactcacgt taagggattt tggtcatgag attatcaaaa aggatcttca cctagatcct
tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa cttggtctga
cagttaccaa tgcttaatca gtgaggcacc tatctcagcg atctgtctat ttcgttcatc
catagttgcc tgactccccg tcgtgtagat aactacgata cgggagggct taccatctgg
ccccagtgct gcaatgatac cgcgagaccc acgctcaccg gctccagatt tatcagcaat
aaaccagcca gccggaaggg ccgagcgcag aagtggtcct gcaactttat ccgcctccat
ccagtctatt aattgttgcc gggaagctag agtaagtagt tcgccagtta atagtttgcg
caacgttgtt gccattgctg caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc
attcagctcc ggttcccaac gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa
agcggttagc tccttcggtc ctccgatcgt tgtcagaagt aagttggccg cagtgttatc
actcatggtt atggcagcac tgcataattc tcttactgtc atgccatccg taagatgctt
ttctgtgact ggtgagtact caaccaagtc attctgagaa tagtgtatgc ggcgaccgag
ttgctcttgc ccggcgtcaa cacgggataa taccgcgcca catagcagaa ctttaaaagt
gctcatcatt ggaaaacgtt cttcggggcg aaaactctca aggatcttac cgctgttgag
atccagttcg atgtaaccca ctcgtgcacc caactgatct tcagcatctt ttactttcac
cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc
gacacggaaa tgttgaatac tcatactctt cctttttcaa tattattgaa gcatttatca
gggttattgt ctcatgagcg gatacatatt tgaatgtatt tagaaaaata aacaaatagg
ggttccgcgc acatttcccc gaaaagtgcg gtaatgactc caacttattg atagtgtttt
atgttcagat aatgcccgat gactttgtca tgcagctcca ccgattttga gaacgacagc
gacttccgtc ccagccgtgc caggtgctgc ctcagattca ggttatgccg ctcaattcgc
tgcgtatatc gcttgctgat tacgtgcagc tttcccttca ggcgtacgca tgaccggcgg
cggatttggc ggctgtatcg tcgcgctgat cccggaagag ctggtgcctg ccgtacagca
acgtgtcgct gaacaatatg aagcaaaaac aggtattaaa gagacttttt acgtttgtaa
accatcacaa ggagcaggac agtgctgaac gaaactcccg cactggcacc cgatggtcag
ccgtaccgac tgttgatctg cctcgctggc ctgccgcagt tcttcaacct cccggcgcag
cttttcgttc tcaatttcag catccctttc ggcataccat tttatgacgg cggcagagtc
ataaagcacc tcattaccct tgccaccgcc tcgcagaacg ggcattccct gttcctgcca
gttctgaatg gtacggatac tcgcaccgaa aatgtcagcc agctgctttt tgttgacttc
cattgttcat tccacggaca aaaacagaga aaggaaacga cagaggccaa aaagcctcgc
tttcagcacc tgtcgtttcc tttcttttca gagggtattt taaataaaaa cattaagtta
tgacgaagaa gaacggaaac gccttaaacc ggaaaatttt cataaatagc gaaaacccgc
g
//