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ID PDO184 preliminary; circular DNA; SYN; 9820 BP.
XX
AC U03460; ATCC87042;
XX
DT 03-FEB-1994 (Rel. 8, Created)
DT 01-JUL-1995 (Rel. 12, Last updated, Version 1)
XX
DE E. coli cosmid vector pDO184 - complete.
XX
KW cloning vector.
XX
OS Cloning vector
OC Artificial sequences; Cloning vehicles.
XX
RN [1]
RP 1-9820
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 1-9820
RC pDO184
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 pDO184 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--10.0; HindIII--10.0; XbaI--9.2, 1.0; BglI--2.8, 2.35 (doublet),
CC 1.55, 0.75, 0.28; PstI--4.0, 3.7, 2.5. (ATCC staff)
CC Plasmid contains the following restriction sites (approximate kb from
CC nt 1): BamHI--4.42, 4.77; BglI--1.96, 4.60, 5.32, 5.56, 7.88, 9.42;
CC EcoRI--4.40; HindIII--4.45; PstI--0.65, 4.44, 8.00; PvuI--0.97, 4.57,
CC 8.13; SmaI--4.41; XbaI--3.45, 4.42. (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 following
CC restriction sites do not cut pDO17: Acc65I AflII ApaI AscI AvrII
CC Ecl136II KpnI NotI PacI PmeI PmlI SacI SfiI SnaBI SpeI SrfI Sse8387I
CC SwaI XbaI. [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: 424108
CC NM (pDO184)
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)(pDO17)(pDO6)(pDO19)
CC OF ()
CC OR ()
XX
FH Key Location/Qualifiers
FH
FT misc_feature 0..0
FT /note="1. pACYC184, p15A ori/kan gene/MCS/lacZ gene
FT 2. pBR322, ori/amp gene
FT -> pDO184 9820bp"
FT misc_binding 0..0
FT /note="MCS EcoRI-SacI-Ecl136II-KpnI-Acc65I-SmaI-XmaI-
FT Sse83871-HindIII"
FT rep_origin 0..0
FT /note="ORI E. coli p15A"
FT CDS 0..0
FT /note="ANT E. coli amp gene"
FT CDS 0..0
FT /note="ANT E. coli kan gene"
FT rep_origin 0..0
FT /note="ORI E. coli pMB1 (ColE1 and pBR322)"
XX
SQ Sequence 9820 BP; 2277 A; 2646 C; 2615 G; 2282 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 catcgtctgc
gcggaaatgg acgaacagtg gggctatgtc ggggctaaat cgcgccagcg ctggctgttt
tacgcgtttt gccacgatcg ggtccagttc atgcgcaaca gcgttgaact cttcaatggt
gacatcacgc agggctggct gctggaagaa acgcgcaccg gtttcgcact gttcacgacg
ggtgttgtat tcgctgccaa ccagggtacg tttgaagtta ctgttgatga tgacgacagc
cacacctttg ggcatggaaa ctgctttggt ccccagtgag cggcaatcga tcagcaaggc
atgatctttc ttgccgagcg cggaaattag ctgatccatg atcccgcagt tacagcctac
aaactggttt tctgcttcct gaccgttaag cgcgatttgt gcgccgtcca gcggcagatg
ataaagctgc tgcaatacgg ttccgaccgc gacttccagt gaagcggaag aacttaaccc
ggcaccctgc ggcacattgc cgtgatcaac catgtccacg ccgccgaagc tgttgttacg
cagttgcaga tgtttcacca cgccacgaac gtagttagcc cattgatagt tttcatgtgc
gacaatgggc gcatcgaggg aaaactcgtc gagctgattt tcataatcgg ctgccatcac
gcgaacttta cggtcatcgc gtggtgcaca actgatcacg gtttgataat caatcgcgca
gggcagaacg aaaccgtcgt tgtagtcggt gtgttcacca atcaaattca cgcggccagg
cgcctgaatg gtgtgagtgg cagggtagcc aaatgcgttg gcaaacagag attgtgtttt
ttctttcaga ctcatttctt acactccgga ttcgcgacgg cctacagcaa cctgggcgat
gtatggcatc aggttattcg gaatgccttg cggatcttcg cccatatcgc ccgacggatg
cgcgccaacc gggttgaagt agcgcacgag ggcaatgctc cagtccggct gggctttttg
cagatcggtg aggatctgtt ccaccatcag cttgcttttg ccgtaagggc tttgcggtgt
gccggtcggg aagctataat gcggtagttt atcacagtta aattgctaac gcagtcaggc
accgtgtatg aaatctaaca atgcgctcat cgtcatcctc ggcaccgtca ccctggatgc
tgtaggcata ggcttggtta tgccggtact gccgggcctc ttgcgggata tcgtccattc
cgacagcatc gccagtcact atggcgtgct gctagcttca cgctgccgca agcactcagg
gcgcaagggc tgctaaagga agcggaacac gtagaaagcc agtccgcaga aacggtgctg
accccggatg aatgtcagct actgggctat ctggacaagg gaaaacgcaa gcgcaaagag
aaagcaggta gcttgcagtg ggcttacatg gcgatagcta gactgggcgg ttttatggac
agcaagcgaa ccggaattgc cagctggggc gccctctggt aaggttggga agccctgcaa
agtaaactgg atggctttct tgccgccaag gatctgatgg cgcaggggat caagatctga
tcaagagaca ggatgaggat cgtttcgcat gattgaacaa gatggattgc acgcaggttc
tccggccgct tgggtggaga ggctattcgg ctatgactgg gcacaacaga caatcggctg
ctctgatgcc gccgtgttcc ggctgtcagc gcaggggcgc ccggttcttt ttgtcaagac
cgacctgtcc ggtgccctga atgaactcca agacgaggca gcgcggctat cgtggctggc
cacgacgggc gttccttgcg cagctgtgct cgacgttgtc actgaagcgg gaagggactg
gctgctattg ggcgaagtgc cggggcagga tctcctgtca tctcaccttg ctcctgccga
gaaagtatcc atcatggctg atgcaatgcg gcggctgcat acgcttgatc cggctacctg
cccattcgac caccaagcga aacatcgcat cgagcgagca cgtactcgga tggaagccgg
tcttgtcgat caggatgatc tggacgaaga gcatcagggg ctcgcgccag ccgaactgtt
cgccaggctc aaggcgcgga tgcccgacgg cgaggatctc gtcgtgaccc atggcgatgc
ctgcttgccg aatatcatgg tggaaaatgg ccgcttttct ggattcatcg actgtggccg
gctgggtgtg gcggaccgct atcaggacat agcgttggct acccgtgata ttgctgaaga
gcttggcggc gaatgggctg accgcttcct cgtgctttac ggtatcgccg ctcccgattc
gcagcgcatc gccttctatc gccttcttga cgagttcttc tgagcgggac tctggggttc
gcgatgataa gctgtcaaac atgagaatta caacttatat cgtatggggc tgacttcagg
tgctacattt gaagagataa attgcactga aatctagaaa tattttatct gattaataag
atgatcttct tgagatcgtt ttggtctgcg cgtaatctct tgctctgaaa acgaaaaaac
cgccttgcag ggcggttttt cgaaggttct ctgagctacc aactctttga accgaggtaa
ctggcttgga ggagcgcagt caccaaaact tgtcctttca gtttagcctt aaccggcgca
tgacttcaag actaactcct ctaaatcaat taccagtggc tgctgccagt ggtgcttttg
catgtctttc cgggttggac tcaagacgat agttaccgga taaggcgcag cggtcggact
gaacgggggg ttcgtgcata cagtccagct tggagcgaac tgcctacccg gaactgagtg
tcaggcgtgg aatgagacaa acgcggccat aacagcggaa tgacaccggt aaaccgaaag
gcaggaacag gagagcgcac gagggagccg ccaggggaaa cgcctggtat ctttatagtc
ctgtcgggtt tcgccaccac tgatttgagc gtcagatttc gtgatgcttg tcaggggggc
ggagcctatg gaaaaacggc tttgccttct ttcctgcgtt atcccctgat tctgtggata
accgtattac cgcctttgag tgagctgata ccgctcgccg cagccgaacg accgagcgca
gcgagtcagt gagcgaggaa gcggaagagc gcccaatacg caaaccgcct ctccccgcgc
gttggccgat tcattaatgc agctggcacg acaggtttcc cgactggaaa gcgggcagtg
agcgcaacgc aattaatgtg agttagctca ctcattaggc accccaggct ttacacttta
tgcttccggc tcgtatgttg tgtggaattg tgagcggata acaatttcac acaggaaaca
gctatgacca tgattacgaa ttcgagctcg gtacccgggg atcctctaga gtcgacctgc
aggcatgcaa gcttggcact ggccgtcgtt ttacaacgtc gtgactggga aaaccctggc
gttacccaac ttaatcgcct tgcagcacat ccccctttcg ccagctggcg taatagcgaa
gaggcccgca ccgatcgccc ttcccaacag ttgcgcagcc tgaatggcga atggcgcgat
aagctagcgc tatatgcgtt gatgcaattt ctatgcgcac ccgttctcgg agcactgtcc
gaccgctttg gccgccgccc agtcctgctc gcttcgctac ttggagccac tatcgactac
gcgatcatgg cgaccacacc cgtcctgtgg atcctctacg ccggacgcat cgtggccggc
atcaccggcg ccacaggtgc ggttgctggc gcctatatcg ccgacatcac cgatggggaa
gatcgggctc gccacttcgg gctcatgagc gcttgtttcg gcgtgggtat ggtggcaggc
cccgtggccg ggggactgtt gggcgccatc tccttgcatg caccattcct tgcggcggcg
gtgctcaacg gcctcaacct actactgggc tgcttcctaa tgcaggagtc gcataaggga
gagcgtcgac cgatgccctt gagagccttc aacccagtca gctccttccg gtgggcgcgg
ggcatgacta tcgtcgccgc acttatgact gtcttcttta tcatgcaact cgtaggacag
gtgccggcag cgctctgggt cattttcggc gaggaccgct ttcgctggag cgcgacgatg
atcggcctgt cgcttgcggt attcggaatc ttgcacgccc tcgctcaagc cttcgtcact
ggtcccgcca ccaaacgttt cggcgagaag caggccatta tcgccggcat ggcggccgac
gcgctgggct acgtcttgct ggcgttcgcg acgcgaggct ggatggcctt ccccattatg
attcttctcg cttccggcgg catcgggatg cccgcgttgc aggccatgct gtccaggcag
gtagatgacg accatcaggg acagcttcaa ggatcgctcg cggctcttac cagcctaact
tcgatcattg gaccgctgat cgtcacggcg atttatgccg cctcggcgag cacatggaac
gggttggcat ggattgtagg cgccgcccta taccttgtct gcctccccgc gttgcgtcgc
ggtgcatgga gccgggccac ctcgacctga atggaagccg gcggcacctc gctaacggat
tcaccactcc aagaattgga gccaatcaat tcttgcggag aactgtgaat gcgcaaacca
acccttggca gaacatatcc atcgcgtccg ccatctccag cagccgcacg cggcgcatct
cgggcagcgt tgggtcctgg ccacgggtgc gcatgatcgt gctcctgtcg ttgaggaccc
ggctaggctg gcggggttgc cttactggtt agcagaatga atcaccgata cgcgagcgaa
cgtgaagcga ctgctgctgc aaaacgtctg cgacctgagc aacaacatga atggtcttcg
gtttccgtgt ttcgtaaagt ctggaaacgc ggaagtcagc gccctgcacc attatgttcc
ggatctgcat cgcaggatgc tgctggctac cctgtggaac acctacatct gtattaacga
agcgctggca ttgaccctga gtgatttttc tctggtcccg ccgcatccat accgccagtt
gtttaccctc acaacgttcc agtaaccggg catgttcatc atcagtaacc cgtatcgtga
gcatcctctc tcgtttcatc ggtatcatta cccccatgaa cagaaattcc cccttacacg
gaggcatcaa gtgaccaaac aggaaaaaac cgcccttaac atggcccgct ttatcagaag
ccagacatta acgcttctgg agaaactcaa cgagctggac gcggatgaac aggcagacat
ctgtgaatcg cttcacgacc acgctgatga gctttaccgc agctgcctcg cgcgtttcgg
tgatgacggt gaaaacctct gacacatgca gctcccggag acggtcacag cttgtctgta
agcggatgcc gggagcagac aagcccgtca gggcgcgtca gcgggtgttg gcgggtgtcg
gggcgcagcc atgacccagt cacgtagcga tagcggagtg tatactggct taactatgcg
gcatcagagc agattgtact gagagtgcac catatgcggt gtgaaatacc gcacagatgc
gtaaggagaa aataccgcat caggcgctct tccgcttcct cgctcactga ctcgctgcgc
tcggtcgttc ggctgcggcg agcggtatca gctcactcaa aggcggtaat acggttatcc
acagaatcag gggataacgc aggaaagaac atgtgagcaa aaggccagca aaaggccagg
aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc tccgcccccc tgacgagcat
cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga caggactata aagataccag
gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga
tacctgtccg cctttctccc ttcgggaagc gtggcgcttt ctcaatgctc acgctgtagg
tatctcagtt cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga accccccgtt
cagcccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc ggtaagacac
gacttatcgc cactggcagc agccactggt aacaggatta gcagagcgag gtatgtaggc
ggtgctacag agttcttgaa gtggtggcct aactacggct acactagaag gacagtattt
ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa gagttggtag ctcttgatcc
ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt gcaagcagca gattacgcgc
agaaaaaaag gatctcaaga agatcctttg atcttttcta cggggtctga cgctcagtgg
aacgaaaact cacgttaagg gattttggtc atgagattat caaaaaggat cttcacctag
atccttttaa attaaaaatg aagttttaaa tcaatctaaa gtatatatga gtaaacttgg
tctgacagtt accaatgctt aatcagtgag gcacctatct cagcgatctg tctatttcgt
tcatccatag ttgcctgact ccccgtcgtg tagataacta cgatacggga gggcttacca
tctggcccca gtgctgcaat gataccgcga gacccacgct caccggctcc agatttatca
gcaataaacc agccagccgg aagggccgag cgcagaagtg gtcctgcaac tttatccgcc
tccatccagt ctattaattg ttgccgggaa gctagagtaa gtagttcgcc agttaatagt
ttgcgcaacg ttgttgccat tgctgcaggc atcgtggtgt cacgctcgtc gtttggtatg
gcttcattca gctccggttc ccaacgatca aggcgagtta catgatcccc catgttgtgc
aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg
ttatcactca tggttatggc agcactgcat aattctctta ctgtcatgcc atccgtaaga
tgcttttctg tgactggtga gtactcaacc aagtcattct gagaatagtg tatgcggcga
ccgagttgct cttgcccggc gtcaacacgg gataataccg cgccacatag cagaacttta
aaagtgctca tcattggaaa acgttcttcg gggcgaaaac tctcaaggat cttaccgctg
ttgagatcca gttcgatgta acccactcgt gcacccaact gatcttcagc atcttttact
ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata
agggcgacac ggaaatgttg aatactcata ctcttccttt ttcaatatta ttgaagcatt
tatcagggtt attgtctcat gagcggatac atatttgaat gtatttagaa aaataaacaa
ataggggttc cgcgcacatt tccccgaaaa gtgcggtaat gactccaact tattgatagt
gttttatgtt cagataatgc ccgatgactt tgtcatgcag ctccaccgat tttgagaacg
acagcgactt ccgtcccagc cgtgccaggt gctgcctcag attcaggtta tgccgctcaa
ttcgctgcgt atatcgcttg ctgattacgt gcagctttcc cttcaggcgg gattcataca
gcggccagcc atccgtcatc catatcacca cgtcaaaggg tgacagcagg ctcataagac
gccccagcgt cgccatagtg cgttcaccga atacgtgcgc aacaaccgtc ttccggagac
tgtcatacgc gtgcgtcata tactgactga aaacgcccgc accgttgaag ctgccagcgc
gctggagcaa ggcgacctga aacgtatggg cgagttgatg gcggagtctc atgcctctat
gcgcgatgat ttcgaaatca ccgtgccgca aattgacact ctggtagaaa tcgtcaaagc
tgtgattggc gacaaaggtg gcgtacgcat gaccggcggc ggatttggcg gctgtatcgt
cgcgctgatc ccggaagagc tggtgcctgc cgtacagcaa cgtgtcgctg aacaatatga
agcaaaaaca ggtattaaag agacttttta cgtttgtaaa ccatcacaag gagcaggaca
gtgctgaacg aaactcccgc actggcaccc gatggtcagc cgtaccgact gttgatctgc
ctcgctggcc tgccgcagtt cttcaacctc ccggcgcagc ttttcgttct caatttcagc
atccctttcg gcataccatt ttatgacggc ggcagagtca taaagcacct cattaccctt
gccaccgcct cgcagaacgg gcattccctg ttcctgccag ttctgaatgg tacggatact
cgcaccgaaa atgtcagcca gctgcttttt gttgacttcc attgttcatt ccacggacaa
aaacagagaa aggaaacgac agaggccaaa aagcctcgct ttcagcacct gtcgtttcct
ttcttttcag agggtatttt aaataaaaac attaagttat gacgaagaag aacggaaacg
ccttaaaccg gaaaattttc ataaatagcg aaaacccgcg
//