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ID PDO2 preliminary; circular DNA; SYN; 9386 BP.
XX
AC U03462; ATCC87038;
XX
DT 03-FEB-1994 (Rel. 8, Created)
DT 01-JUL-1995 (Rel. 12, Last updated, Version 1)
XX
DE E. coli cosmid vector pDO2 - complete.
XX
KW cloning vector.
XX
OS Cloning vector
OC Artificial sequences; Cloning vehicles.
XX
RN [1]
RP 1-9386
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 1-9386
RC pDO2
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 pDO2 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--7.2, 2.2; HindIII--5.6, 4.1; XhoI--9.6. (ATCC staff)
CC Plasmid contains the following restriction sites (approximate kb from
CC nt 1): BamHI--6.03; BglI--5.85, 7.44, 8.98; EcoRI--3.89, 6.00;
CC HindIII--2.31, 6.08; PstI--0.65, 1.78, 6.06, 7.57; PvuI--5.88, 7.69;
CC SmaI--0.99, 4.61, 6.01; XbaI--6.04; XhoI--0.96. (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 restriction sites do not cut pDO2:
CC AflII ApaI AvrII BspDI Bst1107I NheI NotI PmlI SfiI SgrAI SnaBI SpeI
CC SrfI SwaI.
CC The order of the major features in the plasmid is: 5' lacZ -
CC Ecl136II/MCS/Sse8387I - 3' lacZ - ren - P - O - ori lambda - cII -
CC cro - kanR - cosR/L - ampR - oriP. [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: 424110
CC NM (pDO2)
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 (color)
CC FN (cloning)(sequencing)
CC SE ()
CC PA (pBR322)
CC BR (pDO6)(pDO17)(pDO184)(pDO19)
CC OF ()
CC OR ()
XX
FH Key Location/Qualifiers
FH
FT misc_feature 0..0
FT /note="1. lambda 48502bp
FT 2. Tn5 5818bp
FT -> lambda::Tn5 54320bp
FT 1. pUC19 2686bp, ori/amp gene/lacZ fragment
FT 2. lambda::Tn5 7674bp, pR/tR2/cos/kan gene
FT 3. pNEB193 EcoRI-HindIII 78bp 397..475, MCS
FT -> pDO2 9386bp"
FT misc_binding 0..0
FT /note="MCS Ecl136II-SacII-Acc651-KpnI-AscI-BamHI-PacI-
FT XbaI-PmeI-Sse83871"
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 rep_origin 0..0
FT /note="ORI E. coli pMB1 (ColE1 and pBR322)"
XX
SQ Sequence 9386 BP; 2463 A; 2415 C; 2369 G; 2139 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 gtgggtggtg
agcagctcgc cgtcgcgcag caggcgccag cgctccatcc acgcagtgac ctcaggcagc
agctgaacca actcgcgagg ggatcgagcc cggggtgggc gaagaactcc agcatgagat
ccccgcgctg gaggatcatc cagccggcgt cccggaaaac gattccgaag cccaaccttt
catagaaggc ggcggtggaa tcgaaatctc gtgatggcag gttgggcgtc gcttggtcgg
tcatttcgaa ccccagagtc ccgctcagaa gaactcgtca agaaggcgat agaaggcgat
gcgctgcgaa tcgggagcgg cgataccgta aagcacgagg aagcggtcag cccattcgcc
gccaagctct tcagcaatat cacgggtagc caacgctatg tcctgatagc ggtccgccac
acccagccgg ccacagtcga tgaatccaga aaagcggcca ttttccacca tgatattcgg
caagcaggca tcgccatggg tcacgacgag atcctcgccg tcgggcatgc gcgccttgag
cctggcgaac agttcggctg gcgcgagccc ctgatgctct tcgtccagat catcctgatc
gacaagaccg gcttccatcc gagtacgtgc tcgctcgatg cgatgtttcg cttggtggtc
gaatgggcag gtagccggat caagcgtatg cagccgccgc attgcatcag ccatgatgga
tactttctcg gcaggagcaa ggtgagatga caggagatcc tgccccggca cttcgcccaa
tagcagccag tcccttcccg cttcagtgac aacgtcgagc acagctgcgc aaggaacgcc
cgtcgtggcc agccacgata gccgcgctgc ctcgtcctgc agttcattca gggcaccgga
caggtcggtc ttgacaaaaa gaaccgggcg cccctgcgct gacagccgga acacggcggc
atcagagcag ccgattgtct gttgtgccca gtcatagccg aatagcctct ccacccaagc
ggccggagaa cctgcgtgca atccatcttg ttcaatcatg cgaaacgatc ctcatcctgt
ctcttgatca gatcttgatc ccctgcgcca tcagatcctt ggcggcaaga aagccatcca
gtttactttg cagggcttcc caaccttacc agagggcgcc ccagctggca attccggttc
gcttgctgtc cataaaaccg cccagtctag ctatcgccat gtaagcccac tgcaagctac
ctgctttctc tttgcgcttg cgttttccct tgtccagata gcccagtagc tgacattcat
ccggggtcag caccgtttct gcggactggc tttctacgtg ttccgcttcc tttagcagcc
cttgcgccct gagtgcttgc ggcagcgtga agcttcaggt gagaacatcc ctgcctgaac
atgagaaaaa acagggtact catactcact tctaagtgac ggctgcatac taaccgcttc
atacatctcg tagatttctc tggcgattga agggctaaat tcttcaacgc taactttgag
aatttttgca agcaatgcgg cgttataagc atttaatgca ttgatgccat taaataaagc
accaacgcct gactgcccca tccccatctt gtctgcgaca gattcctggg ataagccaag
ttcatttttc tttttttcat aaattgcttt aaggcgacgt gcgtcctcaa gctgctcttg
tgttaatggt ttcttttttg tgctcatacg ttaaatctat caccgcaagg gataaatatc
taacaccgtg cgtgttgact attttacctc tggcggtgat aatggttgca tgtactaagg
aggttgtatg gaacaacgca taaccctgaa agattatgca atgcgctttg ggcaaaccaa
gacagctaaa gatctcggcg tatatcaaag cgcgatcaac aaggccattc atgcaggccg
aaagattttt ttaactataa acgctgatgg aagcgtttat gcggaagagg taaagccctt
cccgagtaac aaaaaaacaa cagcataaat aaccccgctc ttacacattc cagccctgaa
aaagggcatc aaattaaacc acacctatgg tgtatgcatt tatttgcata cattcaatca
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 gtgacgtcta agaaaccatt attatcatga cattaaccta taaaaatagg
cgtatcacga ggccctttcg tctcgcgcgt ttcggtgatg acggtgaaaa cctctgacac
atgcagctcc cggagacggt cacagcttgt ctgtaagcgg atgccgggag cagacaagcc
cgtcagggcg cgtcagcggg tgttggcggg tgtcggggct ggcttaacta tgcggcatca
gagcagattg tactgagagt gcaccatatg cggtgtgaaa taccgcacag atgcgtaagg
agaaaatacc gcatcaggcg ccattcgcca ttcaggctgc gcaactgttg ggaagggcga
tcggtgcggg cctcttcgct attacgccag ctggcgaaag ggggatgtgc tgcaaggcga
ttaagttggg taacgccagg gttttcccag tcacgacgtt gtaaaacgac ggccagtgaa
ttcgagctcg gtacccgggg gcgcgccgga tccttaatta agtctagagt cgactgttta
aacctgcagg catgcaagct tggcgtaatc atggtcatag ctgtttcctg tgtgaaattg
ttatccgctc acaattccac acaacatacg agccggaagc ataaagtgta aagcctgggg
tgcctaatga gtgagctaac tcacattaat tgcgttgcgc tcactgcccg ctttccagtc
gggaaacctg tcgtgccagc tgcattaatg aatcggccaa cgcgcgggga gaggcggttt
gcgtattggg cgctcttccg cttcctcgct cactgactcg ctgcgctcgg tcgttcggct
gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg ttatccacag aatcagggga
taacgcagga aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc
cgcgttgctg gcgtttttcc ataggctccg cccccctgac gagcatcaca aaaatcgacg
ctcaagtcag aggtggcgaa acccgacagg actataaaga taccaggcgt ttccccctgg
aagctccctc gtgcgctctc ctgttccgac cctgccgctt accggatacc tgtccgcctt
tctcccttcg ggaagcgtgg cgctttctca atgctcacgc tgtaggtatc tcagttcggt
gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg
cgccttatcc ggtaactatc gtcttgagtc caacccggta agacacgact tatcgccact
ggcagcagcc actggtaaca ggattagcag agcgaggtat gtaggcggtg ctacagagtt
cttgaagtgg tggcctaact acggctacac tagaaggaca gtatttggta tctgcgctct
gctgaagcca gttaccttcg gaaaaagagt tggtagctct tgatccggca aacaaaccac
cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc
tcaagaagat cctttgatct tttctacggg gtctgacgct cagtggaacg aaaactcacg
ttaagggatt ttggtcatga gattatcaaa aaggatcttc acctagatcc ttttaaatta
aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa acttggtctg acagttacca
atgcttaatc agtgaggcac ctatctcagc gatctgtcta tttcgttcat ccatagttgc
ctgactcccc gtcgtgtaga taactacgat acgggagggc ttaccatctg gccccagtgc
tgcaatgata ccgcgagacc cacgctcacc ggctccagat ttatcagcaa taaaccagcc
agccggaagg gccgagcgca gaagtggtcc tgcaacttta tccgcctcca tccagtctat
taattgttgc cgggaagcta gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt
tgccattgct gcaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc
cggttcccaa cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa aagcggttag
ctccttcggt cctccgatcg ttgtcagaag taagttggcc gcagtgttat cactcatggt
tatggcagca ctgcataatt ctcttactgt catgccatcc gtaagatgct tttctgtgac
tggtgagtac tcaaccaagt cattctgaga atagtgtatg cggcgaccga gttgctcttg
cccggcgtca acacgggata ataccgcgcc acatagcaga actttaaaag tgctcatcat
tggaaaacgt tcttcggggc gaaaactctc aaggatctta ccgctgttga gatccagttc
gatgtaaccc actcgtgcac ccaactgatc ttcagcatct tttactttca ccagcgtttc
tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa
atgttgaata ctcatactct tcctttttca atattattga agcatttatc agggttattg
tctcatgagc ggatacatat ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg
cacatttccc cgaaaagtgc ggtaatgact ccaacttatt gatagtgttt tatgttcaga
taatgcccga tgactttgtc atgcagctcc accgattttg agaacgacag cgacttccgt
cccagccgtg ccaggtgctg cctcagattc aggttatgcc gctcaattcg ctgcgtatat
cgcttgctga ttacgtgcag ctttcccttc aggcgggatt catacagcgg ccagccatcc
gtcatccata tcaccacgtc aaagggtgac agcaggctca taagacgccc cagcgtcgcc
atagtgcgtt caccgaatac gtgcgcaaca accgtcttcc ggagactgtc atacgcgtgc
gtcatatact gactgaaaac gcccgcaccg ttgaagctgc cagcgcgctg gagcaaggcg
acctgaaacg tatgggcgag ttgatggcgg agtctcatgc ctctatgcgc gatgatttcg
aaatcaccgt gccgcaaatt gacactctgg tagaaatcgt caaagctgtg attggcgaca
aaggtggcgt acgcatgacc ggcggcggat ttggcggctg tatcgtcgcg ctgatcccgg
aagagctggt gcctgccgta cagcaacgtg tcgctgaaca atatgaagca aaaacaggta
ttaaagagac tttttacgtt tgtaaaccat cacaaggagc aggacagtgc tgaacgaaac
tcccgcactg gcacccgatg gtcagccgta ccgactgttg atctgcctcg ctggcctgcc
gcagttcttc aacctcccgg cgcagctttt cgttctcaat ttcagcatcc ctttcggcat
accattttat gacggcggca gagtcataaa gcacctcatt acccttgcca ccgcctcgca
gaacgggcat tccctgttcc tgccagttct gaatggtacg gatactcgca ccgaaaatgt
cagccagctg ctttttgttg acttccattg ttcattccac ggacaaaaac agagaaagga
aacgacagag gccaaaaagc ctcgctttca gcacctgtcg tttcctttct tttcagaggg
tattttaaat aaaaacatta agttatgacg aagaagaacg gaaacgcctt aaaccggaaa
attttcataa atagcgaaaa cccgcg
//