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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
//