Vector pbace36

This is the information supplied by BACPAC Resources about the new vector pBACe3.6 that was created by them to improve the BAC vector system:

In the course of our work, we experienced feedback from users of our PAC clones at several major sequencing centers. A minor disadvantage was the size of the PAC vector (16 kb left in recombinants) as compared to the size of the BAC vector (7 to 8 kb, depending on the vector variant). For the use of BACs or PACs with inserts in the range of 100-150 kb, this means that a larger fraction of the clones in shotgun sequence libraries (M13 or pUC) will be derived from the PAC/BAC vector sequence, hence resulting in a few percent increase in the cost per base pair of final sequence.
In view of these observations and because the existing BAC libraries were of insufficient size, we have improved the BAC vector (Shizuya H. et. al. 1992, Proc. Natl. Acad. Sci. USA 89:8794-8797) to include some of the retrofitting options included in the pPAC4 vector. The new vector has been named pBACe3.6. Specifically, we maintained the wildtype loxP site, added an additional mutant loxP511 site, a site for the intron encoded nuclease PI-SceI and the Tn7att site.
In addition, we changed the cloning area to allow positive selection for inserts containing BAC clones through inclusion of the sacBII gene from Nat Sternberg's P1 vector (Pierce et al. 1992, Proc. Natl. Acad. Sci. USA 89:2056-2060) and disrupted the BamHI cloning site with a fragment containing a pUC plasmid.

The presence of this pUC plasmid serves dual functions: high copy number of the vector for preparing large quantities and appropriate disruption of the sacBII gene to increase viability of the vector containing strain. In addition to the BNamHI site, 5 additional sites can be used for preparing BAC libraries: SacI, SacII, MluI, EcoRI, and AvaIII. The EcoRI site is particularly important in view of the use of this site in the RARE cleavage procedure, thus opening the possibility for selective cloning of similar fragments from different DNA donors. pBACe3.6 clones have chloramphenicol antibiotic resistance. Clones should be grown in LB containing 20ug chloramphenicol/ml.


origin:
Roswell Park Cancer Institute, Human Genetics Department, Pieter J. de Jong - Principal Investigator, Elm and Carltin Streets, Buffalo, New York 14263-0001

Related Links:
Bacpac Resources (http://bacpac.chori.org)
pbace36
        1 gatccgcgga attcgagctc acgcgtactg atgcatgatc cgggtttaaa cccagtactc 
       61 tagatcctct agagtcgacc tgcaggcatg caagcttggc gtaatcatgg tcatagctgt 
      121 ttcctgtgtg aaattgttat ccgctcacaa ttccacacaa catacgagcc ggaagcataa 
      181 agtgtaaagc ctggggtgcc taatgagtga gctaactcac attaattgcg ttgcgctcac 
      241 tgcccgcttt ccagtcggga aacctgtcgt gccagctgca ttaatgaatc ggccaacgcg 
      301 cggggagagg cggtttgcgt attgggcgct cttccgcttc ctcgctcact gactcgctgc 
      361 gctcggtcgt tcggctgcgg cgagcggtat cagctcactc aaaggcggta atacggttat 
      421 ccacagaatc aggggataac gcaggaaaga acatgtgagc aaaaggccag caaaaggcca 
      481 ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag gctccgcccc cctgacgagc 
      541 atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc gacaggacta taaagatacc 
      601 aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg 
      661 gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct ttctcaatgc tcacgctgta 
      721 ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg 
      781 ttcagcccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggtaagac 
      841 acgacttatc gccactggca gcagccactg gtaacaggat tagcagagcg aggtatgtag 
      901 gcggtgctac agagttcttg aagtggtggc ctaactacgg ctacactaga aggacagtat 
      961 ttggtatctg cgctctgctg aagccagtta ccttcggaaa aagagttggt agctcttgat 
     1021 ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag cagattacgc 
     1081 gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc tacggggtct gacgctcagt 
     1141 ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt atcaaaaagg atcttcacct 
     1201 agatcctttt aaattaaaaa tgaagtttta aatcaatcta aagtatatat gagtaaactt 
     1261 ggtctgacag ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc tgtctatttc 
     1321 gttcatccat agttgcctga ctccccgtcg tgtagataac tacgatacgg gagggcttac 
     1381 catctggccc cagtgctgca atgataccgc gagacccacg ctcaccggct ccagatttat 
     1441 cagcaataaa ccagccagcc ggaagggccg agcgcagaag tggtcctgca actttatccg 
     1501 cctccatcca gtctattaat tgttgccggg aagctagagt aagtagttcg ccagttaata 
     1561 gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt gtcacgctcg tcgtttggta 
     1621 tggcttcatt cagctccggt tcccaacgat caaggcgagt tacatgatcc cccatgttgt 
     1681 gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag 
     1741 tgttatcact catggttatg gcagcactgc ataattctct tactgtcatg ccatccgtaa 
     1801 gatgcttttc tgtgactggt gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc 
     1861 gaccgagttg ctcttgcccg gcgtcaatac gggataatac cgcgccacat agcagaactt 
     1921 taaaagtgct catcattgga aaacgttctt cggggcgaaa actctcaagg atcttaccgc 
     1981 tgttgagatc cagttcgatg taacccactc gtgcacccaa ctgatcttca gcatctttta 
     2041 ctttcaccag cgtttctggg tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa 
     2101 taagggcgac acggaaatgt tgaatactca tactcttcct ttttcaatat tattgaagca 
     2161 tttatcaggg ttattgtctc atgagcggat acatatttga atgtatttag aaaaataaac 
     2221 aaataggggt tccgcgcaca tttccccgaa aagtgccacc tgacgtctaa gaaaccatta 
     2281 ttatcatgac attaacctat aaaaataggc gtatcacgag gccctttcgt ctcgcgcgtt 
     2341 tcggtgatga cggtgaaaac ctctgacaca tgcagctccc ggagacggtc acagcttgtc 
     2401 tgtaagcgga tgccgggagc agacaagccc gtcagggcgc gtcagcgggt gttggcgggt 
     2461 gtcggggctg gcttaactat gcggcatcag agcagattgt actgagagtg caccatatgc 
     2521 ggtgtgaaat accgcacaga tgcgtaagga gaaaataccg catcaggcgc cattcgccat 
     2581 tcaggctgcg caactgttgg gaagggcgat cggtgcgggc ctcttcgcta ttacgccagc 
     2641 tggcgaaagg gggatgtgct gcaaggcgat taagttgggt aacgccaggg ttttcccagt 
     2701 cacgacgttg taaaacgacg gccagtgaat tcgagctcgg tacccgggga tctagagtac 
     2761 tggttgaaac ccggatcatg catcagtacg cgtgagctcg aattccgcgg atccttctat 
     2821 agtgtcacct aaatgtcgac ggccaggcgg ccgccaggcc tacccactag tcaattcggg 
     2881 aggatcgaaa cggcagatcg caaaaaacag tacatacaga aggagacatg aacatgaaca 
     2941 tcaaaaaaat tgtaaaacaa gccacagttc tgacttttac gactgcactt ctggcaggag 
     3001 gagcgactca agccttcgcg aaagaaaata accaaaaagc atacaaagaa acgtacggcg 
     3061 tctctcatat tacacgccat gatatgctgc agatccctaa acagcagcaa aacgaaaaat 
     3121 accaagtgcc tcaattcgat caatcaacga ttaaaaatat tgagtctgca aaaggacttg 
     3181 atgtgtggga cagctggccg ctgcaaaacg ctgacggaac agtagctgaa tacaacggct 
     3241 atcacgttgt gtttgctctt gcgggaagcc cgaaagacgc tgatgacaca tcaatctaca 
     3301 tgttttatca aaaggtcggc gacaactcaa tcgacagctg gaaaaacgcg ggccgtgtct 
     3361 ttaaagacag cgataagttc gacgccaacg atccgatcct gaaagatcag acgcaagaat 
     3421 ggtccggttc tgcaaccttt acatctgacg gaaaaatccg tttattctac actgactatt 
     3481 ccggtaaaca ttacggcaaa caaagcctga caacagcgca ggtaaatgtg tcaaaatctg 
     3541 atgacacact caaaatcaac ggagtggaag atcacaaaac gatttttgac ggagacggaa 
     3601 aaacatatca gaacgttcag cagtttatcg atgaaggcaa ttatacatcc ggcgacaacc 
     3661 atacgctgag agaccctcac tacgttgaag acaaaggcca taaatacctt gtattcgaag 
     3721 ccaacacggg aacagaaaac ggataccaag gcgaagaatc tttatttaac aaagcgtact 
     3781 acggcggcgg cacgaacttc ttccgtaaag aaagccagaa gcttcagcag agcgctaaaa 
     3841 aacgcgatgc tgagttagcg aacggcgccc tcggtatcat agagttaaat aatgattaca 
     3901 cattgaaaaa agtaatgaag ccgctgatca cttcaaacac ggtaactgat gaaatcgagc 
     3961 gcgcgaatgt tttcaaaatg aacggcaaat ggtacttgtt cactgattca cgcggttcaa 
     4021 aaatgacgat cgatggtatt aactcaaacg atatttacat gcttggttat gtatcaaact 
     4081 ctttaaccgg cccttacaag ccgctgaaca aaacagggct tgtgctgcaa atgggtcttg 
     4141 atccaaacga tgtgacattc acttactctc acttcgcagt gccgcaagcc aaaggcaaca 
     4201 atgtggttat cacaagctac atgacaaaca gaggcttctt cgaggataaa aaggcaacat 
     4261 ttgcgccaag cttcttaatg aacatcaaag gcaataaaac atccgttgtc aaaaacagca 
     4321 tcctggagca aggacagctg acagtcaact aataacagca aaaagaaaat gccgatactt 
     4381 cattggcatt ttcttttatt tctcaacaag atggtgaatt gactagtggg tagatccaca 
     4441 ggacgggtgt ggtcgccatg atcgcgtagt cgatagtggc tccaagtagc gaagcgagca 
     4501 ggactgggcg gcggccaaag cggtcggaca gtgctccgag aacgggtgcg catagaaatt 
     4561 gcatcaacgc atatagcgct agcagcacgc catagtgact ggcgatgctg tcggaatgga 
     4621 cgatatcccg caagaggccc ggcagtaccg gcataaccaa gcctatgcct acagcatcca 
     4681 gggtgacggt gccgaggatg acgatgagcg cattgttaga tttcatacac ggtgcctgac 
     4741 tgcgttagca atttaactgt gataaactac cgcattaaag cttatcgatg ataagctgtc 
     4801 aaacatgaga attgatccgg aacccttaat ataacttcgt ataatgtatg ctatacgaag 
     4861 ttattaggtc cctcgactat agggtcaccg tcgacagcga cacacttgca tcggatgcag 
     4921 cccggttaac gtgccggcac ggcctgggta accaggtatt ttgtccacat aaccgtgcgc 
     4981 aaaatgttgt ggataagcag gacacagcag caatccacag caggcataca accgcacacc 
     5041 gaggttactc cgttctacag gttacgacga catgtcaata cttgcccttg acaggcattg 
     5101 atggaatcgt agtctcacgc tgatagtctg atcgacaata caagtgggac cgtggtccca 
     5161 gaccgataat cagaccgaca acacgagtgg gatcgtggtc ccagactaat aatcagaccg 
     5221 acgatacgag tgggaccgtg gtcccagact aataatcaga ccgacgatac gagtgggacc 
     5281 gtggttccag actaataatc agaccgacga tacgagtggg accgtggtcc cagactaata 
     5341 atcagaccga cgatacgagt gggaccatgg tcccagacta ataatcagac cgacgatacg 
     5401 agtgggaccg tggtcccagt ctgattatca gaccgacgat acgagtggga ccgtggtccc 
     5461 agactaataa tcagaccgac gatacgagtg ggaccgtggt cccagactaa taatcagacc 
     5521 gacgatacga gtgggaccgt ggtcccagtc tgattatcag accgacgata caagtggaac 
     5581 agtgggccca gagagaatat tcaggccagt tatgctttct ggcctgtaac aaaggacatt 
     5641 aagtaaagac agataaacgt agactaaaac gtggtcgcat cagggtgctg gcttttcaag 
     5701 ttccttaaga atggcctcaa ttttctctat acactcagtt ggaacacgag acctgtccag 
     5761 gttaagcacc attttatcgc ccttatacaa tactgtcgct ccaggagcaa actgatgtcg 
     5821 tgagcttaaa ctagttcttg atgcagatga cgttttaagc acagaagtta aaagagtgat 
     5881 aacttcttca gcttcaaata tcaccccagc ttttttctgc tcatgaaggt tagatgcctg 
     5941 ctgcttaagt aattcctctt tatctgtaaa ggctttttga agtgcatcac ctgaccgggc 
     6001 agatagttca ccggggtgag aaaaaagagc aacaactgat ttaggcaatt tggcggtgtt 
     6061 gatacagcgg gtaataatct tacgtgaaat attttccgca tcagccagcg cagaaatatt 
     6121 tccagcaaat tcattctgca atcggcttgc ataacgctga ccacgttcat aagcacttgt 
     6181 tgggcgataa tcgttaccca atctggataa tgcagccatc tgctcatcat ccagctcgcc 
     6241 aaccagaaca cgataatcac tttcggtaag tgcagcagct ttacgacggc gactcccatc 
     6301 ggcaatttct atgacaccag atactcttcg accgaacgcc ggtgtctgtt gaccagtcag 
     6361 tagaaaagaa gggatgagat catccagtgc gtcctcagta agcagctcct ggtcacgttc 
     6421 attacctgac catacccgag aggtcttctc aacactatca ccccggagca cttcaagagt 
     6481 aaacttcaca tcccgaccac atacaggcaa agtaatggca ttaccgcgag ccattactcc 
     6541 tacgcgcgca attaacgaat ccaccatcgg ggcagctggt gtcgataacg aagtatcttc 
     6601 aaccggttga gtattgagcg tatgttttgg aataacaggc gcacgcttca ttatctaatc 
     6661 tcccagcgtg gtttaatcag acgatcgaaa atttcattgc agacaggttc ccaaatagaa 
     6721 agagcatttc tccaggcacc agttgaagag cgttgatcaa tggcctgttc aaaaacagtt 
     6781 ctcatccgga tctgaccttt accaacttca tccgtttcac gtacaacatt ttttagaacc 
     6841 atgcttcccc aggcatcccg aatttgctcc tccatccacg gggactgaga gccattacta 
     6901 ttgctgtatt tggtaagcaa aatacgtaca tcaggctcga accctttaag atcaacgttc 
     6961 ttgagcagat cacgaagcat atcgaaaaac tgcagtgcgg aggtgtagtc aaacaactca 
     7021 gcaggcgtgg gaacaatcag cacatcagca gcacatacga cattaatcgt gccgataccc 
     7081 aggttaggcg cgctgtcaat aactatgaca tcatagtcat gagcaacagt ttcaatggcc 
     7141 agtcggagca tcaggtgtgg atcggtgggc agtttacctt catcaaattt gcccattaac 
     7201 tcagtttcaa tacggtgcag agccagacag gaaggaataa tgtcaagccc cggccagcaa 
     7261 gtgggcttta ttgcataagt gacatcgtcc ttttccccaa gatagaaagg caggagagtg 
     7321 tcttctgcat gaatatgaag atctggtacc catccgtgat acattgaggc tgttccctgg 
     7381 gggtcgttac cttccacgag caaaacacgt agccccttca gagccagatc ctgagcaaga 
     7441 tgaacagaaa ctgaggtttt gtaaacgcca cctttatggg cagcaacccc gatcaccggt 
     7501 ggaaatacgt cttcagcacg tcgcaatcgc gtaccaaaca catcacgcat atgattaatt 
     7561 tgttcaattg tataaccaac acgttgctca acccgtcctc gaatttccat atccgggtgc 
     7621 ggtagtcgcc ctgctttctc ggcatctctg atagcctgag aagaaacccc aactaaatcc 
     7681 gctgcttcac ctattctcca gcgccgggtt attttcctcg cttccgggct gtcatcatta 
     7741 aactgtgcaa tggcgatagc cttcgtcatt tcatgaccag cgtttatgca ctggttaagt 
     7801 gtttccatga gtttcattct gaacatcctt taatcattgc tttgcgtttt tttattaaat 
     7861 cttgcaattt actgcaaagc aacaacaaaa tcgcaaagtc atcaaaaaac cgcaaagttg 
     7921 tttaaaataa gagcaacact acaaaaggag ataagaagag cacatacctc agtcacttat 
     7981 tatcactagc gctcgccgca gccgtgtaac cgagcatagc gagcgaactg gcgaggaagc 
     8041 aaagaagaac tgttctgtca gatagctctt acgctcagcg caagaagaaa tatccaccgt 
     8101 gggaaaaact ccaggtagag gtacacacgc ggatagccaa ttcagagtaa taaactgtga 
     8161 taatcaaccc tcatcaatga tgacgaacta acccccgata tcaggtcaca tgacgaaggg 
     8221 aaagagaagg aaatcaactg tgacaaactg ccctcaaatt tggcttcctt aaaaattaca 
     8281 gttcaaaaag tatgagaaaa tccatgcagg ctgaaggaaa cagcaaaact gtgacaaatt 
     8341 accctcagta ggtcagaaca aatgtgacga accaccctca aatctgtgac agataaccct 
     8401 cagactatcc tgtcgtcatg gaagtgatat cgcggaagga aaatacgata tgagtcgtct 
     8461 ggcggccttt ctttttctca atgtatgaga ggcgcattgg agttctgctg ttgatctcat 
     8521 taacacagac ctgcaggaag cggcggcgga agtcaggcat acgctggtaa ctttgaggca 
     8581 gctggtaacg ctctatgatc cagtcgattt tcagagagac gatgcctgag ccatccggct 
     8641 tacgatactg acacagggat tcgtataaac gcatggcata cggattggtg atttcttttg 
     8701 tttcactaag ccgaaactgc gtaaaccggt tctgtaaccc gataaagaag ggaatgagat 
     8761 atgggttgat atgtacactg taaagccctc tggatggact gtgcgcacgt ttgataaacc 
     8821 aaggaaaaga ttcatagcct ttttcatcgc cggcatcctc ttcagggcga taaaaaacca 
     8881 cttccttccc cgcgaaactc ttcaatgcct gccgtatatc cttactggct tccgcagagg 
     8941 tcaatccgaa tatttcagca tatttagcaa catggatctc gcagataccg tcatgttcct 
     9001 gtagggtgcc atcagatttt ctgatctggt caacgaacag atacagcata cgtttttgat 
     9061 cccgggagag actatatgcc gcctcagtga ggtcgtttga ctggacgatt cgcgggctat 
     9121 ttttacgttt cttgtgattg ataaccgctg tttccgccat gacagatcca tgtgaagtgt 
     9181 gacaagtttt tagattgtca cactaaataa aaaagagtca ataagcaggg ataactttgt 
     9241 gaaaaaacag cttcttctga gggcaatttg tcacagggtt aagggcaatt tgtcacagac 
     9301 aggactgtca tttgagggtg atttgtcaca ctgaaagggc aatttgtcac aacaccttct 
     9361 ctagaaccag catggataaa ggcctacaag gcgctctaaa aaagaagatc taaaaactat 
     9421 aaaaaaaata attataaaaa tatccccgtg gataagtgga taaccccaag ggaagttttt 
     9481 tcaggcatcg tgtgtaagca gaatatataa gtgctgttcc ctggtgcttc ctcgctcact 
     9541 cgagggcttc gccctgtcgc tcaactgcgg cgagcactac tggctgtaaa aggacagacc 
     9601 acatcatggt tctgtgttca ttaggttgtt ctgtccattg ctgacataat ccgctccact 
     9661 tcaacgtaac accgcacgaa gatttctatt gttcctgaag gcatattcaa atcgttttcg 
     9721 ttaccgcttg caggcatcat gacagaacac tacttcctat aaacgctaca caggctcctg 
     9781 agattaataa tgcggatctc tacgataatg ggagattttc ccgactgttt cgttcgcttc 
     9841 tcagtggata acagccagct tctctgttta acagacaaaa acagcatatc cactcagttc 
     9901 cacatttcca tataaaggcc aaggcattta ttctcaggat aattgtttca gcatcgcaac 
     9961 cgcatcagac tccggcatcg caaactgcac ccggtgccgg gcagccacat ccagcgcaaa 
    10021 aaccttcgtg tagacttccg ttgaactgat ggacttatgt cccatcaggc tttgcagaac 
    10081 tttcagcggt ataccggcat acagcatgtg catcgcatag gaatggcgga acgtatgtgg 
    10141 tgtgaccgga acagagaacg tcacaccgtc agcagcagcg gcggcaaccg cctccccaat 
    10201 ccaggtcctg accgttctgt ccgtcacttc ccagatccgc gctttctctg tccttcctgt 
    10261 gcgacggtta cgccgctcca tgagcttatc gcgaataaat acctgtgacg gaagatcact 
    10321 tcgcagaata aataaatcct ggtgtccctg ttgataccgg gaagccctgg gccaactttt 
    10381 ggcgaaaatg agacgttgat cggcacgtaa gaggttccaa ctttcaccat aatgaaataa 
    10441 gatcactacc gggcgtattt tttgagttat cgagattttc aggagctaag gaagctaaaa 
    10501 tggagaaaaa aatcactgga tataccaccg ttgatatatc ccaatggcat cgtaaagaac 
    10561 attttgaggc atttcagtca gttgctcaat gtacctataa ccagaccgtt cagctggata 
    10621 ttacggcctt tttaaagacc gtaaagaaaa ataagcacaa gttttatccg gcctttattc 
    10681 acattcttgc ccgcctgatg aatgctcatc cggagttccg tatggcaatg aaagacggtg 
    10741 agctggtgat atgggatagt gttcaccctt gttacaccgt tttccatgag caaactgaaa 
    10801 cgttttcatc gctctggagt gaataccacg acgatttccg gcagtttcta cacatatatt 
    10861 cgcaagatgt ggcgtgttac ggtgaaaacc tggcctattt ccctaaaggg tttattgaga 
    10921 atatgttttt cgtctcagcc aatccctggg tgagtttcac cagttttgat ttaaacgtgg 
    10981 ccaatatgga caacttcttc gcccccgttt tcaccatggg caaatattat acgcaaggcg 
    11041 acaaggtgct gatgccgctg gcgattcagg ttcatcatgc cgtttgtgat ggcttccatg 
    11101 tcggcagaat gcttaatgaa ttacaacagt actgcgatga gtggcagggc ggggcgtaat 
    11161 ttttttaagg cagttattgg tgcccttaaa cgcctggttg ctacgcctga ataagtgata 
    11221 ataagcggat gaatggcaga aattcgatga taagctgtca aacatgagaa ttggtcgacg 
    11281 gcgcgccaaa gcttgcatgc ctgcagccgc gtaacctggc aaaatcggtt acggttgagt 
    11341 aataaatgga tgccctgcgt aagcggggca catttcatta cctctttctc cgcacccgac 
    11401 atagataata acttcgtata gtatacatta tacgaagtta tctagtagac ttaattaagg 
    11461 atcgatccgg cgcgccaata gtcatgcccc gcgcccaccg gaaggagctg actgggttga 
    11521 aggctctcaa gggcatcggt cgagcttgac attgtaggac tatattgctc taataaattt 
    11581 gcggccgcta atacgactca ctatagggag ag