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ID   PJB8       preliminary; circular DNA; SYN; 5464 BP.
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AC   U00003; ATCC37074;
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DT   02-NOV-1992 (Rel. 7, Created)
DT   01-JUL-1995 (Rel. 12, Last updated, Version 1)
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DE   E. coli cosmid vector pJB8 - incomplete.
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KW   cloning vector.
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OS   Cloning vector
OC   Artificial sequences; Cloning vehicles.
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RN   [1]
RC   pJB8 from HomerI & linker
RA   Ish-Horowicz D., Burke J.F.;
RT   "Rapid and efficient cosmid cloning";
RL   Nucleic Acids Res. 9:2989-2998(1981).
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RN   [2]
RC   pJB1, pJB2, pJB3 from pBR322 & S.typhimurium glnA gene
RC   pJB1-B from pJB1
RC   pJB5 from pJB1
RC   pJB5, pJB6 from pJB1 & pBR322
RC   pJB10 from pJB1-B
RC   pJB8 from pJB10
RC   pJB11, pJB12 from pJB8
RA   Koduri R.K., Bedwell D.M., Brenchley J.E.;
RT   "Characterization of a HindIII-generated DNA fragment carrying the
RT   glutamine synthetase gene of Salmonella typhimurium";
RL   Gene 11:227-237(1980).
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CC   Restriction digests analyzed on agarose gels give the following
CC   sizes (kb): HindIII--5.4; BamHI--5.4; PstI/HindIII--4.6, 0.8;
CC   PstI/SalI--3.3, 2.3. (ATCC staff)
CC   A cosmid vector containing a single cos site. [2]
CC   Cloning into the BamHI or EcoRI site involves 2 preparations of
CC   dephosphorylated vector, one linearized with HindIII, the other
CC   linearized with SalI. [2]
CC   An equimolar ratio of right-hand and left-hand cos containing vector
CC   fragments, digested with BamHI or EcoRI can be ligated to
CC   appropriately digested and dephosphorylated inserts. [2]
CC   Only molecules with 2 cos sites separated by 37-50 kb can be
CC   packaged. [2]
CC   Medium is 1227 LB plus ampicillin.
CC   NM (pJB8)
CC   CM (no)
CC   NA (ds-DNA)
CC   TP (circular)
CC   ST ()
CC   TY (cosmid)
CC   SP (ATCC)
CC   HO (E.coli HB101)(E.coli)
CC   CP ()
CC   FN (cloning 30000 to 40000 bp)
CC   SE ()
CC   PA (Charon 4)(pAT153)
CC   BR ()
CC   OF ()
CC   OR ()
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FH   Key             Location/Qualifiers
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FT   misc_feature    0..0
FT                   /note="1. pAT153 BamHI 3658bp 376..376
FT                   2. Charon 4 BamHI/BstI-BamHI/BstI 1800bp, lambda cos
FT                   -> pJB8 5464bp
FT                   1. HomerI EcoRI 5450bp 5449..5449
FT                   T4 DNA polymerase:T4 DNA polymerase
FT                   BamHI linker 6bp ggatcc:BamHI linker 6bp ggatcc
FT                   remove BamHI-BamHI 6bp
FT                   -> pJB8 5464bp"
FT   rep_origin      604..801
FT                   /note="ORI bacteriophage lambda"
FT   misc_binding    604..801
FT                   /note="SIT bacteriophage lambda cos region"
FT   rep_origin      3694..3694
FT                   /note="ORI E. coli pMB1 (ColE1 and pBR322)"
FT   CDS             complement(4452..5312)
FT                   /note="ANT E. coli beta-lactamase gene (bla)
FT                   ampicillin resistance gene (apr/amp)"
FT   CDS             complement(4452..5312)
FT                   /note="GEN Salmonella typhimurium glutamine
FT                   synthetase gene"
FT   misc_binding    0..0
FT                   /note="SIT EcoRI, from pAT153 (5519..5519)"
FT   misc_binding    0..0
FT                   /note="SIT BamHI, from BamHI linker"
FT   misc_binding    0..0
FT                   /note="SIT EcoRI, from pAT153 (5519..5519)"
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SQ   Sequence 5464 BP; 923 A; 1147 C; 1051 G; 986 T; 1357 other;
     agtcaggcac cgtgtatgaa atctaacaat gcgctcatcg tcatcctcgg caccgtcacc
     ctggatgctg taggcatagg cttggttatg ccggtactgc cgggcctctt gcgggatatc
     gtccattccg acagcatcgc cagtcactat ggcgtgctgc tagcgctata tgcgttgatg
     caatttctat gcgcacccgt tctcggagca ctgtccgacc gctttggccg ccgcccagtc
     ctgctcgctt cgctacttgg agccactatc gactacgcga tcatggcgac cacacccgtc
     ctgtggatcg atctgcctcg ctggcctgcc gcagttcttc aacctcccgg cgcagctttt
     cgttctcaat ttcagcatcc ctttcggccg gcataccatt ttatgacggc ggcagagtca
     taaagcacct cattaccctt gccaccgcct cgcagaacgg gcattccctg ttcctgccag
     ttctgaatgg tacggatact cgcaccgaaa atgtcagcca gctgcttttt gttgacttcc
     attgttcatt ccacggacaa aaacagagaa agcgacagag gccaaaaagc ctcgctttca
     gcacctgtcg tttcctttct tttcagaggg taggtattta aataaaaaca ttaagttatg
     acgaagaaga acggaaacgc cttaaaccgg aaaattttca taaatagcga aaacccgcga
     ggtcgccgcc cnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn
     nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn
     nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn
     nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn
     nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn
     nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn
     nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn
     nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn
     nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn
     nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn
     nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn
     nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn
     nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn
     nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn
     nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn
     nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn
     nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn
     nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn
     nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn
     nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn
     nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn
     nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn
     nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnct 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 cactggaccg 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 tcttccgctt cctcgctcac tgactcgctg cgctcggtcg
     ttcggctgcg gcgagcggta tcagctcact caaaggcggt aatacggtta tccacagaat
     caggggataa cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta
     aaaaggccgc gttgctggcg tttttccata ggctccgccc ccctgacgag catcacaaaa
     atcgacgctc aagtcagagg tggcgaaacc cgacaggact ataaagatac caggcgtttc
     cccctggaag ctccctcgtg cgctctcctg ttccgaccct gccgcttacc ggatacctgt
     ccgcctttct cccttcggga agcgtggcgc tttctcatag ctcacgctgt aggtatctca
     gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc gttcagcccg
     accgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggtaaga cacgacttat
     cgccactggc agcagccact ggtaacagga ttagcagagc gaggtatgta ggcggtgcta
     cagagttctt gaagtggtgg cctaactacg gctacactag aaggacagta tttggtatct
     gcgctctgct gaagccagtt accttcggaa aaagagttgg tagctcttga tccggcaaac
     aaaccaccgc tggtagcggt ggtttttttg tttgcaagca gcagattacg cgcagaaaaa
     aaggatctca agaagatcct ttgatctttt ctacggggtc tgacgctcag tggaacgaaa
     actcacgtta agggattttg gtcatgagat tatcaaaaag gatcttcacc tagatccttt
     taaattaaaa atgaagtttt aaatcaatct aaagtatata tgagtaaact tggtctgaca
     gttaccaatg cttaatcagt gaggcaccta tctcagcgat ctgtctattt cgttcatcca
     tagttgcctg actccccgtc gtgtagataa ctacgatacg ggagggctta ccatctggcc
     ccagtgctgc aatgataccg cgagacccac gctcaccggc tccagattta tcagcaataa
     accagccagc cggaagggcc gagcgcagaa gtggtcctgc aactttatcc gcctccatcc
     agtctattaa ttgttgccgg gaagctagag taagtagttc gccagttaat agtttgcgca
     acgttgttgc cattgctgca ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat
     tcagctccgg ttcccaacga tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag
     cggttagctc cttcggtcct ccgatcgttg tcagaagtaa gttggccgca gtgttatcac
     tcatggttat ggcagcactg cataattctc ttactgtcat gccatccgta agatgctttt
     ctgtgactgg tgagtactca accaagtcat tctgagaata gtgtatgcgg cgaccgagtt
     gctcttgccc ggcgtcaaca cgggataata ccgcgccaca tagcagaact ttaaaagtgc
     tcatcattgg aaaacgttct tcggggcgaa aactctcaag gatcttaccg ctgttgagat
     ccagttcgat gtaacccact cgtgcaccca actgatcttc agcatctttt actttcacca
     gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga ataagggcga
     cacggaaatg ttgaatactc atactcttcc tttttcaata ttattgaagc atttatcagg
     gttattgtct catgagcgga tacatatttg aatgtattta gaaaaataaa caaatagggg
     ttccgcgcac atttccccga aaagtgccac ctgacgtcta agaaaccatt attatcatga
     cattaaccta taaaaatagg cgtatcacga ggccctttcg tcttcaagaa ttccggatcc
     ggaa
//