LOCUS       pOCUS-2      1930 bp    DNA   circular            20-OCT-2003
DEFINITION  Minimal cloning vector pOCUS-2.

REFERENCE   1 (bases 1 to 1930)
            Novagen, Inc.
            pOCUS-2 vector sequence
            http://www.novagen.com/SharedImages/TechnicalLiterature/7_POCUS2S.htm
REFERENCE   2 (sites)
            Novagen, Inc.
            pOCUS-2 vector map (TB100 12/98)
            http://www.novagen.com/SharedImages/TechnicalLiterature/7_TB100.pdf
REFERENCE   3 (sites)
            Novagen, Inc.
            Locus Pocus System Protocols (TB #91 1/94)
            http://www.novagen.com/SharedImages/TechnicalLiterature/7_tb091.pdf
REFERENCE   4 (rep_origin endpoints)
  AUTHORS   Selzer,G., Som,T., Itoh,T. and Tomizawa,J.
  TITLE     The origin of replication of plasmid p15A and comparative studies
            on the nucleotide sequences around the origin of related plasmids
  JOURNAL   Cell 32 (1), 119-129 (1983)
  MEDLINE   83129391
   PUBMED   6186390
            
COMMENT     The pOCUS-2 vector features a minimal plasmid backbone and wide 
            array of cloning sites. It differs from the original pOCUS-1 by 
            the removal of a 158 bp fragment shown to be the occasional target 
            for the gammadelta transposon (R. Novy, unpublished results), the 
            presence of a loxP site, and the replacement of the T3 promoter 
            with 5 additional restriction sites.

FEATURES             Location/Qualifiers
     promoter        6..22
                     /note="T7 promoter; transcription start at 23"
     misc_feature    6..25
                     /note="T7 promoter primer #69348-3"
     misc_feature    29..108
                     /note="multiple cloning region (NotI-PmeI)"
     misc_feature    complement(113..141)
                     /note="POCUSDOWN primer #69528-3"
     CDS             168..1028
                     /gene="bla"
                     /note="ampicillin resistance"
                     /product="beta-lactamase"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGY
                     IELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVE
                     YSPVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRL
                     DRWEPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPL
                     LRSALPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIA
                     EIGASLIKHW"
     rep_origin      1041..1629
                     /standard_name="pMB1 (ColE1-like) plasmid origin of
                     replication"
                     /note="coordinates include the region from the -35 
                     promoter sequence of the RNAII (primer) transcript to the 
                     RNA/DNA switch point"
                     /direction=right
     misc_feature    1810..1843
                     /note="loxP site"
     misc_feature    1895..1920
                     /note="POCUSUP primer #69527-3"
                     
BASE COUNT      481 a    476 c    510 g    463 t
ORIGIN
         1 TTAATTAATA CGACTCACTA TAGGGCGAGC GGCCGCGAAT TCGAGCTCGA TATCAAGCTT
        61 CTCGAGACTA GTTGGGCCCT GCAGGCCTGT CGACAGATCT GTTTAAACGC GCGGACATGA
       121 GACAATAACC CTGATAAATG CTTCAATAAT ATTGAAAAAG GAAGAGTATG AGTATTCAAC
       181 ATTTCCGTGT CGCCCTTATT CCCTTTTTTG CGGCATTTTG CCTTCCTGTT TTTGCTCACC
       241 CAGAAACGCT GGTGAAAGTA AAAGATGCTG AAGATCAGTT GGGTGCACGA GTGGGTTACA
       301 TCGAACTGGA TCTCAACAGC GGTAAGATCC TTGAGAGTTT TCGCCCCGAA GAACGTTTTC
       361 CAATGATGAG CACTTTTAAA GTTCTGCTAT GTGGCGCGGT ATTATCCCGT ATTGACGCCG
       421 GGCAAGAGCA ACTCGGTCGC CGCATACACT ATTCTCAGAA TGACTTGGTT GAGTACTCAC
       481 CAGTCACAGA AAAGCATCTT ACGGATGGCA TGACAGTAAG AGAATTATGC AGTGCTGCCA
       541 TAACCATGAG TGATAACACT GCGGCCAACT TACTTCTGAC AACGATCGGA GGACCGAAGG
       601 AGCTAACCGC TTTTTTGCAC AACATGGGGG ATCATGTAAC TCGCCTTGAT CGTTGGGAAC
       661 CGGAGCTGAA TGAAGCCATA CCAAACGACG AGCGTGACAC CACGATGCCT GTAGCAATGG
       721 CAACAACGTT GCGCAAACTA TTAACTGGCG AACTACTTAC TCTAGCTTCC CGGCAACAAT
       781 TAATAGACTG GATGGAGGCG GATAAAGTTG CAGGACCACT TCTGCGCTCG GCCCTTCCGG
       841 CTGGCTGGTT TATTGCTGAT AAATCTGGAG CCGGTGAGCG TGGGTCTCGC GGTATCATTG
       901 CAGCACTGGG GCCAGATGGT AAGCCCTCCC GTATCGTAGT TATCTACACG ACGGGGAGTC
       961 AGGCAACTAT GGATGAACGA AATAGACAGA TCGCTGAGAT AGGTGCCTCA CTGATTAAGC
      1021 ATTGGTAACA AAGGATCTTC TTGAGATCCT TTTTTTCTGC GCGTAATCTG CTGCTTGCAA
      1081 ACAAAAAAAC CACCGCTACC AGCGGTGGTT TGTTTGCCGG ATCAAGAGCT ACCAACTCTT
      1141 TTTCCGAAGG TAACTGGCTT CAGCAGAGCG CAGATACCAA ATACTGTCCT TCTAGTGTAG
      1201 CCGTAGTTAG GCCACCACTT CAAGAACTCT GTAGCACCGC CTACATACCT CGCTCTGCTA
      1261 ATCCTGTTAC CAGTGGCTGC TGCCAGTGGC GATAAGTCGT GTCTTACCGG GTTGGACTCA
      1321 AGACGATAGT TACCGGATAA GGCGCAGCGG TCGGGCTGAA CGGGGGGTTC GTGCACACAG
      1381 CCCAGCTTGG AGCGAACGAC CTACACCGAA CTGAGATACC TACAGCGTGA GCATTGAGAA
      1441 AGCGCCACGC TTCCCGAAGG GAGAAAGGCG GACAGGTATC CGGTAAGCGG CAGGGTCGGA
      1501 ACAGGAGAGC GCACGAGGGA GCTTCCAGGG GGAAACGCCT GGTATCTTTA TAGTCCTGTC
      1561 GGGTTTCGCC ACCTCTGACT TGAGCGTCGA TTTTTGTGAT GCTCGTCAGG GGGGCGGAGC
      1621 CTATGGAAAA ACGCCAGCAA CGCGGCCTTT TTACGGTTCC TGGCCTTTTG CTGGCCTTTT
      1681 GCTCACATGT TCTTTCCTGC GTTATCCCCT GATTCTGTGG ATAACCGTAT TACCGCCTTT
      1741 GAGTGAGCTG ATACCGCTCG CCGCAGCCGA ACGACCGAGC GCAGCGAGTC AGTGAGCGAG
      1801 GAAGGGGATA TAACTTCGTA TAATGTATGC TATACGAAGT TATTAGGTAC ACAGCTGAGC
      1861 GGAAGAGCGC CCAATACGCA AACCGCCTCT CCCCGCGCGT TGGCCGATTC ATTAATGCAG
      1921 GTTCGCGCAG 
//