LOCUS       PI76KSIV     4098 bp    DNA   Circular  SYN        3-JUN-1997
DEFINITION  Plasmid pI176KSiv - internal PCR standard
ACCESSION   MD88001
KEYWORDS    .
SOURCE      derived from recombinant construct pI176KS.
  ORGANISM  synthetic
REFERENCE   1  (sites)
  AUTHORS   Fristensky, Brian W
  TITLE     Untitled
  JOURNAL   Unpublished (1997) Plant Science, University of Manitoba
  STANDARD  full automatic
COMMENT
            This clone was made Mary DePauw in Brian Fristensky's lab,
            Experiment MD88. It contains the 341bp pUC18/Sau3A1 fragment
            inserted at the BglII site within the cDNA. Amplification
            using primers flanking this insertion gives a PCR fragment
            341bp longer than that generated in RT-PCR from mRNA.
            Corresponding Address for Citation 1:
            Brian W Fristensky
            University of Manitoba
            Winnipeg, MB  R3T 2N2
            Canada
            Phone: 204-474-6085
            Fax:   204-261-5732
            Email: frist@cc.umanitoba.ca
FEATURES            LOCATION/QUALIFIERS
     misc_feature    M18249:1..355
                     /evidence= EXPERIMENTAL
                     /label=PR10.2_cDNA_left
     misc_feature    L08752:complement(1369..1709)
                     /function= "vector"
                     /evidence= EXPERIMENTAL
                     /standard_name= "SYNPUC18V"
                     /label=pUC18_frag
                     /note= "concatenated Sau3A1 digest fragments"
     misc_feature    M18249:356..728
                     /evidence= EXPERIMENTAL
                     /label=PR10.2_cDNA_right
     unsure          "aaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaa" 
                     /label=polyA_tail
                     /note= "About 50bp but precise length unknown"
     misc_feature    "tgcagg"
                     /evidence= EXPERIMENTAL
                     /label=Pst_to_Sal
                     /note= "pUC9 polylinker, from T-tailed PstI site to SalI"
     misc_feature    X52331:join(735..2958,1..719)
                     /function= "vector"
                     /evidence= EXPERIMENTAL
                     /standard_name= "Bluescript KSm13+"
                     /label=KSm13_H3_Sal
                     /note= "Bluescript KSm13+/HindIII/SalI cut"
     misc_feature    "agcttggctgcacccccccccccccccccc"
                     /evidence= EXPERIMENTAL
                     /label=polyC-tail
                     /note= "pUC9 polylinker from HindIII to C-tailed PstI
                     site"
     source          join(PR10.2_cDNA_left,pUC18_frag,PR10.2_cDNA_right,
                     polyA_tail,Pst_to_Sal,KSm13_H3_Sal,polyC-tail)
                     /organism="Escherichia coli"
                     /plasmid="pI176KSiv"
                     /evidence= EXPERIMENTAL
                     /label=pI176KSiv

BASE COUNT     1088 a    980 c    981 g   1049 t
ORIGIN
        1 tacacaacta ggcaagcaat ttcttagttc tttctcacac tttagcaata taaaacaata
       61 tcatcatggg tgtttttaat gttgaagatg aaatcacttc tgttgtagca cctgctatac
      121 tctacaaagc tctagttaca gatgctgaca ctcttactcc aaaggttatt gatgccatca
      181 aaagtatcga aattgttgaa ggaaacggag gtgctggaac catcaagaaa ctcactttcg
      241 ttgaagatgg tgaaaccaaa catgtgttgc acaaagtgga gttagtagat gttgctaact
      301 tggcttacaa ctatagcata gttggtggtg ttggatttcc agacacagtt gagaagatct
      361 gtctatttcg ttcatccata gttgcctgac tccccgtcgt gtagataact acgatacggg
      421 agggcttacc atctggcccc agtgctgcaa tgataccgcg agacccacgc tcaccggctc
      481 cagatttatc agcaataaac cagccagccg gaagggccga gcgcagaagt ggtcctgcaa
      541 ctttatccgc ctccatccag tctattaatt gttgccggga agctagagta agtagttcgc
      601 cagttaatag tttgcgcaac gttgttgcca ttgctacagg catcgtggtg tcacgctcgt
      661 cgtttggtat ggcttcattc agctccggtt cccaacgatc tcatttgagg ctaaactgtc
      721 tgcaggacca aatggaggat ccattgcaaa gctgagtgtg aaatattaca caaaaggtga
      781 tgctgctgct cctactgaag agcaactcaa gagtgacaaa gctaagggtg atggtctttt
      841 caaggctctt gagcgttact gtttggctca tcctgattac aactaaacta tataatcatc
      901 aacaagtgtg tgttgttatg tatactatat aatcatcatg tgtgcttaat ttggctgcca
      961 atgtaatccc cctgttttgt tttttccctt tcccttttct ttgtggacta aaagtgtgag
     1021 attgtaagtc atgtatacct ctccttcaat aaattataat catttatata aaaaaaaaaa
     1081 aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa aaaaaaaaat gcaggtcgac ctcgaggggg
     1141 ggcccggtac ccagcttttg ttccctttag tgagggttaa tttcgagctt ggcgtaatca
     1201 tggtcatagc tgtttcctgt gtgaaattgt tatccgctca caattccaca caacatacga
     1261 gccggaagca taaagtgtaa agcctggggt gcctaatgag tgagctaact cacattaatt
     1321 gcgttgcgct cactgcccgc tttccagtcg ggaaacctgt cgtgccagct gcattaatga
     1381 atcggccaac gcgcggggag aggcggtttg cgtattgggc gctcttccgc ttcctcgctc
     1441 actgactcgc tgcgctcggt cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg
     1501 gtaatacggt tatccacaga atcaggggat aacgcaggaa agaacatgtg agcaaaaggc
     1561 cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca taggctccgc
     1621 ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga
     1681 ctataaagat accaggcgtt tccccctgga agctccctcg tgcgctctcc tgttccgacc
     1741 ctgccgctta ccggatacct gtccgccttt ctcccttcgg gaagcgtggc gctttctcat
     1801 agctcacgct gtaggtatct cagttcggtg taggtcgttc gctccaagct gggctgtgtg
     1861 cacgaacccc ccgttcagcc cgaccgctgc gccttatccg gtaactatcg tcttgagtcc
     1921 aacccggtaa gacacgactt atcgccactg gcagcagcca ctggtaacag gattagcaga
     1981 gcgaggtatg taggcggtgc tacagagttc ttgaagtggt ggcctaacta cggctacact
     2041 agaaggacag tatttggtat ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt
     2101 ggtagctctt gatccggcaa acaaaccacc gctggtagcg gtggtttttt tgtttgcaag
     2161 cagcagatta cgcgcagaaa aaaaggatct caagaagatc ctttgatctt ttctacgggg
     2221 tctgacgctc agtggaacga aaactcacgt taagggattt tggtcatgag attatcaaaa
     2281 aggatcttca cctagatcct tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata
     2341 tatgagtaaa cttggtctga cagttaccaa tgcttaatca gtgaggcacc tatctcagcg
     2401 atctgtctat ttcgttcatc catagttgcc tgactccccg tcgtgtagat aactacgata
     2461 cgggagggct taccatctgg ccccagtgct gcaatgatac cgcgagaccc acgctcaccg
     2521 gctccagatt tatcagcaat aaaccagcca gccggaaggg ccgagcgcag aagtggtcct
     2581 gcaactttat ccgcctccat ccagtctatt aattgttgcc gggaagctag agtaagtagt
     2641 tcgccagtta atagtttgcg caacgttgtt gccattgcta caggcatcgt ggtgtcacgc
     2701 tcgtcgtttg gtatggcttc attcagctcc ggttcccaac gatcaaggcg agttacatga
     2761 tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt tgtcagaagt
     2821 aagttggccg cagtgttatc actcatggtt atggcagcac tgcataattc tcttactgtc
     2881 atgccatccg taagatgctt ttctgtgact ggtgagtact caaccaagtc attctgagaa
     2941 tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca
     3001 catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg aaaactctca
     3061 aggatcttac cgctgttgag atccagttcg atgtaaccca ctcgtgcacc caactgatct
     3121 tcagcatctt ttactttcac cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc
     3181 gcaaaaaagg gaataagggc gacacggaaa tgttgaatac tcatactctt cctttttcaa
     3241 tattattgaa gcatttatca gggttattgt ctcatgagcg gatacatatt tgaatgtatt
     3301 tagaaaaata aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc acctaaattg
     3361 taagcgttaa tattttgtta aaattcgcgt taaatttttg ttaaatcagc tcatttttta
     3421 accaataggc cgaaatcggc aaaatccctt ataaatcaaa agaatagacc gagatagggt
     3481 tgagtgttgt tccagtttgg aacaagagtc cactattaaa gaacgtggac tccaacgtca
     3541 aagggcgaaa aaccgtctat cagggcgatg gcccactacg tgaaccatca ccctaatcaa
     3601 gttttttggg gtcgaggtgc cgtaaagcac taaatcggaa ccctaaaggg agcccccgat
     3661 ttagagcttg acggggaaag ccggcgaacg tggcgagaaa ggaagggaag aaagcgaaag
     3721 gagcgggcgc tagggcgctg gcaagtgtag cggtcacgct gcgcgtaacc accacacccg
     3781 ccgcgcttaa tgcgccgcta cagggcgcgt cccattcgcc attcaggctg cgcaactgtt
     3841 gggaagggcg atcggtgcgg gcctcttcgc tattacgcca gctggcgaaa gggggatgtg
     3901 ctgcaaggcg attaagttgg gtaacgccag ggttttccca gtcacgacgt tgtaaaacga
     3961 cggccagtga attgtaatac gactcactat agggcgaatt ggagctccac cgcggtggcg
     4021 gccgctctag aactagtgga tcccccgggc tgcaggaatt cgatatcaag cttggctgca
     4081 cccccccccc cccccccc
//
