LOCUS       pDGO186NX-m1219-pLT_237      4772 bp    DNA     circular UNA 28-SEP-2010
DEFINITION  
ACCESSION   pMU102
DBSOURCE    GenBank
SOURCE      Cloning vector pNW33N (unknown)
  ORGANISM  Cloning vector pNW33N
            Cloning vector pNW33N
            other sequences; artificial sequences; vectors
FEATURES             Location/Qualifiers
     Replication_ori 1..589
                     /note="Oriinal location: complement(2766..3354)"
                     /note="Escherichia coli origin of replication from
                     plasmid pUC19"
                     /label="E. coli origin of replication"
     primer          complement(22..41)
                     /label="TY112"
     primer          complement(56..73)
                     /label="X03373"
     primer          complement(355..374)
                     /label="X03372"
     primer          complement(646..665)
                     /label="X03371"
     primer          692..716
                     /label="BB R2 int F"
     primer          complement(701..725)
                     /label="BB F int R"
     Replication_ori 826..1280
                     /label="C. therm origin of replication"
     primer          complement(963..982)
                     /label="X03370"
     primer          complement(1252..1274)
                     /label="X03369"
     CDS             1281..2285
                     /gene="repB"
                     /codon_start=1
                     /transl_table=11
                     /product="RepB"
                     /protein_id="AAO86061.1"
                     /db_xref="GI:29650180"
                     /note="Oriinal location: complement(1070..2074)"
                     /note="replication protein; rolling circle replication
                     initiation protein; Bacillus origin of replication from
                     Geobacillus stearothermophilus plasmid pTHT15"
     gene            1281..2285
                     /gene="repB"
                     /note="Oriinal location: complement(1070..2074)"
     primer          1355..1381
                     /label="X07369"
     primer          complement(1855..1878)
                     /label="X03367"
     primer          2075..2095
                     /label="qPCR F"
     primer          complement(2086..2110)
                     /label="X07370"
     primer          complement(2161..2190)
                     /label="TY113R+"
     primer          2161..2190
                     /label="TY113+"
     primer          complement(2165..2184)
                     /label="qPCR R"
     primer          2171..2190
                     /label="TY113"
     primer          complement(2171..2190)
                     /label="TY113R"
     primer          complement(2256..2296)
                     /label="TY1A R"
     primer          complement(2272..2296)
                     /label="184N R"
     primer          2297..2326
                     /label="TY1B F"
     primer          complement(2313..2367)
                     /label="BackboneR1"
     SPACER          2338..3487
                     /label="HA gblock"
     primer          complement(2338..2367)
                     /label="SR"
     primer          complement(2338..2362)
                     /label="SR1"
     gblock          2368..3467
                     /label="1219 Homology Arm"
     primer          2649..2669
                     /label="1219SF"
     primer          2714..2738
                     /label="HRM F PolIII"
     misc_feature    2772
                     /feature_type="silent mutation"
                     /label="silent mutation"
     mutation        2774
                     /label="G to A"
     mutation        2774
                     /label="G to A mutation"
     misc_feature    2781
                     /feature_type="silent mutation"
                     /label="silent mutation"
     primer          complement(2799..2823)
                     /label="HRM R PolIII"
     PAM             3173..3177
     SPACER          3178..3214
     primer          complement(3197..3214)
                     /label="LT_654"
     primer          3468..3527
                     /label="Backbone Forward"
     misc_feature    complement(<3468..4558)
                     /feature_type="Insertion"
                     /note="Region on donor sequence (pMU104): 1..3564"
                     /label="pMU104"
     insulator       3468..3494
     primer          3472..3491
                     /label="SF2"
     promoter        3504..3532
                     /label="CAT promoter"
     primer          complement(3566..3595)
                     /label="TY1B R"
     CDS             3596..4246
                     /gene="cat"
                     /function="chloramphenicol resistance"
                     /codon_start=1
                     /transl_table=11
                     /product="chloramphenicol acetyltransferase"
                     /protein_id="AAO86060.1"
                     /db_xref="GI:29650179"
                     /note="Oriinal location: complement(313..963)"
                     /note="derived from rom Staphylococcus aureus plasmid
                     pC194"
     gene            3596..4246
                     /gene="cat"
                     /note="Oriinal location: complement(313..963)"
     primer          3647..3678
                     /label="TY114R+"
     primer          complement(3647..3678)
                     /label="TY114+"
     primer          complement(3657..3678)
                     /label="TY114"
     primer          3657..3678
                     /label="TY114R"
     primer          complement(3659..3678)
                     /label="X03365"
     primer          3713..3732
                     /label="X07303"
     primer          3885..3903
                     /note="qPCR cat primer forward"
                     /label="X07706"
     primer          3885..3903
                     /label="qPCR F"
     primer          complement(3897..3916)
                     /label="X07304"
     primer          complement(3959..3978)
                     /label="X03364"
     primer          complement(3983..4001)
                     /note="qPCR cat primer reverse"
                     /label="X07707"
     primer          complement(3983..4001)
                     /label="qPCR R"
     primer          4085..4104
                     /label="X07305"
     primer          4174..4195
                     /label="TY111"
     primer          complement(4184..4205)
                     /label="X07302"
     primer          4247..4276
                     /label="TY1A F"
     primer          complement(4271..4291)
                     /label="X03363"
     promoter        4412..4448
                     /label="pLac"
     -35_signal      4412..4417
                     /label="pLac -35"
     -10_signal      4436..4441
                     /label="pLac -10"
     Primer_binding_ 4501..4515
                     /label="PstI in-fusion primer binding site F"
     primer          complement(4508..4532)
                     /label="TC01"
     Primer_binding_ 4517..4531
                     /label="in-fusion XbaI primer binding site"
     Primer_binding_ complement(4528..4542)
                     /label="XbaI in-fusion primer binding site F"
     Primer_binding_ complement(4541..4555)
                     /label="in-fusion SmaI primer binding site"
     primer          4542..4566
                     /label="TC02"
     primer          complement(4580..4602)
                     /label="X03362"
ORIGIN
        1 TTGAGATCCT TTTTTTCTGC GCGTAATCTG CTGCTTGCAA ACAAAAAAAC CACCGCTACC
       61 AGCGGTGGTT TGTTTGCCGG ATCAAGAGCT ACCAACTCTT TTTCCGAAGG TAACTGGCTT
      121 CAGCAGAGCG CAGATACCAA ATACTGTTCT TCTAGTGTAG CCGTAGTTAG GCCACCACTT
      181 CAAGAACTCT GTAGCACCGC CTACATACCT CGCTCTGCTA ATCCTGTTAC CAGTGGCTGC
      241 TGCCAGTGGC GATAAGTCGT GTCTTACCGG GTTGGACTCA AGACGATAGT TACCGGATAA
      301 GGCGCAGCGG TCGGGCTGAA CGGGGGGTTC GTGCACACAG CCCAGCTTGG AGCGAACGAC
      361 CTACACCGAA CTGAGATACC TACAGCGTGA GCTATGAGAA AGCGCCACGC TTCCCGAAGG
      421 GAGAAAGGCG GACAGGTATC CGGTAAGCGG CAGGGTCGGA ACAGGAGAGC GCACGAGGGA
      481 GCTTCCAGGG GGAAACGCCT GGTATCTTTA TAGTCCTGTC GGGTTTCGCC ACCTCTGACT
      541 TGAGCGTCGA TTTTTGTGAT GCTCGTCAGG GGGGCGGAGC CTATGGAAAA ACGCCAGCAA
      601 CGCGGCCTTT TTACGGTTCC TGGCCTTTTG CTGGCCTTTT GCTCACATGT TCTTTCCTGC
      661 GTTATCCCCT GATTCTGTGG ATAACCGTAT TACCGCCTTT GAGTGAGCTG ATACCGCTCG
      721 CCGCAGCCGA ACGACCGAGC GCAGCGAGTC AGTGAGCGAG GAAGCGGAAG AGCGCCCAAT
      781 ACGCAAACCG CCTCTCCCCG CGCGTTGGCC GATTCATTAA TGCAGTATGG GAAACAAAAT
      841 ATTGCGTATG CGACTGTTCA CATGGACGAG AAAACCCCTC ACATGCATTT AGGAGTTGTT
      901 CCTATGCGCC TAGAGGGCTT TTCGTGCGTC AGCATGAGCG ATCGGAAGAA AAGAAGAATA
      961 CTTTTTCGCT TCAAGATGTT TTGCAGCGTG ATCGAGAACT TCGTGAGCAA AGAAAAGCAA
     1021 AGAGGAAAAA ATCGCATGAT TTGGAGCGAT AAGAAAAAGC ACTCGAATGA GTGCTTTTTT
     1081 TGCGTTTTGA GCGTAGCGAA AAACGAGTTC TTTCTATTCT TGATACATAT AGAAATAACG
     1141 TCATTTTTAT TTTAGTTGCT GAAAGGTGCG TTGAAGTGTT GGTATGTATG TGTTTTAAAG
     1201 TATTGAAAAC CCTTAAAATT GGTTGCACAG AAAAACCCCA TCTGTTAAAG TTATAAGTGA
     1261 CCAAACAAAT AACTAAATAG ATGGGGGTTT CTTTTAATAT TATGTGTCCT AATAGTAGCA
     1321 TTTATTCAGA TGAAAAATCA AGGGTTTTAG TGGACAAGAC AAAAAGTGGA AAAGTGAGAC
     1381 CATGGAGAGA AAAGAAAATC GCTAATGTTG ATTACTTTGA ACTTCTGCAT ATTCTTGAAT
     1441 TTAAAAAGGC TGAAAGAGTA AAAGATTGTG CTGAAATATT AGAGTATAAA CAAAATCGTG
     1501 AAACAGGCGA AAGAAAGTTG TATCGAGTGT GGTTTTGTAA ATCCAGGCTT TGTCCAATGT
     1561 GCAACTGGAG GAGAGCAATG AAACATGGCA TTCAGTCACA AAAGGTTGTT GCTGAAGTTA
     1621 TTAAACAAAA GCCAACAGTT CGTTGGTTGT TTCTCACATT AACAGTTAAA AATGTTTATG
     1681 ATGGCGAAGA ATTAAATAAG AGTTTGTCAG ATATGGCTCA AGGATTTCGC CGAATGATGC
     1741 AATATAAAAA AATTAATAAA AATCTTGTTG GTTTTATGCG TGCAACGGAA GTGACAATAA
     1801 ATAATAAAGA TAATTCTTAT AATCAGCACA TGCATGTATT GGTATGTGTG GAACCAACTT
     1861 ATTTTAAGAA TACAGAAAAC TACGTGAATC AAAAACAATG GATTCAATTT TGGAAAAAGG
     1921 CAATGAAATT AGACTATGAT CCAAATGTAA AAGTTCAAAT GATTCGACCG AAAAATAAAT
     1981 ATAAATCGGA TATACAATCG GCAATTGACG AAACTGCAAA ATATCCTGTA AAGGATACGG
     2041 ATTTTATGAC CGATGATGAA GAAAAGAATT TGAAACGTTT GTCTGATTTG GAGGAAGGTT
     2101 TACACCGTAA AAGGTTAATC TCCTATGGTG GTTTGTTAAA AGAAATACAT AAAAAATTAA
     2161 ACCTTGATGA CACAGAAGAA GGCGATTTGA TTCATACAGA TGATGACGAA AAAGCCGATG
     2221 AAGATGGATT TTCTATTATT GCAATGTGGA ATTGGGAACG GAAAAATTAT TTTATTAAAG
     2281 AGTAGTTCAA GGATCCCTCG TATGTTGTGT GGAATTGTGA GCGGATAACA ATTTCACACA
     2341 GGAAACAGCT ATGACCATGA TTACGCCGAC AATCCTGTAA TTGACACTTT GGAACTTTCA
     2401 AGGCAAATGT TTCCGGAGCT TAAAAAACAC AAACTGGATG TGGTTGCAAA GCATCTTGGT
     2461 GTTTCATTGG AAAACCATCA CAGGGCTCTT GACGATGCAA AGGCCTGTGG GGAGATCTTT
     2521 ATCAAATGCC TTGAGATACT GGCTGAAAAG AATGTAAAAA CAATTGATGA CATACAAAAT
     2581 GTTTTTGAGG GTTGTTGGAA TTATCAGAAA GCAAATTCAT ACCATGCCAT TATACTTGTA
     2641 AAAAATTACG TAGGCCTTAG AAATCTGTAC AAGATTGTGT CCAAGACTCA TTTGGAGTAT
     2701 TTTCACAAAA GGCCGAGACT TCCAAGAAAG CTTTTAATGA TGCACAGAGA AGGACTCATT
     2761 TTGGGAAGTG CaTaTGAAGC tGGAGAGCTT TACCGTGCGA TTCTTGAAAA CAAAGATGAG
     2821 GACGAAATTT CAAGGATTGT AAAGTTTTAC GATTATCTTG AAATACAGCC TTTGGGCAAC
     2881 AACCGTTTCC TTGTCGAGAG CGGAAAGGTG AACAGTGAGG AAGACTTAAA GAATATAAAC
     2941 AGAAAAATAG TAGCTTTGGG TGAGAAGTAC AACAAGCCGG TGGTCGCTAC CTGCGATGTT
     3001 CATTTCATGG ATCCGCAGGA CGAAGTTTTC AGAAGAATAC TTATGGCGGG GCAAGGTTTT
     3061 TCCGATGCGG ACAACCAGGC ACCGTTGTAT TTCAGGACCA CTGATGAAAT GCTGGAAGAG
     3121 TTTGAATATC TGGGCAGGGA AAAATGCTAT GAAGTGGTTG TAACAAATAC AAATTTAATT
     3181 GCGGATATGT GCGAGGACAT ACTTCCCATA CCGGAAGGAA CTTTTCCTCC TAAAATTGAA
     3241 GGTGCGGAAG AAGAAATAAG GATGCTTGCC GAAAACAAGG CGAGAGAAAT TTATGGCGAC
     3301 CCTCTTCCTG AGATTGTGGA AAAGCGCATG GAAAAGGAGC TTAACTCAAT AATAAAAAAT
     3361 GGCTTCTCCG TTATGTATAT TATTGCGCAA AAACTTGTAT GGAAATCTTT AAGTGACGGC
     3421 TATCTTGTAG GGTCGAGGGG ATCCGTCGGA TCATCCTTTG TTGCCTATAA TAGAAATAAT
     3481 TTTGTTTAAC TTTACAAACG GGATTGACTT TTAAAAAAGG ATTGATTCTA ATGAAGAAAG
     3541 CAGACAAGTA AGCCTCCTAA ATTCACTTTA GATAAAAATT TAGGAGGCAT ATCAAATGAA
     3601 CTTTAATAAA ATTGATTTAG ACAATTGGAA GAGAAAAGAG ATATTTAATC ATTATTTGAA
     3661 CCAACAAACG ACTTTTAGTA TAACCACAGA AATTGATATT AGTGTTTTAT ACCGAAACAT
     3721 AAAACAAGAA GGATATAAAT TTTACCCTGC ATTTATTTTC TTAGTGACAA GGGTGATAAA
     3781 CTCAAATACA GCTTTTAGAA CTGGTTACAA TAGCGACGGA GAGTTAGGTT ATTGGGATAA
     3841 GTTAGAGCCA CTTTATACAA TTTTTGATGG TGTATCTAAA ACATTCTCTG GTATTTGGAC
     3901 TCCTGTAAAG AATGACTTCA AAGAGTTTTA TGATTTATAC CTTTCTGATG TAGAGAAATA
     3961 TAATGGTTCG GGGAAATTGT TTCCCAAAAC ACCTATACCT GAAAATGCTT TTTCTCTTTC
     4021 TATTATTCCA TGGACTTCAT TTACTGGGTT TAACTTAAAT ATCAATAATA ATAGTAATTA
     4081 CCTTCTACCC ATTATTACAG CAGGAAAATT CATTAATAAA GGTAATTCAA TATATTTACC
     4141 GCTATCTTTA CAGGTACATC ATTCTGTTTG TGATGGTTAT CATGCAGGAT TGTTTATGAA
     4201 CTCTATTCAG GAATTGTCAG ATAGGCCTAA TGACTGGCTT TTATAATATG AGATAATGCC
     4261 GACTGTACTT TTTACAGTCG GTTTTCTAAT GTCACTAGGG CTCGCCTTTG GGAAGTTTGA
     4321 AGGGCTGGCA CGACAGGTTT CCCGACTGGA AAGCGGGCAG TGAGCGCAAC GCAATTAATG
     4381 TGAGTTAGCT CACTCATTAG GCACCCCAGG CTTTACACTT TATGCTTCCG GCTCGTATGT
     4441 TGTGTGGAAT TGTGAGCGGA TAACAATTTC ACACAGGAAA CAGCTATGAC CATGATTACG
     4501 CCAAGCTTGC ATGCCTGCAG GTCGACTCTA GAGGATCCCC GGGTACCGAG CTCGAATTGA
     4561 CAGATCGCTG AGATAGGTGC CTCACTGATT AAGCATTGGT AACTGTCAGA CCAAGTTTAC
     4621 TCATATATAC TTTAGATTGA TTTAAAACTT CATTTTTAAT TTAAAAGGAT CTAGGTGAAG
     4681 ATCCTTTTTG ATAATCTCAT GACCAAAATC CCTTAACGTG AGTTTTCGTT CCACTGAGCG
     4741 TCAGACCCCG TAGAAAAGAT CAAAGGATCT TC
//
