LOCUS       pET-Duet1_EcPPtase_St1      9167 bp    DNA     circular UNA 14-SEP-2013
FEATURES             Location/Qualifiers
     Site            3..27
                     /site_type="binding site"
                     /label="LacO"
     gene            71..700
                     /dnas_title="PPT-ase Ec"
                     /vntifkey="15"
                     /label="PPT-ase Ec"
     PCR_primer      748..767
                     /pair
                     /primer="TGTACACGGCCGCATAATCG"
                     /current="0"
     PCR_primer      complement(749..768)
                     /pair
                     /primer="TCGATTATGCGGCCGTGTAC"
                     /current="1"
     promoter        772..788
                     /label="T7 Gen 2"
     Site            791..815
                     /site_type="binding site"
                     /label="LacO"
     RBS             840..857
     misc_feature    870..887
                     /label="HIStag"
     primer_bind     893..932
                     /label="Gibson overhang CAR_neu_5'"
     primer_bind     complement(909..932)
                     /label="pETDuet1:EcPPTase rev"
     misc_feature    909..929
                     /label="TEV cleavage site"
     CDS             933..4100
                     /label="St1"
     primer_bind     4101..4125
                     /label="pETDuet1:EcPPTase fwd"
     primer_bind     4110..4140
                     /label="3' Gibson overlap"
     PCR_primer      complement(4195..4213)
                     /pair
                     /primer="GCTAGTTATTGCTCAGCGG"
                     /current="0"
     terminator      complement(4209..4256)
                     /label="T7_ter"
     PCR_primer      9043..9062
                     /pair="pETT7up"
                     /primer="CGGTGATGTCGGCGATATAG"
                     /current="0"
     PCR_primer      9105..9122
                     /pair="pETUpstreammod"
                     /primer="ATGCGTCCGGCGTAGAGG"
                     /current="0"
     misc_feature    9151..9167
                     /note="Promoter"
ORIGIN
        1 GGGGAATTGT GAGCGGATAA CAATTCCCCT CTAGAAATAA TTTTGTTTAA CTTTAAGAAG
       61 GAGATATACC ATGGTCGATA TGAAAACTAC GCATACCTCC CTCCCCTTTG CCGGACATAC
      121 GCTGCATTTT GTTGAGTTCG ATCCGGCGAA TTTTTGTGAG CAGGATTTAC TCTGGCTGCC
      181 GCACTACGCA CAACTGCAAC ACGCTGGACG TAAACGTAAA ACAGAGCATT TAGCCGGACG
      241 GATCGCTGCT GTTTATGCTT TGCGGGAATA TGGCTATAAA TGTGTGCCCG CAATCGGCGA
      301 GCTACGCCAA CCTGTCTGGC CTGCGGAGGT ATACGGCAGT ATTAGCCACT GTGGGACTAC
      361 GGCATTAGCC GTGGTATCTC GTCAACCGAT TGGCATTGAT ATAGAAGAAA TTTTTTCTGT
      421 ACAAACCGCA AGAGAATTGA CAGACAACAT TATTACACCA GCGGAACACG AGCGACTCGC
      481 AGACTGCGGT TTAGCCTTTT CTCTGGCGCT GACACTGGCA TTTTCCGCCA AAGAGAGCGC
      541 ATTTAAGGCA AGTGAGATCC AAACTGATGC AGGTTTTCTG GACTATCAGA TAATTAGCTG
      601 GAATAAACAG CAGGTCATCA TTCATCGTGA GAATGAGATG TTTGCTGTGC ACTGGCAGAT
      661 AAAAGAAAAG ATAGTCATAA CGCTGTGCCA ACACGATTAA AAGCTTGCGG CCGCATAATG
      721 CTTAAGTCGA ACAGAAAGTA ATCGTATTGT ACACGGCCGC ATAATCGAAA TTAATACGAC
      781 TCACTATAGG GGAATTGTGA GCGGATAACA ATTCCCCATC TTAGTATATT AGTTAAGTAT
      841 AAGAAGGAGA TATACATATG TCGTACTACC ATCACCATCA CCATCACGAT TACGACATCC
      901 CAACGACCGA AAACCTGTAT TTTCAGGGCG CCATGACAGC AATGAGGCAA AGAACAGACT
      961 ACGGAAAGAG GCTGCTCGTC AACATCGTCG ACGAGCGTGC GGAAACGGAG CCCAATCGAG
     1021 AGTGGGTGTC CATCCCGACC TCCTCCAACC CCAAAGACGG CTGGAAGAAG ATCACGTACC
     1081 GACAAGCCGC CAATGCCGTC AACCGGGTCG CCCACAAACT GGTGAGCTCA ACAGGCCGGC
     1141 CGAAGGAGGG CGAGTTCCCG ACCGTCGCCT ACATCGGCCC CAACGATGTG CGATACGTCG
     1201 TCTTTGCCCT GGGAGCCATC AAGGCGGGGT ACCAGGCGCT CTTCATCTCG CCGCGCAACT
     1261 CGCAGGAGGG CCAGCTCAAC CTCTTCGAGC TGACCAACTG CCGCACCATT TGGTTTGACG
     1321 CCATGTACAA GGACGCGGTC CAGTCGTGGG TGCAAGAGCG GGACATGCAC GCCATCATGA
     1381 CCTTCCCAGT CGCCGCCTGG TTCCCCGACG AGGACGTCGA GCCGTATCCG TACGACAAGA
     1441 CGTTTGAACA AGCCGAGTGG GACCCGCTCA TGGTTCTGCA CACGAGCGGC AGCACCGGCT
     1501 TCCCCAAGCC GGTCGTGGTG AGGCAGGGCA TGTTGGCGAT CGGGGATGGC TACCACAACC
     1561 TGGGCGAGTG GAAAGGGAGG AAGATCTGGC TGGACGAGAT GTCGCGGCGG TCCAAGAGGA
     1621 TGCTGTGCCC AATGCCCCTC TTTCACGCGG CCGCCATGTA CGTCACTCTC CTCCTAGTTC
     1681 AGTACTGGGA TCTCCCCGTG GCTCTGGGCA TCGGCGACCG ACCCCTGTCC GCAGATATGG
     1741 CTCTGGAGTG CTTGAAATAT GCGGAAGTGG ACTCGGTCAT CCTGCCGCCC GCAATCCTGG
     1801 AGGAACTAGG CCAGACGCAA GAAGCGATCG ATGCCCTAAA AGCGCTGTCG TTCGTCGCCT
     1861 TTGGGGGTGG GAACTTGTCG GACGAGGCGG GAGACAAGCT CGCCAAGGCC GGTGTTAAGC
     1921 TCTTCAACGC CATCTCGACA ACCGAGTTTG CTCCCTATCC ACTCTATTGG CAGACAAATC
     1981 CGGAGCTCTG GCGATATTTC ATCTTCAACT CGGAGCTCTT CGGCTGTGAA TGGAGGCCGG
     2041 CGACGGACGA GAACACGTAC GAGCAGGTCA TCGTACGAAA GGACAAGCAC CCGGGATGTC
     2101 AAGGGATCTT CTACACCTTC CCCGAGGCCT GGGAATACAG CACCAAGGAC TTGTTCAGCC
     2161 CACACCCAAC ACTTCCCGAT CATTGGAAGT ATTGTGGGCG ATCGGACAAC ATCATCGTCT
     2221 TCTCCAATGG CGAGAAGCTC AACCCAGCAT CCATTGAAAC GATTCTGATG GGCCATCCTC
     2281 GGGTCAAGGG GGCACTCGTG GTTGGCAGCA ACCGGTTCCA GCCGGCTCTC ATCCTTGAGC
     2341 CCGTGGAGCA CCCGCGTACG GAGCAGGGCG TCAGGGAGTT CATCGACAGC GTGTGGCCGA
     2401 CGGTTGTCAA GGCCAACAAA GAGACTGTAG CCCACGGTCA GATCGGGCGG CAGTTCATCG
     2461 CCATCTCTAA TCCCGACAAG CCGTTCCTGC GAGCGGGCAA GGGCACGATC CAGCGGGCAG
     2521 GTACCATACG CATTTACGAG GACGAGATCG ATCAGATATA CGAACAAGCC GACGGGGTCG
     2581 CGTCCTCGGA GGCCCCGGTG CTGAACCTTG AGTCGACCGA GTCGCTAACC CGGTCCATCG
     2641 AGATCCTCTT CGAGAAGTGG CTGCAGGCGC CGAAGCTTGA GCCGGATACG GACTTCTTCA
     2701 CGGTCGGAAT CGACTCGATG CAGGTCATCA ATGCTTCACG GCTGCTCAGG GCAGGGTTGG
     2761 AAGCCGGCGG CGTCCGGGTA GACTCCGCTT CCCTGGCCAC GCGCGTCATT TATGGACACC
     2821 CGACGGCACG ACGGCTGGCC GAGTATCTCT TCTCGGTGGT GAACCAAAAG GGCCAGGATG
     2881 CCACCTCTGG CGAGCCGCAG CACGAAGATC ATGCCATGGA GGCCATCCTG GAGAAATACA
     2941 CGCGGGATAT GCCCCGAGCC CCGACCGCCG GCAAGCCCGC CCCGGCCGAC GAGGGCCAGG
     3001 TAATTATCAT TACGGGAACA ACGGGCACGC TCGGGTCGTA CACGCTCGAC ATCGCCTCGC
     3061 GCTGCCCCCG AGTCAGGAAA GTCATCTGCC TCAACCGGTC GGACGACGCC GAAGCGCGAC
     3121 AGAGGCGGTC GAATGCGGAA CGCGGTTTGC ACATCGACTT CTCCAAGGCC GAGTTCCTGC
     3181 GCGCCGACAT GTCCAAGCAC GACCTTGGGC TCGGGAGGGA GGCGTACGAG AGACTGCTGC
     3241 ACGAAGTCGA CCGGGTCATC CACAACCAAT GGCCGGTCAA CTTCAACATG CCCGTCGAGT
     3301 CCTTCGAGCC GCACATCCGG GGAGTGCGCA ACCTCGTCGA CTTCAGTTGC AAAGCGACGA
     3361 AGCGGGTTCC CATCGTGTTT ATCTCGTCTA TCGCAACCGT CAACGGGTGG AGGAAGTCAG
     3421 AGGCCGTGCC GGAACGATCG CTGAAGGAGC CGGAAATCGC CGTCGGCGGG TACGGGCGGT
     3481 CCAAACTCGT GAGCAGCCTC ATCCTGGAGA AGGCAACCGA GGTCTCGGGC GTTCCCACCG
     3541 AGATCATGCG TATCGGGCAG ATTGCCGGCC CGAGCAGCGA GAAGGGCCTC TGGAACCGGC
     3601 AGGAGTGGCT CCCCAGCATC ATCGCCAGCT CTCTGTATCT CGGAGTCCTA CCTGACTCCC
     3661 TCGGACACAT GTCCACGATC GACTGGACGC CCATCGAAGG GATCGCAAAC CTCGTGCTCG
     3721 AAGCGTCCGG GGTGACCTCT CAGGTTCCTC TTGAGGACAT CAACGGCTAC TTCCACGGTG
     3781 TCAACCCGGT CACGACGCAG TGGAGACCGC TCGCCGAGGC GGTAAAGGAG TTTTACGGCG
     3841 GGCGCATCCA AAAGCTAGTC CCCCTGGACG AGTGGGTCGA GGCCCTGGAA CAGAGCCAGG
     3901 CCAAGGCCGA AGACATCAGC AAGAACCCCG GCGTCAAGCT GCTGGACACA TATAAGGCTT
     3961 GGGTGGCGGC GGCAAGGGAG GGGCAGAACT ACGTCGCCAT GGACACGTCG CGGACGACGA
     4021 GCCGCAGCAG GACAATGCGC GAGATGCGCG CCATCACGCC CGAGCTGATG AAGAACTGGT
     4081 GCCGCCAGTG GGCGTTTTAA CTCGAGTCTG GTAAAGAAAC CGCTGCTGCG AAATTTGAAC
     4141 GCCAGCACAT GGACTCGTCT ACTAGCGCAG CTTAATTAAC CTAGGCTGCT GCCACCGCTG
     4201 AGCAATAACT AGCATAACCC CTTGGGGCCT CTAAACGGGT CTTGAGGGGT TTTTTGCTGA
     4261 AAGGAGGAAC TATATCCGGA TTGGCGAATG GGACGCGCCC TGTAGCGGCG CATTAAGCGC
     4321 GGCGGGTGTG GTGGTTACGC GCAGCGTGAC CGCTACACTT GCCAGCGCCC TAGCGCCCGC
     4381 TCCTTTCGCT TTCTTCCCTT CCTTTCTCGC CACGTTCGCC GGCTTTCCCC GTCAAGCTCT
     4441 AAATCGGGGG CTCCCTTTAG GGTTCCGATT TAGTGCTTTA CGGCACCTCG ACCCCAAAAA
     4501 ACTTGATTAG GGTGATGGTT CACGTAGTGG GCCATCGCCC TGATAGACGG TTTTTCGCCC
     4561 TTTGACGTTG GAGTCCACGT TCTTTAATAG TGGACTCTTG TTCCAAACTG GAACAACACT
     4621 CAACCCTATC TCGGTCTATT CTTTTGATTT ATAAGGGATT TTGCCGATTT CGGCCTATTG
     4681 GTTAAAAAAT GAGCTGATTT AACAAAAATT TAACGCGAAT TTTAACAAAA TATTAACGTT
     4741 TACAATTTCT GGCGGCACGA TGGCATGAGA TTATCAAAAA GGATCTTCAC CTAGATCCTT
     4801 TTAAATTAAA AATGAAGTTT TAAATCAATC TAAAGTATAT ATGAGTAAAC TTGGTCTGAC
     4861 AGTTACCAAT GCTTAATCAG TGAGGCACCT ATCTCAGCGA TCTGTCTATT TCGTTCATCC
     4921 ATAGTTGCCT GACTCCCCGT CGTGTAGATA ACTACGATAC GGGAGGGCTT ACCATCTGGC
     4981 CCCAGTGCTG CAATGATACC GCGAGACCCA CGCTCACCGG CTCCAGATTT ATCAGCAATA
     5041 AACCAGCCAG CCGGAAGGGC CGAGCGCAGA AGTGGTCCTG CAACTTTATC CGCCTCCATC
     5101 CAGTCTATTA ATTGTTGCCG GGAAGCTAGA GTAAGTAGTT CGCCAGTTAA TAGTTTGCGC
     5161 AACGTTGTTG CCATTGCTAC AGGCATCGTG GTGTCACGCT CGTCGTTTGG TATGGCTTCA
     5221 TTCAGCTCCG GTTCCCAACG ATCAAGGCGA GTTACATGAT CCCCCATGTT GTGCAAAAAA
     5281 GCGGTTAGCT CCTTCGGTCC TCCGATCGTT GTCAGAAGTA AGTTGGCCGC AGTGTTATCA
     5341 CTCATGGTTA TGGCAGCACT GCATAATTCT CTTACTGTCA TGCCATCCGT AAGATGCTTT
     5401 TCTGTGACTG GTGAGTACTC AACCAAGTCA TTCTGAGAAT AGTGTATGCG GCGACCGAGT
     5461 TGCTCTTGCC CGGCGTCAAT ACGGGATAAT ACCGCGCCAC ATAGCAGAAC TTTAAAAGTG
     5521 CTCATCATTG GAAAACGTTC TTCGGGGCGA AAACTCTCAA GGATCTTACC GCTGTTGAGA
     5581 TCCAGTTCGA TGTAACCCAC TCGTGCACCC AACTGATCTT CAGCATCTTT TACTTTCACC
     5641 AGCGTTTCTG GGTGAGCAAA AACAGGAAGG CAAAATGCCG CAAAAAAGGG AATAAGGGCG
     5701 ACACGGAAAT GTTGAATACT CATACTCTTC CTTTTTCAAT CATGATTGAA GCATTTATCA
     5761 GGGTTATTGT CTCATGAGCG GATACATATT TGAATGTATT TAGAAAAATA AACAAATAGG
     5821 TCATGACCAA AATCCCTTAA CGTGAGTTTT CGTTCCACTG AGCGTCAGAC CCCGTAGAAA
     5881 AGATCAAAGG ATCTTCTTGA GATCCTTTTT TTCTGCGCGT AATCTGCTGC TTGCAAACAA
     5941 AAAAACCACC GCTACCAGCG GTGGTTTGTT TGCCGGATCA AGAGCTACCA ACTCTTTTTC
     6001 CGAAGGTAAC TGGCTTCAGC AGAGCGCAGA TACCAAATAC TGTCCTTCTA GTGTAGCCGT
     6061 AGTTAGGCCA CCACTTCAAG AACTCTGTAG CACCGCCTAC ATACCTCGCT CTGCTAATCC
     6121 TGTTACCAGT GGCTGCTGCC AGTGGCGATA AGTCGTGTCT TACCGGGTTG GACTCAAGAC
     6181 GATAGTTACC GGATAAGGCG CAGCGGTCGG GCTGAACGGG GGGTTCGTGC ACACAGCCCA
     6241 GCTTGGAGCG AACGACCTAC ACCGAACTGA GATACCTACA GCGTGAGCTA TGAGAAAGCG
     6301 CCACGCTTCC CGAAGGGAGA AAGGCGGACA GGTATCCGGT AAGCGGCAGG GTCGGAACAG
     6361 GAGAGCGCAC GAGGGAGCTT CCAGGGGGAA ACGCCTGGTA TCTTTATAGT CCTGTCGGGT
     6421 TTCGCCACCT CTGACTTGAG CGTCGATTTT TGTGATGCTC GTCAGGGGGG CGGAGCCTAT
     6481 GGAAAAACGC CAGCAACGCG GCCTTTTTAC GGTTCCTGGC CTTTTGCTGG CCTTTTGCTC
     6541 ACATGTTCTT TCCTGCGTTA TCCCCTGATT CTGTGGATAA CCGTATTACC GCCTTTGAGT
     6601 GAGCTGATAC CGCTCGCCGC AGCCGAACGA CCGAGCGCAG CGAGTCAGTG AGCGAGGAAG
     6661 CGGAAGAGCG CCTGATGCGG TATTTTCTCC TTACGCATCT GTGCGGTATT TCACACCGCA
     6721 TATATGGTGC ACTCTCAGTA CAATCTGCTC TGATGCCGCA TAGTTAAGCC AGTATACACT
     6781 CCGCTATCGC TACGTGACTG GGTCATGGCT GCGCCCCGAC ACCCGCCAAC ACCCGCTGAC
     6841 GCGCCCTGAC GGGCTTGTCT GCTCCCGGCA TCCGCTTACA GACAAGCTGT GACCGTCTCC
     6901 GGGAGCTGCA TGTGTCAGAG GTTTTCACCG TCATCACCGA AACGCGCGAG GCAGCTGCGG
     6961 TAAAGCTCAT CAGCGTGGTC GTGAAGCGAT TCACAGATGT CTGCCTGTTC ATCCGCGTCC
     7021 AGCTCGTTGA GTTTCTCCAG AAGCGTTAAT GTCTGGCTTC TGATAAAGCG GGCCATGTTA
     7081 AGGGCGGTTT TTTCCTGTTT GGTCACTGAT GCCTCCGTGT AAGGGGGATT TCTGTTCATG
     7141 GGGGTAATGA TACCGATGAA ACGAGAGAGG ATGCTCACGA TACGGGTTAC TGATGATGAA
     7201 CATGCCCGGT TACTGGAACG TTGTGAGGGT AAACAACTGG CGGTATGGAT GCGGCGGGAC
     7261 CAGAGAAAAA TCACTCAGGG TCAATGCCAG CGCTTCGTTA ATACAGATGT AGGTGTTCCA
     7321 CAGGGTAGCC AGCAGCATCC TGCGATGCAG ATCCGGAACA TAATGGTGCA GGGCGCTGAC
     7381 TTCCGCGTTT CCAGACTTTA CGAAACACGG AAACCGAAGA CCATTCATGT TGTTGCTCAG
     7441 GTCGCAGACG TTTTGCAGCA GCAGTCGCTT CACGTTCGCT CGCGTATCGG TGATTCATTC
     7501 TGCTAACCAG TAAGGCAACC CCGCCAGCCT AGCCGGGTCC TCAACGACAG GAGCACGATC
     7561 ATGCTAGTCA TGCCCCGCGC CCACCGGAAG GAGCTGACTG GGTTGAAGGC TCTCAAGGGC
     7621 ATCGGTCGAG ATCCCGGTGC CTAATGAGTG AGCTAACTTA CATTAATTGC GTTGCGCTCA
     7681 CTGCCCGCTT TCCAGTCGGG AAACCTGTCG TGCCAGCTGC ATTAATGAAT CGGCCAACGC
     7741 GCGGGGAGAG GCGGTTTGCG TATTGGGCGC CAGGGTGGTT TTTCTTTTCA CCAGTGAGAC
     7801 GGGCAACAGC TGATTGCCCT TCACCGCCTG GCCCTGAGAG AGTTGCAGCA AGCGGTCCAC
     7861 GCTGGTTTGC CCCAGCAGGC GAAAATCCTG TTTGATGGTG GTTAACGGCG GGATATAACA
     7921 TGAGCTGTCT TCGGTATCGT CGTATCCCAC TACCGAGATG TCCGCACCAA CGCGCAGCCC
     7981 GGACTCGGTA ATGGCGCGCA TTGCGCCCAG CGCCATCTGA TCGTTGGCAA CCAGCATCGC
     8041 AGTGGGAACG ATGCCCTCAT TCAGCATTTG CATGGTTTGT TGAAAACCGG ACATGGCACT
     8101 CCAGTCGCCT TCCCGTTCCG CTATCGGCTG AATTTGATTG CGAGTGAGAT ATTTATGCCA
     8161 GCCAGCCAGA CGCAGACGCG CCGAGACAGA ACTTAATGGG CCCGCTAACA GCGCGATTTG
     8221 CTGGTGACCC AATGCGACCA GATGCTCCAC GCCCAGTCGC GTACCGTCTT CATGGGAGAA
     8281 AATAATACTG TTGATGGGTG TCTGGTCAGA GACATCAAGA AATAACGCCG GAACATTAGT
     8341 GCAGGCAGCT TCCACAGCAA TGGCATCCTG GTCATCCAGC GGATAGTTAA TGATCAGCCC
     8401 ACTGACGCGT TGCGCGAGAA GATTGTGCAC CGCCGCTTTA CAGGCTTCGA CGCCGCTTCG
     8461 TTCTACCATC GACACCACCA CGCTGGCACC CAGTTGATCG GCGCGAGATT TAATCGCCGC
     8521 GACAATTTGC GACGGCGCGT GCAGGGCCAG ACTGGAGGTG GCAACGCCAA TCAGCAACGA
     8581 CTGTTTGCCC GCCAGTTGTT GTGCCACGCG GTTGGGAATG TAATTCAGCT CCGCCATCGC
     8641 CGCTTCCACT TTTTCCCGCG TTTTCGCAGA AACGTGGCTG GCCTGGTTCA CCACGCGGGA
     8701 AACGGTCTGA TAAGAGACAC CGGCATACTC TGCGACATCG TATAACGTTA CTGGTTTCAC
     8761 ATTCACCACC CTGAATTGAC TCTCTTCCGG GCGCTATCAT GCCATACCGC GAAAGGTTTT
     8821 GCGCCATTCG ATGGTGTCCG GGATCTCGAC GCTCTCCCTT ATGCGACTCC TGCATTAGGA
     8881 AGCAGCCCAG TAGTAGGTTG AGGCCGTTGA GCACCGCCGC CGCAAGGAAT GGTGCATGCA
     8941 AGGAGATGGC GCCCAACAGT CCCCCGGCCA CGGGGCCTGC CACCATACCC ACGCCGAAAC
     9001 AAGCGCTCAT GAGCCCGAAG TGGCGAGCCC GATCTTCCCC ATCGGTGATG TCGGCGATAT
     9061 AGGCGCCAGC AACCGCACCT GTGGCGCCGG TGATGCCGGC CACGATGCGT CCGGCGTAGA
     9121 GGATCGAGAT CGATCTCGAT CCCGCGAAAT TAATACGACT CACTATA
//
