!!NA_SEQUENCE 1.0 Plasmid pTYB12 Update 3/26/04 Features: 140-1000 beta-lactamase (bla; amp-r) CDS (start 140) 140- 208 beta-lactamase signal peptide CDS (start 140) 1042-1555 M13 origin of replication (- +) 1666-2254 pMB1 origin of replication (clockwise) (RNAII -35 to RNA/DNA switch point) 2814-2623 rop CDS (start 2814, complementary strand) 4453-3371 lacI CDS (start 4453, complementary strand) 4840-4857 T7 promoter (transcript start 4840 clockwise) 4840-4859 T7 universal primer sequence (NEB #S1248S) 4859-4883 lac operator 4913-4919 RBS for expression CDS 4928-6562 expression CDS (start 4928) 4928-4972 malE fragment (translational start) 4973-6502 Sce VMA intein with embedded CBD 4973-5791 Sce VMA intein 5' region 5798-5950 chitin binding domain (CBD) 5963-6502 Sce VMA intein 3' region 6503-6559 multiple cloning site (NdeI-SmaI) 6362-6385 Intein forward primer sequence (NEB #S1263S) 6582-6704 T7 Tphi transcription terminator ptyb12.seq Length: 7415 March 26, 2004 15:01 Type: N Check: 747 .. 1 AGAAGGCCAT CCTGACGGAT GGCCTTTTTG CGTTTCTACA AACTCTTTTT 51 GTTTATTTTT CTAAATACAT TCAAATATGT ATCCGCTCAT GAGACAATAA 101 CCCTGATAAA TGCTTCAATA ATATTGAAAA AGGAAGAGTA TGAGTATTCA 151 ACATTTCCGT GTCGCCCTTA TTCCCTTTTT TGCGGCATTT TGCCTTCCTG 201 TTTTTGCTCA CCCAGAAACG CTGGTGAAAG TAAAAGATGC TGAAGATCAG 251 TTGGGTGCAC GAGTGGGTTA CATCGAACTG GATCTCAACA GCGGTAAGAT 301 CCTTGAGAGT TTTCGCCCCG AAGAACGTTC TCCAATGATG AGCACTTTTA 351 AAGTTCTGCT ATGTGGCGCG GTATTATCCC GTGTTGACGC CGGGCAAGAG 401 CAACTCGGTC GCCGCATACA CTATTCTCAG AATGACTTGG TTGAGTACTC 451 ACCAGTCACA GAAAAGCATC TTACGGATGG CATGACAGTA AGAGAATTAT 501 GCAGTGCTGC CATAACCATG AGTGATAACA CTGCGGCCAA CTTACTTCTG 551 ACAACGATCG GAGGACCGAA GGAGCTAACC GCTTTTTTGC ACAACATGGG 601 GGATCATGTA ACTCGCCTTG ATCGTTGGGA ACCGGAGCTG AATGAAGCCA 651 TACCAAACGA CGAGCGTGAC ACCACGATGC CTGTAGCAAT GGCAACAACG 701 TTGCGCAAAC TATTAACTGG CGAACTACTT ACTCTAGCTT CCCGGCAACA 751 ATTAATAGAC TGGATGGAGG CGGATAAAGT TGCAGGACCA CTTCTGCGCT 801 CGGCCCTTCC GGCTGGCTGG TTTATTGCTG ATAAATCTGG AGCCGGTGAG 851 CGTGGGTCTC GCGGTATCAT TGCAGCACTG GGGCCAGATG GTAAGCCCTC 901 CCGTATCGTA GTTATCTACA CGACGGGGAG TCAGGCAACT ATGGATGAAC 951 GAAATAGACA GATCGCTGAG ATAGGTGCCT CACTGATTAA GCATTGGTAA 1001 CTGTCAGACC AAGTTTACTC ATATATACTT TAGATTGATT TACCCCGGTT 1051 GATAATCAGA AAAGCCCCAA AAACAGGAAG ATTGTATAAG CAAATATTTA 1101 AATTGTAAAC GTTAATATTT TGTTAAAATT CGCGTTAAAT TTTTGTTAAA 1151 TCAGCTCATT TTTTAACCAA TAGGCCGAAA TCGGCAAAAT CCCTTATAAA 1201 TCAAAAGAAT AGCCCGAGAT AGGGTTGAGT GTTGTTCCAG TTTGGAACAA 1251 GAGTCCACTA TTAAAGAACG TGGACTCCAA CGTCAAAGGG CGAAAAACCG 1301 TCTATCAGGG CGATGGCCCA CTACGTGAAC CATCACCCAA ATCAAGTTTT 1351 TTGGGGTCGA GGTGCCGTAA AGCACTAAAT CGGAACCCTA AAGGGAGCCC 1401 CCGATTTAGA GCTTGACGGG GAAAGCCGGC GAACGTGGCG AGAAAGGAAG 1451 GGAAGAAAGC GAAAGGAGCG GGCGCTAGGG CGCTGGCAAG TGTAGCGGTC 1501 ACGCTGCGCG TAACCACCAC ACCCGCCGCG CTTAATGCGC CGCTACAGGG 1551 CGCGTAAAAG GATCTAGGTG AAGATCCTTT TTGATAATCT CATGACCAAA 1601 ATCCCTTAAC GTGAGTTTTC GTTCCACTGA GCGTCAGACC CCGTAGAAAA 1651 GATCAAAGGA TCTTCTTGAG ATCCTTTTTT TCTGCGCGTA ATCTGCTGCT 1701 TGCAAACAAA AAAACCACCG CTACCAGCGG TGGTTTGTTT GCCGGATCAA 1751 GAGCTACCAA CTCTTTTTCC GAAGGTAACT GGCTTCAGCA GAGCGCAGAT 1801 ACCAAATACT GTCCTTCTAG TGTAGCCGTA GTTAGGCCAC CACTTCAAGA 1851 ACTCTGTAGC ACCGCCTACA TACCTCGCTC TGCTAATCCT GTTACCAGTG 1901 GCTGCTGCCA GTGGCGATAA GTCGTGTCTT ACCGGGTTGG ACTCAAGACG 1951 ATAGTTACCG GATAAGGCGC AGCGGTCGGG CTGAACGGGG GGTTCGTGCA 2001 CACAGCCCAG CTTGGAGCGA ACGACCTACA CCGAACTGAG ATACCTACAG 2051 CGTGAGCTAT GAGAAAGCGC CACGCTTCCC GAAGGGAGAA AGGCGGACAG 2101 GTATCCGGTA AGCGGCAGGG TCGGAACAGG AGAGCGCACG AGGGAGCTTC 2151 CAGGGGGAAA CGCCTGGTAT CTTTATAGTC CTGTCGGGTT TCGCCACCTC 2201 TGACTTGAGC GTCGATTTTT GTGATGCTCG TCAGGGGGGC GGAGCCTATG 2251 GAAAAACGCC AGCAACGCGG CCTTTTTACG GTTCCTGGCC TTTTGCTGGC 2301 CTTTTGCTCA CATGTTCTTT CCTGCGTTAT CCCCTGATTC TGTGGATAAC 2351 CGTATTACCG CCTTTGAGTG AGCTGATACC GCTCGCCGCA GCCGAACGAC 2401 CGAGCGCAGC GAGTCAGTGA GCGAGGAAGC TATGGTGCAC TCTCAGTACA 2451 ATCTGCTCTG ATGCCGCATA GTTAAGCCAG TATACACTCC GCTATCGCTA 2501 CGTGACTGGG TCATGGCTGC GCCCCGACAC CCGCCAACAC CCGCTGACGC 2551 GCCCTGACGG GCTTGTCTGC TCCCGGCATC CGCTTACAGA CAAGCTGTGA 2601 CCGTCTCCGG GAGCTGCATG TGTCAGAGGT TTTCACCGTC ATCACCGAAA 2651 CGCGCGAGGC AGCTGCGGTA AAGCTCATCA GCGTGGTCGT GCAGCGATTC 2701 ACAGATGTCT GCCTGTTCAT CCGCGTCCAG CTCGTTGAGT TTCTCCAGAA 2751 GCGTTAATGT CTGGCTTCTG ATAAAGCGGG CCATGTTAAG GGCGGTTTTT 2801 TCCTGTTTGG TCACTGATGC CTCCGTGTAA GGGGGATTTC TGTTCATGGG 2851 GGTAATGATA CCGATGAAAC GAGAGAGGAT GCTCACGATA CGGGTTACTG 2901 ATGATGAACA TGCCCGGTTA CTGGAACGTT GTGAGGGTAA ACAACTGGCG 2951 GTATGGATGC GGCGGGACCA GAGAAAAATC ACTCAGGGTC AATGCCAGCC 3001 GAACGCCAGC AAGACGTAGC CCAGCGCGTC GGCCGCCATG CCGGCGATAA 3051 TGGCCTGCTT CTCGCCGAAA CGTTTGGTGG CGGGACCAGT GACGAAGGCT 3101 TGAGCGAGGG CGTGCAAGAT TCCGAATACC GCAAGCGACA GGCCGATCAT 3151 CGTCGCGCTC CAGCGAAAGC GGTCCTCGCC GAAAATGACC CAGAGCGCTG 3201 CCGGCACCTG TCCTACGAGT TGCATGATAA AGAAGACAGT CATAAGTGCG 3251 GCGACGATAG TCATGCCCCG CGCCCACCGG AAGGAGCTGA CTGGGTTGAA 3301 GGCTCTCAAG GGCATCGGTC GAGATCCCGG TGCCTAATGA GTGAGCTAAC 3351 TTACATTAAT TGCGTTGCGC TCACTGCCCG CTTTCCAGTC GGGAAACCTG 3401 TCGTGCCAGC TGCATTAATG AATCGGCCAA CGCGCGGGGA GAGGCGGTTT 3451 GCGTATTGGG CGCCAGGGTG GTTTTTCTTT TCACCAGTGA GACGGGCAAC 3501 AGCTGATTGC CCTTCACCGC CTGGCCCTGA GAGAGTTGCA GCAAGCGGTC 3551 CACGCTGGTT TGCCCCAGCA GGCGAAAATC CTGTTTGATG GTGGTTAACG 3601 GCGGGATATA ACATGAGCTG TCTTCGGTAT CGTCGTATCC CACTACCGAG 3651 ATATCCGCAC CAACGCGCAG CCCGGACTCG GTAATGGCGC GCATTGCGCC 3701 CAGCGCCATC TGATCGTTGG CAACCAGCAT CGCAGTGGGA ACGATGCCCT 3751 CATTCAGCAT TTGCATGGTT TGTTGAAAAC CGGACATGGC ACTCCAGTCG 3801 CCTTCCCGTT CCGCTATCGG CTGAATTTGA TTGCGAGTGA GATATTTATG 3851 CCAGCCAGCC AGACGCAGAC GCGCCGAGAC AGAACTTAAT GGGCCCGCTA 3901 ACAGCGCGAT TTGCTGGTGA CCCAATGCGA CCAGATGCTC CACGCCCAGT 3951 CGCGTACCGT CTTCATGGGA GAAAATAATA CTGTTGATGG GTGTCTGGTC 4001 AGAGACATCA AGAAATAACG CCGGAACATT AGTGCAGGCA GCTTCCACAG 4051 CAATGGCATC CTGGTCATCC AGCGGATAGT TAATGATCAG CCCACTGACG 4101 CGTTGCGCGA GAAGATTGTG CACCGCCGCT TTACAGGCTT CGACGCCGCT 4151 TCGTTCTACC ATCGACACCA CCACGCTGGC ACCCAGTTGA TCGGCGCGAG 4201 ATTTAATCGC CGCGACAATT TGCGACGGCG CGTGCAGGGC CAGACTGGAG 4251 GTGGCAACGC CAATCAGCAA CGACTGTTTG CCCGCCAGTT GTTGTGCCAC 4301 GCGGTTGGGA ATGTAATTCA GCTCCGCCAT CGCCGCTTCC ACTTTTTCCC 4351 GCGTTTTCGC AGAAACGTGG CTGGCCTGGT TCACCACGCG GGAAACGGTC 4401 TGATAAGAGA CACCGGCATA CTCTGCGACA TCGTATAACG TTACTGGTTT 4451 CACATTCACC ACCCTGAATT GACTCTCTTC CGGGCGCTAT CATGCCATAC 4501 CGCGAAAGGT TTTGCGCCAT TCGATGGTGT CCGGGATCTC GACGCTCTCC 4551 CTTATGCGAC TCCTGCATTA GGAAGCAGCC CAGTAGTAGG TTGAGGCCGT 4601 TGAGCACCGC CGCCGCAAGG AATGGTGCAT GCAAGGAGAT GGCGCCCAAC 4651 AGTCCCCCGG CCACGGGGCC TGCCACCATA CCCACGCCGA AACAAGCGCT 4701 CATGAGCCCG AAGTGGCGAG CCCGATCTTC CCCATCGGTG ATGTCGGCGA 4751 TATAGGCGCC AGCAACCGCA CCTGTGGCGC CGGTGATGCC GGCCACGATG 4801 CGTCCGGCGT AGAGGATCGA GATCTCGATC CCGCGAAATT AATACGACTC 4851 ACTATAGGGG AATTGTGAGC GGATAACAAT TCCCCTCTAG AAATAATTTT 4901 GTTTAACTTT AAGAAGGAGA TATACTTATG AAAATCGAAG AAGGTAAACT 4951 GGTAATCGGT TCCCTGGAGG GTTGCTTTGC CAAGGGTACC AATGTTTTAA 5001 TGGCGGATGG GTCTATTGAA TGTATTGAAA ACATTGAGGT TGGTAATAAG 5051 GTCATGGGTA AAGATGGCAG ACCTCGTGAG GTAATTAAAT TGCCCAGAGG 5101 AAGAGAAACT ATGTACAGCG TCGTGCAGAA AAGTCAGCAC AGAGCCCACA 5151 AAAGTGACTC AAGTCGTGAA GTGCCAGAAT TACTCAAGTT TACGTGTAAT 5201 GCGACCCATG AGTTGGTTGT TAGAACACCT CGTAGTGTCC GCCGTTTGTC 5251 TCGTACCATT AAGGGTGTCG AATATTTTGA AGTTATTACT TTTGAGATGG 5301 GCCAAAAGAA AGCCCCCGAC GGTAGAATTG TTGAGCTTGT CAAGGAAGTT 5351 TCAAAGAGCT ACCCAATATC TGAGGGGCCT GAGAGAGCCA ACGAATTAGT 5401 AGAATCCTAT AGAAAGGCTT CAAATAAAGC TTATTTTGAG TGGACTATTG 5451 AGGCCAGAGA TCTTTCTCTG TTGGGTTCCC ATGTTCGTAA AGCTACCTAC 5501 CAGACTTACG CTCCAATTCT TTATGAGAAT GACCACTTTT TCGACTACAT 5551 GCAAAAAAGT AAGTTTCATC TCACCATTGA AGGTCCAAAA GTACTTGCTT 5601 ATTTACTTGG TTTATGGATT GGTGATGGAT TGTCTGACAG GGCAACTTTT 5651 TCGGTTGATT CCAGAGATAC TTCTTTGATG GAACGTGTTA CTGAATATGC 5701 TGAAAAGTTG AATTTGTGCG CCGAGTATAA GGACAGAAAA GAACCACAAG 5751 TTGCCAAAAC TGTTAATTTG TACTCTAAAG TTGTCAGAGG TGCTAGCACA 5801 AATCCTGGTG TATCCGCTTG GCAGGTCAAC ACAGCTTATA CTGCGGGACA 5851 ATTGGTCACA TATAACGGCA AGACGTATAA ATGTTTGCAG CCCCACACCT 5901 CCTTGGCAGG ATGGGAACCA TCCAACGTTC CTGCCTTGTG GCAGCTTCAA 5951 GGTGGCCATG GTGGTATTCG CAATAATCTT AATACTGAGA ATCCATTATG 6001 GGACGCTATT GTTGGCTTAG GATTCTTGAA GGACGGTGTC AAAAATATTC 6051 CTTCTTTCTT GTCTACGGAC AATATCGGTA CTCGTGAAAC ATTTCTTGCT 6101 GGTCTAATTG ATTCTGATGG CTATGTTACT GATGAGCATG GTATTAAAGC 6151 AACAATAAAG ACAATTCATA CTTCTGTCAG AGATGGTTTG GTTTCCCTTG 6201 CTCGTTCTTT AGGCTTAGTA GTCTCGGTTA ACGCAGAACC TGCTAAGGTT 6251 GACATGAATG TCACCAAACA TAAAATTAGT TATGCTATTT ATATGTCTGG 6301 TGGAGATGTT TTGCTTAACG TTCTTTCGAA GTGTGCCGGC TCTAAAAAAT 6351 TCAGGCCTGC TCCCGCCGCT GCTTTTGCAC GTGAGTGCCG CGGATTTTAT 6401 TTCGAGTTAC AAGAATTGAA GGAAGACGAT TATTATGGGA TTACTTTATC 6451 TGATGATTCT GATCATCAGT TTTTGCTTGG ATCCCAGGTT GTTGTACAGA 6501 ATGCTGGTCA TATGACTAGT TCGCGAGTCG ACGGCGGCCG CGAATTCCTC 6551 GAGCCCGGGT GACTGCAGGA AGGGGATCCG GCTGCTAACA AAGCCCGAAA 6601 GGAAGCTGAG TTGGCTGCTG CCACCGCTGA GCAATAACTA GCATAACCCC 6651 TTGGGGCCTC TAAACGGGTC TTGAGGGGTT TTTTGCTGAA AGGAGGAACT 6701 ATATCCGGAT ATCCCGCAAG AGGCCCGGCA GTACCGGCAT AACCAAGCCT 6751 ATGCCTACAG CATCCAGGGT GACGGTGCCG AGGATGACGA TGAGCGCATT 6801 GTTAGATTTC ATACACGGTG CCTGACTGCG TTAGCAATTT AACTGTGATA 6851 AACTACCGCA TTAAAGCTTG GCACTGGCCG TCGTTTTACA ACGTCGTGAC 6901 TGGGAAAACC CTGGCGTTAC CCAACTTAAT CGCCTTGCAG CACATCCCCC 6951 TTTCGCCAGC TGGCGTAATA GCGAAGAGGC CCGCACCGAT CGCCCTTCCC 7001 AACAGTTGCG CAGCCTGAAT GGCGAATGGC AGCTTGGCTG TTTTGGCGGA 7051 TGAGATAAGA TTTTCAGCCT GATACAGATT AAATCAGAAC GCAGAAGCGG 7101 TCTGATAAAA CAGAATTTGC CTGGCGGCAG TAGCGCGGTG GTCCCACCTG 7151 ACCCCATGCC GAACTCAGAA GTGAAACGCC GTAGCGCCGA TGGTAGTGTG 7201 GGGTCTCCCC ATGCGAGAGT AGGGAACTGC CAGGCATCAA ATAAAACGAA 7251 AGGCTCAGTC GAAAGACTGG GCCTTTCGTT TTATCTGTTG TTTGTCGGTG 7301 AACGCTCTCC TGAGTAGGAC AAATCCGCCG GGAGCGGATT TGAACGTTGC 7351 GAAGCAACGG CCCGGAGGGT GGCGGGCAGG ACGCCCGCCA TAAACTGCCA 7401 GGCATCAAAT TAAGC