!!NA_SEQUENCE 1.0 Plasmid pTWIN-MBP1 Update 7/17/03 Comments: The unique SpeI site at 6462 of pTWIN1 is not present in pTWIN-MBP1. 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) 5637-5654 T7 promoter (transcript start 5654 clockwise) 5637-5656 T7 universal primer sequence (NEB #S1248S) 5656-5680 lac operator 5710-5716 RBS for expression CDS 5725-7227 expression CDS (start 5725) 5725-5901 chitin binding domain (CBD) 5941-6402 Ssp DnaB intein 6403-7587 E. coli malE gene (SapI insert) 7588-8181 Mxe GyrA intein 8212-8370 chitin binding domain (CBD) 8390-8512 T7 Tphi transcription terminator ptwin-mbp1.seq Length: 8518 July 17, 2003 13:46 Type: N Check: 8707 .. 1 AACTACGTCA GGTGGCACTT TTCGGGGAAA TGTGCGCGGA ACCCCTATTT 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 GCCGGCATGC CGCCCTTTCG 4651 TCTTCAAGAA TTAATTCCCA ATTCCCCAGG CATCAAATAA AACGAAAGGC 4701 TCAGTCGAAA GACTGGGCCT TTCGTTTTAT CTGTTGTTTG TCGGTGAACG 4751 CTCTCCTGAG TAGGACAAAT CCGCCGGGAG CGGATTTGAA CGTTGCGAAG 4801 CAACGGCCCG GAGGGTGGCG GGCAGGACGC CCGCCATAAA CTGCCAGGAA 4851 TTAATTCCCC AGGCATCAAA TAAAACGAAA GGCTCAGTCG AAAGACTGGG 4901 CCTTTCGTTT TATCTGTTGT TTGTCGGTGA ACGCTCTCCT GAGTAGGACA 4951 AATCCGCCGG GAGCGGATTT GAACGTTGCG AAGCAACGGC CCGGAGGGTG 5001 GCGGGCAGGA CGCCCGCCAT AAACTGCCAG GAATTAATTC CCCAGGCATC 5051 AAATAAAACG AAAGGCTCAG TCGAAAGACT GGGCCTTTCG TTTTATCTGT 5101 TGTTTGTCGG TGAACGCTCT CCTGAGTAGG ACAAATCCGC CGGGAGCGGA 5151 TTTGAACGTT GCGAAGCAAC GGCCCGGAGG GTGGCGGGCA GGACGCCCGC 5201 CATAAACTGC CAGGAATTAA TTCCCCAGGC ATCAAATAAA ACGAAAGGCT 5251 CAGTCGAAAG ACTGGGCCTT TCGTTTTATC TGTTGTTTGT CGGTGAACGC 5301 TCTCCTGAGT AGGACAAATC CGCCGGGAGC GGATTTGAAC GTTGCGAAGC 5351 AACGGCCCGG AGGGTGGCGG GCAGGACGCC CGCCATAAAC TGCCAGGAAT 5401 TAATTCCCCA GGCATCAAAT AAAACGAAAG GCTCAGTCGA AAGACTGGGC 5451 CTTTCGTTTT ATCTGTTGTT TGTCGGTGAA CGCTCTCCTG AGTAGGACAA 5501 ATCCGCCGGG AGCGGATTTG AACGTTGCGA AGCAACGGCC CGGAGGGTGG 5551 CGGGCAGGAC GCCCGCCATA AACTGCCAGG AATTGGGGAT CGGAATTAAT 5601 TCCCGGTTTA AACCGGGGAT CTCGATCCCG CGAAATTAAT ACGACTCACT 5651 ATAGGGGAAT TGTGAGCGGA TAACAATTCC CCTCTAGAAA TAATTTTGTT 5701 TAACTTTAAG AAGGAGATAT ACATATGAAA ATCGAAGAAG GTAAACTGAC 5751 AAATCCTGGT GTATCCGCTT GGCAGGTCAA CACAGCTTAT ACTGCGGGAC 5801 AATTGGTCAC ATATAACGGC AAGACGTATA AATGTTTGCA GCCCCACACC 5851 TCCTTGGCAG GATGGGAACC ATCCAACGTT CCTGCCTTGT GGCAGCTTCA 5901 AAACAACGGT AACAACGGTC TCGAACTGCG CGAGTCCGGA GCTATCTCTG 5951 GCGATAGTCT GATCAGCCTG GCTAGCACAG GAAAAAGAGT TTCTATTAAA 6001 GATTTGTTAG ATGAAAAAGA TTTTGAAATA TGGGCAATTA ATGAACAGAC 6051 GATGAAGCTA GAATCAGCTA AAGTTAGTCG TGTATTTTGT ACTGGCAAAA 6101 AGCTAGTTTA TATTCTAAAA ACTCGACTAG GTAGAACTAT CAAGGCAACA 6151 GCAAATCATA GATTTTTAAC TATTGATGGT TGGAAAAGAT TAGATGAGCT 6201 ATCTTTAAAA GAGCATATTG CTCTACCCCG TAAACTAGAA AGCTCCTCTT 6251 TACAATTGTC ACCAGAAATA GAAAAGTTGT CTCAGAGTGA TATTTACTGG 6301 GACTCCATCG TTTCTATTAC GGAGACTGGA GTCGAAGAGG TTTTTGATTT 6351 GACTGTGCCA GGACCACATA ACTTTGTCGC GAATGACATC ATTGTACACA 6401 ACTGCCGCGC CATGGGCATC GAAGAAGGTA AACTGGTAAT CTGGATTAAC 6451 GGCGATAAAG GCTATAACGG TCTCGCTGAA GTCGGTAAGA AATTCGAGAA 6501 AGATACCGGA ATTAAAGTCA CCGTTGAGCA TCCGGATAAA CTGGAAGAGA 6551 AATTCCCACA GGTTGCGGCA ACTGGCGATG GCCCTGACAT TATCTTCTGG 6601 GCACACGACC GCTTTGGTGG CTACGCTCAA TCTGGCCTGT TGGCTGAAAT 6651 CACCCCGGAC AAAGCGTTCC AGGACAAGCT GTATCCGTTT ACCTGGGATG 6701 CCGTACGTTA CAACGGCAAG CTGATTGCTT ACCCGATCGC TGTTGAAGCG 6751 TTATCGCTGA TTTATAACAA AGATCTGCTG CCGAACCCGC CAAAAACCTG 6801 GGAAGAGATC CCGGCGCTGG ATAAAGAACT GAAAGCGAAA GGTAAGAGCG 6851 CGCTGATGTT CAACCTGCAA GAACCGTACT TCACCTGGCC GCTGATTGCT 6901 GCTGACGGGG GTTATGCGTT CAAGTATGAA AACGGCAAGT ACGACATTAA 6951 AGACGTGGGC GTGGATAACG CTGGCGCGAA AGCGGGTCTG ACCTTCCTGG 7001 TTGACCTGAT TAAAAACAAA CACATGAATG CAGACACCGA TTACTCCATC 7051 GCAGAAGCTG CCTTTAATAA AGGCGAAACA GCGATGACCA TCAACGGCCC 7101 GTGGGCATGG TCCAACATCG ACACCAGCAA AGTGAATTAT GGTGTAACGG 7151 TACTGCCGAC CTTCAAGGGT CAACCATCCA AACCGTTCGT TGGCGTGCTG 7201 AGCGCAGGTA TTAACGCCGC CAGTCCGAAC AAAGAGCTGG CAAAAGAGTT 7251 CCTCGAAAAC TATCTGCTGA CTGATGAAGG TCTGGAAGCG GTTAATAAAG 7301 ACAAACCGCT GGGTGCCGTA GCGCTGAAGT CTTACGAGGA AGAGTTGGCG 7351 AAAGATCCAC GTATTGCCGC CACTATGGAA AACGCCCAGA AAGGTGAAAT 7401 CATGCCGAAC ATCCCGCAGA TGTCCGCTTT CTGGTATGCC GTGCGTACTG 7451 CGGTGATCAA CGCCGCCAGC GGTCGTCAGA CTGTCGATGA AGCCCTGAAA 7501 GACGCGCAGA CTAATTCGAG CTCGAACAAC AACAACAATA ACAATAACAA 7551 CAACCTCGGG ATCGAGGGAA GGGGTACGCT CGAGATGTGC ATCACGGGAG 7601 ATGCTCTAGT TGCCCTACCC GAGGGCGAGT CGGTACGCAT CGCCGACATC 7651 GTGCCGGGTG CGCGGCCCAA CAGTGACAAC GCCATCGACC TGAAAGTCCT 7701 TGACCGGCAT GGCAATCCCG TGCTCGCCGA CCGGCTGTTC CACTCCGGCG 7751 AGCATCCGGT GTACACGGTG CGTACGGTCG AAGGTCTGCG TGTGACGGGC 7801 ACCGCGAACC ACCCGTTGTT GTGTTTGGTC GACGTCGCCG GGGTGCCGAC 7851 CCTGCTGTGG AAGCTGATCG ACGAAATCAA GCCGGGCGAT TACGCGGTGA 7901 TTCAACGCAG CGCATTCAGC GTCGACTGTG CAGGTTTTGC CCGCGGAAAA 7951 CCCGAATTTG CGCCCACAAC CTACACAGTC GGCGTCCCTG GACTGGTGCG 8001 TTTCTTGGAA GCACACCACC GAGACCCGGA CGCCCAAGCT ATCGCCGACG 8051 AGCTGACCGA CGGGCGGTTC TACTACGCGA AAGTCGCCAG TGTCACCGAC 8101 GCCGGCGTGC AGCCGGTGTA TAGCCTTCGT GTCGACACGG CAGACCACGC 8151 GTTTATCACG AACGGGTTCG TCAGCCACGC TACTGGCCTC ACCGGTCTGA 8201 ACTCAGGCCT CACGACAAAT CCTGGTGTAT CCGCTTGGCA GGTCAACACA 8251 GCTTATACTG CGGGACAATT GGTCACATAT AACGGCAAGA CGTATAAATG 8301 TTTGCAGCCC CACACCTCCT TGGCAGGATG GGAACCATCC AACGTTCCTG 8351 CCTTGTGGCA GCTTCAATGA CTGCAGGAAG GGGATCCGGC TGCTAACAAA 8401 GCCCGAAAGG AAGCTGAGTT GGCTGCTGCC ACCGCTGAGC AATAACTAGC 8451 ATAACCCCTT GGGGCCTCTA AACGGGTCTT GAGGGGTTTT TTGCTGAAAG 8501 GAGGAACTAT ATCCGGAT