!!NA_SEQUENCE 1.0 Plasmid pNEB205A Update 6/11/03 Features: 493- 146 lacZ alpha CDS (start 493, complementary strand) 543- 538 Plac promoter -10 sequence (TATGTT) 567- 562 Plac promoter -35 sequence (TTTACA) 599- 587 CAP protein binding site 393- 476 multiple cloning site (EcoRI-HindIII) 438- 449 top strand deleted by XbaI + N.BbvCIB digestion 430- 441 bottom strand deleted by XbaI + N.BbvCIB digestion 1479- 891 origin of replication (counterclockwise) (RNAII -35 to RNA/DNA switch point): 1297-1302 RNAI transcript promoter -35 sequence (TTGAAG) 1319-1324 RNAI transcript promoter -10 sequence (GCTACA) 1333-1440 RNAI transcript 1443- 891 RNAII transcript (complementary strand) 1458-1453 RNAII transcript promoter -10 sequence (CGTAAT) 1479-1474 RNAII transcript promoter -35 sequence (TTGAGA) 2510-1650 beta-lactamase (bla; amp-r) CDS (start 2510, complementary strand) 2510-2442 beta-lactamase signal peptide CDS (start 2510, complementary strand) pneb205a.seq Length: 2710 June 11, 2003 12:52 Type: N Check: 323 .. 1 TCGCGCGTTT CGGTGATGAC GGTGAAAACC TCTGACACAT GCAGCTCCCG 51 GAGACGGTCA CAGCTTGTCT GTAAGCGGAT GCCGGGAGCA GACAAGCCCG 101 TCAGGGCGCG TCAGCGGGTG TTGGCGGGTG TCGGGGCTGG CTTAACTATG 151 CGGCATCAGA GCAGATTGTA CTGAGAGTGC ACCATATGCG GTGTGAAATA 201 CCGCACAGAT GCGTAAGGAG AAAATACCGC ATCAGGCGCC ATTCGCCATT 251 CAGGCTGCGC AACTGTTGGG AAGGGCGATC GGTGCGGGCC TCTTCGCTAT 301 TACGCCAGCT GGCGAAAGGG GGATGTGCTG CAAGGCGATT AAGTTGGGTA 351 ACGCCAGGGT TTTCCCAGTC ACGACGTTGT AAAACGACGG CCAGTGAATT 401 CGAGCTCAGG CGCGCCTTAA TTAAGCTGAG GGAAAGTCTA GATGTCTCCT 451 CAGCGTTTAA ACCCTGCAGG AAGCTTGGCG TAATCATGGT CATAGCTGTT 501 TCCTGTGTGA AATTGTTATC CGCTCACAAT TCCACACAAC ATACGAGCCG 551 GAAGCATAAA GTGTAAAGCC TGGGGTGCCT AATGAGTGAG CTAACTCACA 601 TTAATTGCGT TGCGCTCACT GCCCGCTTTC CAGTCGGGAA ACCTGTCGTG 651 CCAGCTGCAT TAATGAATCG GCCAACGCGC GGGGAGAGGC GGTTTGCGTA 701 TTGGGCGCTC TTCCGCTTCC TCGCTCACTG ACTCGCTGCG CTCGGTCGTT 751 CGGCTGCGGC GAGCGGTATC AGCTCACTCA AAGGCGGTAA TACGGTTATC 801 CACAGAATCA GGGGATAACG CAGGAAAGAA CATGTGAGCA AAAGGCCAGC 851 AAAAGGCCAG GAACCGTAAA AAGGCCGCGT TGCTGGCGTT TTTCCATAGG 901 CTCCGCCCCC CTGACGAGCA TCACAAAAAT CGACGCTCAA GTCAGAGGTG 951 GCGAAACCCG ACAGGACTAT AAAGATACCA GGCGTTTCCC CCTGGAAGCT 1001 CCCTCGTGCG CTCTCCTGTT CCGACCCTGC CGCTTACCGG ATACCTGTCC 1051 GCCTTTCTCC CTTCGGGAAG CGTGGCGCTT TCTCATAGCT CACGCTGTAG 1101 GTATCTCAGT TCGGTGTAGG TCGTTCGCTC CAAGCTGGGC TGTGTGCACG 1151 AACCCCCCGT TCAGCCCGAC CGCTGCGCCT TATCCGGTAA CTATCGTCTT 1201 GAGTCCAACC CGGTAAGACA CGACTTATCG CCACTGGCAG CAGCCACTGG 1251 TAACAGGATT AGCAGAGCGA GGTATGTAGG CGGTGCTACA GAGTTCTTGA 1301 AGTGGTGGCC TAACTACGGC TACACTAGAA GAACAGTATT TGGTATCTGC 1351 GCTCTGCTGA AGCCAGTTAC CTTCGGAAAA AGAGTTGGTA GCTCTTGATC 1401 CGGCAAACAA ACCACCGCTG GTAGCGGTGG TTTTTTTGTT TGCAAGCAGC 1451 AGATTACGCG CAGAAAAAAA GGATCTCAAG AAGATCCTTT GATCTTTTCT 1501 ACGGGGTCTG ACGCTCAGTG GAACGAAAAC TCACGTTAAG GGATTTTGGT 1551 CATGAGATTA TCAAAAAGGA TCTTCACCTA GATCCTTTTA AATTAAAAAT 1601 GAAGTTTTAA ATCAATCTAA AGTATATATG AGTAAACTTG GTCTGACAGT 1651 TACCAATGCT TAATCAGTGA GGCACCTATC TCAGCGATCT GTCTATTTCG 1701 TTCATCCATA GTTGCCTGAC TCCCCGTCGT GTAGATAACT ACGATACGGG 1751 AGGGCTTACC ATCTGGCCCC AGTGCTGCAA TGATACCGCG AGACCCACGC 1801 TCACCGGCTC CAGATTTATC AGCAATAAAC CAGCCAGCCG GAAGGGCCGA 1851 GCGCAGAAGT GGTCCTGCAA CTTTATCCGC CTCCATCCAG TCTATTAATT 1901 GTTGCCGGGA AGCTAGAGTA AGTAGTTCGC CAGTTAATAG TTTGCGCAAC 1951 GTTGTTGCCA TTGCTACAGG CATCGTGGTG TCACGCTCGT CGTTTGGTAT 2001 GGCTTCATTC AGCTCCGGTT CCCAACGATC AAGGCGAGTT ACATGATCCC 2051 CCATGTTGTG CAAAAAAGCG GTTAGCTCCT TCGGTCCTCC GATCGTTGTC 2101 AGAAGTAAGT TGGCCGCAGT GTTATCACTC ATGGTTATGG CAGCACTGCA 2151 TAATTCTCTT ACTGTCATGC CATCCGTAAG ATGCTTTTCT GTGACTGGTG 2201 AGTACTCAAC CAAGTCATTC TGAGAATAGT GTATGCGGCG ACCGAGTTGC 2251 TCTTGCCCGG CGTCAATACG GGATAATACC GCGCCACATA GCAGAACTTT 2301 AAAAGTGCTC ATCATTGGAA AACGTTCTTC GGGGCGAAAA CTCTCAAGGA 2351 TCTTACCGCT GTTGAGATCC AGTTCGATGT AACCCACTCG TGCACCCAAC 2401 TGATCTTCAG CATCTTTTAC TTTCACCAGC GTTTCTGGGT GAGCAAAAAC 2451 AGGAAGGCAA AATGCCGCAA AAAAGGGAAT AAGGGCGACA CGGAAATGTT 2501 GAATACTCAT ACTCTTCCTT TTTCAATATT ATTGAAGCAT TTATCAGGGT 2551 TATTGTCTCA TGAGCGGATA CATATTTGAA TGTATTTAGA AAAATAAACA 2601 AATAGGGGTT CCGCGCACAT TTCCCCGAAA AGTGCCACCT GACGTCTAAG 2651 AAACCATTAT TATCATGACA TTAACCTATA AAAATAGGCG TATCACGAGG 2701 CCCTTTCGTC