pALTERŪ-Ex2 Vector(a,b) Sequence This vector can be obtained from Promega Corporation, Madison, WI. Call one of the following numbers for order or technical information. Order or Technical 1-800-356-9526 Outside U.S. 608-274-4330 pALTERŪ-Ex2 Vector sequence reference points: Base pairs 5835 T7 RNA polymerase transcription initiation site 1 SP6 RNA polymerase transcription initiation site 177 tac transcription initiation site 113 T7 RNA polymerase promoter (-17 to +3) 5819-3 SP6 RNA polymerase promoter (-17 to +3) 175-194 tac promoter 78-150 T7 translation start codon 37 tac and SP6 translation start codons 66,70 multiple cloning region 5-78 lacZ start codon N\A lac operon sequences N\A lac operator N\A Beta-lactamase (AmpR) coding region N\A TetR gene coding region 719-1906 Chloramphenicol acetyltransferase (CmR) coding region 2938-3596 phage f1 region 5200-5655 binding site of pUC/M13 Forward Sequencing primer 5776-5792 binding site of pUC/M13 Reverse Sequencing primer 212-228 binding site of Ampicillin Repair Oligonucleotide N\A binding site of Tetracycline knockout Oligonucleotide 802-832 binding site of Chloramphenicol Repair Oligonucleotide 3372-3396 NOTE: This vector is tetracycline resistant and chloramphenicol sensitive as supplied by Promega. After mutagenesis with the Chloramphenicol Repair Oligonucleotide it will become chloramphenicol resistant and after mutagenesis with the Tetracycline Repair Oligonucleotide it will become tetracycline sensitive. Use the T7 or pUC/M13 Forward Primer to sequence ssDNA produced by the pALTERŪ-Ex2 Vector. pALTER is a trademark of Promega Corporation and is registered with the U.S. Patent and Trademark Office. (a)U.S Pat. No. 5,955,363 has been issued to Promega Corporation for a vector for in vitro mutagenesis and use thereof. (b)U.S. Pat. No. 4,766,072 has been issued to Promega Corporation for transcription vectors having two different bacteriophage RNA polymerase promoter sequences separated by a series of unique restriction sites into which foreign DNA can be inserted. .. 1 GGGCGAATTC GTTAACTTTA AGAAGGAGAT ATACCCATGG AGGCCTCTAG 51 ACTGCAGCGG CCGCATGCAT ATGGATCCTG TTTCCTGTGT GAAATTGTTA 101 TCCGCTCACA ATTCCACACA TTATACGAGC CGATGATTAA TTGTCAACAG 151 GGGGATGGGG AGTAAGCTTG AGTATTCTAT AGTGTCACCT AAATAGCTTG 201 GCGTAATCAT GGTCATAGCT GTTTCCTGTG TGAAATTGTT ATCCGCTCAC 251 AATTCCACAC AACATACGAG CCGGAAGCAT AAAGTGTAAA GCCTGGGGTG 301 CCTAATGAGT GAGCTAACTC ACATTAATTG CGTTGCGCTC ACTGCCCGCT 351 TTCCAGTCGG GAAACCTGTC GTGCCAGCTG CATTAATGAA TCGGCCAACG 401 CGCGGGGAGA GGCGGTTTGC GTATTGGGCG CTCTTCCGCT TCCTCGCTCA 451 CTGACTCGCT GCGCTCGGTC GTTCGGCTGC GGCGAGCGGT ATCAGCTCAC 501 TCAAAGGCGG TAATACGGTT ATCCACAGAA TCAGGGGATA ACGCAGGAAA 551 GAACATGAAT TAATTCTCAT GTTTGACAGC TTATCATCGG ATAACCCCTT 601 GGGGCCTCTA AACGGGTCTT GAGGGGTTTT TTGTTCTCAT GTTTGACAGC 651 TTATCTTCGA TTAGCTTTAA TGCGGTAGTT TATCACAGTT AAATTGCTAA 701 CGCAGTCAGG CACCGTGTAT GAAATCTAAC AATGCGCTCA TCGTCATCCT 751 CGGCACCGTC ACCCTGGATG CTGTAGGCAT AGGCTTGGTT ATGCCGGTAC 801 TGCCGGGCCT CTTGCGGGAT ATCGTCCATT CCGACAGCAT CGCCAGTCAC 851 TATGGCGTGC TGCTAGCGCT ATATGCGTTG ATGCAATTTC TATGCGCACC 901 CGTTCTCGGA GCACTGTCCG ACCGCTTTGG CCGCCGCCCA GTCCTGCTCG 951 CTTCGCTACT TGGAGCCACT ATCGACTACG CGATCATGGC GACCACACCC 1001 GTCCTGTGGA TTCTCTACGC CGGACGCATC GTGGCCGGCA TCACCGGCGC 1051 CACAGGTGCG GTTGCTGGCG CCTATATCGC CGACATCACC GATGGGGAAG 1101 ATCGGGCTCG CCACTTCGGG CTCATGAGCG CTTGTTTCGG CGTGGGTATG 1151 GTGGCAGGCC CCGTGGCCGG GGGACTGTTG GGCGCCATCT CCTTACATGC 1201 ACCATTCCTT GCGGCGGCGG TGCTCAACGG CCTCAACCTA CTACTGGGCT 1251 GCTTCCTAAT GCAGGAGTCG CATAAGGGAG AGCGCCGACC CATGCCCTTG 1301 AGAGCCTTCA ACCCAGTCAG CTCCTTCCGG TGGGCGCGGG GCATGACTAT 1351 CGTCGCCGCA CTTATGACTG TCTTCTTTAT CATGCAACTC GTAGGACAGG 1401 TGCCGGCAGC GCTCTGGGTC ATTTTCGGCG AGGACCGCTT TCGCTGGAGC 1451 GCGACGATGA TCGGCCTGTC GCTTGCGGTA TTCGGAATCT TGCACGCCCT 1501 CGCTCAAGCC TTCGTCACTG GTCCCGCCAC CAAACGTTTC GGCGAGAAGC 1551 AGGCCATTAT CGCCGGCATG GCGGCCGACG CGCTGGGCTA CGTCTTGCTG 1601 GCGTTCGCGA CGCGAGGCTG GATGGCCTTC CCCATTATGA TTCTTCTCGC 1651 TTCCGGCGGC ATCGGGATGC CCGCGTTGCA GGCCATGCTG TCCAGGCAGG 1701 TAGATGACGA CCATCAGGGA CAGCTTCAAG GATCGCTCGC GGCTCTTACC 1751 AGCCTAACTT CGATCATTGG ACCGCTGATC GTCACGGCGA TTTATGCCGC 1801 CTCGGCGAGC ACATGGAACG GGTTGGCATG GATTGTAGGC GCCGCCCTAT 1851 ACCTTGTCTG CCTCCCCGCG TTGCGTCGCG GTGCATGGAG CCGGGCCACC 1901 TCGACCTGAA TGGAAGCCGG CGGCACCTCG CTAACGGATT CACCACTCCA 1951 AGAATTGGAG CCAATCAATT CTTGCGGAGA ACTGTGAATG CGCAAACCAA 2001 CCCTTGGCAG AACATATCCA TCGCGTCCGC CATCTCCAGC AGCCGCACGC 2051 GGCGCATCTC GGGCAGCGTT GGGTCCTGGC CACGGGTGCG CATGATCGTG 2101 CTCCTGTCGT TGAGGACCCG GCTAGGCTGG CGGGGTTGCC TTACTGGTTA 2151 GCAGAATGAA TCACCGATAC GCGAGCGAAC GTGAAGCGAC TGCTGCTGCA 2201 AAACGTCTGC GACCTGAGCA ACAACATGAA TGGTCTTCGG TTTCCGTGTT 2251 TCGTAAAGTC TGGAAACGCG GAAGTCCCCT ACGTGCTGCT GAAGTTGCCC 2301 GCAACAGAGA GTGGAACCAA CCGGTGATAC CACGATACTA TGACTGAGAG 2351 TCAACGCCAT GAGCGGCCTC ATTTCTTATT CTGAGTTACA ACAGTCCGCA 2401 CCGCTGCCGG TAGCTCCTTC CGGTGGGCGC GGGGCATGAC TATCGTCGCC 2451 GCACTTATGA CTGTCTTCTT TATCATGCAA CTCGTAGGAC AGGTGCCGGC 2501 AGCGCCCAAC AGTCCCCCGG CCACGGGGCC TGCCACCATA CCCACGCCGA 2551 AACAAGCGCC CTGCACCATT ATGTTCCGGA TCTGCATCGC AGGATGCTGC 2601 TGGCTACCCT GTGGAACACC TACATCTGTA TTAACGAAGC GCTAACCGTT 2651 TTTATCAGGC TCTGGGAGGC AGAATAAATG ATCATATCGT CAATTATTAC 2701 CTCCACGGGG AGAGCCTGAG CAAACTGGCC TCAGGCATTT GAGAAGCACA 2751 CGGTCACACT GCTTCCGGTA GTCAATAAAC CGGTAAACCA GCAATAGACA 2801 TAAGCGGCTA TTTAACGACC CTGCCCTGAA CCGACGACCG GGTCGAATTT 2851 GCTTTCGAAT TTCTGCCATT CATCCGCTTA TTATCACTTA TTCAGGCGTA 2901 GCAACCAGGC GTTTAAGGGC ACCAATAACT GCCTTAAAAA AATTACGCCC 2951 CGCCCTGCCA CTCATCGCAG TACTGTTGTA ATTCATTAAG CATTCTGCCG 3001 ACATGGAAGC CATCACAAAC GGCATGATGA ACCTGAATCG CCAGCGGCAT 3051 CAGCACCTTG TCGCCTTGCG TATAATATTT GCCCATCGTG AAAACGGGGG 3101 CGAAGAAGTT GTCCATATTG GCCACGTTTA AATCAAAACT GGTGAAACTC 3151 ACCCAGGGAT TGGCTGAGAC GAAAAACATA TTCTCAATAA ACCCTTTAGG 3201 GAAATAGGCC AGGTTTTCAC CGTAACACGC CACATCTTGC GAATATATGT 3251 GTAGAAACTG CCGGAAATCG TCGTGGTATT CACTCCAGAG CGATGAAAAC 3301 GTTTCAGTTT GCTCATGGAA AACGGTGTAA CAAGGGTGAA CACTATCCCA 3351 TATCACCAGC TCACCGTCTT TCATTGCCAT ACGGAACCGG ATGAGCATTC 3401 ATCAGGCGGG CAAGAATGTG AATAAAGGCC GGATAAAACT TGTGCTTATT 3451 TTTCTTTACG GTCTTTAAAA AGGCCGTAAT ATCCAGCTGA ACGGTCTGGT 3501 TATAGGTACA TTGAGCAACT GACTGAAATG CCTCAAAATG TTCTTTACGA 3551 TGCCATTGGG ATATATCAAC GGTGGTATAT CCAGTGATTT TTTTCTCCAT 3601 TTTAGCTTCC TTAGCTCCTG AAAATCTCGA TAACTCAAAA AATACGCCCG 3651 GTAGTGATCT TATTTCATTA TGGTGAAAGT TGGAACCTCT TACGTGCCGA 3701 TCAACGTCTC ATTTTCGCCA AAAGTTGGCC CAGGGCTTCC CGGTATCAAC 3751 AGGGACACCA GGATTTATTT ATTCTGCGAA GTGATCTTCC GTCACAGGTA 3801 TTTATTCGGC GCAAAGTGCG TCGGGTGATG CTGCCAACTT ACTGATTTAG 3851 TGTATGATGG TGTTTTTGAG GTGCTCCAGT GGCTTCTGTT TCTATCAGCT 3901 GTCCCTCCTG TTCAGCTACT GACGGGGTGG TGCGTAACGG CAAAAGCACC 3951 GCCGGACATC AGCGCTAGCG GAGTGTATAC TGGCTTACTA TGTTGGCACT 4001 GATGAGGGTG TCAGTGAAGT GCTTCATGTG GCAGGAGAAA AAAGGCTGCA 4051 CCGGTGCGTC AGCAGAATAT GTGATACAGG ATATATTCCG CTTCCTCGCT 4101 CACTGACTCG CTACGCTCGG TCGTTCGACT GCGGCGAGCG GAAATGGCTT 4151 ACGAACGGGG CGGAGATTTC CTGGAAGATG CCAGGAAGAT ACTTAACAGG 4201 GAAGTGAGAG GGCCGCGGCA AAGCCGTTTT TCCATAGGCT CCGCCCCCCT 4251 GACAAGCATC ACGAAATCTG ACGCTCAAAT CAGTGGTGGC GAAACCCGAC 4301 AGGACTATAA AGATACCAGG CGTTTCCCCC TGGCGGCTCC CTCGTGCGCT 4351 CTCCTGTTCC TGCCTTTCGG TTTACCGGTG TCATTCCGCT GTTATGGCCG 4401 CGTTTGTCTC ATTCCACGCC TGACACTCAG TTCCGGGTAG GCAGTTCGCT 4451 CCAAGCTGGA CTGTATGCAC GAACCCCCCG TTCAGTCCGA CCGCTGCGCC 4501 TTATCCGGTA ACTATCGTCT TGAGTCCAAC CCGGAAAGAC ATGCAAAAGC 4551 ACCACTGGCA GCAGCCACTG GTAATTGATT TAGAGGAGTT AGTCTTGAAG 4601 TCATGCGCCG GTTAAGGCTA AACTGAAAGG ACAAGTTTTG GTGACTGCGC 4651 TCCTCCAAGC CAGTTACCTC GGTTCAAAGA GTTGGTAGCT CAGAGAACCT 4701 TCGAAAAACC GCCCTGCAAG GCGGTTTTTT CGTTTTCAGA GCAAGAGATT 4751 ACGCGCAGAC CAAAACGATC TCAAGAAGAT CATCTTATTA ATCAGATAAA 4801 ATATTTCTAG ATTTCAGTGC AATTTATCTC TTCAAATGTA GCACCTGAAG 4851 TCAGCCCCAT ACGATATAAG TTGTAATTCT CATGTTTGAC AGCTTATCAT 4901 CTAAGAAACC ATTATTATCA TGACATTAAC CTATAAAAAT AGGCGTATCA 4951 CGAGGCCCTT TCGTCTCGCG CGTTTCGGTG ATGACGGTGA AAACCTCTGA 5001 CACATGCAGC TCCCGGAGAC GGTCACAGCT TGTCTGTAAG CGGATGCCGG 5051 GAGCAGACAA GCCCGTCAGG GCGCGTCAGC GGGTGTTGGC GGGTGTCGGG 5101 GCTGGCTTAA CTATGCGGCA TCAGAGCAGA TTGTAGAGAG TGCACCCTAT 5151 GCGGTGTGAA ATACCGCACA GATGCGTAAG GAGAAAATAC CGCATCAGGA 5201 AATTGTAAGC GTTAATATTT TGTTAAAATT CGCGTTAAAT TTTTGTTAAA 5251 TCAGCTCATT TTTTAACCAA TAGGCCGAAA TCGGCAAAAT CCCTTATAAA 5301 TCAAAAGAAT AGACCGAGAT AGGGTTGAGT GTTGTTCCAG TTTGGAACAA 5351 GAGTCCACTA TTAAAGAACG TGGACTCCAA CGTCAAAGGG CGAAAAACCG 5401 TCTATCAGGG CGATGGCCCA CTACGTGAAC CATCACCCTA ATCAAGTTTT 5451 TTGGGGTCGA GGTGCCGTAA AGCACTAAAT CGGAACCCTA AAGGGAGCCC 5501 CCGATTTAGA GCTTGACGGG GAAAGCCGGC GAACGTGGCG AGAAAGGAAG 5551 GGAAGAAAGC GAAAGGAGCG GGCGCTAGGG CGCTGGCAAG TGTAGCGGTC 5601 ACGCTGCGCG TAACCACCAC ACCCGCCGCG CTTAATGCGC CGCTACAGGG 5651 CGCGTCCATT CGCCATTCAG GCTGCGCAAC TGTTGGGAAG GGCGATCGGT 5701 GCGGGCCTCT TCGCTATTAC GCCAGCTGGC GAAAGGGGGA TGTGCTGCAA 5751 GGCGATTAAG TTGGGTAACG CCAGGGTTTT CCCAGTCACG ACGTTGTAAA 5801 ACGACGGCCA GTGAATTGTA ATACGACTCA CTATA Sequence updated 16-Jul-01.