pGEMEX®-1 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 pGEMEX®-1 Vector sequence reference points: Base pairs 3993 T7 RNA polymerase transcription initiation site 968 SP6 RNA polymerase transcription initiation site 1 T3 RNA polymerase transcriptioninitation site 89 T7 RNA polymerase promoter (-17 to +3) 966-985 SP6 RNA polymerase promoter (-17 to +3) 3977-3 T3 RNA polymerase promoter (-17 to +3) 87-106 Multiple cloning region 9-84 T7 gene 10 start codon 905 T7 terminator region 3839-3929 Beta-lactamase (Ampr) coding region 2007-2867 phage f1 region 3304-3759 (a)Usage restrictions apply to Bacterial Strains JM109(DE3) and BL21(DE3) pLysS, to the following Promega products that include these bacterial strains (pGEMEX®-1 and pGEMEX®-2 Vectors) and to any derivatives thereof. Please read the following statement describing these restrictions before purchasing any of these products. Usage Restrictions for the T7 Expression System The T7 expression system is based on technology developed at Brookhaven National Laboratory under contract with the U.S. Department of Energy and is the subject of patents and patent applications assigned to Brookhaven Science Associates, LLC (BSA). This technology, including bacteria, phage and plasmids that carry the gene for T7 RNA polymerase, is to be used for academic or nonprofit laboratory or licensed commercial research purposes only. By accepting or using the T7 expression technology you agree to be bound by the following conditions set forth by BSA. The initial purchaser may refuse to accept the conditions of this notice by returning this product and the enclosed materials to Promega unused. Academic and NonProfit Laboratories No materials that contain the cloned gene for T7 RNA polymerase may be distributed further to third parties outside of your laboratory unless the recipient receives a copy of this assurance notice and agrees to be bound by its terms. This limitation applies to Bacterial Strains JM109(DE3) and BL21(DE3)pLysS, to the following Promega products that include these bacterial strains (pGEMEX®-1 and pGEMEX®-2 Vectors) and to any derivatives thereof. Commercial Laboratories A license is required for any commercial use of the T7 expression system, including use of the T7 system for research purposes or for production purposes by any commercial entity. Information about commercial licenses may be obtained from the Licensing Office, Brookhaven National Laboratory, Upton, NY 11973, Telephone: 631-344-7134, FAX: 631-344-3729. (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. Note: The sequence for the pGEMEX®-2 Vector is identical to that of pGEMEX®-1 Vector except for the addition of two cytosine bases at positions 65 and 84 (in relation to the numbering and sequence below) in the pGEMEX®-2 Vector. pGEMEX is trademark of Promega Corporation and is registered with the U.S Patent and Trademark Office. .. 1 GAATACTCAA GCTTATGCAT GCGGCCGCAT CTAGAGGGCC CGGATCCCTC 51 GAGGTCGACG AATTCGAGCT CGGCCGACTT GGCCTTCCCT TTAGTGAGGG 101 TTAATAAACT TGGCCAATTC GATCCGGTAC CAGCACCACC AGCGGTGAGG 151 TGCGGAACTT CTACAACCTC AAAGCCCATA ACGTTGCGGA TAGAACCCTT 201 CTCAGGGTCA ATCAGAGCAG CGTAGTTTGC TGCGTTCGGC ATCAGTGCTG 251 CCAGAATCGC AGAGTAGCTA TCTGGGTCAC AGTAGAACAC ACGGTCAGCA 301 GCCGGAACAT AGTTCTTGGT CAGAGCCGCA CGAGCCTTAG TCAGAGCCGC 351 AATAATCTCC TTACCCAGCG CAACTTGGTC GGTAAGTGCG GCCTTGTTCT 401 GAGTGGTCTC AATTACGGTA GCAGTACCTA AGCCCTCGAT GTTCTCATTA 451 TATTTGCTTT CCACGTTACA CAGACCGGCA ATCTCAGCCA GAACCGCACC 501 ATCCGCAGCC ATCGCCAGAG ATTCACCCAA CTGAGAGGTA TACTCAGAGC 551 GAACGTCGTA GTGGTTCATC GCGTCCTCAA TATCATAAAT CAGAACGTCA 601 GCCGTCAGGA GACCGTCAAT GGTGATTACC TTCTCGGTGT GTTTGATGTC 651 CTTACGTTTA TCGTCGAGGT TCTCGCCCGG AGCCAGATAC GCTGCCTGAG 701 TGCGACCCAG AACAGGGAAC TGAGCGGATT TACCGCTGGA GATGGAACGT 751 ACCATGTGGC GAGAAGTGGT CACGGAGGTA CGAGCGAACG CAGTCAGGAC 801 TTCACCGCCA AATACCTTCA AGAACAACGC CAGTTTATCT CCAGCAGCAA 851 CTACACCTTT ACCTTGGTTA GTACCCATTT GCTGTCCACC AGTCATGCTA 901 GCCATATGTA TATCTCCTTC TTAAAGTTAA ACAAAATTAT TTCTAGAGGG 951 AAACCGTTGT GGTCTCCCTA TAGTGAGTCG TATTAATTTC GCGGGATCGA 1001 GATCTCCCGG GCTGCATTAA TGAATCGGCC AACGCGCGGG GAGAGGCGGT 1051 TTGCGTATTG GGCGCTCTTC CGCTTCCTCG CTCACTGACT CGCTGCGCTC 1101 GGTCGTTCGG CTGCGGCGAG CGGTATCAGC TCACTCAAAG GCGGTAATAC 1151 GGTTATCCAC AGAATCAGGG GATAACGCAG GAAAGAACAT GTGAGCAAAA 1201 GGCCAGCAAA AGGCCAGGAA CCGTAAAAAG GCCGCGTTGC TGGCGTTTTT 1251 CCATAGGCTC CGCCCCCCTG ACGAGCATCA CAAAAATCGA CGCTCAAGTC 1301 AGAGGTGGCG AAACCCGACA GGACTATAAA GATACCAGGC GTTTCCCCCT 1351 GGAAGCTCCC TCGTGCGCTC TCCTGTTCCG ACCCTGCCGC TTACCGGATA 1401 CCTGTCCGCC TTTCTCCCTT CGGGAAGCGT GGCGCTTTCT CATAGCTCAC 1451 GCTGTAGGTA TCTCAGTTCG GTGTAGGTCG TTCGCTCCAA GCTGGGCTGT 1501 GTGCACGAAC CCCCCGTTCA GCCCGACCGC TGCGCCTTAT CCGGTAACTA 1551 TCGTCTTGAG TCCAACCCGG TAAGACACGA CTTATCGCCA CTGGCAGCAG 1601 CCACTGGTAA CAGGATTAGC AGAGCGAGGT ATGTAGGCGG TGCTACAGAG 1651 TTCTTGAAGT GGTGGCCTAA CTACGGCTAC ACTAGAAGAA CAGTATTTGG 1701 TATCTGCGCT CTGCTGAAGC CAGTTACCTT CGGAAAAAGA GTTGGTAGCT 1751 CTTGATCCGG CAAACAAACC ACCGCTGGTA GCGGTGGTTT TTTTGTTTGC 1801 AAGCAGCAGA TTACGCGCAG AAAAAAAGGA TCTCAAGAAG ATCCTTTGAT 1851 CTTTTCTACG GGGTCTGACG CTCAGTGGAA CGAAAACTCA CGTTAAGGGA 1901 TTTTGGTCAT GAGATTATCA AAAAGGATCT TCACCTAGAT CCTTTTAAAT 1951 TAAAAATGAA GTTTTAAATC AATCTAAAGT ATATATGAGT AAACTTGGTC 2001 TGACAGTTAC CAATGCTTAA TCAGTGAGGC ACCTATCTCA GCGATCTGTC 2051 TATTTCGTTC ATCCATAGTT GCCTGACTCC CCGTCGTGTA GATAACTACG 2101 ATACGGGAGG GCTTACCATC TGGCCCCAGT GCTGCAATGA TACCGCGAGA 2151 CCCACGCTCA CCGGCTCCAG ATTTATCAGC AATAAACCAG CCAGCCGGAA 2201 GGGCCGAGCG CAGAAGTGGT CCTGCAACTT TATCCGCCTC CATCCAGTCT 2251 ATTAATTGTT GCCGGGAAGC TAGAGTAAGT AGTTCGCCAG TTAATAGTTT 2301 GCGCAACGTT GTTGCCATTG CTACAGGCAT CGTGGTGTCA CGCTCGTCGT 2351 TTGGTATGGC TTCATTCAGC TCCGGTTCCC AACGATCAAG GCGAGTTACA 2401 TGATCCCCCA TGTTGTGCAA AAAAGCGGTT AGCTCCTTCG GTCCTCCGAT 2451 CGTTGTCAGA AGTAAGTTGG CCGCAGTGTT ATCACTCATG GTTATGGCAG 2501 CACTGCATAA TTCTCTTACT GTCATGCCAT CCGTAAGATG CTTTTCTGTG 2551 ACTGGTGAGT ACTCAACCAA GTCATTCTGA GAATAGTGTA TGCGGCGACC 2601 GAGTTGCTCT TGCCCGGCGT CAATACGGGA TAATACCGCG CCACATAGCA 2651 GAACTTTAAA AGTGCTCATC ATTGGAAAAC GTTCTTCGGG GCGAAAACTC 2701 TCAAGGATCT TACCGCTGTT GAGATCCAGT TCGATGTAAC CCACTCGTGC 2751 ACCCAACTGA TCTTCAGCAT CTTTTACTTT CACCAGCGTT TCTGGGTGAG 2801 CAAAAACAGG AAGGCAAAAT GCCGCAAAAA AGGGAATAAG GGCGACACGG 2851 AAATGTTGAA TACTCATACT CTTCCTTTTT CAATATTATT GAAGCATTTA 2901 TCAGGGTTAT TGTCTCATGA GCGGATACAT ATTTGAATGT ATTTAGAAAA 2951 ATAAACAAAT AGGGGTTCCG CGCACATTTC CCCGAAAAGT GCCACCTGAC 3001 GTCTAAGAAA CCATTATTAT CATGACATTA ACCTATAAAA ATAGGCGTAT 3051 CACGAGGCCC TTTCGTCTCG CGCGTTTCGG TGATGACGGT GAAAACCTCT 3101 GACACATGCA GCTCCCGGAG ACGGTCACAG CTTGTCTGTA AGCGGATGCC 3151 GGGAGCAGAC AAGCCCGTCA GGGCGCGTCA GCGGGTGTTG GCGGGTGTCG 3201 GGGCTGGCTT AACTATGCGG CATCAGAGCA GATTGTACTG AGAGTGCACC 3251 ATATGCGGTG TGAAATACCG CACAGATGCG TAAGGAGAAA ATACCGCATC 3301 AGGACGCGCC CTGTAGCGGC GCATTAAGCG CGGCGGGTGT GGTGGTTACG 3351 CGCAGCGTGA CCGCTACACT TGCCAGCGCC CTAGCGCCCG CTCCTTTCGC 3401 TTTCTTCCCT TCCTTTCTCG CCACGTTCGC CGGCTTTCCC CGTCAAGCTC 3451 TAAATCGGGG GCTCCCTTTA GGGTTCCGAT TTAGTGCTTT ACGGCACCTC 3501 GACCCCAAAA AACTTGATTA GGGTGATGGT TCACGTAGTG GGCCATCGCC 3551 CTGATAGACG GTTTTTCGCC CTTTGACGTT GGAGTCCACG TTCTTTAATA 3601 GTGGACTCTT GTTCCAAACT GGAACAACAC TCAACCCTAT CTCGGTCTAT 3651 TCTTTTGATT TATAAGGGAT TTTGCCGATT TCGGCCTATT GGTTAAAAAA 3701 TGAGCTGATT TAACAAAAAT TTAACGCGAA TTTTAACAAA ATATTAACGC 3751 TTACAATTTC CATTCGCCAT TCAGGCTGCG CAACTGTTGG GAAGGGCGAT 3801 CGGTGCGGGC CTCTTCGCTA TTACGCCAGG TTATCCGGAT ATAGTTCCTC 3851 CTTTCAGCAA AAAACCCCTC AAGACCCGTT TAGAGGCCCC AAGGGGTTAT 3901 GCTAGTTATT GCTCAGCGGT GGCAGCAGCC AACTCAGCTT CCTTTCGGGC 3951 TTTGTTAGCA GCCGGATCAA GCTGGGATTT AGGTGACACT ATA Sequence updated 5-Jun-00.