The pET-44 vectors are designed for cloning and high-level expression of peptide sequences fused with the 495 aa Nus?Tag? protein. Compared to the pET-43.1 series, the pET-44 vectors encode an additional N-terminal His?Tag. Unique sites are shown on the circle map. Note that the sequence is numbered by the pBR322 convention, so the T7 expression region is reversed on the circle map. The cloning/expression region of the coding strand transcribed by T7 RNA polymerase is shown below. The f1 origin is oriented so that infection with helper phage will produce virions containing single stranded DNA that corresponds to the coding strand. Therefore, single stranded sequencing should be performed using the COLIDOWN primer (Cat. No. 70845-3). Vector encoded sequence can be completely removed when cloning into the PshA I or Sma I sites (as shown below) by cleaving the Nus?Tag fusion protein with enterokinase or thrombin, respectively.

0001 ATCCGGATAT AGTTCCTCCT TTCAGCAAAA AACCCCTCAA GACCCGTTTA GAGGCCCCAA 
0061 GGGGTTATGC TAGTTATTGC TCAGCGGATA CATATTTGAA TGTATTTAGA AAAATAAACA 
0121 AAAGAGTTTG TAGAAACGCA AAAAGGCCAT CCGTCAGGAT GGCCTTCTGC TTAATTTGAT 
0181 GCCTGGCAGT TTATGGCGGG CGTCCTGCCC GCCACCCTCC GGGCCGTTGC TTCGCAACGT 
0241 TCAAATCCGC TCCCGGCGGA TTTGTCCTAC TCAGGAGAGC GTTCACCGAC AAACAACAGA 
0301 TAAAACGAAA GGCCCAGTCT TTCGACTGAG CCTTTCGTTT TATTTGATGC CTGGCAGTTC 
0361 CCTACTCTCG CATGGGGAGA CCCCACACTA CCATCGGCGC TACGGCGTTT CACTTCTGAG 
0421 TTCGGCATGG GGTCAGGTGG GACCACCGCG CTACTGCCGC CAGGCAAATT CTGTTTTATC 
0481 AGCCTAGGAA CGCCCAACTT AATTAACATT AGTGGTGGTG GTGGTGGTGC TCGAGATCCT 
0541 CTGGGTCTTC AGGAGCGAGT TCTGGCTGGC TTGCACGTGT ATACAGCTGT GCGGCCGCAA 
0601 GCTTCGAACG CGTATCGATG GTACCGTCGA CGTCCTGCAG GCGCGCCTGT ACAGAATTCG 
0661 GATCCACGAG CTCCCGGACT CTTGTCGTCG TCATCAATCG TACCAGAACC CGCGCTGCCC 
0721 CGGGGGACCA GACCAGTTGG CGGTGGCGAG TCCATGTGCT GGCGTTCAAA TTTCGCAGCA 
0781 GCGGTTTCTT TACCCGCGGA GTGATGGTGA TGGTGATGAC CAGAACCACT AGTCGCTTCG 
0841 TCACCGAACC AGCAAATATT ACGGGCAGCC ATAATCAGTG CTCCGGCTTT TTCGTCGGTC 
0901 AACCCTTCGA TATCAGCCAG ATCATCAATG CCCTGTTCGG CGAGATCTTC CAGCGTACAA 
0961 ACGCCACGGG CGGCCAGTTT GAATGCCAAA TCACGATCTA CCCCTTCAAG GTTCAGCAGA 
1021 TCGTCAGCCG GTTTGTTATC ACCGAGGCTT TCTTCCTGGG CCTGTGCAAT GGTGGCCAGT 
1081 GCATTTTTAG CACGCTCGCG CAGTGCTTCA ACGGTCGGCT CATCAAGGCC TTCGATTTCC 
1141 AACAGCTCTT TCATCGGCAC ATAGGCCAAT TCTTCCAGCG TCGAGAAGCC TTCTTCTACC 
1201 AGAACAGTCG CGAAGTCTTC GTCGATGTCG AGATATTTGG TGAAGGTGTC GATCGCTGCG 
1261 TGCGCTTCCG CCTGATGCTT AGCTTGCAGG TCGTCAACGG TCATCACGTT GAGTTCCCAA 
1321 CCGCTCAGTT GCGAAGCCAG ACGCACGTTC TGACCGTTAC GGCCAATCGC CTGCGCCAGA 
1381 TTACCGGCTT CAACGGCGAT GTCCATGGTG TGTTTATCTT CATCCACCAC GATAGAAGCA 
1441 ACGTCTGCCG GTGCCATTGC GTTAATCACG AACTGCGCCG GGTTATCATC CCACAGGACG 
1501 ATATCGATAC GCTCGCCACC CAGTTCAGTA GACACCGCCT GAACACGCGC GCCACGCATA 
1561 CCTACGCAAG CACCTACCGG ATCGATACGT TTATCGTTGG TTTTCACCGC GATTTTCGCA 
1621 CGAGAACCCG GATCGCGAGC CGCTGCTTTA ATTTCAATCA CTTCTTCGCC GATTTCTGGC 
1681 ACTTCAATAC GGAACAGTTC GATCAGCATT TCCGGCTTGG AACGAGTGAC GAACAGTTGC 
1741 GCGCCACGCG CTTCCGGGCG AACGGAATAG AGCACGCCAC GAACGCGGTC GCCAGGGCGG 
1801 AAGTTTTCAC GCGGCAGCAT ATCTTCGCGC AGGATCACGG CTTCAGCGTT GTTGCCCAGA 
1861 TCCAGAGAGA TGTTGTCGCG GTTTACTTTT TTCACCACGC CGGTGATGAT TTCACCTTCG 
1921 TGTTCACGGA ACTGATCAAC CACCATCGCA CGTTCGGCTT CACGCACTTT CTGCACGATA 
1981 ACCTGTTTTG CCGTCTGGGT AGTGATACGG TCAAAGGTAA CAGACTCAAT CTGATCTTCA 
2041 ACGTAATCGC CCAGGTTCAG GCTTTCATCT TCATAACGTG CGGCTTCAAG GGTGATTTCC 
2101 TTGGTCGGCT GGGTGACTTC ATCAACAACT AACCAGCGAC GGAAAGTGTC AAAATCACCG 
2161 CTTTTGCGAT CGATCTGTAC GCGGACGTCG ATCTCTTGTT CATATTTTTT CTTTGTTGCT 
2221 GTCGCCAGCG CGCTTTCCAA TGCTTCGAAA ATCTTCTCGC GAGGTAGCGC CTTTTCATTG 
2281 GATACGGCTT CAACTACAGC CAAAATTTCT TTGTTCATAC TAGAATGGTG ATGGTGGTGA 
2341 TGGCTGCTGC CCATATGTAT ATCTCCTTCT TAAAGTTAAA CAAAATTATT TCTAGAGGGG 
2401 AATTGTTATC CGCTCACAAT TCCCCTATAG TGAGTCGTAT TAATTTCGCG GGATCGAGAT 
2461 CGATCTCGAT CCTCTACGCC GGACGCATCG TGGCCGGCAT CACCGGCGCC ACAGGTGCGG 
2521 TTGCTGGCGC CTATATCGCC GACATCACCG ATGGGGAAGA TCGGGCTCGC CACTTCGGGC 
2581 TCATGAGCGC TTGTTTCGGC GTGGGTATGG TGGCAGGCCC CGTGGCCGGG GGACTGTTGG 
2641 GCGCCATCTC CTTGCATGCA CCATTCCTTG CGGCGGCGGT GCTCAACGGC CTCAACCTAC 
2701 TACTGGGCTG CTTCCTAATG CAGGAGTCGC ATAAGGGAGA GCGTCGAGAT CCCGGACACC 
2761 ATCGAATGGC GCAAAACCTT TCGCGGTATG GCATGATAGC GCCCGGAAGA GAGTCAATTC 
2821 AGGGTGGTGA ATGTGAAACC AGTAACGTTA TACGATGTCG CAGAGTATGC CGGTGTCTCT 
2881 TATCAGACCG TTTCCCGCGT GGTGAACCAG GCCAGCCACG TTTCTGCGAA AACGCGGGAA 
2941 AAAGTGGAAG CGGCGATGGC GGAGCTGAAT TACATTCCCA ACCGCGTGGC ACAACAACTG 
3001 GCGGGCAAAC AGTCGTTGCT GATTGGCGTT GCCACCTCCA GTCTGGCCCT GCACGCGCCG 
3061 TCGCAAATTG TCGCGGCGAT TAAATCTCGC GCCGATCAAC TGGGTGCCAG CGTGGTGGTG 
3121 TCGATGGTAG AACGAAGCGG CGTCGAAGCC TGTAAAGCGG CGGTGCACAA TCTTCTCGCG 
3181 CAACGCGTCA GTGGGCTGAT CATTAACTAT CCGCTGGATG ACCAGGATGC CATTGCTGTG 
3241 GAAGCTGCCT GCACTAATGT TCCGGCGTTA TTTCTTGATG TCTCTGACCA GACACCCATC 
3301 AACAGTATTA TTTTCTCCCA TGAAGACGGT ACGCGACTGG GCGTGGAGCA TCTGGTCGCA 
3361 TTGGGTCACC AGCAAATCGC GCTGTTAGCG GGCCCATTAA GTTCTGTCTC GGCGCGTCTG 
3421 CGTCTGGCTG GCTGGCATAA ATATCTCACT CGCAATCAAA TTCAGCCGAT AGCGGAACGG 
3481 GAAGGCGACT GGAGTGCCAT GTCCGGTTTT CAACAAACCA TGCAAATGCT GAATGAGGGC 
3541 ATCGTTCCCA CTGCGATGCT GGTTGCCAAC GATCAGATGG CGCTGGGCGC AATGCGCGCC 
3601 ATTACCGAGT CCGGGCTGCG CGTTGGTGCG GACATCTCGG TAGTGGGATA CGACGATACC 
3661 GAAGACAGCT CATGTTATAT CCCGCCGTTA ACCACCATCA AACAGGATTT TCGCCTGCTG 
3721 GGGCAAACCA GCGTGGACCG CTTGCTGCAA CTCTCTCAGG GCCAGGCGGT GAAGGGCAAT 
3781 CAGCTGTTGC CCGTCTCACT GGTGAAAAGA AAAACCACCC TGGCGCCCAA TACGCAAACC 
3841 GCCTCTCCCC GCGCGTTGGC CGATTCATTA ATGCAGCTGG CACGACAGGT TTCCCGACTG 
3901 GAAAGCGGGC AGTGAGCGCA ACGCAATTAA TGTAAGTTAG CTCACTCATT AGGCACCGGG 
3961 ATCTCGACCG ATGCCCTTGA GAGCCTTCAA CCCAGTCAGC TCCTTCCGGT GGGCGCGGGG 
4021 CATGACTAGC ATGATCGTGC TCCTGTCGTT GAGGACCCGG CTAGGCTGGC GGGGTTGCCT 
4081 TACTGGTTAG CAGAATGAAT CACCGATACG CGAGCGAACG TGAAGCGACT GCTGCTGCAA 
4141 AACGTCTGCG ACCTGAGCAA CAACATGAAT GGTCTTCGGT TTCCGTGTTT CGTAAAGTCT 
4201 GGAAACGCGG AAGTCAGCGC CCTGCACCAT TATGTTCCGG ATCTGCATCG CAGGATGCTG 
4261 CTGGCTACCC TGTGGAACAC CTACATCTGT ATTAACGAAG CGCTGGCATT GACCCTGAGT 
4321 GATTTTTCTC TGGTCCCGCC GCATCCATAC CGCCAGTTGT TTACCCTCAC AACGTTCCAG 
4381 TAACCGGGCA TGTTCATCAT CAGTAACCCG TATCGTGAGC ATCCTCTCTC GTTTCATCGG 
4441 TATCATTACC CCCATGAACA GAAATCCCCC TTACACGGAG GCATCAGTGA CCAAACAGGA 
4501 AAAAACCGCC CTTAACATGG CCCGCTTTAT CAGAAGCCAG ACATTAACGC TTCTGGAGAA 
4561 ACTCAACGAG CTGGACGCGG ATGAACAGGC AGACATCTGT GAATCGCTTC ACGACCACGC 
4621 TGATGAGCTT TACCGCAGCT GCCTCGCGCG TTTCGGTGAT GACGGTGAAA ACCTCTGACA 
4681 CATGCAGCTC CCGGAGACGG TCACAGCTTG TCTGTAAGCG GATGCCGGGA GCAGACAAGC 
4741 CCGTCAGGGC GCGTCAGCGG GTGTTGGCGG GTGTCGGGGC GCAGCCATGA CCCAGTCACG 
4801 TAGCGATAGC GGAGTGTATA CTGGCTTAAC TATGCGGCAT CAGAGCAGAT TGTACTGAGA 
4861 GTGCACCATA TATGCGGTGT GAAATACCGC ACAGATGCGT AAGGAGAAAA TACCGCATCA 
4921 GGCGCTCTTC CGCTTCCTCG CTCACTGACT CGCTGCGCTC GGTCGTTCGG CTGCGGCGAG 
4981 CGGTATCAGC TCACTCAAAG GCGGTAATAC GGTTATCCAC AGAATCAGGG GATAACGCAG 
5041 GAAAGAACAT GTGAGCAAAA GGCCAGCAAA AGGCCAGGAA CCGTAAAAAG GCCGCGTTGC 
5101 TGGCGTTTTT CCATAGGCTC CGCCCCCCTG ACGAGCATCA CAAAAATCGA CGCTCAAGTC 
5161 AGAGGTGGCG AAACCCGACA GGACTATAAA GATACCAGGC GTTTCCCCCT GGAAGCTCCC 
5221 TCGTGCGCTC TCCTGTTCCG ACCCTGCCGC TTACCGGATA CCTGTCCGCC TTTCTCCCTT 
5281 CGGGAAGCGT GGCGCTTTCT CATAGCTCAC GCTGTAGGTA TCTCAGTTCG GTGTAGGTCG 
5341 TTCGCTCCAA GCTGGGCTGT GTGCACGAAC CCCCCGTTCA GCCCGACCGC TGCGCCTTAT 
5401 CCGGTAACTA TCGTCTTGAG TCCAACCCGG TAAGACACGA CTTATCGCCA CTGGCAGCAG 
5461 CCACTGGTAA CAGGATTAGC AGAGCGAGGT ATGTAGGCGG TGCTACAGAG TTCTTGAAGT 
5521 GGTGGCCTAA CTACGGCTAC ACTAGAAGGA CAGTATTTGG TATCTGCGCT CTGCTGAAGC 
5581 CAGTTACCTT CGGAAAAAGA GTTGGTAGCT CTTGATCCGG CAAACAAACC ACCGCTGGTA 
5641 GCGGTGGTTT TTTTGTTTGC AAGCAGCAGA TTACGCGCAG AAAAAAAGGA TCTCAAGAAG 
5701 ATCCTTTGAT CTTTTCTACG GGGTCTGACG CTCAGTGGAA CGAAAACTCA CGTTAAGGGA 
5761 TTTTGGTCAT GACCTATTTG TTTATTTTTC TAAATACATT CAAATATGTA TCCGCTCATG 
5821 AGACAATAAC CCTGATAAAT GCTTCAATCA TGATTGAAAA AGGAAGAGTA TGAGTATTCA 
5881 ACATTTCCGT GTCGCCCTTA TTCCCTTTTT TGCGGCATTT TGCCTTCCTG TTTTTGCTCA 
5941 CCCAGAAACG CTGGTGAAAG TAAAAGATGC TGAAGATCAG TTGGGTGCAC GAGTGGGTTA 
6001 CATCGAACTG GATCTCAACA GCGGTAAGAT CCTTGAGAGT TTTCGCCCCG AAGAACGTTT 
6061 TCCAATGATG AGCACTTTTA AAGTTCTGCT ATGTGGCGCG GTATTATCCC GTATTGACGC 
6121 CGGGCAAGAG CAACTCGGTC GCCGCATACA CTATTCTCAG AATGACTTGG TTGAGTACTC 
6181 ACCAGTCACA GAAAAGCATC TTACGGATGG CATGACAGTA AGAGAATTAT GCAGTGCTGC 
6241 CATAACCATG AGTGATAACA CTGCGGCCAA CTTACTTCTG ACAACGATCG GAGGACCGAA 
6301 GGAGCTAACC GCTTTTTTGC ACAACATGGG GGATCATGTA ACTCGCCTTG ATCGTTGGGA 
6361 ACCGGAGCTG AATGAAGCCA TACCAAACGA CGAGCGTGAC ACCACGATGC CTGTAGCAAT 
6421 GGCAACAACG TTGCGCAAAC TATTAACTGG CGAACTACTT ACTCTAGCTT CCCGGCAACA 
6481 ATTAATAGAC TGGATGGAGG CGGATAAAGT TGCAGGACCA CTTCTGCGCT CGGCCCTTCC 
6541 GGCTGGCTGG TTTATTGCTG ATAAATCTGG AGCCGGTGAG CGTGGGTCTC GCGGTATCAT 
6601 TGCAGCACTG GGGCCAGATG GTAAGCCCTC CCGTATCGTA GTTATCTACA CGACGGGGAG 
6661 TCAGGCAACT ATGGATGAAC GAAATAGACA GATCGCTGAG ATAGGTGCCT CACTGATTAA 
6721 GCATTGGTAA CTGTCAGACC AAGTTTACTC ATATATACTT TAGATTGATT TAAAACTTCA 
6781 TTTTTAATTT AAAAGGATCT AGGTGAAGAT CCTTTTTGAT AATCTCATGC CATCGTGCCG 
6841 CCAGAAATTG TAAACGTTAA TATTTTGTTA AAATTCGCGT TAAATTTTTG TTAAATCAGC 
6901 TCATTTTTTA ACCAATAGGC CGAAATCGGC AAAATCCCTT ATAAATCAAA AGAATAGACC 
6961 GAGATAGGGT TGAGTGTTGT TCCAGTTTGG AACAAGAGTC CACTATTAAA GAACGTGGAC 
7021 TCCAACGTCA AAGGGCGAAA AACCGTCTAT CAGGGCGATG GCCCACTACG TGAACCATCA 
7081 CCCTAATCAA GTTTTTTGGG GTCGAGGTGC CGTAAAGCAC TAAATCGGAA CCCTAAAGGG 
7141 AGCCCCCGAT TTAGAGCTTG ACGGGGAAAG CCGGCGAACG TGGCGAGAAA GGAAGGGAAG 
7201 AAAGCGAAAG GAGCGGGCGC TAGGGCGCTG GCAAGTGTAG CGGTCACGCT GCGCGTAACC 
7261 ACCACACCCG CCGCGCTTAA TGCGCCGCTA CAGGGCGCGT CCCATTCGCC A