LOCUS Exported 3998 bp ds-DNA circular SYN 05-12-2013 DEFINITION Bacterial vector for expressing DHFR fusion proteins with a C-terminal 6xHis tag. Similar to pQE-16 and pQE-19, but with a different reading frame after the DHFR. ACCESSION . VERSION . KEYWORDS pQE-22 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 3998) AUTHORS Qiagen TITLE Direct Submission COMMENT Discontinued pQE vector. FEATURES Location/Qualifiers source 1..3998 /organism="synthetic DNA construct" /lab_host="Escherichia coli" /mol_type="other DNA" promoter 10..54 /note="T5 promoter" /note="bacteriophage T5 promoter for E. coli RNA polymerase, with embedded lac operator" protein_bind 30..46 /bound_moiety="lac repressor encoded by lacI" /note="lac operator" /note="The lac repressor binds to the lac operator to inhibit transcription in E. coli. This inhibition can be relieved by adding lactose or isopropyl-beta-D-thiogalactopyranoside (IPTG)." protein_bind 62..78 /bound_moiety="lac repressor encoded by lacI" /note="lac operator" /note="The lac repressor binds to the lac operator to inhibit transcription in E. coli. This inhibition can be relieved by adding lactose or isopropyl-beta-D-thiogalactopyranoside (IPTG)." RBS 101..106 /note="ribosome binding site" CDS 115..117 /codon_start=1 /product="start codon" /note="ATG" /translation="M" CDS 133..690 /codon_start=1 /gene="Mus musculus Dhfr" /product="mouse dihydrofolate reductase" /note="DHFR" /translation="MVRPLNSIVAVSQNMGIGKNGDLPWPPLRNEFKYFQRMTTTSSVE GKQNLVIMGRKTWFSIPEKNRPLKDRINIVLSRELKEPPRGAHFLAKSLDDALRLIEQP ELASKVDMVWIVGGSSVYQEAMNQPGHLRLFVTRIMQEFESDTFFPEIDLGKYKLLPEY PGVLSEVQEEKGIKYKFEVYEKK" CDS 704..721 /codon_start=1 /product="6xHis affinity tag" /note="6xHis" /translation="HHHHHH" misc_feature 728..738 /note="stop codons" /note="stop codons in all three reading frames" terminator 744..838 /gene=" " /note="lambda t0 terminator" /note="transcription terminator from phage lambda" CDS 882..1541 /codon_start=1 /gene="cat" /product="chloramphenicol acetyltransferase" /note="CmR" /note="confers resistance to chloramphenicol" /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA" terminator 1606..1692 /gene="Escherichia coli rrnB" /note="rrnB T1 terminator" /note="transcription terminator T1 from the E. coli rrnB gene" rep_origin complement(2174..2762) /direction=LEFT /note="ori" /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" CDS complement(2933..3793) /codon_start=1 /gene="bla" /product="beta-lactamase" /note="AmpR" /note="confers resistance to ampicillin, carbenicillin, and related antibiotics" /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQAAMDERNRQIAEIGAS LIKHW" promoter complement(3794..3898) /gene="bla" /note="AmpR promoter" ORIGIN 1 ctcgagaaat cataaaaaat ttatttgctt tgtgagcgga taacaattat aatagattca 61 attgtgagcg gataacaatt tcacacagaa ttcattaaag aggagaaatt aactatgaga 121 ggatccggca tcatggttcg accattgaac tcgatcgtcg ccgtgtccca aaatatgggg 181 attggcaaga acggagacct accctggcct ccgctcagga acgagttcaa gtacttccaa 241 agaatgacca caacctcttc agtggaaggt aaacagaatc tggtgattat gggtaggaaa 301 acctggttct ccattcctga gaagaatcga cctttaaagg acagaattaa tatagttctc 361 agtagagaac tcaaagaacc accacgagga gctcattttc ttgccaaaag tttggatgat 421 gccttaagac ttattgaaca accggaattg gcaagtaaag tagacatggt ttggatagtc 481 ggaggcagtt ctgtttacca ggaagccatg aatcaaccag gccaccttag actctttgtg 541 acaaggatca tgcaggaatt tgaaagtgac acgtttttcc cagaaattga tttggggaaa 601 tataaacttc tcccagaata cccaggcgtc ctctctgagg tccaggagga aaaaggcatc 661 aagtataagt ttgaagtcta cgagaagaaa ggttggaaga tctcatcacc atcaccatca 721 ctaagcttaa ttagctgagc ttggactcct gttgatagat ccagtaatga cctcagaact 781 ccatctggat ttgttcagaa cgctcggttg ccgccgggcg ttttttattg gtgagaatcc 841 aagctagctt ggcgagattt tcaggagcta aggaagctaa aatggagaaa aaaatcactg 901 gatataccac cgttgatata tcccaatggc atcgtaaaga acattttgag gcatttcagt 961 cagttgctca atgtacctat aaccagaccg ttcagctgga tattacggcc tttttaaaga 1021 ccgtaaagaa aaataagcac aagttttatc cggcctttat tcacattctt gcccgcctga 1081 tgaatgctca tccggaattt cgtatggcaa tgaaagacgg tgagctggtg atatgggata 1141 gtgttcaccc ttgttacacc gttttccatg agcaaactga aacgttttca tcgctctgga 1201 gtgaatacca cgacgatttc cggcagtttc tacacatata ttcgcaagat gtggcgtgtt 1261 acggtgaaaa cctggcctat ttccctaaag ggtttattga gaatatgttt ttcgtctcag 1321 ccaatccctg ggtgagtttc accagttttg atttaaacgt ggccaatatg gacaacttct 1381 tcgcccccgt tttcaccatg ggcaaatatt atacgcaagg cgacaaggtg ctgatgccgc 1441 tggcgattca ggttcatcat gccgtctgtg atggcttcca tgtcggcaga atgcttaatg 1501 aattacaaca gtactgcgat gagtggcagg gcggggcgta atttttttaa ggcagttatt 1561 ggtgccctta aacgcctggg gtaatgactc tctagcttga ggcatcaaat aaaacgaaag 1621 gctcagtcga aagactgggc ctttcgtttt atctgttgtt tgtcggtgaa cgctctcctg 1681 agtaggacaa atccgccgct ctagagctgc ctcgcgcgtt tcggtgatga cggtgaaaac 1741 ctctgacaca tgcagctccc ggagacggtc acagcttgtc tgtaagcgga tgccgggagc 1801 agacaagccc gtcagggcgc gtcagcgggt gttggcgggt gtcggggcgc agccatgacc 1861 cagtcacgta gcgatagcgg agtgtatact ggcttaacta tgcggcatca gagcagattg 1921 tactgagagt gcaccatatg cggtgtgaaa taccgcacag atgcgtaagg agaaaatacc 1981 gcatcaggcg ctcttccgct tcctcgctca ctgactcgct gcgctcggtc tgtcggctgc 2041 ggcgagcggt atcagctcac tcaaaggcgg taatacggtt atccacagaa tcaggggata 2101 acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg 2161 cgttgctggc gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct 2221 caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa 2281 gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc 2341 tcccttcggg aagcgtggcg ctttctcaat gctcacgctg taggtatctc agttcggtgt 2401 aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg 2461 ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg 2521 cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct 2581 tgaagtggtg gcctaactac ggctacacta gaaggacagt atttggtatc tgcgctctgc 2641 tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg 2701 ctggtagcgg tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc 2761 aagaagatcc tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt 2821 aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa 2881 aatgaagttt taaatcaatc taaagtatat atgagtaaac ttggtctgac agttaccaat 2941 gcttaatcag tgaggcacct atctcagcga tctgtctatt tcgttcatcc atagctgcct 3001 gactccccgt cgtgtagata actacgatac gggagggctt accatctggc cccagtgctg 3061 caatgatacc gcgagaccca cgctcaccgg ctccagattt atcagcaata aaccagccag 3121 ccggaagggc cgagcgcaga agtggtcctg caactttatc cgcctccatc cagtctatta 3181 attgttgccg ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg 3241 ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg 3301 gttcccaacg atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa gcggttagct 3361 ccttcggtcc tccgatcgtt gtcagaagta agttggccgc agtgttatca ctcatggtta 3421 tggcagcact gcataattct cttactgtca tgccatccgt aagatgcttt tctgtgactg 3481 gtgagtactc aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc 3541 cggcgtcaat acgggataat accgcgccac atagcagaac tttaaaagtg ctcatcattg 3601 gaaaacgttc ttcggggcga aaactctcaa ggatcttacc gctgttgaga tccagttcga 3661 tgtaacccac tcgtgcaccc aactgatctt cagcatcttt tactttcacc agcgtttctg 3721 ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg aataagggcg acacggaaat 3781 gttgaatact catactcttc ctttttcaat attattgaag catttatcag ggttattgtc 3841 tcatgagcgg atacatattt gaatgtattt agaaaaataa acaaataggg gttccgcgca 3901 catttccccg aaaagtgcca cctgacgtct aagaaaccat tattatcatg acattaacct 3961 ataaaaatag gcgtatcacg aggccctttc gtcttcac //