LOCUS Exported 3504 bp ds-DNA circular SYN 09-1-2013 DEFINITION Bacterial expression and 6xHis-tagging vector for UA cloning of a PCR product. ACCESSION . VERSION . KEYWORDS pQE-30 UA SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 3504) AUTHORS Qiagen TITLE Direct Submission COMMENT This vector is supplied in a linearized form, with bases 174-175 replaced with 3'-U overhangs. FEATURES Location/Qualifiers source 1..3504 /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 127..144 /codon_start=1 /product="6xHis affinity tag" /note="6xHis" /translation="HHHHHH" misc_feature 145..235 /note="MCS" /note="multiple cloning site" misc_feature 235..245 /note="stop codons" /note="stop codons in all three reading frames" terminator 251..345 /gene=" " /note="lambda t0 terminator" /note="transcription terminator from phage lambda" CDS 389..1048 /codon_start=1 /gene="cat" /product="chloramphenicol acetyltransferase" /note="CmR" /note="confers resistance to chloramphenicol" /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA" terminator 1113..1199 /gene="Escherichia coli rrnB" /note="rrnB T1 terminator" /note="transcription terminator T1 from the E. coli rrnB gene" rep_origin complement(1680..2268) /direction=LEFT /note="ori" /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" CDS complement(2439..3299) /codon_start=1 /gene="bla" /product="beta-lactamase" /note="AmpR" /note="confers resistance to ampicillin, carbenicillin, and related antibiotics" /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS LIKHW" promoter complement(3300..3404) /gene="bla" /note="AmpR promoter" ORIGIN 1 ctcgagaaat cataaaaaat ttatttgctt tgtgagcgga taacaattat aatagattca 61 attgtgagcg gataacaatt tcacacagaa ttcattaaag aggagaaatt aactatgaga 121 ggatcgcatc accatcacca tcacggatcc cacgtgatat cctcaatcgc ttctagaagc 181 gattgaggag atctgagctc ggtacccggg tcgacaggcc tctgcagcca agcttaatta 241 gctgagcttg gactcctgtt gatagatcca gtaatgacct cagaactcca tctggatttg 301 ttcagaacgc tcggttgccg ccgggcgttt tttattggtg agaatccaag ctagcttggc 361 gagattttca ggagctaagg aagctaaaat ggagaaaaaa atcactggat ataccaccgt 421 tgatatatcc caatggcatc gtaaagaaca ttttgaggca tttcagtcag ttgctcaatg 481 tacctataac cagaccgttc agctggatat tacggccttt ttaaagaccg taaagaaaaa 541 taagcacaag ttttatccgg cctttattca cattcttgcc cgcctgatga atgctcatcc 601 ggaatttcgt atggcaatga aagacggtga gctggtgata tgggatagtg ttcacccttg 661 ttacaccgtt ttccatgagc aaactgaaac gttttcatcg ctctggagtg aataccacga 721 cgatttccgg cagtttctac acatatattc gcaagatgtg gcgtgttacg gtgaaaacct 781 ggcctatttc cctaaagggt ttattgagaa tatgtttttc gtctcagcca atccctgggt 841 gagtttcacc agttttgatt taaacgtggc caatatggac aacttcttcg cccccgtttt 901 caccatgggc aaatattata cgcaaggcga caaggtgctg atgccgctgg cgattcaggt 961 tcatcatgcc gtttgtgatg gcttccatgt cggcagaatg cttaatgaat tacaacagta 1021 ctgcgatgag tggcagggcg gggcgtaatt tttttaaggc agttattggt gcccttaaac 1081 gcctggggta atgactctct agcttgaggc atcaaataaa acgaaaggct cagtcgaaag 1141 actgggcctt tcgttttatc tgttgtttgt cggtgaacgc tctcctgagt aggacaaatc 1201 cgccctctag agctgcctcg cgcgtttcgg tgatgacggt gaaaacctct gacacatgca 1261 gctcccggag acggtcacag cttgtctgta agcggatgcc gggagcagac aagcccgtca 1321 gggcgcgtca gcgggtgttg gcgggtgtcg gggcgcagcc atgacccagt cacgtagcga 1381 tagcggagtg tatactggct taactatgcg gcatcagagc agattgtact gagagtgcac 1441 catatgcggt gtgaaatacc gcacagatgc gtaaggagaa aataccgcat caggcgctct 1501 tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca 1561 gctcactcaa aggcggtaat acggttatcc acagaatcag gggataacgc aggaaagaac 1621 atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt 1681 ttccataggc tccgcccccc tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg 1741 cgaaacccga caggactata aagataccag gcgtttcccc ctggaagctc cctcgtgcgc 1801 tctcctgttc cgaccctgcc gcttaccgga tacctgtccg cctttctccc ttcgggaagc 1861 gtggcgcttt ctcatagctc acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc 1921 aagctgggct gtgtgcacga accccccgtt cagcccgacc gctgcgcctt atccggtaac 1981 tatcgtcttg agtccaaccc ggtaagacac gacttatcgc cactggcagc agccactggt 2041 aacaggatta gcagagcgag gtatgtaggc ggtgctacag agttcttgaa gtggtggcct 2101 aactacggct acactagaag gacagtattt ggtatctgcg ctctgctgaa gccagttacc 2161 ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt 2221 ttttttgttt gcaagcagca gattacgcgc agaaaaaaag gatctcaaga agatcctttg 2281 atcttttcta cggggtctga cgctcagtgg aacgaaaact cacgttaagg gattttggtc 2341 atgagattat caaaaaggat cttcacctag atccttttaa attaaaaatg aagttttaaa 2401 tcaatctaaa gtatatatga gtaaacttgg tctgacagtt accaatgctt aatcagtgag 2461 gcacctatct cagcgatctg tctatttcgt tcatccatag ttgcctgact ccccgtcgtg 2521 tagataacta cgatacggga gggcttacca tctggcccca gtgctgcaat gataccgcga 2581 gacccacgct caccggctcc agatttatca gcaataaacc agccagccgg aagggccgag 2641 cgcagaagtg gtcctgcaac tttatccgcc tccatccagt ctattaattg ttgccgggaa 2701 gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat tgctacaggc 2761 atcgtggtgt cacgctcgtc gtttggtatg gcttcattca gctccggttc ccaacgatca 2821 aggcgagtta catgatcccc catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg 2881 atcgttgtca gaagtaagtt ggccgcagtg ttatcactca tggttatggc agcactgcat 2941 aattctctta ctgtcatgcc atccgtaaga tgcttttctg tgactggtga gtactcaacc 3001 aagtcattct gagaatagtg tatgcggcga ccgagttgct cttgcccggc gtcaatacgg 3061 gataataccg cgccacatag cagaacttta aaagtgctca tcattggaaa acgttcttcg 3121 gggcgaaaac tctcaaggat cttaccgctg ttgagatcca gttcgatgta acccactcgt 3181 gcacccaact gatcttcagc atcttttact ttcaccagcg tttctgggtg agcaaaaaca 3241 ggaaggcaaa atgccgcaaa aaagggaata agggcgacac ggaaatgttg aatactcata 3301 ctcttccttt ttcaatatta ttgaagcatt tatcagggtt attgtctcat gagcggatac 3361 atatttgaat gtatttagaa aaataaacaa ataggggttc cgcgcacatt tccccgaaaa 3421 gtgccacctg acgtctaaga aaccattatt atcatgacat taacctataa aaataggcgt 3481 atcacgaggc cctttcgtct tcac //