LOCUS Exported 4263 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION Bacterial expression plasmid for PURE system components. ACCESSION . VERSION . KEYWORDS pQE60-EFTs-His SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 4263) AUTHORS Shimizu Y, Inoue A, Tomari Y, Suzuki T, Yokogawa T, Nishikawa K, Ueda T TITLE Cell-free translation reconstituted with purified components. JOURNAL Nat Biotechnol. 2001 Aug;19(8):751-5. doi: 10.1038/90802. PUBMED 11479568 REFERENCE 2 (bases 1 to 4263) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..4263 /organism="synthetic DNA construct" /mol_type="other DNA" CDS complement(257..1117) /codon_start=1 /gene="bla" /product="beta-lactamase" /label=AmpR /note="confers resistance to ampicillin, carbenicillin, and related antibiotics" /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS LIKHW" primer_bind 880..899 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(1118..1222) /gene="bla" /label=AmpR promoter primer_bind 1290..1308 /label=pBRforEco /note="pBR322 vectors, upsteam of EcoRI site, forward primer" promoter 1332..1376 /label=T5 promoter /note="bacteriophage T5 promoter for E. coli RNA polymerase, with embedded lac operator" protein_bind 1352..1368 /label=lac operator /bound_moiety="lac repressor encoded by lacI" /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 1384..1400 /label=lac operator /bound_moiety="lac repressor encoded by lacI" /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)." CDS 2288..2305 /codon_start=1 /product="6xHis affinity tag" /label=6xHis /translation="HHHHHH" terminator 2328..2422 /label=lambda t0 terminator /note="transcription terminator from phage lambda" primer_bind complement(2532..2551) /label=CAT-R /note="Chloramphenicol resistance gene, reverse primer" terminator 3194..3280 /gene="Escherichia coli rrnB" /label=rrnB T1 terminator /note="transcription terminator T1 from the E. coli rrnB gene" primer_bind complement(3327..3349) /label=pGEX 3' /note="pGEX vectors, reverse primer" misc_feature 3435..3575 /label=bom /note="basis of mobility region from pBR322" primer_bind 3490..3509 /label=pRS-marker /note="pRS vectors, use to sequence yeast selectable marker" primer_bind complement(3590..3607) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(3761..86) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(3841..3860) /label=pBR322ori-F /note="pBR322 origin, forward primer" ORIGIN 1 cttgatccgg caaacaaacc accgctggta gcggtggttt ttttgtttgc aagcagcaga 61 ttacgcgcag aaaaaaagga tctcaagaag atcctttgat cttttctacg gggtctgacg 121 ctcagtggaa cgaaaactca cgttaaggga ttttggtcat gagattatca aaaaggatct 181 tcacctagat ccttttaaat taaaaatgaa gttttaaatc aatctaaagt atatatgagt 241 aaacttggtc tgacagttac caatgcttaa tcagtgaggc acctatctca gcgatctgtc 301 tatttcgttc atccatagtt gcctgactcc ccgtcgtgta gataactacg atacgggagg 361 gcttaccatc tggccccagt gctgcaatga taccgcgaga cccacgctca ccggctccag 421 atttatcagc aataaaccag ccagccggaa gggccgagcg cagaagtggt cctgcaactt 481 tatccgcctc catccagtct attaattgtt gccgggaagc tagagtaagt agttcgccag 541 ttaatagttt gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt 601 ttggtatggc ttcattcagc tccggttccc aacgatcaag gcgagttaca tgatccccca 661 tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat cgttgtcaga agtaagttgg 721 ccgcagtgtt atcactcatg gttatggcag cactgcataa ttctcttact gtcatgccat 781 ccgtaagatg cttttctgtg actggtgagt actcaaccaa gtcattctga gaatagtgta 841 tgcggcgacc gagttgctct tgcccggcgt caatacggga taataccgcg ccacatagca 901 gaactttaaa agtgctcatc attggaaaac gttcttcggg gcgaaaactc tcaaggatct 961 taccgctgtt gagatccagt tcgatgtaac ccactcgtgc acccaactga tcttcagcat 1021 cttttacttt caccagcgtt tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa 1081 agggaataag ggcgacacgg aaatgttgaa tactcatact cttccttttt caatattatt 1141 gaagcattta tcagggttat tgtctcatga gcggatacat atttgaatgt atttagaaaa 1201 ataaacaaat aggggttccg cgcacatttc cccgaaaagt gccacctgac gtctaagaaa 1261 ccattattat catgacatta acctataaaa ataggcgtat cacgaggccc tttcgtcttc 1321 acctcgagaa atcataaaaa atttatttgc tttgtgagcg gataacaatt ataatagatt 1381 caattgtgag cggataacaa tttcacacag aattgatctc cgaggatttt agatggctga 1441 aattaccgca tccctggtaa aagagctgcg tgagcgtact ggcgcaggca tgatggattg 1501 caaaaaagca ctgactgaag ctaacggcga catcgagctg gcaatcgaaa acatgcgtaa 1561 gtccggtgct attaaagcag cgaaaaaagc aggcaacgtt gctgctgacg gcgtgatcaa 1621 aaccaaaatc gacggcaact acggcatcat tctggaagtt aactgccaga ctgacttcgt 1681 tgcaaaagac gctggtttcc aggcgttcgc agacaaagtt ctggacgcag ctgttgctgg 1741 caaaatcact gacgttgaag ttctgaaagc acagttcgaa gaagaacgtg ttgcgctggt 1801 agcgaaaatt ggtgaaaaca tcaacattcg ccgcgttgct gcgctggaag gcgacgttct 1861 gggttcttat cagcacggtg cgcgtatcgg cgttctggtt gctgctaaag gcgctgacga 1921 agagctggtt aaacacatcg ctatgcacgt tgctgcaagc aagccagaat tcatcaaacc 1981 ggaagacgta tccgctgaag tggtagaaaa agaataccag gtacagctgg atatcgcgat 2041 gcagtctggt aagccgaaag aaatcgcaga gaaaatggtt gaaggccgca tgaagaaatt 2101 caccggcgaa gtttctctga ccggtcagcc gttcgttatg gaaccaagca aaactgttgg 2161 tcagctgctg aaagagcata acgctgaagt gactggcttc atccgcttcg aagtgggtga 2221 aggcatcgag aaagttgaga ctgactttgc agcagaagtt gctgcgatgt ccaagcagtc 2281 tagatctcat caccatcacc atcactaagc ttaattagct gagcttggac tcctgttgat 2341 agatccagta atgacctcag aactccatct ggatttgttc agaacgctcg gttgccgccg 2401 ggcgtttttt attggtgaga atccaagcta gcttggcgag attttcagga gctaaggaag 2461 ctaaaatgga gaaaaaaatc actggatata ccaccgttga tatatcccaa tggcatcgta 2521 aagaacattt tgaggcattt cagtcagttg ctcaatgtac ctataaccag accgttcagc 2581 tggatattac ggccttttta aagaccgtaa agaaaaataa gcacaagttt tatccggcct 2641 ttattcacat tcttgcccgc ctgatgaatg ctcatccgga atttcgtatg gcaatgaaag 2701 acggtgagct ggtgatatgg gatagtgttc acccttgtta caccgttttc catgagcaaa 2761 ctgaaacgtt ttcatcgctc tggagtgaat accacgacga tttccggcag tttctacaca 2821 tatattcgca agatgtggcg tgttacggtg aaaacctggc ctatttccct aaagggttta 2881 ttgagaatat gtttttcgtc tcagccaatc cctgggtgag tttcaccagt tttgatttaa 2941 acgtggccaa tatggacaac ttcttcgccc ccgttttcac catgcatggg caaatattat 3001 acgcaaggcg acaaggtgct gatgccgctg gcgattcagg ttcatcatgc cgtttgtgat 3061 ggcttccatg tcggcagaat gcttaatgaa ttacaacagt actgcgatga gtggcagggc 3121 ggggcgtaat ttttttaagg cagttattgg tgcccttaaa cgcctggggt aatgactctc 3181 tagcttgagg catcaaataa aacgaaaggc tcagtcgaaa gactgggcct ttcgttttat 3241 ctgttgtttg tcggtgaacg ctctcctgag taggacaaat ccgccctcta gagctgcctc 3301 gcgcgtttcg gtgatgacgg tgaaaacctc tgacacatgc agctcccgga gacggtcaca 3361 gcttgtctgt aagcggatgc cgggagcaga caagcccgtc agggcgcgtc agcgggtgtt 3421 ggcgggtgtc ggggcgcagc catgacccag tcacgtagcg atagcggagt gtatactggc 3481 ttaactatgc ggcatcagag cagattgtac tgagagtgca ccatatgcgg tgtgaaatac 3541 cgcacagatg cgtaaggaga aaataccgca tcaggcgctc ttccgcttcc tcgctcactg 3601 actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc agctcactca aaggcggtaa 3661 tacggttatc cacagaatca ggggataacg caggaaagaa catgtgagca aaaggccagc 3721 aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg ctccgccccc 3781 ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg acaggactat 3841 aaagatacca ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc 3901 cgcttaccgg atacctgtcc gcctttctcc cttcgggaag cgtggcgctt tctcatagct 3961 cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg 4021 aaccccccgt tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc 4081 cggtaagaca cgacttatcg ccactggcag cagccactgg taacaggatt agcagagcga 4141 ggtatgtagg cggtgctaca gagttcttga agtggtggcc taactacggc tacactagaa 4201 ggacagtatt tggtatctgc gctctgctga agccagttac cttcggaaaa agagttggta 4261 gct //