LOCUS Exported 1284 bp ds-DNA linear SYN 17-MAY-2021 DEFINITION Expresses SpyTag-SpyCatcher mediated cyclized version of TEM-1 beta-lactamase (BLA). Cyclization of BLA led to an increase in thermal resilience of >60 degrees celsius.. ACCESSION . VERSION . KEYWORDS SpyTag-beta-Lactamase-SpyCatcher SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 1284) AUTHORS Schoene C, Fierer JO, Bennett SP, Howarth M TITLE SpyTag/SpyCatcher cyclization confers resilience to boiling on a mesophilic enzyme. JOURNAL Angew Chem Int Ed Engl. 2014 Jun 10;53(24):6101-4. doi: 10.1002/anie.201402519. Epub 2014 May 9. PUBMED 24817566 REFERENCE 2 (bases 1 to 1284) AUTHORS . TITLE Direct Submission JOURNAL Exported May 17, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..1284 /organism="synthetic DNA construct" /mol_type="other DNA" CDS 13..30 /codon_start=1 /product="6xHis affinity tag" /label=6xHis /translation="HHHHHH" CDS 40..57 /codon_start=1 /product="thrombin recognition and cleavage site" /label=thrombin site /translation="LVPRGS" CDS 127..912 /codon_start=1 /gene="bla(M)" /product="beta-lactamase lacking the signal sequence" /label=bla(M) /note="allows cytosolic expression of beta-lactamase" /translation="PETLVKVKDAEDQLGARVGYIELDLNSGKILESFRPEERFPMMST FKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYSPVTEKHLTDGMTVRELCSAAITMS DNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRWEPELNEAIPNDERDTTMPAAMATT LRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSALPAGWFIADKSGAGERGSRGIIAA LGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGASLIKHW" primer_bind complement(273..292) /label=Amp-R /note="Ampicillin resistance gene, reverse primer" ORIGIN 1 atgggcagca gccatcatca tcatcatcac agcagcggcc tggtgccgcg cggcagccat 61 atgggagccc acatcgtgat ggtggacgcc tacaagccga cgaagggttc agggggttcc 121 ggtcacccag aaacgctggt gaaagtaaaa gatgctgaag atcagttggg tgcacgagtg 181 ggttacatcg aactggatct caacagcggt aagatccttg agagttttcg ccccgaagaa 241 cgttttccaa tgatgagcac ttttaaagtt ctgctatgtg gcgcggtatt atcccgtatt 301 gacgccgggc aagagcaact cggtcgccgc atacactatt ctcagaatga cttggttgag 361 tactcaccag tcacagaaaa gcatcttacg gatggcatga cagtaagaga attatgcagt 421 gctgccataa ccatgagtga taacactgcg gccaacttac ttctgacaac gatcggagga 481 ccgaaggagc taaccgcttt tttgcacaac atgggggatc atgtaactcg ccttgatcgt 541 tgggaaccgg agctgaatga agccatacca aacgacgagc gtgacaccac gatgcctgca 601 gcaatggcaa caacgttgcg caaactatta actggcgaac tacttactct agcttcccgg 661 caacaattaa tagactggat ggaggcggat aaagttgcag gaccacttct gcgctcggcc 721 cttccggctg gctggtttat tgctgataaa tctggagccg gtgagcgtgg gtctcgcggt 781 atcattgcag cactggggcc agatggtaag ccctcccgta tcgtagttat ctacacgacg 841 gggagtcagg caactatgga tgaacgaaat agacagatcg ctgagatagg tgcctcactg 901 attaagcatt gggggagtgg tggcagcgga ggcgccatgg ttgatacctt atcaggttta 961 tcaagtgagc aaggtcagtc cggtgatatg acaattgaag aagatagtgc tacccatatt 1021 aaattctcaa aacgtgatga ggacggcaaa gagttagctg gtgcaactat ggagttgcgt 1081 gattcatctg gtaaaactat tagtacatgg atttcagatg gacaagtgaa agatttctac 1141 ctgtatccag gaaaatatac atttgtcgaa accgcagcac cagacggtta tgaggtagca 1201 actgctatta cctttacagt taatgagcaa ggtcaggtta ctgtaaatgg caaagcaact 1261 aaaggtgacg ctcatattta atga //