LOCUS Exported 1563 bp ds-DNA linear SYN 21-MAY-2021 DEFINITION Expresses AviTag-DogCatcher-MBP in bacterial cytoplasm. ACCESSION . VERSION . KEYWORDS pET28-AviTag-DogCatcher-MBP SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 1563) TITLE DogCatcher allows Loop-friendly Protein-Protein Ligation JOURNAL Unpublished REFERENCE 2 (bases 1 to 1563) AUTHORS . TITLE Direct Submission JOURNAL Exported May 21, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..1563 /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 64..108 /codon_start=1 /product="peptide tag that allows for enzymatic biotinylation" /label=AviTag(TM) /translation="GLNDIFEAQKIEWHE" CDS 451..1548 /codon_start=1 /gene="malE (mutated)" /product="maltose binding protein from E. coli" /label=MBP /note="This version of the gene does not encode a signal sequence, so MBP will remain in the cytosol." /translation="KIEEGKLVIWINGDKGYNGLAEVGKKFEKDTGIKVTVEHPDKLEE KFPQVAATGDGPDIIFWAHDRFGGYAQSGLLAEITPDKAFQDKLYPFTWDAVRYNGKLI AYPIAVEALSLIYNKDLLPNPPKTWEEIPALDKELKAKGKSALMFNLQEPYFTWPLIAA DGGYAFKYENGKYDIKDVGVDNAGAKAGLTFLVDLIKNKHMNADTDYSIAEAAFNKGET AMTINGPWAWSNIDTSKVNYGVTVLPTFKGQPSKPFVGVLSAGINAASPNKELAKEFLE NYLLTDEGLEAVNKDKPLGAVALKSYEEELAKDPRIAATMENAQKGEIMPNIPQMSAFW YAVRTAVINAASGRQTVDEALKDAQT" primer_bind 1522..1545 /label=MBP-F /note="Maltose binding protein, forward primer" ORIGIN 1 atgggcagca gccatcatca tcatcatcac agcagcggcc tggtgccgcg cggcagccat 61 atgggtctga atgatatttt cgaagcgcag aaaattgaat ggcatgaagg tagcggaatg 121 aaactgggcg agattgaatt tattaaagtg gacaaaaccg ataaaaagcc gctgcgtggt 181 gccgtgttta gcctgcagaa acagcatccc gactatcccg atatctatgg cgcgattgat 241 cagaatggga cctatcaaga tgtgcgtacc ggcgaagatg gtaaactgac ctttacgaat 301 ctgagcgatg gcaaatatcg cctgattgaa aatagcgaac ccccgggcta taaaccggtg 361 cagaataagc cgattgtgag ctttcgtatt gtggatggcg aagtgcgtga tgtgaccagt 421 attgtgccgc agggtagtgg tgaaagtggt aaaatcgaag aaggtaaact ggtaatctgg 481 attaacggcg ataaaggcta taacggtctc gctgaagtcg gtaagaaatt cgagaaagat 541 accggaatta aagtcaccgt tgagcatccg gataaactgg aagagaaatt cccacaggtt 601 gcggcaactg gcgatggccc tgacattatc ttctgggcac acgaccgctt tggtggctac 661 gctcaatctg gcctgttggc tgaaatcacc ccggacaaag cgttccagga caagctgtat 721 ccgtttacct gggatgccgt acgttacaac ggcaagctga ttgcttaccc gatcgctgtt 781 gaagcgttat cgctgattta taacaaagat ctgctgccga acccgccaaa aacctgggaa 841 gagatcccgg cgctggataa agaactgaaa gcgaaaggta agagcgcgct gatgttcaac 901 ctgcaagaac cgtacttcac ctggccgctg attgctgctg acgggggtta tgcgttcaag 961 tatgaaaacg gcaagtacga cattaaagac gtgggcgtgg ataacgctgg cgcgaaagcg 1021 ggtctgacct tcctggttga cctgattaaa aacaaacaca tgaatgcaga caccgattac 1081 tccatcgcag aagctgcctt taataaaggc gaaacagcga tgaccatcaa cggcccgtgg 1141 gcatggtcca acatcgacac cagcaaagtg aattatggtg taacggtact gccgaccttc 1201 aagggtcaac catccaaacc gttcgttggc gtgctgagcg caggtattaa cgccgccagt 1261 ccgaacaaag agctggcaaa agagttcctc gaaaactatc tgctgactga tgaaggtctg 1321 gaagcggtta ataaagacaa accgctgggt gccgtagcgc tgaagtctta cgaggaagag 1381 ttggcgaaag atccacgtat tgccgccact atggaaaacg cccagaaagg tgaaatcatg 1441 ccgaacatcc cgcagatgtc cgctttctgg tatgccgtgc gtactgcggt gatcaacgcc 1501 gccagcggtc gtcagactgt cgatgaagcc ctgaaagacg cgcagactaa ttcgagctcg 1561 taa //