LOCUS Exported 4055 bp ds-DNA circular SYN 12-MAY-2022 DEFINITION Expresses SpySwitch in the bacterial cytoplasm. SpySwitch binds SpyTag-, SpyTag002- or SpyTag003-fusions non-covalently for affinity purification, allowing gentle pH or temperature release.. ACCESSION . VERSION . KEYWORDS SpySwitch SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 4055) TITLE SpySwitch enables pH- or heat-responsive capture and release for plug-and-display nanoassembly JOURNAL Unpublished REFERENCE 2 (bases 1 to 4055) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2022 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..4055 /organism="synthetic DNA construct" /mol_type="other DNA" RBS 41..63 /note="efficient ribosome binding site from bacteriophage T7 gene 10 (Olins and Rangwala, 1989)" CDS 83..100 /codon_start=1 /product="6xHis affinity tag" /label=6xHis /translation="HHHHHH" CDS 546..563 /codon_start=1 /product="6xHis affinity tag" /label=6xHis /translation="HHHHHH" primer_bind complement(616..634) /label=T7 Term /note="T7 terminator, reverse primer" terminator 630..677 /label=T7 terminator /note="transcription terminator for bacteriophage T7 RNA polymerase" rep_origin 714..1169 /direction=RIGHT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(801..820) /label=F1ori-R /note="F1 origin, reverse primer" primer_bind 1011..1032 /label=F1ori-F /note="F1 origin, forward primer" promoter 1195..1299 /gene="bla" /label=AmpR promoter CDS 1300..2160 /codon_start=1 /gene="bla" /product="beta-lactamase" /label=AmpR /note="confers resistance to ampicillin, carbenicillin, and related antibiotics" /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS LIKHW" primer_bind complement(1518..1537) /label=Amp-R /note="Ampicillin resistance gene, reverse primer" rep_origin 2331..2919 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 2820..2839 /label=pBR322ori-F /note="pBR322 origin, forward primer" primer_bind 3073..3090 /label=L4440 /note="L4440 vector, forward primer" misc_feature 3105..3244 /label=bom /note="basis of mobility region from pBR322" primer_bind complement(3170..3189) /label=pRS-marker /note="pRS vectors, use to sequence yeast selectable marker" primer_bind 3330..3352 /label=pGEX 3' /note="pGEX vectors, reverse primer" CDS complement(3346..3537) /codon_start=1 /gene="rop" /product="Rop protein, which maintains plasmids at low copy number" /label=rop /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA DELYRSCLARFGDDGENL" primer_bind 4046..10 /label=T7 /note="T7 promoter, forward primer" promoter 4046..9 /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" ORIGIN 1 cactataggg agaccacaac ggtttccctc tagaaataat tttgtttaac tttaagaagg 61 agatatacat atgtcgtact accatcacca tcaccatcac ggtagcggcg gtagcggcgc 121 catggtaacc accttatcag gtttatcagg tgagcaaggt ccgtccggtg atatgacaac 181 tgaagaagat agtgctaccc atattcactt ccataaacat gatgaggacg gccgtgagtt 241 agctggtgca actatggagt tgcgtgattg ctctggtaaa actattagta catggattac 301 agatggacat gtgaaggatt tctacctgta tccaggaaaa tatacatttg tcgcgaccgc 361 agcaccagac ggtcatcacg tagcaactcc aattgaattt acaattaatg aggacggtca 421 ggttactgta gatggtgaag caactgaagg tgaccctcat actggatcca gtggtagcta 481 ataaatggtt gatgcttgag gatccgaatt cgagctccgt cgacaagctt gcggccgcac 541 tcgagcacca ccaccaccac cactgagatc cggctgctaa caaagcccga aaggaagctg 601 agttggctgc tgccaccgct gagcaataac tagcataacc ccttggggcc tctaaacggg 661 tcttgagggg ttttttgctg aaaggaggaa ctatatccgg attggcgaat gggacgcgcc 721 ctgtagcggc gcattaagcg cggcgggtgt ggtggttacg cgcagcgtga ccgctacact 781 tgccagcgcc ctagcgcccg ctcctttcgc tttcttccct tcctttctcg ccacgttcgc 841 cggctttccc cgtcaagctc taaatcgggg gctcccttta gggttccgat ttagtgcttt 901 acggcacctc gaccccaaaa aacttgatta gggtgatggt tcacgtagtg ggccatcgcc 961 ctgatagacg gtttttcgcc ctttgacgtt ggagtccacg ttctttaata gtggactctt 1021 gttccaaact ggaacaacac tcaaccctat ctcggtctat tcttttgatt tataagggat 1081 tttgccgatt tcggcctatt ggttaaaaaa tgagctgatt taacaaaaat ttaacgcgaa 1141 ttttaacaaa atattaacgc ttacaattta ggtggcactt ttcggggaaa tgtgcgcgga 1201 acccctattt gtttattttt ctaaatacat tcaaatatgt atccgctcat gagacaataa 1261 ccctggtaaa tgcttcaata atattgaaaa aggaagagta tgagtattca acatttccgt 1321 gtcgccctta ttcccttttt tgcggcattt tgccttcctg tttttgctca cccagaaacg 1381 ctggtgaaag taaaagatgc tgaagatcag ttgggtgcac gagtgggtta catcgaactg 1441 gatctcaaca gcggtaagat ccttgagagt tttcgccccg aagaacgttt tccaatgatg 1501 agcactttta aagttctgct atgtggcgcg gtattatccc gtattgacgc cgggcaagag 1561 caactcggtc gccgcataca ctattctcag aatgacttgg ttgagtactc accagtcaca 1621 gaaaagcatc ttacggatgg catgacagta agagaattat gcagtgctgc cataaccatg 1681 agtgataaca ctgcggccaa cttacttctg acaacgatcg gaggaccgaa ggagctaacc 1741 gcttttttgc acaacatggg ggatcatgta actcgccttg atcgttggga accggagctg 1801 aatgaagcca taccaaacga cgagcgtgac accacgatgc ctgcagcaat ggcaacaacg 1861 ttgcgcaaac tattaactgg cgaactactt actctagctt cccggcaaca attaatagac 1921 tggatggagg cggataaagt tgcaggacca cttctgcgct cggcccttcc ggctggctgg 1981 tttattgctg ataaatctgg agccggtgag cgtgggtctc gcggtatcat tgcagcactg 2041 gggccagatg gtaagccctc ccgtatcgta gttatctaca cgacggggag tcaggcaact 2101 atggatgaac gaaatagaca gatcgctgag ataggtgcct cactgattaa gcattggtaa 2161 ctgtcagacc aagtttactc atatatactt tagattgatt taaaacttca tttttaattt 2221 aaaaggatct aggtgaagat cctttttgat aatctcatga ccaaaatccc ttaacgtgag 2281 ttttcgttcc actgagcgtc agaccccgta gaaaagatca aaggatcttc ttgagatcct 2341 ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt 2401 tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt cagcagagcg 2461 cagataccaa atactgtcct tctagtgtag ccgtagttag gccaccactt caagaactct 2521 gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc tgccagtggc 2581 gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa ggcgcagcgg 2641 tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac ctacaccgaa 2701 ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg 2761 gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga gcttccaggg 2821 ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact tgagcgtcga 2881 tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa cgcggccttt 2941 ttacggttcc tggccttttg ctggcctttt gctcacatgt tctttcctgc gttatcccct 3001 gattctgtgg ataaccgtat taccgccttt gagtgagctg ataccgctcg ccgcagccga 3061 acgaccgagc gcagcgagtc agtgagcgag gaagcggaag agcgcctgat gcggtatttt 3121 ctccttacgc atctgtgcgg tatttcacac cgcaatggtg cactctcagt acaatctgct 3181 ctgatgccgc atagttaagc cagtatacac tccgctatcg ctacgtgact gggtcatggc 3241 tgcgccccga cacccgccaa cacccgctga cgcgccctga cgggcttgtc tgctcccggc 3301 atccgcttac agacaagctg tgaccgtctc cgggagctgc atgtgtcaga ggttttcacc 3361 gtcatcaccg aaacgcgcga ggcagctgcg gtaaagctca tcagcgtggt cgtgaagcga 3421 ttcacagatg tctgcctgtt catccgcgtc cagctcgttg agcttctcca gaagcgttaa 3481 tgtctggctt ctgataaagc gggccatgtt aagggcggtt ttttcctgtt tggtcactga 3541 tgcctccgtg taagggggat ttctgttcat gggggtaatg ataccgatga aacgagagag 3601 gatgctcacg atacgggtta ctgatgatga acatgcccgg ttactggaac gttgtgaggg 3661 taaacaactg gcggtatgga tgcggcggga ccagagaaaa atcactcagg gtcaatgcca 3721 gcgcttcgtt aatacagatg taggtgttcc acagggtagc cagcagcatc ctgcgatgca 3781 gatccggaac ataatggtgc agggcgctga cttccgcgtt tccagacttt acgaaacacg 3841 gaaaccgaag accattcatg ttgttgctca ggtcgcagac gttttgcagc agcagtcgct 3901 tcacgttcgc tcgcgtatcg gtgattcatt ctgctaacca gtaaggcaac cccgccagcc 3961 tagccgggtc ctcaacgaca ggagcacaat catgcgcacc cgtggccagg acccaacgct 4021 gcccgagatc tcgatcccgc gaaattaata cgact //