LOCUS Exported 5653 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION Bacterial expression of SnoopCatcher, an engineered protein forming a spontaneous covalent bond with its cognate peptide partner SnoopTag.. ACCESSION . VERSION . KEYWORDS pET28a-SnoopCatcher SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 5653) AUTHORS Veggiani G, Nakamura T, Brenner MD, Gayet RV, Yan J, Robinson CV, Howarth M TITLE Programmable polyproteams built using twin peptide superglues. JOURNAL Proc Natl Acad Sci U S A. 2016 Jan 19. pii: 201519214. PUBMED 26787909 REFERENCE 2 (bases 1 to 5653) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..5653 /organism="synthetic DNA construct" /mol_type="other DNA" terminator 26..73 /label=T7 terminator /note="transcription terminator for bacteriophage T7 RNA polymerase" primer_bind 69..87 /label=T7 Term /note="T7 terminator, reverse primer" CDS complement(527..544) /codon_start=1 /product="thrombin recognition and cleavage site" /label=thrombin site /translation="LVPRGS" CDS complement(554..571) /codon_start=1 /product="6xHis affinity tag" /label=6xHis /translation="HHHHHH" RBS 590..612 /note="efficient ribosome binding site from bacteriophage T7 gene 10 (Olins and Rangwala, 1989)" protein_bind 627..651 /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)." primer_bind complement(651..670) /label=T7 /note="T7 promoter, forward primer" promoter complement(652..670) /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" primer_bind complement(754..773) /label=pBRrevBam /note="pBR322 vectors, tet region, downstream of BamHI, reverse primer" promoter 979..1056 /gene="lacI" /label=lacI promoter CDS 1057..2139 /codon_start=1 /gene="lacI" /product="lac repressor" /label=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)." /translation="MKPVTLYDVAEYAGVSYQTVSRVVNQASHVSAKTREKVEAAMAEL NYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSRADQLGASVVVSMVERSGV EACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSIIFSH EDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREGDWSA MSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTEDSSC YIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR ALADSLMQLARQVSRLESGQ" primer_bind complement(1076..1095) /label=LacI-R /note="LacI, reverse primer" CDS 2948..3139 /codon_start=1 /gene="rop" /product="Rop protein, which maintains plasmids at low copy number" /label=rop /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA DELYRSCLARFGDDGENL" primer_bind complement(3133..3155) /label=pGEX 3' /note="pGEX vectors, reverse primer" misc_feature 3241..3383 /label=bom /note="basis of mobility region from pBR322" primer_bind 3296..3315 /label=pRS-marker /note="pRS vectors, use to sequence yeast selectable marker" primer_bind complement(3398..3415) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(3569..4157) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(3649..3668) /label=pBR322ori-F /note="pBR322 origin, forward primer" primer_bind complement(4268..4287) /label=pENTR-R /note="pENTR vectors, reverse primer" CDS 4279..5094 /codon_start=1 /gene="aph(3')-Ia" /product="aminoglycoside phosphotransferase" /label=KanR /note="confers resistance to kanamycin in bacteria or G418 (Geneticin(R)) in eukaryotes" /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF" primer_bind complement(4352..4371) /label=Kan-R /note="Kanamycin resistance gene, reverse primer" rep_origin complement(5187..5642) /direction=LEFT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(5324..5345) /label=F1ori-F /note="F1 origin, forward primer" primer_bind 5536..5555 /label=F1ori-R /note="F1 origin, reverse primer" ORIGIN 1 atccggatat agttcctcct ttcagcaaaa aacccctcaa gacccgttta gaggccccaa 61 ggggttatgc tagttattgc tcagcggtgg cagcagccaa ctcagcttcc tttcgggctt 121 tgttagcagc cggatctcag tggtggtggt ggtagtgctc gagtgcggcc gcaagctttt 181 attatttcgg cggtatcggt tcattggtga tataatgttt accgttggta aattcatatg 241 tagccggaat atcctgcggc acaatgctgg tcacatcacg cacttcgcca ttcacaatct 301 gaaacgccac aatcggctta ttctgcaccg gtttatagcc agcgggttcg ctattttcaa 361 acaggcgata tttgccatcg ctcagattct taaaggtcag tttaccatct tcgccggtac 421 gcacattttg ataggtccca ttctgatcaa tcgcgccata gatatcggga tagtcgggat 481 gctgtttctg caggctaaac acggcaccac gcagcggctt catatggctg ccgcgcggca 541 ccaggccgct gctgtgatga tgatgatgat ggctgctgcc catggtatat ctccttctta 601 aagttaaaca aaattatttc tagaggggaa ttgttatccg ctcacaattc ccctatagtg 661 agtcgtatta atttcgcggg atcgagatct cgatcctcta cgccggacgc atcgtggccg 721 gcatcaccgg cgccacaggt gcggttgctg gcgcctatat cgccgacatc accgatgggg 781 aagatcgggc tcgccacttc gggctcatga gcgcttgttt cggcgtgggt atggtggcag 841 gccccgtggc cgggggactg ttgggcgcca tctccttgca tgcaccattc cttgcggcgg 901 cggtgctcaa cggcctcaac ctactactgg gctgcttcct aatgcaggag tcgcataagg 961 gagagcgtcg agatcccgga caccatcgaa tggcgcaaaa cctttcgcgg tatggcatga 1021 tagcgcccgg aagagagtca attcagggtg gtgaatgtga aaccagtaac gttatacgat 1081 gtcgcagagt atgccggtgt ctcttatcag accgtttccc gcgtggtgaa ccaggccagc 1141 cacgtttctg cgaaaacgcg ggaaaaagtg gaagcggcga tggcggagct gaattacatt 1201 cccaaccgcg tggcacaaca actggcgggc aaacagtcgt tgctgattgg cgttgccacc 1261 tccagtctgg ccctgcacgc gccgtcgcaa attgtcgcgg cgattaaatc tcgcgccgat 1321 caactgggtg ccagcgtggt ggtgtcgatg gtagaacgaa gcggcgtcga agcctgtaaa 1381 gcggcggtgc acaatcttct cgcgcaacgc gtcagtgggc tgatcattaa ctatccgctg 1441 gatgaccagg atgccattgc tgtggaagct gcctgcacta atgttccggc gttatttctt 1501 gatgtctctg accagacacc catcaacagt attattttct cccatgaaga cggtacgcga 1561 ctgggcgtgg agcatctggt cgcattgggt caccagcaaa tcgcgctgtt agcgggccca 1621 ttaagttctg tctcggcgcg tctgcgtctg gctggctggc ataaatatct cactcgcaat 1681 caaattcagc cgatagcgga acgggaaggc gactggagtg ccatgtccgg ttttcaacaa 1741 accatgcaaa tgctgaatga gggcatcgtt cccactgcga tgctggttgc caacgatcag 1801 atggcgctgg gcgcaatgcg cgccattacc gagtccgggc tgcgcgttgg tgcggatatc 1861 tcggtagtgg gatacgacga taccgaagac agctcatgtt atatcccgcc gttaaccacc 1921 atcaaacagg attttcgcct gctggggcaa accagcgtgg accgcttgct gcaactctct 1981 cagggccagg cggtgaaggg caatcagctg ttgcccgtct cactggtgaa aagaaaaacc 2041 accctggcgc ccaatacgca aaccgcctct ccccgcgcgt tggccgattc attaatgcag 2101 ctggcacgac aggtttcccg actggaaagc gggcagtgag cgcaacgcaa ttaatgtaag 2161 ttagctcact cattaggcac cgggatctcg accgatgccc ttgagagcct tcaacccagt 2221 cagctccttc cggtgggcgc ggggcatgac tatcgtcgcc gcacttatga ctgtcttctt 2281 tatcatgcaa ctcgtaggac aggtgccggc agcgctctgg gtcattttcg gcgaggaccg 2341 ctttcgctgg agcgcgacga tgatcggcct gtcgcttgcg gtattcggaa tcttgcacgc 2401 cctcgctcaa gccttcgtca ctggtcccgc caccaaacgt ttcggcgaga agcaggccat 2461 tatcgccggc atggcggccc cacgggtgcg catgatcgtg ctcctgtcgt tgaggacccg 2521 gctaggctgg cggggttgcc ttactggtta gcagaatgaa tcaccgatac gcgagcgaac 2581 gtgaagcgac tgctgctgca aaacgtctgc gacctgagca acaacatgaa tggtcttcgg 2641 tttccgtgtt tcgtaaagtc tggaaacgcg gaagtcagcg ccctgcacca ttatgttccg 2701 gatctgcatc gcaggatgct gctggctacc ctgtggaaca cctacatctg tattaacgaa 2761 gcgctggcat tgaccctgag tgatttttct ctggtcccgc cgcatccata ccgccagttg 2821 tttaccctca caacgttcca gtaaccgggc atgttcatca tcagtaaccc gtatcgtgag 2881 catcctctct cgtttcatcg gtatcattac ccccatgaac agaaatcccc cttacacgga 2941 ggcatcagtg accaaacagg aaaaaaccgc ccttaacatg gcccgcttta tcagaagcca 3001 gacattaacg cttctggaga aactcaacga gctggacgcg gatgaacagg cagacatctg 3061 tgaatcgctt cacgaccacg ctgatgagct ttaccgcagc tgcctcgcgc gtttcggtga 3121 tgacggtgaa aacctctgac acatgcagct cccggagacg gtcacagctt gtctgtaagc 3181 ggatgccggg agcagacaag cccgtcaggg cgcgtcagcg ggtgttggcg ggtgtcgggg 3241 cgcagccatg acccagtcac gtagcgatag cggagtgtat actggcttaa ctatgcggca 3301 tcagagcaga ttgtactgag agtgcaccat atatgcggtg tgaaataccg cacagatgcg 3361 taaggagaaa ataccgcatc aggcgctctt ccgcttcctc gctcactgac tcgctgcgct 3421 cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa ggcggtaata cggttatcca 3481 cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga 3541 accgtaaaaa ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc 3601 acaaaaatcg acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg 3661 cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat 3721 acctgtccgc ctttctccct tcgggaagcg tggcgctttc tcatagctca cgctgtaggt 3781 atctcagttc ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc 3841 agcccgaccg ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg 3901 acttatcgcc actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg 3961 gtgctacaga gttcttgaag tggtggccta actacggcta cactagaagg acagtatttg 4021 gtatctgcgc tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg 4081 gcaaacaaac caccgctggt agcggtggtt tttttgtttg caagcagcag attacgcgca 4141 gaaaaaaagg atctcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga 4201 acgaaaactc acgttaaggg attttggtca tgaacaataa aactgtctgc ttacataaac 4261 agtaatacaa ggggtgttat gagccatatt caacgggaaa cgtcttgctc taggccgcga 4321 ttaaattcca acatggatgc tgatttatat gggtataaat gggctcgcga taatgtcggg 4381 caatcaggtg cgacaatcta tcgattgtat gggaagcccg atgcgccaga gttgtttctg 4441 aaacatggca aaggtagcgt tgccaatgat gttacagatg agatggtcag actaaactgg 4501 ctgacggaat ttatgcctct tccgaccatc aagcatttta tccgtactcc tgatgatgca 4561 tggttactca ccactgcgat ccccgggaaa acagcattcc aggtattaga agaatatcct 4621 gattcaggtg aaaatattgt tgatgcgctg gcagtgttcc tgcgccggtt gcattcgatt 4681 cctgtttgta attgtccttt taacagcgat cgcgtatttc gtctcgctca ggcgcaatca 4741 cgaatgaata acggtttggt tgatgcgagt gattttgatg acgagcgtaa tggctggcct 4801 gttgaacaag tctggaaaga aatgcataaa cttttgccat tctcaccgga ttcagtcgtc 4861 actcatggtg atttctcact tgataacctt atttttgacg aggggaaatt aataggttgt 4921 attgatgttg gacgagtcgg aatcgcagac cgataccagg atcttgccat cctatggaac 4981 tgcctcggtg agttttctcc ttcattacag aaacggcttt ttcaaaaata tggtattgat 5041 aatcctgata tgaataaatt gcagtttcat ttgatgctcg atgagttttt ctaagaatta 5101 attcatgagc ggatacatat ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg 5161 cacatttccc cgaaaagtgc cacctgaaat tgtaaacgtt aatattttgt taaaattcgc 5221 gttaaatttt tgttaaatca gctcattttt taaccaatag gccgaaatcg gcaaaatccc 5281 ttataaatca aaagaataga ccgagatagg gttgagtgtt gttccagttt ggaacaagag 5341 tccactatta aagaacgtgg actccaacgt caaagggcga aaaaccgtct atcagggcga 5401 tggcccacta cgtgaaccat caccctaatc aagttttttg gggtcgaggt gccgtaaagc 5461 actaaatcgg aaccctaaag ggagcccccg atttagagct tgacggggaa agccggcgaa 5521 cgtggcgaga aaggaaggga agaaagcgaa aggagcgggc gctagggcgc tggcaagtgt 5581 agcggtcacg ctgcgcgtaa ccaccacacc cgccgcgctt aatgcgccgc tacagggcgc 5641 gtcccattcg cca //