LOCUS Exported 5677 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION wildtype Fgß peptide with ddFLN4 fingerprint domain with C-terminal ybbr tag for covalent immobilization - resulting in high forces. ACCESSION . VERSION . KEYWORDS pET28a-Fgß-ddFLN4-HIS-ybbr SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 5677) AUTHORS Milles LF, Schulten K, Gaub HE, Bernardi RC, TITLE Molecular mechanism of extreme mechanostability in a pathogen adhesin JOURNAL Science 30 Mar 2018: Vol. 359, Issue 6383, pp. 1527-1533 REFERENCE 2 (bases 1 to 5677) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..5677 /organism="synthetic DNA construct" /mol_type="other DNA" primer_bind 34..53 /label=T7 /note="T7 promoter, forward primer" promoter 34..52 /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" protein_bind 53..77 /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)." RBS 92..114 /note="efficient ribosome binding site from bacteriophage T7 gene 10 (Olins and Rangwala, 1989)" CDS 527..544 /codon_start=1 /product="6xHis affinity tag" /label=6xHis /translation="HHHHHH" primer_bind complement(645..663) /label=T7 Term /note="T7 terminator, reverse primer" terminator 659..706 /label=T7 terminator /note="transcription terminator for bacteriophage T7 RNA polymerase" rep_origin 743..1198 /direction=RIGHT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(830..849) /label=F1ori-R /note="F1 origin, reverse primer" primer_bind 1040..1061 /label=F1ori-F /note="F1 origin, forward primer" CDS complement(1290..2105) /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 2013..2032 /label=Kan-R /note="Kanamycin resistance gene, reverse primer" primer_bind 2097..2116 /label=pENTR-R /note="pENTR vectors, reverse primer" rep_origin 2227..2815 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 2716..2735 /label=pBR322ori-F /note="pBR322 origin, forward primer" primer_bind 2969..2986 /label=L4440 /note="L4440 vector, forward primer" misc_feature 3001..3140 /label=bom /note="basis of mobility region from pBR322" primer_bind complement(3066..3085) /label=pRS-marker /note="pRS vectors, use to sequence yeast selectable marker" primer_bind 3226..3248 /label=pGEX 3' /note="pGEX vectors, reverse primer" CDS complement(3242..3433) /codon_start=1 /gene="rop" /product="Rop protein, which maintains plasmids at low copy number" /label=rop /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA DELYRSCLARFGDDGENL" CDS complement(4242..5324) /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 5286..5305 /label=LacI-R /note="LacI, reverse primer" promoter complement(5325..5402) /gene="lacI" /label=lacI promoter primer_bind 5608..5627 /label=pBRrevBam /note="pBR322 vectors, tet region, downstream of BamHI, reverse primer" ORIGIN 1 gcgtagagga tcgagatctc gatcccgcga aattaatacg actcactata ggggaattgt 61 gagcggataa caattcccct gtagaaataa ttttgtttaa ctttaagaag gagatataca 121 tatgggtacc aatgaagagg gctttttcag cgcacgtggt catcgtccgc tggatggtag 181 cggtagcggt tcaggtagcg caggtacagg tagcggtgca gatccggaaa aaagctatgc 241 cgaaggtccg ggtctggatg gtggtgaaag ttttcagccg agcaaattca aaattcatgc 301 agttgatccg gatggtgttc atcgtaccga tggcggtgat ggttttgttg ttaccattga 361 aggtcctgca ccggtggatc cggttatggt tgataatggt gatggcacct atgatgttga 421 atttgaaccg aaagaagcag gcgattacgt tattaatctg accctggatg gcgataatgt 481 taatggcttt ccgaaaaccg ttaccgttaa accggcaccg agcggtcatc atcatcacca 541 tcatggtagc gactctctgg aattcatcgc ttctaaactg gcttaactcg agtaagatcc 601 ggctgctaac aaagcccgaa aggaagctga gttggctgct gccaccgctg agcaataact 661 agcataaccc cttggggcct ctaaacgggt cttgaggggt tttttgctga aaggaggaac 721 tatatccgga ttggcgaatg ggacgcgccc tgtagcggcg cattaagcgc ggcgggtgtg 781 gtggttacgc gcagcgtgac cgctacactt gccagcgccc tagcgcccgc tcctttcgct 841 ttcttccctt cctttctcgc cacgttcgcc ggctttcccc gtcaagctct aaatcggggg 901 ctccctttag ggttccgatt tagtgcttta cggcacctcg accccaaaaa acttgattag 961 ggtgatggtt cacgtagtgg gccatcgccc tgatagacgg tttttcgccc tttgacgttg 1021 gagtccacgt tctttaatag tggactcttg ttccaaactg gaacaacact caaccctatc 1081 tcggtctatt cttttgattt ataagggatt ttgccgattt cggcctattg gttaaaaaat 1141 gagctgattt aacaaaaatt taacgcgaat tttaacaaaa tattaacgct tacaatttag 1201 gtggcacttt tcggggaaat gtgcgcggaa cccctatttg tttatttttc taaatacatt 1261 caaatatgta tccgctcatg aattaattct tagaaaaact catcgagcat caaatgaaac 1321 tgcaatttat tcatatcagg attatcaata ccatattttt gaaaaagccg tttctgtaat 1381 gaaggagaaa actcaccgag gcagttccat aggatggcaa gatcctggta tcggtctgcg 1441 attccgactc gtccaacatc aatacaacct attaatttcc cctcgtcaaa aataaggtta 1501 tcaagtgaga aatcaccatg agtgacgact gaatccggtg agaatggcaa aagtttatgc 1561 atttctttcc agacttgttc aacaggccag ccattacgct cgtcatcaaa atcactcgca 1621 tcaaccaaac cgttattcat tcgtgattgc gcctgagcga gacgaaatac gcgatcgctg 1681 ttaaaaggac aattacaaac aggaatcgaa tgcaaccggc gcaggaacac tgccagcgca 1741 tcaacaatat tttcacctga atcaggatat tcttctaata cctggaatgc tgttttcccg 1801 gggatcgcag tggtgagtaa ccatgcatca tcaggagtac ggataaaatg cttgatggtc 1861 ggaagaggca taaattccgt cagccagttt agtctgacca tctcatctgt aacatcattg 1921 gcaacgctac ctttgccatg tttcagaaac aactctggcg catcgggctt cccatacaat 1981 cgatagattg tcgcacctga ttgcccgaca ttatcgcgag cccatttata cccatataaa 2041 tcagcatcca tgttggaatt taatcgcggc ctagagcaag acgtttcccg ttgaatatgg 2101 ctcataacac cccttgtatt actgtttatg taagcagaca gttttattgt tcatgaccaa 2161 aatcccttaa cgtgagtttt cgttccactg agcgtcagac cccgtagaaa agatcaaagg 2221 atcttcttga gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc 2281 gctaccagcg gtggtttgtt tgccggatca agagctacca actctttttc cgaaggtaac 2341 tggcttcagc agagcgcaga taccaaatac tgtccttcta gtgtagccgt agttaggcca 2401 ccacttcaag aactctgtag caccgcctac atacctcgct ctgctaatcc tgttaccagt 2461 ggctgctgcc agtggcgata agtcgtgtct taccgggttg gactcaagac gatagttacc 2521 ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc acacagccca gcttggagcg 2581 aacgacctac accgaactga gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc 2641 cgaagggaga aaggcggaca ggtatccggt aagcggcagg gtcggaacag gagagcgcac 2701 gagggagctt ccagggggaa acgcctggta tctttatagt cctgtcgggt ttcgccacct 2761 ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc 2821 cagcaacgcg gcctttttac ggttcctggc cttttgctgg ccttttgctc acatgttctt 2881 tcctgcgtta tcccctgatt ctgtggataa ccgtattacc gcctttgagt gagctgatac 2941 cgctcgccgc agccgaacga ccgagcgcag cgagtcagtg agcgaggaag cggaagagcg 3001 cctgatgcgg tattttctcc ttacgcatct gtgcggtatt tcacaccgca atggtgcact 3061 ctcagtacaa tctgctctga tgccgcatag ttaagccagt atacactccg ctatcgctac 3121 gtgactgggt catggctgcg ccccgacacc cgccaacacc cgctgacgcg ccctgacggg 3181 cttgtctgct cccggcatcc gcttacagac aagctgtgac cgtctccggg agctgcatgt 3241 gtcagaggtt ttcaccgtca tcaccgaaac gcgcgaggca gctgcggtaa agctcatcag 3301 cgtggtcgtg aagcgattca cagatgtctg cctgttcatc cgcgtccagc tcgttgagtt 3361 tctccagaag cgttaatgtc tggcttctga taaagcgggc catgttaagg gcggtttttt 3421 cctgtttggt cactgatgcc tccgtgtaag ggggatttct gttcatgggg gtaatgatac 3481 cgatgaaacg agagaggatg ctcacgatac gggttactga tgatgaacat gcccggttac 3541 tggaacgttg tgagggtaaa caactggcgg tatggatgcg gcgggaccag agaaaaatca 3601 ctcagggtca atgccagcgc ttcgttaata cagatgtagg tgttccacag ggtagccagc 3661 agcatcctgc gatgcagatc cggaacataa tggtgcaggg cgctgacttc cgcgtttcca 3721 gactttacga aacacggaaa ccgaagacca ttcatgttgt tgctcaggtc gcagacgttt 3781 tgcagcagca gtcgcttcac gttcgctcgc gtatcggtga ttcattctgc taaccagtaa 3841 ggcaaccccg ccagcctagc cgggtcctca acgacaggag cacgatcatg cgcacccgtg 3901 gggccgccat gccggcgata atggcctgct tctcgccgaa acgtttggtg gcgggaccag 3961 tgacgaaggc ttgagcgagg gcgtgcaaga ttccgaatac cgcaagcgac aggccgatca 4021 tcgtcgcgct ccagcgaaag cggtcctcgc cgaaaatgac ccagagcgct gccggcacct 4081 gtcctacgag ttgcatgata aagaagacag tcataagtgc ggcgacgata gtcatgcccc 4141 gcgcccaccg gaaggagctg actgggttga aggctctcaa gggcatcggt cgagatcccg 4201 gtgcctaatg agtgagctaa cttacattaa ttgcgttgcg ctcactgccc gctttccagt 4261 cgggaaacct gtcgtgccag ctgcattaat gaatcggcca acgcgcgggg agaggcggtt 4321 tgcgtattgg gcgccagggt ggtttttctt ttcaccagtg agacgggcaa cagctgattg 4381 cccttcaccg cctggccctg agagagttgc agcaagcggt ccacgctggt ttgccccagc 4441 aggcgaaaat cctgtttgat ggtggttaac ggcgggatat aacatgagct gtcttcggta 4501 tcgtcgtatc ccactaccga gatatccgca ccaacgcgca gcccggactc ggtaatggcg 4561 cgcattgcgc ccagcgccat ctgatcgttg gcaaccagca tcgcagtggg aacgatgccc 4621 tcattcagca tttgcatggt ttgttgaaaa ccggacatgg cactccagtc gccttcccgt 4681 tccgctatcg gctgaatttg attgcgagtg agatatttat gccagccagc cagacgcaga 4741 cgcgccgaga cagaacttaa tgggcccgct aacagcgcga tttgctggtg acccaatgcg 4801 accagatgct ccacgcccag tcgcgtaccg tcttcatggg agaaaataat actgttgatg 4861 ggtgtctggt cagagacatc aagaaataac gccggaacat tagtgcaggc agcttccaca 4921 gcaatggcat cctggtcatc cagcggatag ttaatgatca gcccactgac gcgttgcgcg 4981 agaagattgt gcaccgccgc tttacaggct tcgacgccgc ttcgttctac catcgacacc 5041 accacgctgg cacccagttg atcggcgcga gatttaatcg ccgcgacaat ttgcgacggc 5101 gcgtgcaggg ccagactgga ggtggcaacg ccaatcagca acgactgttt gcccgccagt 5161 tgttgtgcca cgcggttggg aatgtaattc agctccgcca tcgccgcttc cactttttcc 5221 cgcgttttcg cagaaacgtg gctggcctgg ttcaccacgc gggaaacggt ctgataagag 5281 acaccggcat actctgcgac atcgtataac gttactggtt tcacattcac caccctgaat 5341 tgactctctt ccgggcgcta tcatgccata ccgcgaaagg ttttgcgcca ttcgatggtg 5401 tccgggatct cgacgctctc ccttatgcga ctcctgcatt aggaagcagc ccagtagtag 5461 gttgaggccg ttgagcaccg ccgccgcaag gaatggtgca tgcaaggaga tggcgcccaa 5521 cagtcccccg gccacggggc ctgccaccat acccacgccg aaacaagcgc tcatgagccc 5581 gaagtggcga gcccgatctt ccccatcggt gatgtcggcg atataggcgc cagcaaccgc 5641 acctgtggcg ccggtgatgc cggccacgat gcgtccg //