LOCUS       sequence-199088-        5677 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  wildtype Fgß peptide with ddFLN4 fingerprint domain with C-terminal 
            ybbr tag for covalent immobilization - resulting in high forces.
ACCESSION   .
VERSION     .
KEYWORDS    .
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. 2018 Mar 30;359(6383):1527-1533. doi: 
            10.1126/science.aar2094.
  PUBMED    29599244
REFERENCE   2  (bases 1 to 5677)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; doi: "10.1126/science"; 
            journalName: "Science"; date: "2018-03-30- 30"; volume: "359"; 
            issue: "6383"; pages: "1527-1533"
COMMENT     Sequence Label: pET28a-Fgß-ddFLN4-HIS-ybbr
FEATURES             Location/Qualifiers
     source          1..5677
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     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
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     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
//
