LOCUS       sequence-199090-        5650 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  minimal SdrG binding peptide FFSARG 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 5650)
  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 5650)
  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-FFSARG-ddFLN4-HIS-ybbr
FEATURES             Location/Qualifiers
     source          1..5650
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    13..37
                     /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             52..74
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             460..477
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(578..596)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      592..639
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      676..1131
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(763..782)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     973..994
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(1223..2038)
                     /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(1223..2038)
                     /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     1946..1965
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     2030..2049
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     rep_origin      2160..2748
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2649..2668
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2902..2919
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    2934..3073
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(2999..3018)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3159..3181
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(3175..3366)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(4175..5257)
                     /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     5219..5238
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(5258..5335)
                     /gene="lacI"
                     /label=lacI promoter
     primer_bind     5541..5560
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     join(5644..5650,1..13)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        join(5644..5650,1..12)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
ORIGIN
        1 actcactata ggggaattgt gagcggataa caattcccct gtagaaataa ttttgtttaa
       61 ctttaagaag gagatataca tatgggtacc ttcttctctg ctcgtggtgg tagcggtagc
      121 ggttcaggta gcgcaggtac aggtagcggt gcagatccgg aaaaaagcta tgccgaaggt
      181 ccgggtctgg atggtggtga aagttttcag ccgagcaaat tcaaaattca tgcagttgat
      241 ccggatggtg ttcatcgtac cgatggcggt gatggttttg ttgttaccat tgaaggtcct
      301 gcaccggtgg atccggttat ggttgataat ggtgatggca cctatgatgt tgaatttgaa
      361 ccgaaagaag caggcgatta cgttattaat ctgaccctgg atggcgataa tgttaatggc
      421 tttccgaaaa ccgttaccgt taaaccggca ccgagcggtc atcatcatca ccatcatggt
      481 agcgactctc tggaattcat cgcttctaaa ctggcttaac tcgagtaaga tccggctgct
      541 aacaaagccc gaaaggaagc tgagttggct gctgccaccg ctgagcaata actagcataa
      601 ccccttgggg cctctaaacg ggtcttgagg ggttttttgc tgaaaggagg aactatatcc
      661 ggattggcga atgggacgcg ccctgtagcg gcgcattaag cgcggcgggt gtggtggtta
      721 cgcgcagcgt gaccgctaca cttgccagcg ccctagcgcc cgctcctttc gctttcttcc
      781 cttcctttct cgccacgttc gccggctttc cccgtcaagc tctaaatcgg gggctccctt
      841 tagggttccg atttagtgct ttacggcacc tcgaccccaa aaaacttgat tagggtgatg
      901 gttcacgtag tgggccatcg ccctgataga cggtttttcg ccctttgacg ttggagtcca
      961 cgttctttaa tagtggactc ttgttccaaa ctggaacaac actcaaccct atctcggtct
     1021 attcttttga tttataaggg attttgccga tttcggccta ttggttaaaa aatgagctga
     1081 tttaacaaaa atttaacgcg aattttaaca aaatattaac gcttacaatt taggtggcac
     1141 ttttcgggga aatgtgcgcg gaacccctat ttgtttattt ttctaaatac attcaaatat
     1201 gtatccgctc atgaattaat tcttagaaaa actcatcgag catcaaatga aactgcaatt
     1261 tattcatatc aggattatca ataccatatt tttgaaaaag ccgtttctgt aatgaaggag
     1321 aaaactcacc gaggcagttc cataggatgg caagatcctg gtatcggtct gcgattccga
     1381 ctcgtccaac atcaatacaa cctattaatt tcccctcgtc aaaaataagg ttatcaagtg
     1441 agaaatcacc atgagtgacg actgaatccg gtgagaatgg caaaagttta tgcatttctt
     1501 tccagacttg ttcaacaggc cagccattac gctcgtcatc aaaatcactc gcatcaacca
     1561 aaccgttatt cattcgtgat tgcgcctgag cgagacgaaa tacgcgatcg ctgttaaaag
     1621 gacaattaca aacaggaatc gaatgcaacc ggcgcaggaa cactgccagc gcatcaacaa
     1681 tattttcacc tgaatcagga tattcttcta atacctggaa tgctgttttc ccggggatcg
     1741 cagtggtgag taaccatgca tcatcaggag tacggataaa atgcttgatg gtcggaagag
     1801 gcataaattc cgtcagccag tttagtctga ccatctcatc tgtaacatca ttggcaacgc
     1861 tacctttgcc atgtttcaga aacaactctg gcgcatcggg cttcccatac aatcgataga
     1921 ttgtcgcacc tgattgcccg acattatcgc gagcccattt atacccatat aaatcagcat
     1981 ccatgttgga atttaatcgc ggcctagagc aagacgtttc ccgttgaata tggctcataa
     2041 caccccttgt attactgttt atgtaagcag acagttttat tgttcatgac caaaatccct
     2101 taacgtgagt tttcgttcca ctgagcgtca gaccccgtag aaaagatcaa aggatcttct
     2161 tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca
     2221 gcggtggttt gtttgccgga tcaagagcta ccaactcttt ttccgaaggt aactggcttc
     2281 agcagagcgc agataccaaa tactgtcctt ctagtgtagc cgtagttagg ccaccacttc
     2341 aagaactctg tagcaccgcc tacatacctc gctctgctaa tcctgttacc agtggctgct
     2401 gccagtggcg ataagtcgtg tcttaccggg ttggactcaa gacgatagtt accggataag
     2461 gcgcagcggt cgggctgaac ggggggttcg tgcacacagc ccagcttgga gcgaacgacc
     2521 tacaccgaac tgagatacct acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg
     2581 agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa caggagagcg cacgagggag
     2641 cttccagggg gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca cctctgactt
     2701 gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac
     2761 gcggcctttt tacggttcct ggccttttgc tggccttttg ctcacatgtt ctttcctgcg
     2821 ttatcccctg attctgtgga taaccgtatt accgcctttg agtgagctga taccgctcgc
     2881 cgcagccgaa cgaccgagcg cagcgagtca gtgagcgagg aagcggaaga gcgcctgatg
     2941 cggtattttc tccttacgca tctgtgcggt atttcacacc gcaatggtgc actctcagta
     3001 caatctgctc tgatgccgca tagttaagcc agtatacact ccgctatcgc tacgtgactg
     3061 ggtcatggct gcgccccgac acccgccaac acccgctgac gcgccctgac gggcttgtct
     3121 gctcccggca tccgcttaca gacaagctgt gaccgtctcc gggagctgca tgtgtcagag
     3181 gttttcaccg tcatcaccga aacgcgcgag gcagctgcgg taaagctcat cagcgtggtc
     3241 gtgaagcgat tcacagatgt ctgcctgttc atccgcgtcc agctcgttga gtttctccag
     3301 aagcgttaat gtctggcttc tgataaagcg ggccatgtta agggcggttt tttcctgttt
     3361 ggtcactgat gcctccgtgt aagggggatt tctgttcatg ggggtaatga taccgatgaa
     3421 acgagagagg atgctcacga tacgggttac tgatgatgaa catgcccggt tactggaacg
     3481 ttgtgagggt aaacaactgg cggtatggat gcggcgggac cagagaaaaa tcactcaggg
     3541 tcaatgccag cgcttcgtta atacagatgt aggtgttcca cagggtagcc agcagcatcc
     3601 tgcgatgcag atccggaaca taatggtgca gggcgctgac ttccgcgttt ccagacttta
     3661 cgaaacacgg aaaccgaaga ccattcatgt tgttgctcag gtcgcagacg ttttgcagca
     3721 gcagtcgctt cacgttcgct cgcgtatcgg tgattcattc tgctaaccag taaggcaacc
     3781 ccgccagcct agccgggtcc tcaacgacag gagcacgatc atgcgcaccc gtggggccgc
     3841 catgccggcg ataatggcct gcttctcgcc gaaacgtttg gtggcgggac cagtgacgaa
     3901 ggcttgagcg agggcgtgca agattccgaa taccgcaagc gacaggccga tcatcgtcgc
     3961 gctccagcga aagcggtcct cgccgaaaat gacccagagc gctgccggca cctgtcctac
     4021 gagttgcatg ataaagaaga cagtcataag tgcggcgacg atagtcatgc cccgcgccca
     4081 ccggaaggag ctgactgggt tgaaggctct caagggcatc ggtcgagatc ccggtgccta
     4141 atgagtgagc taacttacat taattgcgtt gcgctcactg cccgctttcc agtcgggaaa
     4201 cctgtcgtgc cagctgcatt aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat
     4261 tgggcgccag ggtggttttt cttttcacca gtgagacggg caacagctga ttgcccttca
     4321 ccgcctggcc ctgagagagt tgcagcaagc ggtccacgct ggtttgcccc agcaggcgaa
     4381 aatcctgttt gatggtggtt aacggcggga tataacatga gctgtcttcg gtatcgtcgt
     4441 atcccactac cgagatatcc gcaccaacgc gcagcccgga ctcggtaatg gcgcgcattg
     4501 cgcccagcgc catctgatcg ttggcaacca gcatcgcagt gggaacgatg ccctcattca
     4561 gcatttgcat ggtttgttga aaaccggaca tggcactcca gtcgccttcc cgttccgcta
     4621 tcggctgaat ttgattgcga gtgagatatt tatgccagcc agccagacgc agacgcgccg
     4681 agacagaact taatgggccc gctaacagcg cgatttgctg gtgacccaat gcgaccagat
     4741 gctccacgcc cagtcgcgta ccgtcttcat gggagaaaat aatactgttg atgggtgtct
     4801 ggtcagagac atcaagaaat aacgccggaa cattagtgca ggcagcttcc acagcaatgg
     4861 catcctggtc atccagcgga tagttaatga tcagcccact gacgcgttgc gcgagaagat
     4921 tgtgcaccgc cgctttacag gcttcgacgc cgcttcgttc taccatcgac accaccacgc
     4981 tggcacccag ttgatcggcg cgagatttaa tcgccgcgac aatttgcgac ggcgcgtgca
     5041 gggccagact ggaggtggca acgccaatca gcaacgactg tttgcccgcc agttgttgtg
     5101 ccacgcggtt gggaatgtaa ttcagctccg ccatcgccgc ttccactttt tcccgcgttt
     5161 tcgcagaaac gtggctggcc tggttcacca cgcgggaaac ggtctgataa gagacaccgg
     5221 catactctgc gacatcgtat aacgttactg gtttcacatt caccaccctg aattgactct
     5281 cttccgggcg ctatcatgcc ataccgcgaa aggttttgcg ccattcgatg gtgtccggga
     5341 tctcgacgct ctcccttatg cgactcctgc attaggaagc agcccagtag taggttgagg
     5401 ccgttgagca ccgccgccgc aaggaatggt gcatgcaagg agatggcgcc caacagtccc
     5461 ccggccacgg ggcctgccac catacccacg ccgaaacaag cgctcatgag cccgaagtgg
     5521 cgagcccgat cttccccatc ggtgatgtcg gcgatatagg cgccagcaac cgcacctgtg
     5581 gcgccggtga tgccggccac gatgcgtccg gcgtagagga tcgagatctc gatcccgcga
     5641 aattaatacg
//
