LOCUS       sequence-199092-        5647 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  wildtype Fgß peptide with ddFLN4 fingerprint domain with N-terminal 
            ybbr tag for covalent immobilization - resulting in weak forces.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5647)
  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 5647)
  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-ybbr-HIS-ddFLN4-Fgß
FEATURES             Location/Qualifiers
     source          1..5647
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(328..346)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      342..389
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      426..881
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(513..532)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     723..744
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(973..1788)
                     /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(973..1788)
                     /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     1696..1715
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     1780..1799
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     rep_origin      1910..2498
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2399..2418
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2652..2669
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    2684..2823
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(2749..2768)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     2909..2931
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(2925..3116)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(3925..5007)
                     /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
                     NYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSRADQLGASVVVVVVERSGV
                     EACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSIIFSH
                     EDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREGDWSA
                     MSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTEDSSC
                     YIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR
                     ALADSLMQLARQVSRLESGQ"
     primer_bind     4969..4988
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(5008..5085)
                     /gene="lacI"
                     /label=lacI promoter
     primer_bind     5291..5310
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     5394..5413
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        5394..5412
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    5413..5437
                     /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             5452..5474
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             5518..5535
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
ORIGIN
        1 gtaccgacgg tggtgacggt ttcgttgtta ccatcgaagg tccggctccg gttgacccgg
       61 ttatggttga caacggtgac ggtacctacg acgttgaatt cgaaccgaaa gaagctggtg
      121 actacgttat caacctgacc ctggacggtg acaacgttaa cggtttcccg aaaaccgtta
      181 ccgttaaacc ggctccgggc tccggttctg gttccggctc taatgaagag ggctttttca
      241 gcgcacgtgg tcatcgtccg ctggattaac tcgagtaaga tccggctgct aacaaagccc
      301 gaaaggaagc tgagttggct gctgccaccg ctgagcaata actagcataa ccccttgggg
      361 cctctaaacg ggtcttgagg ggttttttgc tgaaaggagg aactatatcc ggattggcga
      421 atgggacgcg ccctgtagcg gcgcattaag cgcggcgggt gtggtggtta cgcgcagcgt
      481 gaccgctaca cttgccagcg ccctagcgcc cgctcctttc gctttcttcc cttcctttct
      541 cgccacgttc gccggctttc cccgtcaagc tctaaatcgg gggctccctt tagggttccg
      601 atttagtgct ttacggcacc tcgaccccaa aaaacttgat tagggtgatg gttcacgtag
      661 tgggccatcg ccctgataga cggtttttcg ccctttgacg ttggagtcca cgttctttaa
      721 tagtggactc ttgttccaaa ctggaacaac actcaaccct atctcggtct attcttttga
      781 tttataaggg attttgccga tttcggccta ttggttaaaa aatgagctga tttaacaaaa
      841 atttaacgcg aattttaaca aaatattaac gcttacaatt taggtggcac ttttcgggga
      901 aatgtgcgcg gaacccctat ttgtttattt ttctaaatac attcaaatat gtatccgctc
      961 atgaattaat tcttagaaaa actcatcgag catcaaatga aactgcaatt tattcatatc
     1021 aggattatca ataccatatt tttgaaaaag ccgtttctgt aatgaaggag aaaactcacc
     1081 gaggcagttc cataggatgg caagatcctg gtatcggtct gcgattccga ctcgtccaac
     1141 atcaatacaa cctattaatt tcccctcgtc aaaaataagg ttatcaagtg agaaatcacc
     1201 atgagtgacg actgaatccg gtgagaatgg caaaagttta tgcatttctt tccagacttg
     1261 ttcaacaggc cagccattac gctcgtcatc aaaatcactc gcatcaacca aaccgttatt
     1321 cattcgtgat tgcgcctgag cgagacgaaa tacgcgatcg ctgttaaaag gacaattaca
     1381 aacaggaatc gaatgcaacc ggcgcaggaa cactgccagc gcatcaacaa tattttcacc
     1441 tgaatcagga tattcttcta atacctggaa tgctgttttc ccggggatcg cagtggtgag
     1501 taaccatgca tcatcaggag tacggataaa atgcttgatg gtcggaagag gcataaattc
     1561 cgtcagccag tttagtctga ccatctcatc tgtaacatca ttggcaacgc tacctttgcc
     1621 atgtttcaga aacaactctg gcgcatcggg cttcccatac aatcgataga ttgtcgcacc
     1681 tgattgcccg acattatcgc gagcccattt atacccatat aaatcagcat ccatgttgga
     1741 atttaatcgc ggcctagagc aagacgtttc ccgttgaata tggctcataa caccccttgt
     1801 attactgttt atgtaagcag acagttttat tgttcatgac caaaatccct taacgtgagt
     1861 tttcgttcca ctgagcgtca gaccccgtag aaaagatcaa aggatcttct tgagatcctt
     1921 tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca gcggtggttt
     1981 gtttgccgga tcaagagcta ccaactcttt ttccgaaggt aactggcttc agcagagcgc
     2041 agataccaaa tactgtcctt ctagtgtagc cgtagttagg ccaccacttc aagaactctg
     2101 tagcaccgcc tacatacctc gctctgctaa tcctgttacc agtggctgct gccagtggcg
     2161 ataagtcgtg tcttaccggg ttggactcaa gacgatagtt accggataag gcgcagcggt
     2221 cgggctgaac ggggggttcg tgcacacagc ccagcttgga gcgaacgacc tacaccgaac
     2281 tgagatacct acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg
     2341 acaggtatcc ggtaagcggc agggtcggaa caggagagcg cacgagggag cttccagggg
     2401 gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat
     2461 ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac gcggcctttt
     2521 tacggttcct ggccttttgc tggccttttg ctcacatgtt ctttcctgcg ttatcccctg
     2581 attctgtgga taaccgtatt accgcctttg agtgagctga taccgctcgc cgcagccgaa
     2641 cgaccgagcg cagcgagtca gtgagcgagg aagcggaaga gcgcctgatg cggtattttc
     2701 tccttacgca tctgtgcggt atttcacacc gcaatggtgc actctcagta caatctgctc
     2761 tgatgccgca tagttaagcc agtatacact ccgctatcgc tacgtgactg ggtcatggct
     2821 gcgccccgac acccaccaac acccgctgac gcgccctgac gggcttgtct gctcccggca
     2881 tccgcttaca gacaagctgt gaccgtctcc gggagctgca tgtgtcagag gttttcaccg
     2941 tcatcaccga aacgcgcgag gcagctgcgg taaagctcat cagcgtggtc gtgaagcgat
     3001 tcacagatgt ctgcctgttc atccgcgtcc agctcgttga gtttctccag aagcgttaat
     3061 gtctggcttc tgataaagcg ggccatgtta agggcggttt tttcctgttt ggtcactgat
     3121 gcctccgtgt aagggggatt tctgttcatg ggggtaatga taccgatgaa acgagagagg
     3181 atgctcacga tacgggttac tgatgatgaa catgcccggt tactggaacg ttgtgagggt
     3241 aaacaactgg cggtatggat gcggcgggac cagagaaaaa tcactcaggg tcaatgccag
     3301 cgcttcgtta atacagatgt aggtgttcca cagggtagcc agcagcatcc tgcgatgcag
     3361 atccggaaca taatggtgca gggcgctgac ttccgcgttt ccagacttta cgaaacacgg
     3421 aaaccgaaga ccattcatgt tgttgctcag gtcgcagacg ttttgcagca gcagtcgctt
     3481 cacgttcgct cgcgtatcgg tgattcattc tgctaaccag taaggcaacc ccgccagcct
     3541 agccgggtcc tcaacgacag gagcacgatc atgcgcaccc gtggggccgc catgccggcg
     3601 ataatggcct gcttctcgcc gaaacgtttg gtggcgggac cagtgacgaa ggcttgagcg
     3661 agggcgtgca agattccgaa taccgcaagc gacaggccga tcatcgtcgc gctccagcga
     3721 aagcggtcct cgccgaaaat gacccagagc gctgccggca cctgtcctac gagttgcatg
     3781 ataaagaaga cagtcataag tgcggcgacg atagtcatgc cccgcgccca ccggaaggag
     3841 ctgactgggt tgaaggctct caagggcatc ggtcgagatc ccggtgccta atgagtgagc
     3901 taacttacat taattgcgtt gcgctcactg cccgctttcc agtcgggaaa cctgtcgtgc
     3961 cagctgcatt aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat tgggcgccag
     4021 ggtggttttt cttttcacca gtgagacggg caacagctga ttgcccttca ccgcctggcc
     4081 ctgagagagt tgcagcaagc ggtccacgct ggtttgcccc agcaggcgaa aatcctgttt
     4141 gatggtggtt aacggcggga tataacatga gctgtcttcg gtatcgtcgt atcccactac
     4201 cgagatatcc gcaccaacgc gcagcccgga ctcggtaatg gcgcgcattg cgcccagcgc
     4261 catctgatcg ttggcaacca gcatcgcagt gggaacgatg ccctcattca gcatttgcat
     4321 ggtttgttga aaaccggaca tggcactcca gtcgccttcc cgttccgcta tcggctgaat
     4381 ttgattgcga gtgagatatt tatgccagcc agccagacgc agacgcgccg agacagaact
     4441 taatgggccc gctaacagcg cgatttgctg gtgacccaat gcgaccagat gctccacgcc
     4501 cagtcgcgta ccgtcttcat gggagaaaat aatactgttg atgggtgtct ggtcagagac
     4561 atcaagaaat aacgccggaa cattagtgca ggcagcttcc acagcaatgg catcctggtc
     4621 atccagcgga tagttaatga tcagcccact gacgcgttgc gcgagaagat tgtgcaccgc
     4681 cgctttacag gcttcgacgc cgcttcgttc taccaccacc accaccacgc tggcacccag
     4741 ttgatcggcg cgagatttaa tcgccgcgac aatttgcgac ggcgcgtgca gggccagact
     4801 ggaggtggca acgccaatca gcaacgactg tttgcccgcc agttgttgtg ccacgcggtt
     4861 gggaatgtaa ttcagctccg ccatcgccgc ttccactttt tcccgcgttt tcgcagaaac
     4921 gtggctggcc tggttcacca cgcgggaaac ggtctgataa gagacaccgg catactctgc
     4981 gacatcgtat aacgttactg gtttcacatt caccaccctg aattgactct cttccgggcg
     5041 ctatcatgcc ataccgcgaa aggttttgcg ccattcgatg gtgtccggga tctcgacgct
     5101 ctcccttatg cgactcctgc attaggaagc agcccagtag taggttgagg ccgttgagca
     5161 ccgccgccgc aaggaatggt gcatgcaagg agatggcgcc caacagtccc ccggccacgg
     5221 ggcctgccac catacccacg ccgaaacaag cgctcatgag cccgaagtgg cgagcccgat
     5281 cttccccatc ggtgatgtcg gcgatatagg cgccagcaac cgcacctgtg gcgccggtga
     5341 tgccggccac gatgcgtccg gcgtagagga tcgagatctc gatcccgcga aattaatacg
     5401 actcactata ggggaattgt gagcggataa caattcccct gtagaaataa ttttgtttaa
     5461 ctttaagaag gagatataca tatggactct ctggaattca tcgcttctaa actggctcac
     5521 caccaccacc accacggttc tgctgacccg gaaaaatctt acgctgaagg tccgggtctg
     5581 gacggtggtg aaagtttcca gccgtctaaa ttcaaaatcc acgctgttga cccggacggt
     5641 gttcacc
//
