LOCUS       sequence-98100-e        6194 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Characteristic fingerprint molecule fused to dockerin type III 
            domain to perform single molecule force spectroscopy experiments; 
            covalently immobilized via introduced Cystein in Xylanase T120C.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6194)
  AUTHORS   Schoeler C, Malinowska KH, Bernardi RC, Milles LF, Jobst MA, Durner 
            E, Ott W, Fried DB, Bayer EA, Schulten K, Gaub HE, Nash MA
  TITLE     Ultrastable cellulosome-adhesion complex tightens under load.
  JOURNAL   Nat Commun. 2014 Dec 8;5:5635. doi: 10.1038/ncomms6635.
  PUBMED    25482395
REFERENCE   2  (bases 1 to 6194)
  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"; journalName: "Nat 
            Commun."; date: "2014-12-8"; volume: "5:5635. doi"; pages: " 
            10.1038/ncomms6635"
COMMENT     Sequence Label: pET28a-HIS-Xyl-Xmod-DocIII
FEATURES             Location/Qualifiers
     source          1..6194
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(4..819)
                     /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(4..819)
                     /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     727..746
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     811..830
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     rep_origin      941..1529
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1430..1449
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     1683..1700
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    1715..1857
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(1783..1802)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     1943..1965
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(1959..2150)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     primer_bind     3608..3627
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     3711..3730
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        3711..3729
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     RBS             3761..3783
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             3796..3813
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(5730..5748)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      5744..5791
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     promoter        complement(join(6157..6162,6163..6179,6180..6185))
                     /label=tet promoter
                     /note="E. coli promoter for tetracycline efflux protein 
                     gene"
ORIGIN
        1 ttcttagaaa aactcatcga gcatcaaatg aaactgcaat ttattcatat caggattatc
       61 aataccatat ttttgaaaaa gccgtttctg taatgaagga gaaaactcac cgaggcagtt
      121 ccataggatg gcaagatcct ggtatcggtc tgcgattccg actcgtccaa catcaataca
      181 acctattaat ttcccctcgt caaaaataag gttatcaagt gagaaatcac catgagtgac
      241 gactgaatcc ggtgagaatg gcaaaagctt atgcatttct ttccagactt gttcaacagg
      301 ccagccatta cgctcgtcat caaaatcact cgcatcaacc aaaccgttat tcattcgtga
      361 ttgcgcctga gcgagacgaa atacgcgatc gctgttaaaa ggacaattac aaacaggaat
      421 cgaatgcaac cggcgcagga acactgccag cgcatcaaca atattttcac ctgaatcagg
      481 atattcttct aatacctgga atgctgtttt cccggggatc gcagtggtga gtaaccatgc
      541 atcatcagga gtacggataa aatgcttgat ggtcggaaga ggcataaatt ccgtcagcca
      601 gtttagtctg accatctcat ctgtaacatc attggcaacg ctacctttgc catgtttcag
      661 aaacaactct ggcgcatcgg gcttcccata caatcgatag attgtcgcac ctgattgccc
      721 gacattatcg cgagcccatt tatacccata taaatcagca tccatgttgg aatttaatcg
      781 cggcctcgag caagacgttt cccgttgaat atggctcata acaccccttg tattactgtt
      841 tatgtaagca gacagtttta ttgttcatga ccaaaatccc ttaacgtgag ttttcgttcc
      901 actgagcgtc agaccccgta gaaaagatca aaggatcttc ttgagatcct ttttttctgc
      961 gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt tgtttgccgg
     1021 atcaagagct accaactctt tttccgaagg taactggctt cagcagagcg cagataccaa
     1081 atactgtcct tctagtgtag ccgtagttag gccaccactt caagaactct gtagcaccgc
     1141 ctacatacct cgctctgcta atcctgttac cagtggctgc tgccagtggc gataagtcgt
     1201 gtcttaccgg gttggactca agacgatagt taccggataa ggcgcagcgg tcgggctgaa
     1261 cggggggttc gtgcacacag cccagcttgg agcgaacgac ctacaccgaa ctgagatacc
     1321 tacagcgtga gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc
     1381 cggtaagcgg cagggtcgga acaggagagc gcacgaggga gcttccaggg ggaaacgcct
     1441 ggtatcttta tagtcctgtc gggtttcgcc acctctgact tgagcgtcga tttttgtgat
     1501 gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa cgcggccttt ttacggttcc
     1561 tggccttttg ctggcctttt gctcacatgt tctttcctgc gttatcccct gattctgtgg
     1621 ataaccgtat taccgccttt gagtgagctg ataccgctcg ccgcagccga acgaccgagc
     1681 gcagcgagtc agtgagcgag gaagcggaag agcgcctgat gcggtatttt ctccttacgc
     1741 atctgtgcgg tatttcacac cgcatatatg gtgcactctc agtacaatct gctctgatgc
     1801 cgcatagtta agccagtata cactccgcta tcgctacgtg actgggtcat ggctgcgccc
     1861 cgacacccgc caacacccgc tgacgcgccc tgacgggctt gtctgctccc ggcatccgct
     1921 tacagacaag ctgtgaccgt ctccgggagc tgcatgtgtc agaggttttc accgtcatca
     1981 ccgaaacgcg cgaggcagct gcggtaaagc tcatcagcgt ggtcgtgaag cgattcacag
     2041 atgtctgcct gttcatccgc gtccagctcg ttgagtttct ccagaagcgt taatgtctgg
     2101 cttctgataa agcgggccat gttaagggcg gttttttcct gtttggtcac tgatgcctcc
     2161 gtgtaagggg gatttctgtt catgggggta atgataccga tgaaacgaga gaggatgctc
     2221 acgatacggg ttactgatga tgaacatgcc cggttactgg aacgttgtga gggtaaacaa
     2281 ctggcggtat ggatgcggcg ggaccagaga aaaatcactc agggtcaatg ccagcgcttc
     2341 gttaatacag atgtaggtgt tccacagggt agccagcagc atcctgcgat gcagatccgg
     2401 aacataatgg tgcagggcgc tgacttccgc gtttccagac tttacgaaac acggaaaccg
     2461 aagaccattc atgttgttgc tcaggtcgca gacgttttgc agcagcagtc gcttcacgtt
     2521 cgctcgcgta tcggtgattc attctgctaa ccagtaaggc aaccccgcca gcctagccgg
     2581 gtcctcaacg acaggagcac gatcatgcgc acccgtggcc aggacccaac gctgcccgag
     2641 atgcgccgcg tgcggctgct ggagatggcg gacgcgatgg atatgttctg ccaagggttg
     2701 gtttgcgcat tcacagttct ccgcaagaat tgattggctc caattcttgg agtggtgaat
     2761 ccgttagcga ggtgccgccg gcttccattc aggtcgaggt ggcccggctc catgcaccgc
     2821 gacgcaacgc ggggaggcag acaaggtata gggcggcgcc tacaatccat gccaacccgt
     2881 tccatgtgct cgccgaggcg gcataaatcg ccgtgacgat cagcggtcca gtgatcgaag
     2941 ttaggctggt aagagccgcg agcgatcctt gaagctgtcc ctgatggtcg tcatctacct
     3001 gcctggacag catggcctgc aacgcgggca tcccgatgcc gccggaagcg agaagaatca
     3061 taatggggaa ggccatccag cctcgcgtcg cgaacgccag caagacgtag cccagcgcgt
     3121 cggccgccat gccggcgata atggcctgct tctcgccgaa acgtttggtg gcgggaccag
     3181 tgacgaaggc ttgagcgagg gcgtgcaaga ttccgaatac cgcaagcgac aggccgatca
     3241 tcgtcgcgct ccagcgaaag cggtcctcgc cgaaaatgac ccagagcgct gccggcacct
     3301 gtcctacgag ttgcatgata aagaagacag tcataagtgc ggcgacgata gtcatgcccc
     3361 gcgcccaccg gaaggagctg actgggttga aggctctcaa gggcatcggt cgacgctctc
     3421 ccttatgcga ctcctgcatt aggaagcagc ccagtagtag gttgaggccg ttgagcaccg
     3481 ccgccgcaag gaatggtgca tgcaaggaga tggcgcccaa cagtcccccg gccacggggc
     3541 ctgccaccat acccacgccg aaacaagcgc tcatgagccc gaagtggcga gcccgatctt
     3601 ccccatcggt gatgtcggcg atataggcgc cagcaaccgc acctgtggcg ccggtgatgc
     3661 cggccacgat gcgtccggcg tagaggatcg agatctcgat cccgcgaaat taatacgact
     3721 cactataggg agaccacaac ggtttccctc tagaaataat tttgtttaac tttaagaagg
     3781 agatatacca tgagccacca tcaccatcac cataagaatg cagattccta tgcgaaaaaa
     3841 cctcacatca gcgcattgaa tgccccacaa ttggatcaac gctacaaaaa cgagttcacg
     3901 attggtgcgg cagtagaacc ttatcaacta caaaatgaaa aagacgtaca aatgctaaag
     3961 cgccacttca acagcattgt tgccgagaac gtaatgaaac cgatcagcat tcaacctgag
     4021 gaaggaaaat tcaattttga acaagcggat cgaattgtga agttcgctaa ggcaaatggc
     4081 atggatattc gcttccatac actcgtttgg cacagccaag tacctcaatg gttctttctt
     4141 gacaaggaag gtaagccaat ggttaatgaa tgcgatccag tgaaacgtga acaaaataaa
     4201 caactgctgt taaaacgact tgaaactcat attaaaacga tcgtcgagcg gtacaaagat
     4261 gacattaagt actgggacgt tgtaaatgag gttgtggggg acgacggaaa actgcgcaac
     4321 tctccatggt atcaaatcgc cggcatcgat tatattaaag tggcattcca agcagctaga
     4381 aaatatggcg gagacaacat taagctttac atgaatgatt acaatacaga agtcgaaccg
     4441 aagcgaaccg ctctttacaa tttagtcaaa caactgaaag aagagggtgt tccgatcgac
     4501 ggcatcggcc atcaatccca catccaaatc ggctggcctt ctgaagcaga aatcgagaaa
     4561 acgattaaca tgttcgccgc tctcggttta gacaaccaaa tcactgagct tgatgtgagc
     4621 atgtacggtt ggccgccgcg cgcttacccg acgtatgacg ccattccaaa acaaaagttt
     4681 ttggatcagg cagcgcgcta tgatcgtttg ttcaaactgt atgaaaagtt gagcgataaa
     4741 attagcaacg tcaccttctg gggcatcgcc gacaatcata cgtggctcga cagccgtgcg
     4801 gatgtgtact atgacgccaa cgggaatgtt gtggttgacc cgaacgctcc gtacgcaaaa
     4861 gtggaaaaag ggaaaggaaa agatgcgccg ttcgtttttg gaccggatta caaagtcaaa
     4921 cccgcatatt gggctattat cgaccacaaa gtggtaccaa acactgttac atcagctgta
     4981 aagacacagt atgttgagat cgaatctgtt gacggcttct acttcaacac agaagacaag
     5041 ttcgatacag ctcagatcaa gaaggctgtt ctccacacag tttataacga aggctacaca
     5101 ggtgatgacg gcgtggctgt agttctcaga gaatatgaga gcgagcctgt tgatatcaca
     5161 gctgagttaa cattcggtga tgcaacacct gctaacacat acaaggctgt tgaaaacaag
     5221 ttcgattacg agatccctgt ttactacaac aacgctacac tcaaggatgc tgagggcaac
     5281 gatgctacag taactgttta catcggtctc aagggcgata cagaccttaa caacatcgtt
     5341 gacggtcgtg acgctacagc aacacttact tactacgctg caacatctac agacggcaag
     5401 gatgctacaa cagttgcact ttcaccaagc acactggttg gcggcaaccc tgagtcagta
     5461 tatgatgact tctctgcatt ccttagcgat gttaaggtag acgctggtaa ggaactcaca
     5521 agattcgcta agaaggctga gagacttatc gacggtagag atgcttcaag catactcaca
     5581 ttctacacta agtcttctgt agatcagtac aaggatatgg ctgctaacga gccaaacaag
     5641 ctttgggata tcgttacagg cgatgctgaa gaagaataag gatccggctg ctaacaaagc
     5701 ccgaaaggaa gctgagttgg ctgctgccac cgctgagcaa taactagcat aaccccttgg
     5761 ggcctctaaa cgggtcttga ggggtttttt gctgaaagga ggaactatat ccggatatcc
     5821 acaggacggg tgtggtcgcc atgatcgcgt agtcgatagt ggctccaagt agcgaagcga
     5881 gcaggactgg gcggcggcca aagcggtcgg acagtgctcc gagaacgggt gcgcatagaa
     5941 attgcatcaa cgcatatagc gctagcagca cgccatagtg actggcgatg ctgtcggaat
     6001 ggacgatatc ccgcaagagg cccggcagta ccggcataac caagcctatg cctacagcat
     6061 ccagggtgac ggtgccgagg atgacgatga gcgcattgtt agatttcata cacggtgcct
     6121 gactgcgtta gcaatttaac tgtgataaac taccgcatta aagcttatcg atgataagct
     6181 gtcaaacatg agaa
//
