LOCUS       sequence-310821-        7072 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  E. coli expression of Rc. XDocB wild-type construct containing ybbr 
            tag, ELP linker and ddFLN4 and I27 fingerprint domains. This 
            construct was designed for AFM measurements..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7072)
  AUTHORS   Liu Z, Liu H, Vera AM, Bernardi RC, Tinnefeld P, Nash MA
  TITLE     High force catch bond mechanism of bacterial adhesion in the human 
            gut.
  JOURNAL   Nat Commun. 2020 Aug 28;11(1):4321. doi: 10.1038/s41467-020-18063-x.
  PUBMED    32859904
REFERENCE   2  (bases 1 to 7072)
  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: "2020-08-28"; volume: "11(1):4321. doi"; pages: " 
            10.1038/s41467-020-18063-x"
COMMENT     Sequence Label: pET28a-ybbr-HIS-ELP-ddFLN4-I27-XMod-Doc (wild ...
FEATURES             Location/Qualifiers
     source          1..7072
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     1..20
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1..19
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    20..44
                     /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             59..81
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             131..148
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(2007..2025)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      2021..2068
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      2105..2560
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2192..2211)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2402..2423
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(2652..3467)
                     /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(2652..3467)
                     /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     3375..3394
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     3459..3478
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     rep_origin      3589..4177
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4078..4097
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4331..4348
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    4363..4502
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(4428..4447)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4588..4610
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(4604..4795)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(5604..6686)
                     /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     6648..6667
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(6687..6764)
                     /gene="lacI"
                     /label=lacI promoter
     primer_bind     6970..6989
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
ORIGIN
        1 taatacgact cactataggg gaattgtgag cggataacaa ttcccctgta gaaataattt
       61 tgtttaactt taagaaggag atatacatat gggtaccgac tctctggaat tcatcgcttc
      121 taaactggct catcatcatc accatcactg gggatccggc cacggcgtgg gtgttccagg
      181 tatgggtgtg cctggtgttg gtgttcctgg cgttggcgtg ccaggtgtgg gagtaccagg
      241 cgtaggcgtt cccggtgtag gcgtgcctgg cgtcggtgta cccggtgtcg gagtccctgg
      301 tgaaggtgtg ccaggcgaag gtgtgccggg cgtgggtgtt ccaggtatgg gtgtgcctgg
      361 tgttggtgtt cctggcgttg gcgtgccagg tgtgggagta ccaggcgtag gcgttcccgg
      421 tgtaggcgtg cctggcgtcg gtgtacccgg tgtcggagtc cctggtgaag gtgtgccagg
      481 cgaaggtgtg ccgggcgtgg gtgttccagg tatgggtgtg cctggtgttg gtgttcctgg
      541 cgttggcgtg ccaggtgtgg gagtaccagg cgtaggcgtt cccggtgtag gcgtgcctgg
      601 cgtcggtgta cccggtgtcg gagtccctgg tgaaggtgtg ccaggcgaag gtgtgccggg
      661 ctggccgtca ggttctgctg acccggaaaa atcttacgct gaaggtccgg gtctggacgg
      721 tggtgaatgc ttccagccgt ctaaattcaa aatccacgct gttgacccgg acggtgttca
      781 ccgtaccgac ggtggtgacg gtttcgttgt taccatcgaa ggtccggctc cggttgaccc
      841 ggttatggtt gacaacggtg acggtaccta cgacgttgaa ttcgaaccga aagaagctgg
      901 tgactacgtt atcaacctga ccctggacgg tgacaacgtt aacggtttcc cgaaaaccgt
      961 taccgttaaa ccggctccgc taatagaagt ggaaaagcct ctgtacggag tagaggtgtt
     1021 tgttggtgaa acagcccact ttgaaattga actttctgaa cctgatgttc acggccagtg
     1081 gaagctgaaa ggacagcctt tggcagcttc ccctgactgt gaaatcattg aggatggaaa
     1141 gaagcatatt ctgatccttc ataactgtca gctgggtatg acaggagagg tttccttcca
     1201 ggctgctaat accaaatctg cagccaatct gaaagtgaaa gaattgggct ccggttctgg
     1261 ttccggctct agcggttccg aaacaactac cgagacttct gtgtctcaat ctttctcctt
     1321 cacagagccc actcgcgtat gctactggtc ccatgacaac cgtcctttcg acctggaggg
     1381 cttgaaagtc attcttcagg agacgatctt tatcatccac gaggacggca cagtcgtcga
     1441 agcagaagag caacccgctg ccgtagacat cacagagcag tgtatgacta tcttctgtaa
     1501 cgataagcaa gtggcgacgc cagaagatac ctacgaaacg cagatctata agtacatgct
     1561 ggagtttgca tatgaaggca acccggacgt tattgtgggg actcatcaag tcttgattgg
     1621 tgtgaaagga gatacaaatc tgagcggcga cgtaacagtt gcagatgctg ttgagatcct
     1681 gatgtaccgt gctgacatga tggctaatcc caacaaccca acatacgtat ttaacgaaga
     1741 ccccgatctg cataaattag ggtatttttt gggtaatgta gattgtatca aagagggtga
     1801 cgatttaaac gttgccgacg ccgtcaatat cttaatgttt cgtgcggagg tcttagccaa
     1861 cggctccgca acgtgggagc agattgttgg ctatgacctt attgacccaa gcgacgagag
     1921 tacctttcgt actattgaga agctgtaact cgagtaagat ccggctgcta acaaagcccg
     1981 aaaggaagct gagttggctg ctgccaccgc tgagcaataa ctagcataac cccttggggc
     2041 ctctaaacgg gtcttgaggg gttttttgct gaaaggagga actatatccg gattggcgaa
     2101 tgggacgcgc cctgtagcgg cgcattaagc gcggcgggtg tggtggttac gcgcagcgtg
     2161 accgctacac ttgccagcgc cctagcgccc gctcctttcg ctttcttccc ttcctttctc
     2221 gccacgttcg ccggctttcc ccgtcaagct ctaaatcggg ggctcccttt agggttccga
     2281 tttagtgctt tacggcacct cgaccccaaa aaacttgatt agggtgatgg ttcacgtagt
     2341 gggccatcgc cctgatagac ggtttttcgc cctttgacgt tggagtccac gttctttaat
     2401 agtggactct tgttccaaac tggaacaaca ctcaacccta tctcggtcta ttcttttgat
     2461 ttataaggga ttttgccgat ttcggcctat tggttaaaaa atgagctgat ttaacaaaaa
     2521 tttaacgcga attttaacaa aatattaacg cttacaattt aggtggcact tttcggggaa
     2581 atgtgcgcgg aacccctatt tgtttatttt tctaaataca ttcaaatatg tatccgctca
     2641 tgaattaatt cttagaaaaa ctcatcgagc atcaaatgaa actgcaattt attcatatca
     2701 ggattatcaa taccatattt ttgaaaaagc cgtttctgta atgaaggaga aaactcaccg
     2761 aggcagttcc ataggatggc aagatcctgg tatcggtctg cgattccgac tcgtccaaca
     2821 tcaatacaac ctattaattt cccctcgtca aaaataaggt tatcaagtga gaaatcacca
     2881 tgagtgacga ctgaatccgg tgagaatggc aaaagtttat gcatttcttt ccagacttgt
     2941 tcaacaggcc agccattacg ctcgtcatca aaatcactcg catcaaccaa accgttattc
     3001 attcgtgatt gcgcctgagc gagacgaaat acgcgatcgc tgttaaaagg acaattacaa
     3061 acaggaatcg aatgcaaccg gcgcaggaac actgccagcg catcaacaat attttcacct
     3121 gaatcaggat attcttctaa tacctggaat gctgttttcc cggggatcgc agtggtgagt
     3181 aaccatgcat catcaggagt acggataaaa tgcttgatgg tcggaagagg cataaattcc
     3241 gtcagccagt ttagtctgac catctcatct gtaacatcat tggcaacgct acctttgcca
     3301 tgtttcagaa acaactctgg cgcatcgggc ttcccataca atcgatagat tgtcgcacct
     3361 gattgcccga cattatcgcg agcccattta tacccatata aatcagcatc catgttggaa
     3421 tttaatcgcg gcctagagca agacgtttcc cgttgaatat ggctcataac accccttgta
     3481 ttactgttta tgtaagcaga cagttttatt gttcatgacc aaaatccctt aacgtgagtt
     3541 ttcgttccac tgagcgtcag accccgtaga aaagatcaaa ggatcttctt gagatccttt
     3601 ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg
     3661 tttgccggat caagagctac caactctttt tccgaaggta actggcttca gcagagcgca
     3721 gataccaaat actgtccttc tagtgtagcc gtagttaggc caccacttca agaactctgt
     3781 agcaccgcct acatacctcg ctctgctaat cctgttacca gtggctgctg ccagtggcga
     3841 taagtcgtgt cttaccgggt tggactcaag acgatagtta ccggataagg cgcagcggtc
     3901 gggctgaacg gggggttcgt gcacacagcc cagcttggag cgaacgacct acaccgaact
     3961 gagataccta cagcgtgagc tatgagaaag cgccacgctt cccgaaggga gaaaggcgga
     4021 caggtatccg gtaagcggca gggtcggaac aggagagcgc acgagggagc ttccaggggg
     4081 aaacgcctgg tatctttata gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt
     4141 tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac gccagcaacg cggccttttt
     4201 acggttcctg gccttttgct ggccttttgc tcacatgttc tttcctgcgt tatcccctga
     4261 ttctgtggat aaccgtatta ccgcctttga gtgagctgat accgctcgcc gcagccgaac
     4321 gaccgagcgc agcgagtcag tgagcgagga agcggaagag cgcctgatgc ggtattttct
     4381 ccttacgcat ctgtgcggta tttcacaccg caatggtgca ctctcagtac aatctgctct
     4441 gatgccgcat agttaagcca gtatacactc cgctatcgct acgtgactgg gtcatggctg
     4501 cgccccgaca cccgccaaca cccgctgacg cgccctgacg ggcttgtctg ctcccggcat
     4561 ccgcttacag acaagctgtg accgtctccg ggagctgcat gtgtcagagg ttttcaccgt
     4621 catcaccgaa acgcgcgagg cagctgcggt aaagctcatc agcgtggtcg tgaagcgatt
     4681 cacagatgtc tgcctgttca tccgcgtcca gctcgttgag tttctccaga agcgttaatg
     4741 tctggcttct gataaagcgg gccatgttaa gggcggtttt ttcctgtttg gtcactgatg
     4801 cctccgtgta agggggattt ctgttcatgg gggtaatgat accgatgaaa cgagagagga
     4861 tgctcacgat acgggttact gatgatgaac atgcccggtt actggaacgt tgtgagggta
     4921 aacaactggc ggtatggatg cggcgggacc agagaaaaat cactcagggt caatgccagc
     4981 gcttcgttaa tacagatgta ggtgttccac agggtagcca gcagcatcct gcgatgcaga
     5041 tccggaacat aatggtgcag ggcgctgact tccgcgtttc cagactttac gaaacacgga
     5101 aaccgaagac cattcatgtt gttgctcagg tcgcagacgt tttgcagcag cagtcgcttc
     5161 acgttcgctc gcgtatcggt gattcattct gctaaccagt aaggcaaccc cgccagccta
     5221 gccgggtcct caacgacagg agcacgatca tgcgcacccg tggggccgcc atgccggcga
     5281 taatggcctg cttctcgccg aaacgtttgg tggcgggacc agtgacgaag gcttgagcga
     5341 gggcgtgcaa gattccgaat accgcaagcg acaggccgat catcgtcgcg ctccagcgaa
     5401 agcggtcctc gccgaaaatg acccagagcg ctgccggcac ctgtcctacg agttgcatga
     5461 taaagaagac agtcataagt gcggcgacga tagtcatgcc ccgcgcccac cggaaggagc
     5521 tgactgggtt gaaggctctc aagggcatcg gtcgagatcc cggtgcctaa tgagtgagct
     5581 aacttacatt aattgcgttg cgctcactgc ccgctttcca gtcgggaaac ctgtcgtgcc
     5641 agctgcatta atgaatcggc caacgcgcgg ggagaggcgg tttgcgtatt gggcgccagg
     5701 gtggtttttc ttttcaccag tgagacgggc aacagctgat tgcccttcac cgcctggccc
     5761 tgagagagtt gcagcaagcg gtccacgctg gtttgcccca gcaggcgaaa atcctgtttg
     5821 atggtggtta acggcgggat ataacatgag ctgtcttcgg tatcgtcgta tcccactacc
     5881 gagatatccg caccaacgcg cagcccggac tcggtaatgg cgcgcattgc gcccagcgcc
     5941 atctgatcgt tggcaaccag catcgcagtg ggaacgatgc cctcattcag catttgcatg
     6001 gtttgttgaa aaccggacat ggcactccag tcgccttccc gttccgctat cggctgaatt
     6061 tgattgcgag tgagatattt atgccagcca gccagacgca gacgcgccga gacagaactt
     6121 aatgggcccg ctaacagcgc gatttgctgg tgacccaatg cgaccagatg ctccacgccc
     6181 agtcgcgtac cgtcttcatg ggagaaaata atactgttga tgggtgtctg gtcagagaca
     6241 tcaagaaata acgccggaac attagtgcag gcagcttcca cagcaatggc atcctggtca
     6301 tccagcggat agttaatgat cagcccactg acgcgttgcg cgagaagatt gtgcaccgcc
     6361 gctttacagg cttcgacgcc gcttcgttct accatcgaca ccaccacgct ggcacccagt
     6421 tgatcggcgc gagatttaat cgccgcgaca atttgcgacg gcgcgtgcag ggccagactg
     6481 gaggtggcaa cgccaatcag caacgactgt ttgcccgcca gttgttgtgc cacgcggttg
     6541 ggaatgtaat tcagctccgc catcgccgct tccacttttt cccgcgtttt cgcagaaacg
     6601 tggctggcct ggttcaccac gcgggaaacg gtctgataag agacaccggc atactctgcg
     6661 acatcgtata acgttactgg tttcacattc accaccctga attgactctc ttccgggcgc
     6721 tatcatgcca taccgcgaaa ggttttgcgc cattcgatgg tgtccgggat ctcgacgctc
     6781 tcccttatgc gactcctgca ttaggaagca gcccagtagt aggttgaggc cgttgagcac
     6841 cgccgccgca aggaatggtg catgcaagga gatggcgccc aacagtcccc cggccacggg
     6901 gcctgccacc atacccacgc cgaaacaagc gctcatgagc ccgaagtggc gagcccgatc
     6961 ttccccatcg gtgatgtcgg cgatataggc gccagcaacc gcacctgtgg cgccggtgat
     7021 gccggccacg atgcgtccgg cgtagaggat cgagatctcg atcccgcgaa at
//
