LOCUS       sequence-306440-        6811 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  E. coli expression of Rc. XDocB binding mode A knock-out construct 
            containing ybbr tag, ELP linker and ddFLN4 fingerprint domain. This 
            construct was designed for AFM measurements..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6811)
  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 6811)
  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-ELP-ddFLN4-XMod-Doc-HIS (BMA-KO)
FEATURES             Location/Qualifiers
     source          1..6811
                     /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             1667..1684
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(1746..1764)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      1760..1807
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      1844..2299
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1931..1950)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2141..2162
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(2391..3206)
                     /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(2391..3206)
                     /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     3114..3133
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     3198..3217
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     rep_origin      3328..3916
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3817..3836
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4070..4087
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    4102..4241
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(4167..4186)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4327..4349
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(4343..4534)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(5343..6425)
                     /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     6387..6406
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(6426..6503)
                     /gene="lacI"
                     /label=lacI promoter
     primer_bind     6709..6728
                     /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 tggggatccg gccacggcgt gggtgttcca ggtatgggtg tgcctggtgt
      181 tggtgttcct ggcgttggcg tgccaggtgt gggagtacca ggcgtaggcg ttcccggtgt
      241 aggcgtgcct ggcgtcggtg tacccggtgt cggagtccct ggtgaaggtg tgccaggcga
      301 aggtgtgccg ggcgtgggtg ttccaggtat gggtgtgcct ggtgttggtg ttcctggcgt
      361 tggcgtgcca ggtgtgggag taccaggcgt aggcgttccc ggtgtaggcg tgcctggcgt
      421 cggtgtaccc ggtgtcggag tccctggtga aggtgtgcca ggcgaaggtg tgccgggcgt
      481 gggtgttcca ggtatgggtg tgcctggtgt tggtgttcct ggcgttggcg tgccaggtgt
      541 gggagtacca ggcgtaggcg ttcccggtgt aggcgtgcct ggcgtcggtg tacccggtgt
      601 cggagtccct ggtgaaggtg tgccaggcga aggtgtgccg ggctggccgt caggttctgc
      661 tgacccggaa aaatcttacg ctgaaggtcc gggtctggac ggtggtgaat gcttccagcc
      721 gtctaaattc aaaatccacg ctgttgaccc ggacggtgtt caccgtaccg acggtggtga
      781 cggtttcgtt gttaccatcg aaggtccggc tccggttgac ccggttatgg ttgacaacgg
      841 tgacggtacc tacgacgttg aattcgaacc gaaagaagct ggtgactacg ttatcaacct
      901 gaccctggac ggtgacaacg ttaacggttt cccgaaaacc gttaccgtta aaccggctcc
      961 gggctccggt tctggttccg gctctagcgg ttccgaaaca actaccgaga cttctgtgtc
     1021 tcaatctttc tccttcacag agcccactcg cgtatgctac tggtcccatg acaaccgtcc
     1081 tttcgacctg gagggcttga aagtcattct tcaggagacg atctttatca tccacgagga
     1141 cggcacagtc gtcgaagcag aagagcaacc cgctgccgta gacatcacag agcagtgtat
     1201 gactatcttc tgtaacgata agcaagtggc gacgccagaa gatacctacg aaacgcagat
     1261 ctataagtac atgctggagt ttgcatatga aggcaacccg gacgttattg tggggactca
     1321 tcaagtcttg attggtgtga aaggagatac aaatctgagc ggcgacgtaa cagttgcaga
     1381 tgctgttgag atcctgatgt accgtgctga catgatggct aatcccaaca acccaacata
     1441 cgtatttaac gaagaccccg atctgcataa attagggtat tttttgggta atgtagattg
     1501 tatcaaagag ggtgacgatt taaacgttgc cgacgccgtc aatatcttaa tgtttgctgc
     1561 ggaggtcgaa gccaacggct ccgcaacgtg ggagcagatt gttggctatg accttattga
     1621 cccaagcgac gagagtacct ttcgtactat tgagaagctg ggctctcatc atcatcacca
     1681 tcactaactc gagtaagatc cggctgctaa caaagcccga aaggaagctg agttggctgc
     1741 tgccaccgct gagcaataac tagcataacc ccttggggcc tctaaacggg tcttgagggg
     1801 ttttttgctg aaaggaggaa ctatatccgg attggcgaat gggacgcgcc ctgtagcggc
     1861 gcattaagcg cggcgggtgt ggtggttacg cgcagcgtga ccgctacact tgccagcgcc
     1921 ctagcgcccg ctcctttcgc tttcttccct tcctttctcg ccacgttcgc cggctttccc
     1981 cgtcaagctc taaatcgggg gctcccttta gggttccgat ttagtgcttt acggcacctc
     2041 gaccccaaaa aacttgatta gggtgatggt tcacgtagtg ggccatcgcc ctgatagacg
     2101 gtttttcgcc ctttgacgtt ggagtccacg ttctttaata gtggactctt gttccaaact
     2161 ggaacaacac tcaaccctat ctcggtctat tcttttgatt tataagggat tttgccgatt
     2221 tcggcctatt ggttaaaaaa tgagctgatt taacaaaaat ttaacgcgaa ttttaacaaa
     2281 atattaacgc ttacaattta ggtggcactt ttcggggaaa tgtgcgcgga acccctattt
     2341 gtttattttt ctaaatacat tcaaatatgt atccgctcat gaattaattc ttagaaaaac
     2401 tcatcgagca tcaaatgaaa ctgcaattta ttcatatcag gattatcaat accatatttt
     2461 tgaaaaagcc gtttctgtaa tgaaggagaa aactcaccga ggcagttcca taggatggca
     2521 agatcctggt atcggtctgc gattccgact cgtccaacat caatacaacc tattaatttc
     2581 ccctcgtcaa aaataaggtt atcaagtgag aaatcaccat gagtgacgac tgaatccggt
     2641 gagaatggca aaagtttatg catttctttc cagacttgtt caacaggcca gccattacgc
     2701 tcgtcatcaa aatcactcgc atcaaccaaa ccgttattca ttcgtgattg cgcctgagcg
     2761 agacgaaata cgcgatcgct gttaaaagga caattacaaa caggaatcga atgcaaccgg
     2821 cgcaggaaca ctgccagcgc atcaacaata ttttcacctg aatcaggata ttcttctaat
     2881 acctggaatg ctgttttccc ggggatcgca gtggtgagta accatgcatc atcaggagta
     2941 cggataaaat gcttgatggt cggaagaggc ataaattccg tcagccagtt tagtctgacc
     3001 atctcatctg taacatcatt ggcaacgcta cctttgccat gtttcagaaa caactctggc
     3061 gcatcgggct tcccatacaa tcgatagatt gtcgcacctg attgcccgac attatcgcga
     3121 gcccatttat acccatataa atcagcatcc atgttggaat ttaatcgcgg cctagagcaa
     3181 gacgtttccc gttgaatatg gctcataaca ccccttgtat tactgtttat gtaagcagac
     3241 agttttattg ttcatgacca aaatccctta acgtgagttt tcgttccact gagcgtcaga
     3301 ccccgtagaa aagatcaaag gatcttcttg agatcctttt tttctgcgcg taatctgctg
     3361 cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc aagagctacc
     3421 aactcttttt ccgaaggtaa ctggcttcag cagagcgcag ataccaaata ctgtccttct
     3481 agtgtagccg tagttaggcc accacttcaa gaactctgta gcaccgccta catacctcgc
     3541 tctgctaatc ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt
     3601 ggactcaaga cgatagttac cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg
     3661 cacacagccc agcttggagc gaacgaccta caccgaactg agatacctac agcgtgagct
     3721 atgagaaagc gccacgcttc ccgaagggag aaaggcggac aggtatccgg taagcggcag
     3781 ggtcggaaca ggagagcgca cgagggagct tccaggggga aacgcctggt atctttatag
     3841 tcctgtcggg tttcgccacc tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg
     3901 gcggagccta tggaaaaacg ccagcaacgc ggccttttta cggttcctgg ccttttgctg
     3961 gccttttgct cacatgttct ttcctgcgtt atcccctgat tctgtggata accgtattac
     4021 cgcctttgag tgagctgata ccgctcgccg cagccgaacg accgagcgca gcgagtcagt
     4081 gagcgaggaa gcggaagagc gcctgatgcg gtattttctc cttacgcatc tgtgcggtat
     4141 ttcacaccgc aatggtgcac tctcagtaca atctgctctg atgccgcata gttaagccag
     4201 tatacactcc gctatcgcta cgtgactggg tcatggctgc gccccgacac ccgccaacac
     4261 ccgctgacgc gccctgacgg gcttgtctgc tcccggcatc cgcttacaga caagctgtga
     4321 ccgtctccgg gagctgcatg tgtcagaggt tttcaccgtc atcaccgaaa cgcgcgaggc
     4381 agctgcggta aagctcatca gcgtggtcgt gaagcgattc acagatgtct gcctgttcat
     4441 ccgcgtccag ctcgttgagt ttctccagaa gcgttaatgt ctggcttctg ataaagcggg
     4501 ccatgttaag ggcggttttt tcctgtttgg tcactgatgc ctccgtgtaa gggggatttc
     4561 tgttcatggg ggtaatgata ccgatgaaac gagagaggat gctcacgata cgggttactg
     4621 atgatgaaca tgcccggtta ctggaacgtt gtgagggtaa acaactggcg gtatggatgc
     4681 ggcgggacca gagaaaaatc actcagggtc aatgccagcg cttcgttaat acagatgtag
     4741 gtgttccaca gggtagccag cagcatcctg cgatgcagat ccggaacata atggtgcagg
     4801 gcgctgactt ccgcgtttcc agactttacg aaacacggaa accgaagacc attcatgttg
     4861 ttgctcaggt cgcagacgtt ttgcagcagc agtcgcttca cgttcgctcg cgtatcggtg
     4921 attcattctg ctaaccagta aggcaacccc gccagcctag ccgggtcctc aacgacagga
     4981 gcacgatcat gcgcacccgt ggggccgcca tgccggcgat aatggcctgc ttctcgccga
     5041 aacgtttggt ggcgggacca gtgacgaagg cttgagcgag ggcgtgcaag attccgaata
     5101 ccgcaagcga caggccgatc atcgtcgcgc tccagcgaaa gcggtcctcg ccgaaaatga
     5161 cccagagcgc tgccggcacc tgtcctacga gttgcatgat aaagaagaca gtcataagtg
     5221 cggcgacgat agtcatgccc cgcgcccacc ggaaggagct gactgggttg aaggctctca
     5281 agggcatcgg tcgagatccc ggtgcctaat gagtgagcta acttacatta attgcgttgc
     5341 gctcactgcc cgctttccag tcgggaaacc tgtcgtgcca gctgcattaa tgaatcggcc
     5401 aacgcgcggg gagaggcggt ttgcgtattg ggcgccaggg tggtttttct tttcaccagt
     5461 gagacgggca acagctgatt gcccttcacc gcctggccct gagagagttg cagcaagcgg
     5521 tccacgctgg tttgccccag caggcgaaaa tcctgtttga tggtggttaa cggcgggata
     5581 taacatgagc tgtcttcggt atcgtcgtat cccactaccg agatatccgc accaacgcgc
     5641 agcccggact cggtaatggc gcgcattgcg cccagcgcca tctgatcgtt ggcaaccagc
     5701 atcgcagtgg gaacgatgcc ctcattcagc atttgcatgg tttgttgaaa accggacatg
     5761 gcactccagt cgccttcccg ttccgctatc ggctgaattt gattgcgagt gagatattta
     5821 tgccagccag ccagacgcag acgcgccgag acagaactta atgggcccgc taacagcgcg
     5881 atttgctggt gacccaatgc gaccagatgc tccacgccca gtcgcgtacc gtcttcatgg
     5941 gagaaaataa tactgttgat gggtgtctgg tcagagacat caagaaataa cgccggaaca
     6001 ttagtgcagg cagcttccac agcaatggca tcctggtcat ccagcggata gttaatgatc
     6061 agcccactga cgcgttgcgc gagaagattg tgcaccgccg ctttacaggc ttcgacgccg
     6121 cttcgttcta ccatcgacac caccacgctg gcacccagtt gatcggcgcg agatttaatc
     6181 gccgcgacaa tttgcgacgg cgcgtgcagg gccagactgg aggtggcaac gccaatcagc
     6241 aacgactgtt tgcccgccag ttgttgtgcc acgcggttgg gaatgtaatt cagctccgcc
     6301 atcgccgctt ccactttttc ccgcgttttc gcagaaacgt ggctggcctg gttcaccacg
     6361 cgggaaacgg tctgataaga gacaccggca tactctgcga catcgtataa cgttactggt
     6421 ttcacattca ccaccctgaa ttgactctct tccgggcgct atcatgccat accgcgaaag
     6481 gttttgcgcc attcgatggt gtccgggatc tcgacgctct cccttatgcg actcctgcat
     6541 taggaagcag cccagtagta ggttgaggcc gttgagcacc gccgccgcaa ggaatggtgc
     6601 atgcaaggag atggcgccca acagtccccc ggccacgggg cctgccacca tacccacgcc
     6661 gaaacaagcg ctcatgagcc cgaagtggcg agcccgatct tccccatcgg tgatgtcggc
     6721 gatataggcg ccagcaaccg cacctgtggc gccggtgatg ccggccacga tgcgtccggc
     6781 gtagaggatc gagatctcga tcccgcgaaa t
//
