LOCUS       sequence-227012-        5716 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  SdrD B1 domain with N-terminal ybbr and sortase tags for covalent 
            immobilization and C-terminal DK peptide for thethering with ClfB.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5716)
  AUTHORS   Milles LF, Unterauer EM, Nicolaus T, Gaub HE
  TITLE     Calcium stabilizes the strongest protein fold.
  JOURNAL   Nat Commun. 2018 Nov 12;9(1):4764. doi: 10.1038/s41467-018-07145-6.
  PUBMED    30420680
REFERENCE   2  (bases 1 to 5716)
  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: "2018-11-12"; volume: "9(1):4764. doi"; pages: " 
            10.1038/s41467-018-07145-6"
COMMENT     Sequence Label: pET28a-MGGG-ybbr-HIS-SdrD_B1-DK
FEATURES             Location/Qualifiers
     source          1..5716
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     54..73
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        54..72
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    73..97
                     /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             112..134
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             187..204
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(704..722)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      718..765
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      802..1257
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(889..908)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1099..1120
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(1349..2164)
                     /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(1349..2164)
                     /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     2072..2091
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     2156..2175
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     rep_origin      2286..2874
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2775..2794
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3028..3045
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    3060..3199
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(3125..3144)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3285..3307
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(3301..3492)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(4301..5383)
                     /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     5345..5364
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(5384..5461)
                     /gene="lacI"
                     /label=lacI promoter
     primer_bind     5667..5686
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
ORIGIN
        1 tgccggccac gatgcgtccg gcgtagagga tcgagatctc gatcccgcga aattaatacg
       61 actcactata ggggaattgt gagcggataa caattcccct gtagaaataa ttttgtttaa
      121 ctttaagaag gagatataca tatggggggc ggtgactctc tggaattcat cgcttctaaa
      181 ctggctcacc accaccacca ccacggttct gctagcggtg gtgcaggtca agaagtgtat
      241 aaaatcggta attacgtgtg ggaagatacc aataaaaacg gtgttcagga tctgggtgaa
      301 gttggtgtta aaggtgttac cgttgtggcc tatgataaca agaccaataa agaagttggt
      361 cgcaccatca ccgatgataa aggtggttat ctgattccga atctgccgaa tggtgattat
      421 cgtgttgaat tttcaaatct gccgcagggt tatgaagtta ccccgagcaa acagggtaat
      481 aatgaagaac tggatagcaa tggtgttagc agcgttatta ccgtgaatgg caaagataat
      541 ctgagcgcag atctgggtat ctacaaaccg aaatataacc tgggtgacta tgttggttcc
      601 ggctctcagt ctggttcttc tggttctggt tctaacggtg actaactcga gtaagatccg
      661 gctgctaaca aagcccgaaa ggaagctgag ttggctgctg ccaccgctga gcaataacta
      721 gcataacccc ttggggcctc taaacgggtc ttgaggggtt ttttgctgaa aggaggaact
      781 atatccggat tggcgaatgg gacgcgccct gtagcggcgc attaagcgcg gcgggtgtgg
      841 tggttacgcg cagcgtgacc gctacacttg ccagcgccct agcgcccgct cctttcgctt
      901 tcttcccttc ctttctcgcc acgttcgccg gctttccccg tcaagctcta aatcgggggc
      961 tccctttagg gttccgattt agtgctttac ggcacctcga ccccaaaaaa cttgattagg
     1021 gtgatggttc acgtagtggg ccatcgccct gatagacggt ttttcgccct ttgacgttgg
     1081 agtccacgtt ctttaatagt ggactcttgt tccaaactgg aacaacactc aaccctatct
     1141 cggtctattc ttttgattta taagggattt tgccgatttc ggcctattgg ttaaaaaatg
     1201 agctgattta acaaaaattt aacgcgaatt ttaacaaaat attaacgctt acaatttagg
     1261 tggcactttt cggggaaatg tgcgcggaac ccctatttgt ttatttttct aaatacattc
     1321 aaatatgtat ccgctcatga attaattctt agaaaaactc atcgagcatc aaatgaaact
     1381 gcaatttatt catatcagga ttatcaatac catatttttg aaaaagccgt ttctgtaatg
     1441 aaggagaaaa ctcaccgagg cagttccata ggatggcaag atcctggtat cggtctgcga
     1501 ttccgactcg tccaacatca atacaaccta ttaatttccc ctcgtcaaaa ataaggttat
     1561 caagtgagaa atcaccatga gtgacgactg aatccggtga gaatggcaaa agtttatgca
     1621 tttctttcca gacttgttca acaggccagc cattacgctc gtcatcaaaa tcactcgcat
     1681 caaccaaacc gttattcatt cgtgattgcg cctgagcgag acgaaatacg cgatcgctgt
     1741 taaaaggaca attacaaaca ggaatcgaat gcaaccggcg caggaacact gccagcgcat
     1801 caacaatatt ttcacctgaa tcaggatatt cttctaatac ctggaatgct gttttcccgg
     1861 ggatcgcagt ggtgagtaac catgcatcat caggagtacg gataaaatgc ttgatggtcg
     1921 gaagaggcat aaattccgtc agccagttta gtctgaccat ctcatctgta acatcattgg
     1981 caacgctacc tttgccatgt ttcagaaaca actctggcgc atcgggcttc ccatacaatc
     2041 gatagattgt cgcacctgat tgcccgacat tatcgcgagc ccatttatac ccatataaat
     2101 cagcatccat gttggaattt aatcgcggcc tagagcaaga cgtttcccgt tgaatatggc
     2161 tcataacacc ccttgtatta ctgtttatgt aagcagacag ttttattgtt catgaccaaa
     2221 atcccttaac gtgagttttc gttccactga gcgtcagacc ccgtagaaaa gatcaaagga
     2281 tcttcttgag atcctttttt tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg
     2341 ctaccagcgg tggtttgttt gccggatcaa gagctaccaa ctctttttcc gaaggtaact
     2401 ggcttcagca gagcgcagat accaaatact gtccttctag tgtagccgta gttaggccac
     2461 cacttcaaga actctgtagc accgcctaca tacctcgctc tgctaatcct gttaccagtg
     2521 gctgctgcca gtggcgataa gtcgtgtctt accgggttgg actcaagacg atagttaccg
     2581 gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca cacagcccag cttggagcga
     2641 acgacctaca ccgaactgag atacctacag cgtgagctat gagaaagcgc cacgcttccc
     2701 gaagggagaa aggcggacag gtatccggta agcggcaggg tcggaacagg agagcgcacg
     2761 agggagcttc cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc
     2821 tgacttgagc gtcgattttt gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc
     2881 agcaacgcgg cctttttacg gttcctggcc ttttgctggc cttttgctca catgttcttt
     2941 cctgcgttat cccctgattc tgtggataac cgtattaccg cctttgagtg agctgatacc
     3001 gctcgccgca gccgaacgac cgagcgcagc gagtcagtga gcgaggaagc ggaagagcgc
     3061 ctgatgcggt attttctcct tacgcatctg tgcggtattt cacaccgcaa tggtgcactc
     3121 tcagtacaat ctgctctgat gccgcatagt taagccagta tacactccgc tatcgctacg
     3181 tgactgggtc atggctgcgc cccgacaccc accaacaccc gctgacgcgc cctgacgggc
     3241 ttgtctgctc ccggcatccg cttacagaca agctgtgacc gtctccggga gctgcatgtg
     3301 tcagaggttt tcaccgtcat caccgaaacg cgcgaggcag ctgcggtaaa gctcatcagc
     3361 gtggtcgtga agcgattcac agatgtctgc ctgttcatcc gcgtccagct cgttgagttt
     3421 ctccagaagc gttaatgtct ggcttctgat aaagcgggcc atgttaaggg cggttttttc
     3481 ctgtttggtc actgatgcct ccgtgtaagg gggatttctg ttcatggggg taatgatacc
     3541 gatgaaacga gagaggatgc tcacgatacg ggttactgat gatgaacatg cccggttact
     3601 ggaacgttgt gagggtaaac aactggcggt atggatgcgg cgggaccaga gaaaaatcac
     3661 tcagggtcaa tgccagcgct tcgttaatac agatgtaggt gttccacagg gtagccagca
     3721 gcatcctgcg atgcagatcc ggaacataat ggtgcagggc gctgacttcc gcgtttccag
     3781 actttacgaa acacggaaac cgaagaccat tcatgttgtt gctcaggtcg cagacgtttt
     3841 gcagcagcag tcgcttcacg ttcgctcgcg tatcggtgat tcattctgct aaccagtaag
     3901 gcaaccccgc cagcctagcc gggtcctcaa cgacaggagc acgatcatgc gcacccgtgg
     3961 ggccgccatg ccggcgataa tggcctgctt ctcgccgaaa cgtttggtgg cgggaccagt
     4021 gacgaaggct tgagcgaggg cgtgcaagat tccgaatacc gcaagcgaca ggccgatcat
     4081 cgtcgcgctc cagcgaaagc ggtcctcgcc gaaaatgacc cagagcgctg ccggcacctg
     4141 tcctacgagt tgcatgataa agaagacagt cataagtgcg gcgacgatag tcatgccccg
     4201 cgcccaccgg aaggagctga ctgggttgaa ggctctcaag ggcatcggtc gagatcccgg
     4261 tgcctaatga gtgagctaac ttacattaat tgcgttgcgc tcactgcccg ctttccagtc
     4321 gggaaacctg tcgtgccagc tgcattaatg aatcggccaa cgcgcgggga gaggcggttt
     4381 gcgtattggg cgccagggtg gtttttcttt tcaccagtga gacgggcaac agctgattgc
     4441 ccttcaccgc ctggccctga gagagttgca gcaagcggtc cacgctggtt tgccccagca
     4501 ggcgaaaatc ctgtttgatg gtggttaacg gcgggatata acatgagctg tcttcggtat
     4561 cgtcgtatcc cactaccgag atatccgcac caacgcgcag cccggactcg gtaatggcgc
     4621 gcattgcgcc cagcgccatc tgatcgttgg caaccagcat cgcagtggga acgatgccct
     4681 cattcagcat ttgcatggtt tgttgaaaac cggacatggc actccagtcg ccttcccgtt
     4741 ccgctatcgg ctgaatttga ttgcgagtga gatatttatg ccagccagcc agacgcagac
     4801 gcgccgagac agaacttaat gggcccgcta acagcgcgat ttgctggtga cccaatgcga
     4861 ccagatgctc cacgcccagt cgcgtaccgt cttcatggga gaaaataata ctgttgatgg
     4921 gtgtctggtc agagacatca agaaataacg ccggaacatt agtgcaggca gcttccacag
     4981 caatggcatc ctggtcatcc agcggatagt taatgatcag cccactgacg cgttgcgcga
     5041 gaagattgtg caccgccgct ttacaggctt cgacgccgct tcgttctacc accaccacca
     5101 ccacgctggc acccagttga tcggcgcgag atttaatcgc cgcgacaatt tgcgacggcg
     5161 cgtgcagggc cagactggag gtggcaacgc caatcagcaa cgactgtttg cccgccagtt
     5221 gttgtgccac gcggttggga atgtaattca gctccgccat cgccgcttcc actttttccc
     5281 gcgttttcgc agaaacgtgg ctggcctggt tcaccacgcg ggaaacggtc tgataagaga
     5341 caccggcata ctctgcgaca tcgtataacg ttactggttt cacattcacc accctgaatt
     5401 gactctcttc cgggcgctat catgccatac cgcgaaaggt tttgcgccat tcgatggtgt
     5461 ccgggatctc gacgctctcc cttatgcgac tcctgcatta ggaagcagcc cagtagtagg
     5521 ttgaggccgt tgagcaccgc cgccgcaagg aatggtgcat gcaaggagat ggcgcccaac
     5581 agtcccccgg ccacggggcc tgccaccata cccacgccga aacaagcgct catgagcccg
     5641 aagtggcgag cccgatcttc cccatcggtg atgtcggcga tataggcgcc agcaaccgca
     5701 cctgtggcgc cggtga
//
