LOCUS       sequence-227015-        5683 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  SdrG 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 5683)
  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 5683)
  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-SdrG_B1-DK
FEATURES             Location/Qualifiers
     source          1..5683
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     RBS             12..34
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             87..104
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     terminator      585..632
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      669..1124
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(756..775)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     966..987
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(1216..2031)
                     /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(1216..2031)
                     /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     1939..1958
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     2023..2042
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     rep_origin      2153..2741
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2642..2661
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2895..2912
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    2927..3066
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(2992..3011)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3152..3174
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(3168..3359)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(4168..5250)
                     /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     5212..5231
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(5251..5328)
                     /gene="lacI"
                     /label=lacI promoter
     primer_bind     5534..5553
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     5637..5656
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        5637..5655
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    5656..5680
                     /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)."
ORIGIN
        1 gtagaaataa ttttgtttaa ctttaagaag gagatataca tatggggggc ggtgactctc
       61 tggaattcat cgcttctaaa ctggctcacc accaccacca ccacggttct gctccggaaa
      121 aaacctataa aatcggtgat tatgtgtggg aggatgtgga taaagatggc attcagaata
      181 ccaacgataa cgaaaaaccg ctgagcaatg ttctggttac cctgacctat ccggatggca
      241 ccagcaaaag cgttcgtacc gatgaagaag gtaaatatca gtttgatggc ctgaaaaatg
      301 gcctgaccta caaaatcacc tttgaaacac cggaaggtta taccccgacc ctgaaacata
      361 gcggcaccaa tccggcactg gatagcgaag gtaatagcgt ttgggttaca attaatggcc
      421 aggatgatat gaccattgat agcggctttt atcagacccc gggttccggc tctcagtctg
      481 gttcttctgg ttctggttct aacggtgact aactcgagta agatccggct gctaacaaag
      541 cccgaaagga agctgagttg gctgctgcca ccgctgagca ataactagca taaccccttg
      601 gggcctctaa acgggtcttg aggggttttt tgctgaaagg aggaactata tccggattgg
      661 cgaatgggac gcgccctgta gcggcgcatt aagcgcggcg ggtgtggtgg ttacgcgcag
      721 cgtgaccgct acacttgcca gcgccctagc gcccgctcct ttcgctttct tcccttcctt
      781 tctcgccacg ttcgccggct ttccccgtca agctctaaat cgggggctcc ctttagggtt
      841 ccgatttagt gctttacggc acctcgaccc caaaaaactt gattagggtg atggttcacg
      901 tagtgggcca tcgccctgat agacggtttt tcgccctttg acgttggagt ccacgttctt
      961 taatagtgga ctcttgttcc aaactggaac aacactcaac cctatctcgg tctattcttt
     1021 tgatttataa gggattttgc cgatttcggc ctattggtta aaaaatgagc tgatttaaca
     1081 aaaatttaac gcgaatttta acaaaatatt aacgcttaca atttaggtgg cacttttcgg
     1141 ggaaatgtgc gcggaacccc tatttgttta tttttctaaa tacattcaaa tatgtatccg
     1201 ctcatgaatt aattcttaga aaaactcatc gagcatcaaa tgaaactgca atttattcat
     1261 atcaggatta tcaataccat atttttgaaa aagccgtttc tgtaatgaag gagaaaactc
     1321 accgaggcag ttccatagga tggcaagatc ctggtatcgg tctgcgattc cgactcgtcc
     1381 aacatcaata caacctatta atttcccctc gtcaaaaata aggttatcaa gtgagaaatc
     1441 accatgagtg acgactgaat ccggtgagaa tggcaaaagt ttatgcattt ctttccagac
     1501 ttgttcaaca ggccagccat tacgctcgtc atcaaaatca ctcgcatcaa ccaaaccgtt
     1561 attcattcgt gattgcgcct gagcgagacg aaatacgcga tcgctgttaa aaggacaatt
     1621 acaaacagga atcgaatgca accggcgcag gaacactgcc agcgcatcaa caatattttc
     1681 acctgaatca ggatattctt ctaatacctg gaatgctgtt ttcccgggga tcgcagtggt
     1741 gagtaaccat gcatcatcag gagtacggat aaaatgcttg atggtcggaa gaggcataaa
     1801 ttccgtcagc cagtttagtc tgaccatctc atctgtaaca tcattggcaa cgctaccttt
     1861 gccatgtttc agaaacaact ctggcgcatc gggcttccca tacaatcgat agattgtcgc
     1921 acctgattgc ccgacattat cgcgagccca tttataccca tataaatcag catccatgtt
     1981 ggaatttaat cgcggcctag agcaagacgt ttcccgttga atatggctca taacacccct
     2041 tgtattactg tttatgtaag cagacagttt tattgttcat gaccaaaatc ccttaacgtg
     2101 agttttcgtt ccactgagcg tcagaccccg tagaaaagat caaaggatct tcttgagatc
     2161 ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg
     2221 tttgtttgcc ggatcaagag ctaccaactc tttttccgaa ggtaactggc ttcagcagag
     2281 cgcagatacc aaatactgtc cttctagtgt agccgtagtt aggcaaccac ttcaagaact
     2341 ctgtagcacc gcctacatac ctcgctctgc taatcctgtt accagtggct gctgccagtg
     2401 gcgataagtc gtgtcttacc gggttggact caagacgata gttaccggat aaggcgcagc
     2461 ggtcgggctg aacggggggt tcgtgcacac agcccagctt ggagcgaacg acctacaccg
     2521 aactgagata cctacagcgt gagctatgag aaagcgccac gcttcccgaa gggagaaagg
     2581 cggacaggta tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg gagcttccag
     2641 ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg ccacctctga cttgagcgtc
     2701 gatttttgtg atgctcgtca ggggggcgga gcctatggaa aaacgccagc aacgcggcct
     2761 ttttacggtt cctggccttt tgctggcctt ttgctcacat gttctttcct gcgttatccc
     2821 ctgattctgt ggataaccgt attaccgcct ttgagtgagc tgataccgct cgccgcagcc
     2881 gaacgaccga gcgcagcgag tcagtgagcg aggaagcgga agagcgcctg atgcggtatt
     2941 ttctccttac gcatctgtgc ggtatttcac accgcaatgg tgcactctca gtacaatctg
     3001 ctctgatgcc gcatagttaa gccagtatac actccgctat cgctacgtga ctgggtcatg
     3061 gctgcgcccc gacacccacc aacacccgct gacgcgccct gacgggcttg tctgctcccg
     3121 gcatccgctt acagacaagc tgtgaccgtc tccgggagct gcatgtgtca gaggttttca
     3181 ccgtcatcac cgaaacgcgc gaggcagctg cggtaaagct catcagcgtg gtcgtgaagc
     3241 gattcacaga tgtctgcctg ttcatccgcg tccagctcgt tgagtttctc cagaagcgtt
     3301 aatgtctggc ttctgataaa gcgggccatg ttaagggcgg ttttttcctg tttggtcact
     3361 gatgcctccg tgtaaggggg atttctgttc atgggggtaa tgataccgat gaaacgagag
     3421 aggatgctca cgatacgggt tactgatgat gaacatgccc ggttactgga acgttgtgag
     3481 ggtaaacaac tggcggtatg gatgcggcgg gaccagagaa aaatcactca gggtcaatgc
     3541 cagcgcttcg ttaatacaga tgtaggtgtt ccacagggta gccagcagca tcctgcgatg
     3601 cagatccgga acataatggt gcagggcgct gacttccgcg tttccagact ttacgaaaca
     3661 cggaaaccga agaccattca tgttgttgct caggtcgcag acgttttgca gcagcagtcg
     3721 cttcacgttc gctcgcgtat cggtgattca ttctgctaac cagtaaggca accccgccag
     3781 cctagccggg tcctcaacga caggagcacg atcatgcgca cccgtggggc cgccatgccg
     3841 gcgataatgg cctgcttctc gccgaaacgt ttggtggcgg gaccagtgac gaaggcttga
     3901 gcgagggcgt gcaagattcc gaataccgca agcgacaggc cgatcatcgt cgcgctccag
     3961 cgaaagcggt cctcgccgaa aatgacccag agcgctgccg gcacctgtcc tacgagttgc
     4021 atgataaaga agacagtcat aagtgcggcg acgatagtca tgccccgcgc ccaccggaag
     4081 gagctgactg ggttgaaggc tctcaagggc atcggtcgag atcccggtgc ctaatgagtg
     4141 agctaactta cattaattgc gttgcgctca ctgcccgctt tccagtcggg aaacctgtcg
     4201 tgccagctgc attaatgaat cggccaacgc gcggggagag gcggtttgcg tattgggcgc
     4261 cagggtggtt tttcttttca ccagtgagac gggcaacagc tgattgccct tcaccgcctg
     4321 gccctgagag agttgcagca agcggtccac gctggtttgc cccagcaggc gaaaatcctg
     4381 tttgatggtg gttaacggcg ggatataaca tgagctgtct tcggtatcgt cgtatcccac
     4441 taccgagata tccgcaccaa cgcgcagccc ggactcggta atggcgcgca ttgcgcccag
     4501 cgccatctga tcgttggcaa ccagcatcgc agtgggaacg atgccctcat tcagcatttg
     4561 catggtttgt tgaaaaccgg acatggcact ccagtcgcct tcccgttccg ctatcggctg
     4621 aatttgattg cgagtgagat atttatgcca gccagccaga cgcagacgcg ccgagacaga
     4681 acttaatggg cccgctaaca gcgcgatttg ctggtgaccc aatgcgacca gatgctccac
     4741 gcccagtcgc gtaccgtctt catgggagaa aataatactg ttgatgggtg tctggtcaga
     4801 gacatcaaga aataacgccg gaacattagt gcaggcagct tccacagcaa tggcatcctg
     4861 gtcatccagc ggatagttaa tgatcagccc actgacgcgt tgcgcgagaa gattgtgcac
     4921 cgccgcttta caggcttcga cgccgcttcg ttctaccacc accaccacca cgctggcacc
     4981 cagttgatcg gcgcgagatt taatcgccgc gacaatttgc gacggcgcgt gcagggccag
     5041 actggaggtg gcaacgccaa tcagcaacga ctgtttgccc gccagttgtt gtgccacgcg
     5101 gttgggaatg taattcagct ccgccatcgc cgcttccact ttttcccgcg ttttcgcaga
     5161 aacgtggctg gcctggttca ccacgcggga aacggtctga taagagacac cggcatactc
     5221 tgcgacatcg tataacgtta ctggtttcac attcaccacc ctgaattgac tctcttccgg
     5281 gcgctatcat gccataccgc gaaaggtttt gcgccattcg atggtgtccg ggatctcgac
     5341 gctctccctt atgcgactcc tgcattagga agcagcccag tagtaggttg aggccgttga
     5401 gcaccgccgc cgcaaggaat ggtgcatgca aggagatggc gcccaacagt cccccggcca
     5461 cggggcctgc caccataccc acgccgaaac aagcgctcat gagcccgaag tggcgagccc
     5521 gatcttcccc atcggtgatg tcggcgatat aggcgccagc aaccgcacct gtggcgccgg
     5581 tgatgccggc cacgatgcgt ccggcgtaga ggatcgagat ctcgatcccg cgaaattaat
     5641 acgactcact ataggggaat tgtgagcgga taacaattcc cct
//
