LOCUS       sequence-171505-        7893 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  E. coli MutS expression plasmid.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7893)
  AUTHORS   Liu J, Hanne J, Britton BM, Bennett J, Kim D, Lee JB, Fishel R
  TITLE     Cascading MutS and MutL sliding clamps control DNA diffusion to 
            activate mismatch repair.
  JOURNAL   Nature. 2016 Nov 16;539(7630):583-587. doi: 10.1038/nature20562.
  PUBMED    27851738
REFERENCE   2  (bases 1 to 7893)
  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"; doi: "10"; journalName: 
            "Nature"; date: "2016-11-16- 16"; volume: "539"; issue: "7630"; 
            pages: "583-587"
COMMENT     Sequence Label: pRAF-MutS
FEATURES             Location/Qualifiers
     source          1..7893
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             568..579
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     CDS             2572..2589
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             2663..2680
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(2733..2751)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      2747..2794
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      2831..3286
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2918..2937)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3128..3149
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(3378..4193)
                     /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(3378..4193)
                     /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     4101..4120
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     4185..4204
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     rep_origin      4315..4903
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4804..4823
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5057..5074
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    5089..5228
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(5154..5173)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5314..5336
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(5330..5521)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(6330..7412)
                     /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     7374..7393
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(7413..7490)
                     /gene="lacI"
                     /label=lacI promoter
     primer_bind     7696..7715
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     7803..7822
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        7803..7821
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    7822..7846
                     /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             7861..7883
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
ORIGIN
        1 atgagtgcaa tagaaaattt cgacgcccat acgcccatga tgcagcagta tctcaggctg
       61 aaagcccagc atcccgagat cctgctgttt taccggatgg gtgattttta tgaactgttt
      121 tatgacgacg caaaacgcgc gtcgcaactg ctggatattt cactgaccaa acgcggtgct
      181 tcggcgggag agccgatccc gatggcgggg attccctacc atgcggtgga aaactatctc
      241 gccaaactgg tgaatcaggg agagtccgtt gccatctgcg aacaaattgg cgatccggcg
      301 accagcaaag gtccggttga gcgcaaagtt gtgcgtatcg ttacgccagg caccatcagc
      361 gatgaagccc tgttgcagga gcgtcaggac aacctgctgg cggctatctg gcaggacagc
      421 aaaggtttcg gctacgcgac gctggatatc agttccgggc gttttcgcct gagcgaaccg
      481 gctgaccgcg aaacgatggc ggcagaactg caacgcacta atcctgcgga actgctgtat
      541 gcagaagatt ttgctgaaat gtcgttaatt gaaggccgtc gcggcctgcg ccgtcgcccg
      601 ctgtgggagt ttgaaatcga caccgcgcgc cagcagttga atctgcaatt tgggacccgc
      661 gatctggtcg gttttggcgt cgagaacgcg ccgcgcggac tttgtgctgc cggttgtctg
      721 ttgcagtatg cgaaagatac ccaacgtacg actctgccgc atattcgttc catcaccatg
      781 gaacgtgagc aggacagcat cattatggat gccgcgacgc gtcgtaatct ggaaatcacc
      841 cagaacctgg cgggtggtgc ggaaaatacg ctggcttctg tgctcgactg caccgtcacg
      901 ccgatgggca gccgtatgct gaaacgctgg ctgcatatgc cagtgcgcga tacccgcgtg
      961 ttgcttgagc gccagcaaac tattggcgca ttgcaggatt tcaccgccgg gctacagccg
     1021 gtactgcgtc aggtcggcga cctggaacgt attctggcac gtctggcttt acgaactgct
     1081 cgcccacgcg atctggcccg tatgcgccac gctttccagc aactgccgga gctgcgtgcg
     1141 cagttagaaa ctgtcgatag tgcaccggta caggcgctac gtgagaagat gggcgagttt
     1201 gccgagctgc gcgatctgct ggagcgagca atcatcgaca caccgccggt gctggtacgc
     1261 gacggtggtg ttatcgcatc gggctataac gaagagctgg atgagtggcg cgcgctggct
     1321 gacggcgcga ccgattatct ggagcgtctg gaagtccgcg agcgtgaacg taccggcctg
     1381 gacacgctga aagttggctt taatgcggtg cacggctact acattcaaat cagccgtggg
     1441 caaagccatc tggcacccat caactacatg cgtcgccaga cgctgaaaaa cgccgagcgc
     1501 tacatcattc cagagctaaa agagtacgaa gataaagttc tcacctcaaa aggcaaagca
     1561 ctggcactgg aaaaacagct ttatgaagag ctgttcgacc tgctgttgcc gcatctggaa
     1621 gcgttgcaac agagcgcgag cgcgctggcg gaactcgacg tgctggttaa cctggcggaa
     1681 cgggcctata ccctgaacta cacctgcccg accttcattg ataaaccggg cattcgcatt
     1741 accgaaggtc gccatccggt agttgaacaa gtactgaatg agccatttat cgccaacccg
     1801 ctgaatctgt cgccgcagcg ccgcatgttg atcatcaccg gtccgaacat gggcggtaaa
     1861 agtacctata tgcgccagac cgcactgatt gcgctgatgg cctacatcgg cagctatgta
     1921 ccggcacaaa aagtcgagat tggacctatc gatcgcatct ttacccgcgt aggcgcggca
     1981 gatgacctgg cgtccgggcg ctcaaccttt atggtggaga tgactgaaac cgccaatatt
     2041 ttacataacg ccaccgaata cagtctggtg ttaatggatg agatcgggcg tggaacgtcc
     2101 acctacgatg gtctgtcgct ggcgtgggcg tgcgcggaaa atctggcgaa taagattaag
     2161 gcattgacgt tatttgctac ccactatttc gagctgaccc agttaccgga gaaaatggaa
     2221 ggcgtcgcta acgtgcatct cgatgcactg gagcacggcg acaccattgc ctttatgcac
     2281 agcgtgcagg atggcgcggc gagcaaaagc tacggcctgg cggttgcagc tctggcaggc
     2341 gtgccaaaag aggttattaa gcgcgcacgg caaaagctgc gtgagctgga aagcatttcg
     2401 ccgaacgccg ccgctacgca agtggatggt acgcaaatgt ctttgctgtc agtaccagaa
     2461 gaaacttcgc ctgcggtcga agctctggaa aatcttgatc cgcgttcact caccccggaa
     2521 caggcgctgg agtggattta tcgcttgaag agcctggtgg gagggggtgg ccatcaccat
     2581 caccatcata gggggctctg cacaccgtcg cgttaaggat ccgaattcga gctccgtcga
     2641 caagcttgcg gccgcactcg agcaccacca ccaccaccac tgagatccgg ctgctaacaa
     2701 agcccgaaag gaagctgagt tggctgctgc caccgctgag caataactag cataacccct
     2761 tggggcctct aaacgggtct tgaggggttt tttgctgaaa ggaggaacta tatccggatt
     2821 ggcgaatggg acgcgccctg tagcggcgca ttaagcgcgg cgggtgtggt ggttacgcgc
     2881 agcgtgaccg ctacacttgc cagcgcccta gcgcccgctc ctttcgcttt cttcccttcc
     2941 tttctcgcca cgttcgccgg ctttccccgt caagctctaa atcgggggct ccctttaggg
     3001 ttccgattta gtgctttacg gcacctcgac cccaaaaaac ttgattaggg tgatggttca
     3061 cgtagtgggc catcgccctg atagacggtt tttcgccctt tgacgttgga gtccacgttc
     3121 tttaatagtg gactcttgtt ccaaactgga acaacactca accctatctc ggtctattct
     3181 tttgatttat aagggatttt gccgatttcg gcctattggt taaaaaatga gctgatttaa
     3241 caaaaattta acgcgaattt taacaaaata ttaacgctta caatttaggt ggcacttttc
     3301 ggggaaatgt gcgcggaacc cctatttgtt tatttttcta aatacattca aatatgtatc
     3361 cgctcatgaa ttaattctta gaaaaactca tcgagcatca aatgaaactg caatttattc
     3421 atatcaggat tatcaatacc atatttttga aaaagccgtt tctgtaatga aggagaaaac
     3481 tcaccgaggc agttccatag gatggcaaga tcctggtatc ggtctgcgat tccgactcgt
     3541 ccaacatcaa tacaacctat taatttcccc tcgtcaaaaa taaggttatc aagtgagaaa
     3601 tcaccatgag tgacgactga atccggtgag aatggcaaaa gtttatgcat ttctttccag
     3661 acttgttcaa caggccagcc attacgctcg tcatcaaaat cactcgcatc aaccaaaccg
     3721 ttattcattc gtgattgcgc ctgagcgaga cgaaatacgc gatcgctgtt aaaaggacaa
     3781 ttacaaacag gaatcgaatg caaccggcgc aggaacactg ccagcgcatc aacaatattt
     3841 tcacctgaat caggatattc ttctaatacc tggaatgctg ttttcccggg gatcgcagtg
     3901 gtgagtaacc atgcatcatc aggagtacgg ataaaatgct tgatggtcgg aagaggcata
     3961 aattccgtca gccagtttag tctgaccatc tcatctgtaa catcattggc aacgctacct
     4021 ttgccatgtt tcagaaacaa ctctggcgca tcgggcttcc catacaatcg atagattgtc
     4081 gcacctgatt gcccgacatt atcgcgagcc catttatacc catataaatc agcatccatg
     4141 ttggaattta atcgcggcct agagcaagac gtttcccgtt gaatatggct cataacaccc
     4201 cttgtattac tgtttatgta agcagacagt tttattgttc atgaccaaaa tcccttaacg
     4261 tgagttttcg ttccactgag cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga
     4321 tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt
     4381 ggtttgtttg ccggatcaag agctaccaac tctttttccg aaggtaactg gcttcagcag
     4441 agcgcagata ccaaatactg tccttctagt gtagccgtag ttaggccacc acttcaagaa
     4501 ctctgtagca ccgcctacat acctcgctct gctaatcctg ttaccagtgg ctgctgccag
     4561 tggcgataag tcgtgtctta ccgggttgga ctcaagacga tagttaccgg ataaggcgca
     4621 gcggtcgggc tgaacggggg gttcgtgcac acagcccagc ttggagcgaa cgacctacac
     4681 cgaactgaga tacctacagc gtgagctatg agaaagcgcc acgcttcccg aagggagaaa
     4741 ggcggacagg tatccggtaa gcggcagggt cggaacagga gagcgcacga gggagcttcc
     4801 agggggaaac gcctggtatc tttatagtcc tgtcgggttt cgccacctct gacttgagcg
     4861 tcgatttttg tgatgctcgt caggggggcg gagcctatgg aaaaacgcca gcaacgcggc
     4921 ctttttacgg ttcctggcct tttgctggcc ttttgctcac atgttctttc ctgcgttatc
     4981 ccctgattct gtggataacc gtattaccgc ctttgagtga gctgataccg ctcgccgcag
     5041 ccgaacgacc gagcgcagcg agtcagtgag cgaggaagcg gaagagcgcc tgatgcggta
     5101 ttttctcctt acgcatctgt gcggtatttc acaccgcaat ggtgcactct cagtacaatc
     5161 tgctctgatg ccgcatagtt aagccagtat acactccgct atcgctacgt gactgggtca
     5221 tggctgcgcc ccgacacccg ccaacacccg ctgacgcgcc ctgacgggct tgtctgctcc
     5281 cggcatccgc ttacagacaa gctgtgaccg tctccgggag ctgcatgtgt cagaggtttt
     5341 caccgtcatc accgaaacgc gcgaggcagc tgcggtaaag ctcatcagcg tggtcgtgaa
     5401 gcgattcaca gatgtctgcc tgttcatccg cgtccagctc gttgagtttc tccagaagcg
     5461 ttaatgtctg gcttctgata aagcgggcca tgttaagggc ggttttttcc tgtttggtca
     5521 ctgatgcctc cgtgtaaggg ggatttctgt tcatgggggt aatgataccg atgaaacgag
     5581 agaggatgct cacgatacgg gttactgatg atgaacatgc ccggttactg gaacgttgtg
     5641 agggtaaaca actggcggta tggatgcggc gggaccagag aaaaatcact cagggtcaat
     5701 gccagcgctt cgttaataca gatgtaggtg ttccacaggg tagccagcag catcctgcga
     5761 tgcagatccg gaacataatg gtgcagggcg ctgacttccg cgtttccaga ctttacgaaa
     5821 cacggaaacc gaagaccatt catgttgttg ctcaggtcgc agacgttttg cagcagcagt
     5881 cgcttcacgt tcgctcgcgt atcggtgatt cattctgcta accagtaagg caaccccgcc
     5941 agcctagccg ggtcctcaac gacaggagca cgatcatgcg cacccgtggg gccgccatgc
     6001 cggcgataat ggcctgcttc tcgccgaaac gtttggtggc gggaccagtg acgaaggctt
     6061 gagcgagggc gtgcaagatt ccgaataccg caagcgacag gccgatcatc gtcgcgctcc
     6121 agcgaaagcg gtcctcgccg aaaatgaccc agagcgctgc cggcacctgt cctacgagtt
     6181 gcatgataaa gaagacagtc ataagtgcgg cgacgatagt catgccccgc gcccaccgga
     6241 aggagctgac tgggttgaag gctctcaagg gcatcggtcg agatcccggt gcctaatgag
     6301 tgagctaact tacattaatt gcgttgcgct cactgcccgc tttccagtcg ggaaacctgt
     6361 cgtgccagct gcattaatga atcggccaac gcgcggggag aggcggtttg cgtattgggc
     6421 gccagggtgg tttttctttt caccagtgag acgggcaaca gctgattgcc cttcaccgcc
     6481 tggccctgag agagttgcag caagcggtcc acgctggttt gccccagcag gcgaaaatcc
     6541 tgtttgatgg tggttaacgg cgggatataa catgagctgt cttcggtatc gtcgtatccc
     6601 actaccgaga tgtccgcacc aacgcgcagc ccggactcgg taatggcgcg cattgcgccc
     6661 agcgccatct gatcgttggc aaccagcatc gcagtgggaa cgatgccctc attcagcatt
     6721 tgcatggttt gttgaaaacc ggacatggca ctccagtcgc cttcccgttc cgctatcggc
     6781 tgaatttgat tgcgagtgag atatttatgc cagccagcca gacgcagacg cgccgagaca
     6841 gaacttaatg ggcccgctaa cagcgcgatt tgctggtgac ccaatgcgac cagatgctcc
     6901 acgcccagtc gcgtaccgtc ttcatgggag aaaataatac tgttgatggg tgtctggtca
     6961 gagacatcaa gaaataacgc cggaacatta gtgcaggcag cttccacagc aatggcatcc
     7021 tggtcatcca gcggatagtt aatgatcagc ccactgacgc gttgcgcgag aagattgtgc
     7081 accgccgctt tacaggcttc gacgccgctt cgttctacca tcgacaccac cacgctggca
     7141 cccagttgat cggcgcgaga tttaatcgcc gcgacaattt gcgacggcgc gtgcagggcc
     7201 agactggagg tggcaacgcc aatcagcaac gactgtttgc ccgccagttg ttgtgccacg
     7261 cggttgggaa tgtaattcag ctccgccatc gccgcttcca ctttttcccg cgttttcgca
     7321 gaaacgtggc tggcctggtt caccacgcgg gaaacggtct gataagagac accggcatac
     7381 tctgcgacat cgtataacgt tactggtttc acattcacca ccctgaattg actctcttcc
     7441 gggcgctatc atgccatacc gcgaaaggtt ttgcgccatt cgatggtgtc cgggatctcg
     7501 acgctctccc ttatgcgact cctgcattag gaagcagccc agtagtaggt tgaggccgtt
     7561 gagcaccgcc gccgcaagga atggtgcatg caaggagatg gcgcccaaca gtcccccggc
     7621 cacggggcct gccaccatac ccacgccgaa acaagcgctc atgagcccga agtggcgagc
     7681 ccgatcttcc ccatcggtga tgtcggcgat ataggcgcca gcaaccgcac ctgtggcgcc
     7741 ggtgatgccg gccacgatgc gtccggcgta gaggatcgag atcgatctcg atcccgcgaa
     7801 attaatacga ctcactatag gggaattgtg agcggataac aattcccctc tagaaataat
     7861 tttgtttaac tttaagaagg agatatacat aat
//
