LOCUS       sequence-263305-        4173 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses tagless S3-M129C mutant ribosomal protein in E.coli.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4173)
  AUTHORS   Duss O, Stepanyuk GA, Puglisi JD, Williamson JR
  TITLE     Transient Protein-RNA Interactions Guide Nascent Ribosomal RNA 
            Folding.
  JOURNAL   Cell. 2019 Nov 27;179(6):1357-1369.e16. doi: 
            10.1016/j.cell.2019.10.035. Epub 2019 Nov 21.
  PUBMED    31761533
REFERENCE   2  (bases 1 to 4173)
  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: 
            "10.1016/j.cell.2019.10.035"; journalName: "Cell"; date: 
            "2019-11-27- 27"; volume: "179"; issue: "6"; pages: "1357-1369.e16"
COMMENT     Sequence Label: pRSF-1b-S3-M129C
FEATURES             Location/Qualifiers
     source          1..4173
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    3..27
                     /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)."
     primer_bind     complement(792..810)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      806..853
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     CDS             complement(1083..1898)
                     /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(1083..1898)
                     /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     1806..1825
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     promoter        complement(1899..1990)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(2006..2755)
                     /direction=LEFT
                     /label=RSF ori
                     /note="Plasmids containing the RSF 1030 origin of 
                     replication can be propagated in E. coli cells that contain
                     additional plasmids with compatible origins."
     primer_bind     complement(2016..2033)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     primer_bind     complement(2256..2275)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(2951..4033)
                     /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     3995..4014
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(4034..4111)
                     /gene="lacI"
                     /label=lacI promoter
     primer_bind     join(4157..4173,1..3)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        join(4157..4173,1..2)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
ORIGIN
        1 ggggaattgt gagcggataa caattcccct gtagaaataa ttttgtttaa ctttaataag
       61 gagatatacc atgggtcaga aagtacatcc taatggtatt cgcctgggta ttgtaaaacc
      121 atggaactct acctggtttg cgaacaccaa agaattcgct gacaacctgg acagcgattt
      181 taaagtacgt cagtacctga ctaaggaact ggctaaagcg tccgtatctc gtatcgttat
      241 cgagcgtccg gctaagagca tccgtgtaac cattcacact gctcgcccgg gtatcgttat
      301 cggtaaaaaa ggtgaagacg tagaaaaact gcgtaaggtc gtagcggaca tcgctggcgt
      361 tcctgcacag atcaacatcg ccgaagttcg taagcctgaa ctggacgcaa aactggttgc
      421 tgacagcatc acttctcagc tggaacgtcg cgtttgcttc cgtcgtgcta tgaagcgtgc
      481 tgtacagaac gcaatgcgtc tgggcgctaa aggtattaaa gttgaagtta gcggccgtct
      541 gggcggcgcg gaaatcgcac gtaccgaatg gtaccgcgaa ggtcgcgtac cgctgcacac
      601 tctgcgtgct gacatcgact acaacacctc tgaagcgcac accacttacg gtgtaatcgg
      661 cgttaaagtg tggatcttca aaggcgagat cctgggtggt atggctgctg ttgaacaacc
      721 ggaaaaaccg gctgctcagc ctaaaaagca gcagcgtaaa ggccgtaaat aattaaccta
      781 ggctgctgcc accgctgagc aataactagc ataacccctt ggggcctcta aacgggtctt
      841 gaggggtttt ttgctgaaac ctcaggcatt tgagaagcac acggtcacac tgcttccggt
      901 agtcaataaa ccggtaaacc agcaatagac ataagcggct atttaacgac cctgccctga
      961 accgacgaca agctgacgac cgggtctccg caagtggcac ttttcgggga aatgtgcgcg
     1021 gaacccctat ttgtttattt ttctaaatac attcaaatat gtatccgctc atgaattaat
     1081 tcttagaaaa actcatcgag catcaaatga aactgcaatt tattcatatc aggattatca
     1141 ataccatatt tttgaaaaag ccgtttctgt aatgaaggag aaaactcacc gaggcagttc
     1201 cataggatgg caagatcctg gtatcggtct gcgattccga ctcgtccaac atcaatacaa
     1261 cctattaatt tcccctcgtc aaaaataagg ttatcaagtg agaaatcacc atgagtgacg
     1321 actgaatccg gtgagaatgg caaaagttta tgcatttctt tccagacttg ttcaacaggc
     1381 cagccattac gctcgtcatc aaaatcactc gcatcaacca aaccgttatt cattcgtgat
     1441 tgcgcctgag cgagacgaaa tacgcggtcg ctgttaaaag gacaattaca aacaggaatc
     1501 gaatgcaacc ggcgcaggaa cactgccagc gcatcaacaa tattttcacc tgaatcagga
     1561 tattcttcta atacctggaa tgctgttttc ccggggatcg cagtggtgag taaccatgca
     1621 tcatcaggag tacggataaa atgcttgatg gtcggaagag gcataaattc cgtcagccag
     1681 tttagtctga ccatctcatc tgtaacatca ttggcaacgc tacctttgcc atgtttcaga
     1741 aacaactctg gcgcatcggg cttcccatac aatcgataga ttgtcgcacc tgattgcccg
     1801 acattatcgc gagcccattt atacccatat aaatcagcat ccatgttgga atttaatcgc
     1861 ggcctagagc aagacgtttc ccgttgaata tggctcatac tcttcctttt tcaatattat
     1921 tgaagcattt atcagggtta ttgtctcatg agcggataca tatttgaatg tatttagaaa
     1981 aataaacaaa taggcatgca gcgctcttcc gcttcctcgc tcactgactc gctacgctcg
     2041 gtcgttcgac tgcggcgagc ggtgtcagct cactcaaaag cggtaatacg gttatccaca
     2101 gaatcagggg ataaagccgg aaagaacatg tgagcaaaaa gcaaagcacc ggaagaagcc
     2161 aacgccgcag gcgtttttcc ataggctccg cccccctgac gagcatcaca aaaatcgacg
     2221 ctcaagccag aggtggcgaa acccgacagg actataaaga taccaggcgt ttccccctgg
     2281 aagctccctc gtgcgctctc ctgttccgac cctgccgctt accggatacc tgtccgcctt
     2341 tctcccttcg ggaagcgtgg cgctttctca tagctcacgc tgttggtatc tcagttcggt
     2401 gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg
     2461 cgccttatcc ggtaactatc gtcttgagtc caacccggta agacacgact tatcgccact
     2521 ggcagcagcc attggtaact gatttagagg actttgtctt gaagttatgc acctgttaag
     2581 gctaaactga aagaacagat tttggtgagt gcggtcctcc aacccactta ccttggttca
     2641 aagagttggt agctcagcga accttgagaa aaccaccgtt ggtagcggtg gtttttcttt
     2701 atttatgaga tgatgaatca atcggtctat caagtcaacg aacagctatt ccgttactct
     2761 agatttcagt gcaatttatc tcttcaaatg tagcacctga agtcagcccc atacgatata
     2821 agttgtaatt ctcatgttag tcatgccccg cgcccaccgg aaggagctga ctgggttgaa
     2881 ggctctcaag ggcatcggtc gagatcccgg tgcctaatga gtgagctaac ttacattaat
     2941 tgcgttgcgc tcactgcccg ctttccagtc gggaaacctg tcgtgccagc tgcattaatg
     3001 aatcggccaa cgcgcgggga gaggcggttt gcgtattggg cgccagggtg gtttttcttt
     3061 tcaccagtga gacgggcaac agctgattgc ccttcaccgc ctggccctga gagagttgca
     3121 gcaagcggtc cacgctggtt tgccccagca ggcgaaaatc ctgtttgatg gtggttaacg
     3181 gcgggatata acatgagctg tcttcggtat cgtcgtatcc cactaccgag atgtccgcac
     3241 caacgcgcag cccggactcg gtaatggcgc gcattgcgcc cagcgccatc tgatcgttgg
     3301 caaccagcat cgcagtggga acgatgccct cattcagcat ttgcatggtt tgttgaaaac
     3361 cggacatggc actccagtcg ccttcccgtt ccgctatcgg ctgaatttga ttgcgagtga
     3421 gatatttatg ccagccagcc agacgcagac gcgccgagac agaacttaat gggcccgcta
     3481 acagcgcgat ttgctggtga cccaatgcga ccagatgctc cacgcccagt cgcgtaccgt
     3541 cttcatggga gaaaataata ctgttgatgg gtgtctggtc agagacatca agaaataacg
     3601 ccggaacatt agtgcaggca gcttccacag caatggcatc ctggtcatcc agcggatagt
     3661 taatgatcag cccactgacg cgttgcgcga gaagattgtg caccgccgct ttacaggctt
     3721 cgacgccgct tcgttctacc atcgacacca ccacgctggc acccagttga tcggcgcgag
     3781 atttaatcgc cgcgacaatt tgcgacggcg cgtgcagggc cagactggag gtggcaacgc
     3841 caatcagcaa cgactgtttg cccgccagtt gttgtgccac gcggttggga atgtaattca
     3901 gctccgccat cgccgcttcc actttttccc gcgttttcgc agaaacgtgg ctggcctggt
     3961 tcaccacgcg ggaaacggtc tgataagaga caccggcata ctctgcgaca tcgtataacg
     4021 ttactggttt cacattcacc accctgaatt gactctcttc cgggcgctat catgccatac
     4081 cgcgaaaggt tttgcgccat tcgatggtgt ccgggatctc gacgctctcc cttatgcgac
     4141 tcctgcatta ggaaattaat acgactcact ata
//
