LOCUS       sequence-263306-        3741 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses tagless S15-T79C mutant ribosomal protein in E.coli.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3741)
  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 3741)
  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-S15-T79C
FEATURES             Location/Qualifiers
     source          1..3741
                     /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(360..378)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      374..421
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     CDS             complement(651..1466)
                     /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(651..1466)
                     /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     1374..1393
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     promoter        complement(1467..1558)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(1574..2323)
                     /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(1584..1601)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     primer_bind     complement(1824..1843)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(2519..3601)
                     /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     3563..3582
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(3602..3679)
                     /gene="lacI"
                     /label=lacI promoter
     primer_bind     join(3725..3741,1..3)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        join(3725..3741,1..2)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
ORIGIN
        1 ggggaattgt gagcggataa caattcccct gtagaaataa ttttgtttaa ctttaataag
       61 gagatatacc atgtctctaa gtactgaagc aacagctaaa atcgtttctg agtttggtcg
      121 tgacgcaaac gacaccggtt ctaccgaagt tcaggtagca ctgctgactg cacagatcaa
      181 ccacctgcag ggccactttg cagagcacaa aaaagatcac cacagccgtc gtggtctgct
      241 gcgcatggtt tctcagcgtc gtaaactgct cgactacctg aaacgtaaag acgtagcacg
      301 ttactgccag ctcatcgagc gcctgggtct gcgtcgctaa ttaacctagg ctgctgccac
      361 cgctgagcaa taactagcat aaccccttgg ggcctctaaa cgggtcttga ggggtttttt
      421 gctgaaacct caggcatttg agaagcacac ggtcacactg cttccggtag tcaataaacc
      481 ggtaaaccag caatagacat aagcggctat ttaacgaccc tgccctgaac cgacgacaag
      541 ctgacgaccg ggtctccgca agtggcactt ttcggggaaa tgtgcgcgga acccctattt
      601 gtttattttt ctaaatacat tcaaatatgt atccgctcat gaattaattc ttagaaaaac
      661 tcatcgagca tcaaatgaaa ctgcaattta ttcatatcag gattatcaat accatatttt
      721 tgaaaaagcc gtttctgtaa tgaaggagaa aactcaccga ggcagttcca taggatggca
      781 agatcctggt atcggtctgc gattccgact cgtccaacat caatacaacc tattaatttc
      841 ccctcgtcaa aaataaggtt atcaagtgag aaatcaccat gagtgacgac tgaatccggt
      901 gagaatggca aaagtttatg catttctttc cagacttgtt caacaggcca gccattacgc
      961 tcgtcatcaa aatcactcgc atcaaccaaa ccgttattca ttcgtgattg cgcctgagcg
     1021 agacgaaata cgcggtcgct gttaaaagga caattacaaa caggaatcga atgcaaccgg
     1081 cgcaggaaca ctgccagcgc atcaacaata ttttcacctg aatcaggata ttcttctaat
     1141 acctggaatg ctgttttccc ggggatcgca gtggtgagta accatgcatc atcaggagta
     1201 cggataaaat gcttgatggt cggaagaggc ataaattccg tcagccagtt tagtctgacc
     1261 atctcatctg taacatcatt ggcaacgcta cctttgccat gtttcagaaa caactctggc
     1321 gcatcgggct tcccatacaa tcgatagatt gtcgcacctg attgcccgac attatcgcga
     1381 gcccatttat acccatataa atcagcatcc atgttggaat ttaatcgcgg cctagagcaa
     1441 gacgtttccc gttgaatatg gctcatactc ttcctttttc aatattattg aagcatttat
     1501 cagggttatt gtctcatgag cggatacata tttgaatgta tttagaaaaa taaacaaata
     1561 ggcatgcagc gctcttccgc ttcctcgctc actgactcgc tacgctcggt cgttcgactg
     1621 cggcgagcgg tgtcagctca ctcaaaagcg gtaatacggt tatccacaga atcaggggat
     1681 aaagccggaa agaacatgtg agcaaaaagc aaagcaccgg aagaagccaa cgccgcaggc
     1741 gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagccagag
     1801 gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt
     1861 gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg
     1921 aagcgtggcg ctttctcata gctcacgctg ttggtatctc agttcggtgt aggtcgttcg
     1981 ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg
     2041 taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccat
     2101 tggtaactga tttagaggac tttgtcttga agttatgcac ctgttaaggc taaactgaaa
     2161 gaacagattt tggtgagtgc ggtcctccaa cccacttacc ttggttcaaa gagttggtag
     2221 ctcagcgaac cttgagaaaa ccaccgttgg tagcggtggt ttttctttat ttatgagatg
     2281 atgaatcaat cggtctatca agtcaacgaa cagctattcc gttactctag atttcagtgc
     2341 aatttatctc ttcaaatgta gcacctgaag tcagccccat acgatataag ttgtaattct
     2401 catgttagtc atgccccgcg cccaccggaa ggagctgact gggttgaagg ctctcaaggg
     2461 catcggtcga gatcccggtg cctaatgagt gagctaactt acattaattg cgttgcgctc
     2521 actgcccgct ttccagtcgg gaaacctgtc gtgccagctg cattaatgaa tcggccaacg
     2581 cgcggggaga ggcggtttgc gtattgggcg ccagggtggt ttttcttttc accagtgaga
     2641 cgggcaacag ctgattgccc ttcaccgcct ggccctgaga gagttgcagc aagcggtcca
     2701 cgctggtttg ccccagcagg cgaaaatcct gtttgatggt ggttaacggc gggatataac
     2761 atgagctgtc ttcggtatcg tcgtatccca ctaccgagat gtccgcacca acgcgcagcc
     2821 cggactcggt aatggcgcgc attgcgccca gcgccatctg atcgttggca accagcatcg
     2881 cagtgggaac gatgccctca ttcagcattt gcatggtttg ttgaaaaccg gacatggcac
     2941 tccagtcgcc ttcccgttcc gctatcggct gaatttgatt gcgagtgaga tatttatgcc
     3001 agccagccag acgcagacgc gccgagacag aacttaatgg gcccgctaac agcgcgattt
     3061 gctggtgacc caatgcgacc agatgctcca cgcccagtcg cgtaccgtct tcatgggaga
     3121 aaataatact gttgatgggt gtctggtcag agacatcaag aaataacgcc ggaacattag
     3181 tgcaggcagc ttccacagca atggcatcct ggtcatccag cggatagtta atgatcagcc
     3241 cactgacgcg ttgcgcgaga agattgtgca ccgccgcttt acaggcttcg acgccgcttc
     3301 gttctaccat cgacaccacc acgctggcac ccagttgatc ggcgcgagat ttaatcgccg
     3361 cgacaatttg cgacggcgcg tgcagggcca gactggaggt ggcaacgcca atcagcaacg
     3421 actgtttgcc cgccagttgt tgtgccacgc ggttgggaat gtaattcagc tccgccatcg
     3481 ccgcttccac tttttcccgc gttttcgcag aaacgtggct ggcctggttc accacgcggg
     3541 aaacggtctg ataagagaca ccggcatact ctgcgacatc gtataacgtt actggtttca
     3601 cattcaccac cctgaattga ctctcttccg ggcgctatca tgccataccg cgaaaggttt
     3661 tgcgccattc gatggtgtcc gggatctcga cgctctccct tatgcgactc ctgcattagg
     3721 aaattaatac gactcactat a
//
