LOCUS       sequence-263279-        3749 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses tagless S19-T48C mutant 30S ribosomal protein in E.coli
             .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3749)
  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 3749)
  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-S19-T48C
FEATURES             Location/Qualifiers
     source          1..3749
                     /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(368..386)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      382..429
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     CDS             complement(659..1474)
                     /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(659..1474)
                     /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     1382..1401
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     promoter        complement(1475..1566)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(1582..2331)
                     /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(1592..1609)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     primer_bind     complement(1832..1851)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(2527..3609)
                     /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     3571..3590
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(3610..3687)
                     /gene="lacI"
                     /label=lacI promoter
     primer_bind     join(3733..3749,1..3)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        join(3733..3749,1..2)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
ORIGIN
        1 ggggaattgt gagcggataa caattcccct gtagaaataa ttttgtttaa ctttaataag
       61 gagatatacc atgccacgtt ctctcaagaa aggtcctttt attgacctgc acttgctgaa
      121 gaaggtagag aaagcggtgg aaagcggaga caagaagccc ctgcgcactt ggtcccgtcg
      181 ttcaacgatc tttcctaaca tgatcggttt gtgcatcgct gtccataatg gtcgtcagca
      241 cgttccggta tttgtaaccg acgaaatggt tggtcacaaa ctgggtgaat tcgcaccgac
      301 tcgtacttat cgcggccacg ctgctgataa aaaagcgaag aagaaataat tacctaggct
      361 gctgccaccg ctgagcaata actagcataa ccccttgggg cctctaaacg ggtcttgagg
      421 ggttttttgc tgaaacctca ggcatttgag aagcacacgg tcacactgct tccggtagtc
      481 aataaaccgg taaaccagca atagacataa gcggctattt aacgaccctg ccctgaaccg
      541 acgacaagct gacgaccggg tctccgcaag tggcactttt cggggaaatg tgcgcggaac
      601 ccctatttgt ttatttttct aaatacattc aaatatgtat ccgctcatga attaattctt
      661 agaaaaactc atcgagcatc aaatgaaact gcaatttatt catatcagga ttatcaatac
      721 catatttttg aaaaagccgt ttctgtaatg aaggagaaaa ctcaccgagg cagttccata
      781 ggatggcaag atcctggtat cggtctgcga ttccgactcg tccaacatca atacaaccta
      841 ttaatttccc ctcgtcaaaa ataaggttat caagtgagaa atcaccatga gtgacgactg
      901 aatccggtga gaatggcaaa agtttatgca tttctttcca gacttgttca acaggccagc
      961 cattacgctc gtcatcaaaa tcactcgcat caaccaaacc gttattcatt cgtgattgcg
     1021 cctgagcgag acgaaatacg cggtcgctgt taaaaggaca attacaaaca ggaatcgaat
     1081 gcaaccggcg caggaacact gccagcgcat caacaatatt ttcacctgaa tcaggatatt
     1141 cttctaatac ctggaatgct gttttcccgg ggatcgcagt ggtgagtaac catgcatcat
     1201 caggagtacg gataaaatgc ttgatggtcg gaagaggcat aaattccgtc agccagttta
     1261 gtctgaccat ctcatctgta acatcattgg caacgctacc tttgccatgt ttcagaaaca
     1321 actctggcgc atcgggcttc ccatacaatc gatagattgt cgcacctgat tgcccgacat
     1381 tatcgcgagc ccatttatac ccatataaat cagcatccat gttggaattt aatcgcggcc
     1441 tagagcaaga cgtttcccgt tgaatatggc tcatactctt cctttttcaa tattattgaa
     1501 gcatttatca gggttattgt ctcatgagcg gatacatatt tgaatgtatt tagaaaaata
     1561 aacaaatagg catgcagcgc tcttccgctt cctcgctcac tgactcgcta cgctcggtcg
     1621 ttcgactgcg gcgagcggtg tcagctcact caaaagcggt aatacggtta tccacagaat
     1681 caggggataa agccggaaag aacatgtgag caaaaagcaa agcaccggaa gaagccaacg
     1741 ccgcaggcgt ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca
     1801 agccagaggt ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc
     1861 tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc
     1921 ccttcgggaa gcgtggcgct ttctcatagc tcacgctgtt ggtatctcag ttcggtgtag
     1981 gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc
     2041 ttatccggta actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca
     2101 gcagccattg gtaactgatt tagaggactt tgtcttgaag ttatgcacct gttaaggcta
     2161 aactgaaaga acagattttg gtgagtgcgg tcctccaacc cacttacctt ggttcaaaga
     2221 gttggtagct cagcgaacct tgagaaaacc accgttggta gcggtggttt ttctttattt
     2281 atgagatgat gaatcaatcg gtctatcaag tcaacgaaca gctattccgt tactctagat
     2341 ttcagtgcaa tttatctctt caaatgtagc acctgaagtc agccccatac gatataagtt
     2401 gtaattctca tgttagtcat gccccgcgcc caccggaagg agctgactgg gttgaaggct
     2461 ctcaagggca tcggtcgaga tcccggtgcc taatgagtga gctaacttac attaattgcg
     2521 ttgcgctcac tgcccgcttt ccagtcggga aacctgtcgt gccagctgca ttaatgaatc
     2581 ggccaacgcg cggggagagg cggtttgcgt attgggcgcc agggtggttt ttcttttcac
     2641 cagtgagacg ggcaacagct gattgccctt caccgcctgg ccctgagaga gttgcagcaa
     2701 gcggtccacg ctggtttgcc ccagcaggcg aaaatcctgt ttgatggtgg ttaacggcgg
     2761 gatataacat gagctgtctt cggtatcgtc gtatcccact accgagatgt ccgcaccaac
     2821 gcgcagcccg gactcggtaa tggcgcgcat tgcgcccagc gccatctgat cgttggcaac
     2881 cagcatcgca gtgggaacga tgccctcatt cagcatttgc atggtttgtt gaaaaccgga
     2941 catggcactc cagtcgcctt cccgttccgc tatcggctga atttgattgc gagtgagata
     3001 tttatgccag ccagccagac gcagacgcgc cgagacagaa cttaatgggc ccgctaacag
     3061 cgcgatttgc tggtgaccca atgcgaccag atgctccacg cccagtcgcg taccgtcttc
     3121 atgggagaaa ataatactgt tgatgggtgt ctggtcagag acatcaagaa ataacgccgg
     3181 aacattagtg caggcagctt ccacagcaat ggcatcctgg tcatccagcg gatagttaat
     3241 gatcagccca ctgacgcgtt gcgcgagaag attgtgcacc gccgctttac aggcttcgac
     3301 gccgcttcgt tctaccatcg acaccaccac gctggcaccc agttgatcgg cgcgagattt
     3361 aatcgccgcg acaatttgcg acggcgcgtg cagggccaga ctggaggtgg caacgccaat
     3421 cagcaacgac tgtttgcccg ccagttgttg tgccacgcgg ttgggaatgt aattcagctc
     3481 cgccatcgcc gcttccactt tttcccgcgt tttcgcagaa acgtggctgg cctggttcac
     3541 cacgcgggaa acggtctgat aagagacacc ggcatactct gcgacatcgt ataacgttac
     3601 tggtttcaca ttcaccaccc tgaattgact ctcttccggg cgctatcatg ccataccgcg
     3661 aaaggttttg cgccattcga tggtgtccgg gatctcgacg ctctccctta tgcgactcct
     3721 gcattaggaa attaatacga ctcactata
//
