LOCUS       sequence-263223-        4197 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses tagless S2 ribosomal protein in E.coli.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4197)
  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 4197)
  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-S2
FEATURES             Location/Qualifiers
     source          1..4197
                     /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(816..834)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      830..877
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     CDS             complement(1107..1922)
                     /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(1107..1922)
                     /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     1830..1849
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     promoter        complement(1923..2014)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(2030..2779)
                     /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(2040..2057)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     primer_bind     complement(2280..2299)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(2975..4057)
                     /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     4019..4038
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(4058..4135)
                     /gene="lacI"
                     /label=lacI promoter
     primer_bind     join(4181..4197,1..3)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        join(4181..4197,1..2)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
ORIGIN
        1 ggggaattgt gagcggataa caattcccct gtagaaataa ttttgtttaa ctttaataag
       61 gagatatacc atggcaactg tttccatgcg cgacatgctc aaggctggtg ttcacttcgg
      121 tcaccagacc cgttactgga acccgaaaat gaagccgttc atcttcggtg cgcgtaacaa
      181 agttcacatc atcaaccttg agaaaactgt accgatgttc aacgaagctc tggctgaact
      241 gaacaagatt gcttctcgca aaggtaaaat ccttttcgtt ggtactaaac gcgctgcaag
      301 cgaagcggtg aaagacgctg ctctgagctg cgaccagttc ttcgtgaacc atcgctggct
      361 gggcggtatg ctgactaact ggaaaaccgt tcgtcagtcc atcaaacgtc tgaaagacct
      421 ggaaactcag tctcaggacg gtactttcga caagctgacc aagaaagaag cgctgatgcg
      481 cactcgtgag ctggagaaac tggaaaacag cctgggcggt atcaaagaca tgggcggtct
      541 gccggacgct ctgtttgtaa tcgatgctga ccacgaacac attgctatca aagaagcaaa
      601 caacctgggt attccggtat ttgctatcgt tgataccaac tctgatccgg acggtgttga
      661 cttcgttatc ccgggtaacg acgacgcaat ccgtgctgtg accctgtacc tgggcgctgt
      721 tgctgcaacc gtacgtgaag gccgttctca ggatctggct tcccaggcgg aagaaagctt
      781 cgtagaagct gagtaattaa cctaggctgc tgccaccgct gagcaataac tagcataacc
      841 ccttggggcc tctaaacggg tcttgagggg ttttttgctg aaacctcagg catttgagaa
      901 gcacacggtc acactgcttc cggtagtcaa taaaccggta aaccagcaat agacataagc
      961 ggctatttaa cgaccctgcc ctgaaccgac gacaagctga cgaccgggtc tccgcaagtg
     1021 gcacttttcg gggaaatgtg cgcggaaccc ctatttgttt atttttctaa atacattcaa
     1081 atatgtatcc gctcatgaat taattcttag aaaaactcat cgagcatcaa atgaaactgc
     1141 aatttattca tatcaggatt atcaatacca tatttttgaa aaagccgttt ctgtaatgaa
     1201 ggagaaaact caccgaggca gttccatagg atggcaagat cctggtatcg gtctgcgatt
     1261 ccgactcgtc caacatcaat acaacctatt aatttcccct cgtcaaaaat aaggttatca
     1321 agtgagaaat caccatgagt gacgactgaa tccggtgaga atggcaaaag tttatgcatt
     1381 tctttccaga cttgttcaac aggccagcca ttacgctcgt catcaaaatc actcgcatca
     1441 accaaaccgt tattcattcg tgattgcgcc tgagcgagac gaaatacgcg gtcgctgtta
     1501 aaaggacaat tacaaacagg aatcgaatgc aaccggcgca ggaacactgc cagcgcatca
     1561 acaatatttt cacctgaatc aggatattct tctaatacct ggaatgctgt tttcccgggg
     1621 atcgcagtgg tgagtaacca tgcatcatca ggagtacgga taaaatgctt gatggtcgga
     1681 agaggcataa attccgtcag ccagtttagt ctgaccatct catctgtaac atcattggca
     1741 acgctacctt tgccatgttt cagaaacaac tctggcgcat cgggcttccc atacaatcga
     1801 tagattgtcg cacctgattg cccgacatta tcgcgagccc atttataccc atataaatca
     1861 gcatccatgt tggaatttaa tcgcggccta gagcaagacg tttcccgttg aatatggctc
     1921 atactcttcc tttttcaata ttattgaagc atttatcagg gttattgtct catgagcgga
     1981 tacatatttg aatgtattta gaaaaataaa caaataggca tgcagcgctc ttccgcttcc
     2041 tcgctcactg actcgctacg ctcggtcgtt cgactgcggc gagcggtgtc agctcactca
     2101 aaagcggtaa tacggttatc cacagaatca ggggataaag ccggaaagaa catgtgagca
     2161 aaaagcaaag caccggaaga agccaacgcc gcaggcgttt ttccataggc tccgcccccc
     2221 tgacgagcat cacaaaaatc gacgctcaag ccagaggtgg cgaaacccga caggactata
     2281 aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc
     2341 gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt ctcatagctc
     2401 acgctgttgg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga
     2461 accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc
     2521 ggtaagacac gacttatcgc cactggcagc agccattggt aactgattta gaggactttg
     2581 tcttgaagtt atgcacctgt taaggctaaa ctgaaagaac agattttggt gagtgcggtc
     2641 ctccaaccca cttaccttgg ttcaaagagt tggtagctca gcgaaccttg agaaaaccac
     2701 cgttggtagc ggtggttttt ctttatttat gagatgatga atcaatcggt ctatcaagtc
     2761 aacgaacagc tattccgtta ctctagattt cagtgcaatt tatctcttca aatgtagcac
     2821 ctgaagtcag ccccatacga tataagttgt aattctcatg ttagtcatgc cccgcgccca
     2881 ccggaaggag ctgactgggt tgaaggctct caagggcatc ggtcgagatc ccggtgccta
     2941 atgagtgagc taacttacat taattgcgtt gcgctcactg cccgctttcc agtcgggaaa
     3001 cctgtcgtgc cagctgcatt aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat
     3061 tgggcgccag ggtggttttt cttttcacca gtgagacggg caacagctga ttgcccttca
     3121 ccgcctggcc ctgagagagt tgcagcaagc ggtccacgct ggtttgcccc agcaggcgaa
     3181 aatcctgttt gatggtggtt aacggcggga tataacatga gctgtcttcg gtatcgtcgt
     3241 atcccactac cgagatgtcc gcaccaacgc gcagcccgga ctcggtaatg gcgcgcattg
     3301 cgcccagcgc catctgatcg ttggcaacca gcatcgcagt gggaacgatg ccctcattca
     3361 gcatttgcat ggtttgttga aaaccggaca tggcactcca gtcgccttcc cgttccgcta
     3421 tcggctgaat ttgattgcga gtgagatatt tatgccagcc agccagacgc agacgcgccg
     3481 agacagaact taatgggccc gctaacagcg cgatttgctg gtgacccaat gcgaccagat
     3541 gctccacgcc cagtcgcgta ccgtcttcat gggagaaaat aatactgttg atgggtgtct
     3601 ggtcagagac atcaagaaat aacgccggaa cattagtgca ggcagcttcc acagcaatgg
     3661 catcctggtc atccagcgga tagttaatga tcagcccact gacgcgttgc gcgagaagat
     3721 tgtgcaccgc cgctttacag gcttcgacgc cgcttcgttc taccatcgac accaccacgc
     3781 tggcacccag ttgatcggcg cgagatttaa tcgccgcgac aatttgcgac ggcgcgtgca
     3841 gggccagact ggaggtggca acgccaatca gcaacgactg tttgcccgcc agttgttgtg
     3901 ccacgcggtt gggaatgtaa ttcagctccg ccatcgccgc ttccactttt tcccgcgttt
     3961 tcgcagaaac gtggctggcc tggttcacca cgcgggaaac ggtctgataa gagacaccgg
     4021 catactctgc gacatcgtat aacgttactg gtttcacatt caccaccctg aattgactct
     4081 cttccgggcg ctatcatgcc ataccgcgaa aggttttgcg ccattcgatg gtgtccggga
     4141 tctcgacgct ctcccttatg cgactcctgc attaggaaat taatacgact cactata
//
