LOCUS       sequence-263268-        3783 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses tagless S10 ribosomal protein in E.coli.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3783)
  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 3783)
  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-S10
FEATURES             Location/Qualifiers
     source          1..3783
                     /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(402..420)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      416..463
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     CDS             complement(693..1508)
                     /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(693..1508)
                     /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     1416..1435
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     promoter        complement(1509..1600)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(1616..2365)
                     /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(1626..1643)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     primer_bind     complement(1866..1885)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(2561..3643)
                     /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     3605..3624
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(3644..3721)
                     /gene="lacI"
                     /label=lacI promoter
     primer_bind     join(3767..3783,1..3)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        join(3767..3783,1..2)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
ORIGIN
        1 ggggaattgt gagcggataa caattcccct gtagaaataa ttttgtttaa ctttaataag
       61 gagatatacc atgcagaacc aaagaatccg tatccgcctg aaagcgtttg atcatcgtct
      121 gatcgatcaa gcaaccgcgg aaatcgtcga gactgccaag cgcactggtg cgcaggtccg
      181 tggtccgatc ccgctgccga cacgcaaaga gcgcttcact gttctgatct ccccgcacgt
      241 caacaaagac gcgcgcgatc agtacgaaat ccgtactcac ttgcgtctgg ttgacatcgt
      301 tgagccaacc gagaaaaccg ttgatgctct gatgcgtctg gatctggctg ccggtgtaga
      361 cgtgcagatc agcctgggtt aattaaccta ggctgctgcc accgctgagc aataactagc
      421 ataacccctt ggggcctcta aacgggtctt gaggggtttt ttgctgaaac ctcaggcatt
      481 tgagaagcac acggtcacac tgcttccggt agtcaataaa ccggtaaacc agcaatagac
      541 ataagcggct atttaacgac cctgccctga accgacgaca agctgacgac cgggtctccg
      601 caagtggcac ttttcgggga aatgtgcgcg gaacccctat ttgtttattt ttctaaatac
      661 attcaaatat gtatccgctc atgaattaat tcttagaaaa actcatcgag catcaaatga
      721 aactgcaatt tattcatatc aggattatca ataccatatt tttgaaaaag ccgtttctgt
      781 aatgaaggag aaaactcacc gaggcagttc cataggatgg caagatcctg gtatcggtct
      841 gcgattccga ctcgtccaac atcaatacaa cctattaatt tcccctcgtc aaaaataagg
      901 ttatcaagtg agaaatcacc atgagtgacg actgaatccg gtgagaatgg caaaagttta
      961 tgcatttctt tccagacttg ttcaacaggc cagccattac gctcgtcatc aaaatcactc
     1021 gcatcaacca aaccgttatt cattcgtgat tgcgcctgag cgagacgaaa tacgcggtcg
     1081 ctgttaaaag gacaattaca aacaggaatc gaatgcaacc ggcgcaggaa cactgccagc
     1141 gcatcaacaa tattttcacc tgaatcagga tattcttcta atacctggaa tgctgttttc
     1201 ccggggatcg cagtggtgag taaccatgca tcatcaggag tacggataaa atgcttgatg
     1261 gtcggaagag gcataaattc cgtcagccag tttagtctga ccatctcatc tgtaacatca
     1321 ttggcaacgc tacctttgcc atgtttcaga aacaactctg gcgcatcggg cttcccatac
     1381 aatcgataga ttgtcgcacc tgattgcccg acattatcgc gagcccattt atacccatat
     1441 aaatcagcat ccatgttgga atttaatcgc ggcctagagc aagacgtttc ccgttgaata
     1501 tggctcatac tcttcctttt tcaatattat tgaagcattt atcagggtta ttgtctcatg
     1561 agcggataca tatttgaatg tatttagaaa aataaacaaa taggcatgca gcgctcttcc
     1621 gcttcctcgc tcactgactc gctacgctcg gtcgttcgac tgcggcgagc ggtgtcagct
     1681 cactcaaaag cggtaatacg gttatccaca gaatcagggg ataaagccgg aaagaacatg
     1741 tgagcaaaaa gcaaagcacc ggaagaagcc aacgccgcag gcgtttttcc ataggctccg
     1801 cccccctgac gagcatcaca aaaatcgacg ctcaagccag aggtggcgaa acccgacagg
     1861 actataaaga taccaggcgt ttccccctgg aagctccctc gtgcgctctc ctgttccgac
     1921 cctgccgctt accggatacc tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca
     1981 tagctcacgc tgttggtatc tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt
     2041 gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc
     2101 caacccggta agacacgact tatcgccact ggcagcagcc attggtaact gatttagagg
     2161 actttgtctt gaagttatgc acctgttaag gctaaactga aagaacagat tttggtgagt
     2221 gcggtcctcc aacccactta ccttggttca aagagttggt agctcagcga accttgagaa
     2281 aaccaccgtt ggtagcggtg gtttttcttt atttatgaga tgatgaatca atcggtctat
     2341 caagtcaacg aacagctatt ccgttactct agatttcagt gcaatttatc tcttcaaatg
     2401 tagcacctga agtcagcccc atacgatata agttgtaatt ctcatgttag tcatgccccg
     2461 cgcccaccgg aaggagctga ctgggttgaa ggctctcaag ggcatcggtc gagatcccgg
     2521 tgcctaatga gtgagctaac ttacattaat tgcgttgcgc tcactgcccg ctttccagtc
     2581 gggaaacctg tcgtgccagc tgcattaatg aatcggccaa cgcgcgggga gaggcggttt
     2641 gcgtattggg cgccagggtg gtttttcttt tcaccagtga gacgggcaac agctgattgc
     2701 ccttcaccgc ctggccctga gagagttgca gcaagcggtc cacgctggtt tgccccagca
     2761 ggcgaaaatc ctgtttgatg gtggttaacg gcgggatata acatgagctg tcttcggtat
     2821 cgtcgtatcc cactaccgag atgtccgcac caacgcgcag cccggactcg gtaatggcgc
     2881 gcattgcgcc cagcgccatc tgatcgttgg caaccagcat cgcagtggga acgatgccct
     2941 cattcagcat ttgcatggtt tgttgaaaac cggacatggc actccagtcg ccttcccgtt
     3001 ccgctatcgg ctgaatttga ttgcgagtga gatatttatg ccagccagcc agacgcagac
     3061 gcgccgagac agaacttaat gggcccgcta acagcgcgat ttgctggtga cccaatgcga
     3121 ccagatgctc cacgcccagt cgcgtaccgt cttcatggga gaaaataata ctgttgatgg
     3181 gtgtctggtc agagacatca agaaataacg ccggaacatt agtgcaggca gcttccacag
     3241 caatggcatc ctggtcatcc agcggatagt taatgatcag cccactgacg cgttgcgcga
     3301 gaagattgtg caccgccgct ttacaggctt cgacgccgct tcgttctacc atcgacacca
     3361 ccacgctggc acccagttga tcggcgcgag atttaatcgc cgcgacaatt tgcgacggcg
     3421 cgtgcagggc cagactggag gtggcaacgc caatcagcaa cgactgtttg cccgccagtt
     3481 gttgtgccac gcggttggga atgtaattca gctccgccat cgccgcttcc actttttccc
     3541 gcgttttcgc agaaacgtgg ctggcctggt tcaccacgcg ggaaacggtc tgataagaga
     3601 caccggcata ctctgcgaca tcgtataacg ttactggttt cacattcacc accctgaatt
     3661 gactctcttc cgggcgctat catgccatac cgcgaaaggt tttgcgccat tcgatggtgt
     3721 ccgggatctc gacgctctcc cttatgcgac tcctgcatta ggaaattaat acgactcact
     3781 ata
//
