LOCUS       sequence-263258-        3941 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses tagless S7-S83C, Delta-157-178 mutant 30S ribosomal 
            protein in E.coli.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3941)
  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 3941)
  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-S7-S83C/Delta-157-178
FEATURES             Location/Qualifiers
     source          1..3941
                     /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(560..578)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      574..621
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     CDS             complement(851..1666)
                     /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(851..1666)
                     /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     1574..1593
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     promoter        complement(1667..1758)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(1774..2523)
                     /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(1784..1801)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     primer_bind     complement(2024..2043)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(2719..3801)
                     /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     3763..3782
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(3802..3879)
                     /gene="lacI"
                     /label=lacI promoter
     primer_bind     join(3925..3941,1..3)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        join(3925..3941,1..2)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
ORIGIN
        1 ggggaattgt gagcggataa caattcccct gtagaaataa ttttgtttaa ctttaataag
       61 gagatatacc atgccacgtc gtcgcgtcat tggtcagcgt aaaattctgc cggatccgaa
      121 gttcggatca gaactgctgg ctaaatttgt aaatatcctg atggtagatg gtaaaaaatc
      181 tactgctgaa tctatcgtat acagcgcgct ggagaccctg gctcagcgct ctggtaaatc
      241 tgaactggaa gcattcgaag tagctctcga aaacgtgcgc ccgactgtag aagttaagtc
      301 tcgccgcgtt ggtggttgta cttatcaggt accagttgaa gtccgtccgg ttcgtcgtaa
      361 tgctctggca atgcgttgga tcgttgaagc tgctcgaaaa cgcggtgata aatccatggc
      421 tctgcgcctg gcgaacgaac tttctgatgc tgcagaaaac aaaggtactg cagttaagaa
      481 acgtgaagac gttcaccgta tggccgaagc caacaaggcg ttcgcacact accgttggtg
      541 attaacctag gtgctgccac cgctgagcaa taactagcat aaccccttgg ggcctctaaa
      601 cgggtcttga ggggtttttt gctgaaacct caggcatttg agaagcacac ggtcacactg
      661 cttccggtag tcaataaacc ggtaaaccag caatagacat aagcggctat ttaacgaccc
      721 tgccctgaac cgacgacaag ctgacgaccg ggtctccgca agtggcactt ttcggggaaa
      781 tgtgcgcgga acccctattt gtttattttt ctaaatacat tcaaatatgt atccgctcat
      841 gaattaattc ttagaaaaac tcatcgagca tcaaatgaaa ctgcaattta ttcatatcag
      901 gattatcaat accatatttt tgaaaaagcc gtttctgtaa tgaaggagaa aactcaccga
      961 ggcagttcca taggatggca agatcctggt atcggtctgc gattccgact cgtccaacat
     1021 caatacaacc tattaatttc ccctcgtcaa aaataaggtt atcaagtgag aaatcaccat
     1081 gagtgacgac tgaatccggt gagaatggca aaagtttatg catttctttc cagacttgtt
     1141 caacaggcca gccattacgc tcgtcatcaa aatcactcgc atcaaccaaa ccgttattca
     1201 ttcgtgattg cgcctgagcg agacgaaata cgcggtcgct gttaaaagga caattacaaa
     1261 caggaatcga atgcaaccgg cgcaggaaca ctgccagcgc atcaacaata ttttcacctg
     1321 aatcaggata ttcttctaat acctggaatg ctgttttccc ggggatcgca gtggtgagta
     1381 accatgcatc atcaggagta cggataaaat gcttgatggt cggaagaggc ataaattccg
     1441 tcagccagtt tagtctgacc atctcatctg taacatcatt ggcaacgcta cctttgccat
     1501 gtttcagaaa caactctggc gcatcgggct tcccatacaa tcgatagatt gtcgcacctg
     1561 attgcccgac attatcgcga gcccatttat acccatataa atcagcatcc atgttggaat
     1621 ttaatcgcgg cctagagcaa gacgtttccc gttgaatatg gctcatactc ttcctttttc
     1681 aatattattg aagcatttat cagggttatt gtctcatgag cggatacata tttgaatgta
     1741 tttagaaaaa taaacaaata ggcatgcagc gctcttccgc ttcctcgctc actgactcgc
     1801 tacgctcggt cgttcgactg cggcgagcgg tgtcagctca ctcaaaagcg gtaatacggt
     1861 tatccacaga atcaggggat aaagccggaa agaacatgtg agcaaaaagc aaagcaccgg
     1921 aagaagccaa cgccgcaggc gtttttccat aggctccgcc cccctgacga gcatcacaaa
     1981 aatcgacgct caagccagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt
     2041 ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg
     2101 tccgcctttc tcccttcggg aagcgtggcg ctttctcata gctcacgctg ttggtatctc
     2161 agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc
     2221 gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta
     2281 tcgccactgg cagcagccat tggtaactga tttagaggac tttgtcttga agttatgcac
     2341 ctgttaaggc taaactgaaa gaacagattt tggtgagtgc ggtcctccaa cccacttacc
     2401 ttggttcaaa gagttggtag ctcagcgaac cttgagaaaa ccaccgttgg tagcggtggt
     2461 ttttctttat ttatgagatg atgaatcaat cggtctatca agtcaacgaa cagctattcc
     2521 gttactctag atttcagtgc aatttatctc ttcaaatgta gcacctgaag tcagccccat
     2581 acgatataag ttgtaattct catgttagtc atgccccgcg cccaccggaa ggagctgact
     2641 gggttgaagg ctctcaaggg catcggtcga gatcccggtg cctaatgagt gagctaactt
     2701 acattaattg cgttgcgctc actgcccgct ttccagtcgg gaaacctgtc gtgccagctg
     2761 cattaatgaa tcggccaacg cgcggggaga ggcggtttgc gtattgggcg ccagggtggt
     2821 ttttcttttc accagtgaga cgggcaacag ctgattgccc ttcaccgcct ggccctgaga
     2881 gagttgcagc aagcggtcca cgctggtttg ccccagcagg cgaaaatcct gtttgatggt
     2941 ggttaacggc gggatataac atgagctgtc ttcggtatcg tcgtatccca ctaccgagat
     3001 gtccgcacca acgcgcagcc cggactcggt aatggcgcgc attgcgccca gcgccatctg
     3061 atcgttggca accagcatcg cagtgggaac gatgccctca ttcagcattt gcatggtttg
     3121 ttgaaaaccg gacatggcac tccagtcgcc ttcccgttcc gctatcggct gaatttgatt
     3181 gcgagtgaga tatttatgcc agccagccag acgcagacgc gccgagacag aacttaatgg
     3241 gcccgctaac agcgcgattt gctggtgacc caatgcgacc agatgctcca cgcccagtcg
     3301 cgtaccgtct tcatgggaga aaataatact gttgatgggt gtctggtcag agacatcaag
     3361 aaataacgcc ggaacattag tgcaggcagc ttccacagca atggcatcct ggtcatccag
     3421 cggatagtta atgatcagcc cactgacgcg ttgcgcgaga agattgtgca ccgccgcttt
     3481 acaggcttcg acgccgcttc gttctaccat cgacaccacc acgctggcac ccagttgatc
     3541 ggcgcgagat ttaatcgccg cgacaatttg cgacggcgcg tgcagggcca gactggaggt
     3601 ggcaacgcca atcagcaacg actgtttgcc cgccagttgt tgtgccacgc ggttgggaat
     3661 gtaattcagc tccgccatcg ccgcttccac tttttcccgc gttttcgcag aaacgtggct
     3721 ggcctggttc accacgcggg aaacggtctg ataagagaca ccggcatact ctgcgacatc
     3781 gtataacgtt actggtttca cattcaccac cctgaattga ctctcttccg ggcgctatca
     3841 tgccataccg cgaaaggttt tgcgccattc gatggtgtcc gggatctcga cgctctccct
     3901 tatgcgactc ctgcattagg aaattaatac gactcactat a
//
