LOCUS       sequence-263278-        3777 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses tagless 30S ribosomal subunit protein S14 in E.coli.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3777)
  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 3777)
  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-S14
FEATURES             Location/Qualifiers
     source          1..3777
                     /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(396..414)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      410..457
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     CDS             complement(687..1502)
                     /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(687..1502)
                     /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     1410..1429
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     promoter        complement(1503..1594)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(1610..2359)
                     /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(1620..1637)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     primer_bind     complement(1860..1879)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(2555..3637)
                     /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     3599..3618
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(3638..3715)
                     /gene="lacI"
                     /label=lacI promoter
     primer_bind     join(3761..3777,1..3)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        join(3761..3777,1..2)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
ORIGIN
        1 ggggaattgt gagcggataa caattcccct gtagaaataa ttttgtttaa ctttaataag
       61 gagatatacc atggctaagc aatcaatgaa agcacgcgaa gtaaaacgcg tagctttagc
      121 tgataaatac ttcgcgaaac gcgctgaact gaaagcgatc atctctgatg tgaacgcttc
      181 cgacgaagat cgttggaacg ctgttctcaa gctgcagact ctgccgcgtg attccagccc
      241 gtctcgtcag cgtaaccgct gccgtcaaac aggtcgtccg catggtttcc tgcggaagtt
      301 cgggttgagc cgtattaagg tccgtgaagc cgctatgcgc ggtgaaatcc cgggtctgaa
      361 aaaggctagc tggtaattaa cctaggctgc tgccaccgct gagcaataac tagcataacc
      421 ccttggggcc tctaaacggg tcttgagggg ttttttgctg aaacctcagg catttgagaa
      481 gcacacggtc acactgcttc cggtagtcaa taaaccggta aaccagcaat agacataagc
      541 ggctatttaa cgaccctgcc ctgaaccgac gacaagctga cgaccgggtc tccgcaagtg
      601 gcacttttcg gggaaatgtg cgcggaaccc ctatttgttt atttttctaa atacattcaa
      661 atatgtatcc gctcatgaat taattcttag aaaaactcat cgagcatcaa atgaaactgc
      721 aatttattca tatcaggatt atcaatacca tatttttgaa aaagccgttt ctgtaatgaa
      781 ggagaaaact caccgaggca gttccatagg atggcaagat cctggtatcg gtctgcgatt
      841 ccgactcgtc caacatcaat acaacctatt aatttcccct cgtcaaaaat aaggttatca
      901 agtgagaaat caccatgagt gacgactgaa tccggtgaga atggcaaaag tttatgcatt
      961 tctttccaga cttgttcaac aggccagcca ttacgctcgt catcaaaatc actcgcatca
     1021 accaaaccgt tattcattcg tgattgcgcc tgagcgagac gaaatacgcg gtcgctgtta
     1081 aaaggacaat tacaaacagg aatcgaatgc aaccggcgca ggaacactgc cagcgcatca
     1141 acaatatttt cacctgaatc aggatattct tctaatacct ggaatgctgt tttcccgggg
     1201 atcgcagtgg tgagtaacca tgcatcatca ggagtacgga taaaatgctt gatggtcgga
     1261 agaggcataa attccgtcag ccagtttagt ctgaccatct catctgtaac atcattggca
     1321 acgctacctt tgccatgttt cagaaacaac tctggcgcat cgggcttccc atacaatcga
     1381 tagattgtcg cacctgattg cccgacatta tcgcgagccc atttataccc atataaatca
     1441 gcatccatgt tggaatttaa tcgcggccta gagcaagacg tttcccgttg aatatggctc
     1501 atactcttcc tttttcaata ttattgaagc atttatcagg gttattgtct catgagcgga
     1561 tacatatttg aatgtattta gaaaaataaa caaataggca tgcagcgctc ttccgcttcc
     1621 tcgctcactg actcgctacg ctcggtcgtt cgactgcggc gagcggtgtc agctcactca
     1681 aaagcggtaa tacggttatc cacagaatca ggggataaag ccggaaagaa catgtgagca
     1741 aaaagcaaag caccggaaga agccaacgcc gcaggcgttt ttccataggc tccgcccccc
     1801 tgacgagcat cacaaaaatc gacgctcaag ccagaggtgg cgaaacccga caggactata
     1861 aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc
     1921 gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt ctcatagctc
     1981 acgctgttgg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga
     2041 accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc
     2101 ggtaagacac gacttatcgc cactggcagc agccattggt aactgattta gaggactttg
     2161 tcttgaagtt atgcacctgt taaggctaaa ctgaaagaac agattttggt gagtgcggtc
     2221 ctccaaccca cttaccttgg ttcaaagagt tggtagctca gcgaaccttg agaaaaccac
     2281 cgttggtagc ggtggttttt ctttatttat gagatgatga atcaatcggt ctatcaagtc
     2341 aacgaacagc tattccgtta ctctagattt cagtgcaatt tatctcttca aatgtagcac
     2401 ctgaagtcag ccccatacga tataagttgt aattctcatg ttagtcatgc cccgcgccca
     2461 ccggaaggag ctgactgggt tgaaggctct caagggcatc ggtcgagatc ccggtgccta
     2521 atgagtgagc taacttacat taattgcgtt gcgctcactg cccgctttcc agtcgggaaa
     2581 cctgtcgtgc cagctgcatt aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat
     2641 tgggcgccag ggtggttttt cttttcacca gtgagacggg caacagctga ttgcccttca
     2701 ccgcctggcc ctgagagagt tgcagcaagc ggtccacgct ggtttgcccc agcaggcgaa
     2761 aatcctgttt gatggtggtt aacggcggga tataacatga gctgtcttcg gtatcgtcgt
     2821 atcccactac cgagatgtcc gcaccaacgc gcagcccgga ctcggtaatg gcgcgcattg
     2881 cgcccagcgc catctgatcg ttggcaacca gcatcgcagt gggaacgatg ccctcattca
     2941 gcatttgcat ggtttgttga aaaccggaca tggcactcca gtcgccttcc cgttccgcta
     3001 tcggctgaat ttgattgcga gtgagatatt tatgccagcc agccagacgc agacgcgccg
     3061 agacagaact taatgggccc gctaacagcg cgatttgctg gtgacccaat gcgaccagat
     3121 gctccacgcc cagtcgcgta ccgtcttcat gggagaaaat aatactgttg atgggtgtct
     3181 ggtcagagac atcaagaaat aacgccggaa cattagtgca ggcagcttcc acagcaatgg
     3241 catcctggtc atccagcgga tagttaatga tcagcccact gacgcgttgc gcgagaagat
     3301 tgtgcaccgc cgctttacag gcttcgacgc cgcttcgttc taccatcgac accaccacgc
     3361 tggcacccag ttgatcggcg cgagatttaa tcgccgcgac aatttgcgac ggcgcgtgca
     3421 gggccagact ggaggtggca acgccaatca gcaacgactg tttgcccgcc agttgttgtg
     3481 ccacgcggtt gggaatgtaa ttcagctccg ccatcgccgc ttccactttt tcccgcgttt
     3541 tcgcagaaac gtggctggcc tggttcacca cgcgggaaac ggtctgataa gagacaccgg
     3601 catactctgc gacatcgtat aacgttactg gtttcacatt caccaccctg aattgactct
     3661 cttccgggcg ctatcatgcc ataccgcgaa aggttttgcg ccattcgatg gtgtccggga
     3721 tctcgacgct ctcccttatg cgactcctgc attaggaaat taatacgact cactata
//
