LOCUS       sequence-252258-        5790 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Production of recombinant human ribosomal protein S15.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5790)
  AUTHORS   Correddu D, Montano Lopez JJ, Vadakkedath PG, Lai A, Pernes JI, 
            Watson PR, Leung IKH
  TITLE     An improved method for the heterologous production of soluble human 
            ribosomal proteins in Escherichia coli.
  JOURNAL   Sci Rep. 2019 Jun 20;9(1):8884. doi: 10.1038/s41598-019-45323-8.
  PUBMED    31222068
REFERENCE   2  (bases 1 to 5790)
  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"; journalName: "Sci Rep."; 
            date: "2019-06-20"; volume: "9(1):8884. doi"; pages: " 
            10.1038/s41598-019-45323-8"
COMMENT     Sequence Label: pNIC28-BSA4-S15
FEATURES             Location/Qualifiers
     source          1..5790
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     1..20
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1..19
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    20..44
                     /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)."
     RBS             59..81
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             92..109
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             134..154
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQS"
     CDS             655..672
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(725..743)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      739..786
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      823..1278
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(910..929)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1120..1141
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(1370..2185)
                     /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(1370..2185)
                     /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     2093..2112
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     2177..2196
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     rep_origin      2307..2895
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2796..2815
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3049..3066
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    3081..3220
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(3146..3165)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3306..3328
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(3322..3513)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(4322..5404)
                     /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     5366..5385
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(5405..5482)
                     /gene="lacI"
                     /label=lacI promoter
     primer_bind     5688..5707
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
ORIGIN
        1 taatacgact cactataggg gaattgtgag cggataacaa ttcccctcta gaaataattt
       61 tgtttaactt taagaaggag atatacatat gcaccatcat catcatcatt cttctggtgt
      121 agatctgggt accgagaacc tgtacttcca atccatggca gaagtagagc agaagaagaa
      181 gcggaccttc cgcaagttca cctaccgcgg cgtggacctc gaccagctgc tggacatgtc
      241 ctacgagcag ctgatgcagc tgtacagtgc gcgccagcgg cggcggctga accggggcct
      301 gcggcggaag cagcactccc tgctgaagcg cctgcgcaag gccaagaagg aggcgccgcc
      361 catggagaag ccggaagtgg tgaagacgca cctgcgggac atgatcatcc tacccgagat
      421 ggtgggcagc atggtgggcg tctacaacgg caagaccttc aaccaggtgg agatcaagcc
      481 cgagatgatc ggccactacc tgggcgagtt ctccatcacc tacaagcccg taaagcatgg
      541 ccggcccggc atcggggcca cccactcctc ccgcttcatc cctctcaagt aacagtaaag
      601 gtggatacgg atccgaattc gagctccgtc gacaagcttg cggccgcact cgagcaccac
      661 caccaccacc actgagatcc ggctgctaac aaagcccgaa aggaagctga gttggctgct
      721 gccaccgctg agcaataact agcataaccc cttggggcct ctaaacgggt cttgaggggt
      781 tttttgctga aaggaggaac tatatccgga ttggcgaatg ggacgcgccc tgtagcggcg
      841 cattaagcgc ggcgggtgtg gtggttacgc gcagcgtgac cgctacactt gccagcgccc
      901 tagcgcccgc tcctttcgct ttcttccctt cctttctcgc cacgttcgcc ggctttcccc
      961 gtcaagctct aaatcggggg ctccctttag ggttccgatt tagtgcttta cggcacctcg
     1021 accccaaaaa acttgattag ggtgatggtt cacgtagtgg gccatcgccc tgatagacgg
     1081 tttttcgccc tttgacgttg gagtccacgt tctttaatag tggactcttg ttccaaactg
     1141 gaacaacact caaccctatc tcggtctatt cttttgattt ataagggatt ttgccgattt
     1201 cggcctattg gttaaaaaat gagctgattt aacaaaaatt taacgcgaat tttaacaaaa
     1261 tattaacgct tacaatttag gtggcacttt tcggggaaat gtgcgcggaa cccctatttg
     1321 tttatttttc taaatacatt caaatatgta tccgctcatg aattaattct tagaaaaact
     1381 catcgagcat caaatgaaac tgcaatttat tcatatcagg attatcaata ccatattttt
     1441 gaaaaagccg tttctgtaat gaaggagaaa actcaccgag gcagttccat aggatggcaa
     1501 gatcctggta tcggtctgcg attccgactc gtccaacatc aatacaacct attaatttcc
     1561 cctcgtcaaa aataaggtta tcaagtgaga aatcaccatg agtgacgact gaatccggtg
     1621 agaatggcaa aagtttatgc atttctttcc agacttgttc aacaggccag ccattacgct
     1681 cgtcatcaaa atcactcgca tcaaccaaac cgttattcat tcgtgattgc gcctgagcga
     1741 gacgaaatac gcgatcgctg ttaaaaggac aattacaaac aggaatcgaa tgcaaccggc
     1801 gcaggaacac tgccagcgca tcaacaatat tttcacctga atcaggatat tcttctaata
     1861 cctggaatgc tgttttcccg gggatcgcag tggtgagtaa ccatgcatca tcaggagtac
     1921 ggataaaatg cttgatggtc ggaagaggca taaattccgt cagccagttt agtctgacca
     1981 tctcatctgt aacatcattg gcaacgctac ctttgccatg tttcagaaac aactctggcg
     2041 catcgggctt cccatacaat cgatagattg tcgcacctga ttgcccgaca ttatcgcgag
     2101 cccatttata cccatataaa tcagcatcca tgttggaatt taatcgcggc ctagagcaag
     2161 acgtttcccg ttgaatatgg ctcataacac cccttgtatt actgtttatg taagcagaca
     2221 gttttattgt tcatgaccaa aatcccttaa cgtgagtttt cgttccactg agcgtcagac
     2281 cccgtagaaa agatcaaagg atcttcttga gatccttttt ttctgcgcgt aatctgctgc
     2341 ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt tgccggatca agagctacca
     2401 actctttttc cgaaggtaac tggcttcagc agagcgcaga taccaaatac tgtccttcta
     2461 gtgtagccgt agttaggcca ccacttcaag aactctgtag caccgcctac atacctcgct
     2521 ctgctaatcc tgttaccagt ggctgctgcc agtggcgata agtcgtgtct taccgggttg
     2581 gactcaagac gatagttacc ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc
     2641 acacagccca gcttggagcg aacgacctac accgaactga gatacctaca gcgtgagcta
     2701 tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca ggtatccggt aagcggcagg
     2761 gtcggaacag gagagcgcac gagggagctt ccagggggaa acgcctggta tctttatagt
     2821 cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg
     2881 cggagcctat ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc cttttgctgg
     2941 ccttttgctc acatgttctt tcctgcgtta tcccctgatt ctgtggataa ccgtattacc
     3001 gcctttgagt gagctgatac cgctcgccgc agccgaacga ccgagcgcag cgagtcagtg
     3061 agcgaggaag cggaagagcg cctgatgcgg tattttctcc ttacgcatct gtgcggtatt
     3121 tcacaccgca atggtgcact ctcagtacaa tctgctctga tgccgcatag ttaagccagt
     3181 atacactccg ctatcgctac gtgactgggt catggctgcg ccccgacacc cgccaacacc
     3241 cgctgacgcg ccctgacggg cttgtctgct cccggcatcc gcttacagac aagctgtgac
     3301 cgtctccggg agctgcatgt gtcagaggtt ttcaccgtca tcaccgaaac gcgcgaggca
     3361 gctgcggtaa agctcatcag cgtggtcgtg aagcgattca cagatgtctg cctgttcatc
     3421 cgcgtccagc tcgttgagtt tctccagaag cgttaatgtc tggcttctga taaagcgggc
     3481 catgttaagg gcggtttttt cctgtttggt cactgatgcc tccgtgtaag ggggatttct
     3541 gttcatgggg gtaatgatac cgatgaaacg agagaggatg ctcacgatac gggttactga
     3601 tgatgaacat gcccggttac tggaacgttg tgagggtaaa caactggcgg tatggatgcg
     3661 gcgggaccag agaaaaatca ctcagggtca atgccagcgc ttcgttaata cagatgtagg
     3721 tgttccacag ggtagccagc agcatcctgc gatgcagatc cggaacataa tggtgcaggg
     3781 cgctgacttc cgcgtttcca gactttacga aacacggaaa ccgaagacca ttcatgttgt
     3841 tgctcaggtc gcagacgttt tgcagcagca gtcgcttcac gttcgctcgc gtatcggtga
     3901 ttcattctgc taaccagtaa ggcaaccccg ccagcctagc cgggtcctca acgacaggag
     3961 cacgatcatg cgcacccgtg gggccgccat gccggcgata atggcctgct tctcgccgaa
     4021 acgtttggtg gcgggaccag tgacgaaggc ttgagcgagg gcgtgcaaga ttccgaatac
     4081 cgcaagcgac aggccgatca tcgtcgcgct ccagcgaaag cggtcctcgc cgaaaatgac
     4141 ccagagcgct gccggcacct gtcctacgag ttgcatgata aagaagacag tcataagtgc
     4201 ggcgacgata gtcatgcccc gcgcccaccg gaaggagctg actgggttga aggctctcaa
     4261 gggcatcggt cgagatcccg gtgcctaatg agtgagctaa cttacattaa ttgcgttgcg
     4321 ctcactgccc gctttccagt cgggaaacct gtcgtgccag ctgcattaat gaatcggcca
     4381 acgcgcgggg agaggcggtt tgcgtattgg gcgccagggt ggtttttctt ttcaccagtg
     4441 agacgggcaa cagctgattg cccttcaccg cctggccctg agagagttgc agcaagcggt
     4501 ccacgctggt ttgccccagc aggcgaaaat cctgtttgat ggtggttaac ggcgggatat
     4561 aacatgagct gtcttcggta tcgtcgtatc ccactaccga gatatccgca ccaacgcgca
     4621 gcccggactc ggtaatggcg cgcattgcgc ccagcgccat ctgatcgttg gcaaccagca
     4681 tcgcagtggg aacgatgccc tcattcagca tttgcatggt ttgttgaaaa ccggacatgg
     4741 cactccagtc gccttcccgt tccgctatcg gctgaatttg attgcgagtg agatatttat
     4801 gccagccagc cagacgcaga cgcgccgaga cagaacttaa tgggcccgct aacagcgcga
     4861 tttgctggtg acccaatgcg accagatgct ccacgcccag tcgcgtaccg tcttcatggg
     4921 agaaaataat actgttgatg ggtgtctggt cagagacatc aagaaataac gccggaacat
     4981 tagtgcaggc agcttccaca gcaatggcat cctggtcatc cagcggatag ttaatgatca
     5041 gcccactgac gcgttgcgcg agaagattgt gcaccgccgc tttacaggct tcgacgccgc
     5101 ttcgttctac catcgacacc accacgctgg cacccagttg atcggcgcga gatttaatcg
     5161 ccgcgacaat ttgcgacggc gcgtgcaggg ccagactgga ggtggcaacg ccaatcagca
     5221 acgactgttt gcccgccagt tgttgtgcca cgcggttggg aatgtaattc agctccgcca
     5281 tcgccgcttc cactttttcc cgcgttttcg cagaaacgtg gctggcctgg ttcaccacgc
     5341 gggaaacggt ctgataagag acaccggcat actctgcgac atcgtataac gttactggtt
     5401 tcacattcac caccctgaat tgactctctt ccgggcgcta tcatgccata ccgcgaaagg
     5461 ttttgcgcca ttcgatggtg tccgggatct cgacgctctc ccttatgcga ctcctgcatt
     5521 aggaagcagc ccagtagtag gttgaggccg ttgagcaccg ccgccgcaag gaatggtgca
     5581 tgcaaggaga tggcgcccaa cagtcccccg gccacggggc ctgccaccat acccacgccg
     5641 aaacaagcgc tcatgagccc gaagtggcga gcccgatctt ccccatcggt gatgtcggcg
     5701 atataggcgc cagcaaccgc acctgtggcg ccggtgatgc cggccacgat gcgtccggcg
     5761 tagaggatcg agatctcgat cccgcgaaat
//
