LOCUS       sequence-252256-        6117 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 6117)
  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 6117)
  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: pNH-TrxT-G-S15
FEATURES             Location/Qualifiers
     source          1..6117
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    1..25
                     /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             40..62
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             73..90
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             100..426
                     /codon_start=1
                     /gene="trxA"
                     /product="E. coli thioredoxin"
                     /label=TrxA
                     /translation="MSDKIIHLTDDSFDTDVLKADGAILVDFWAEWCGPCKMIAPILDE
                     IADEYQGKLTVAKLNIDQNPGTAPKYGIRGIPTLLLFKNGEVAATKVGALSKGQLKEFL
                     DANLA"
     CDS             433..453
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQS"
     CDS             963..980
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(1033..1051)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      1047..1094
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      1131..1586
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1218..1237)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1428..1449
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(1678..2493)
                     /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(1678..2493)
                     /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     2401..2420
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     2485..2504
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     rep_origin      2615..3203
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3104..3123
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3357..3374
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    3389..3528
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(3454..3473)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3614..3636
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(3630..3821)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(4630..5712)
                     /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     5674..5693
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(5713..5790)
                     /gene="lacI"
                     /label=lacI promoter
     primer_bind     5996..6015
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     join(6099..6117,1)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        6099..6117
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
ORIGIN
        1 ggaattgtga gcggataaca attcccctct agaaataatt ttgtttaact ttaagaagga
       61 gatatacata tgcaccatca tcatcatcat tcttctggta tgagcgataa aattattcac
      121 ctgactgacg acagttttga cacggatgta ctcaaagcgg acggggcgat cctcgtcgat
      181 ttctgggcag agtggtgcgg tccgtgcaaa atgatcgccc cgattctgga tgaaatcgct
      241 gacgaatatc agggcaaact gaccgttgca aaactgaaca tcgatcaaaa ccctggcact
      301 gcgccgaaat atggcatccg tggtatcccg actctgctgc tgttcaaaaa cggtgaagtg
      361 gcggcaacca aagtgggcgc actgtctaaa ggtcagttga aagagttcct cgacgctaac
      421 ctggccggta ccgagaactt gtacttccaa tccggcggcg gcatggcaga agtagagcag
      481 aagaagaagc ggaccttccg caagttcacc taccgcggcg tggacctcga ccagctgctg
      541 gacatgtcct acgagcagct gatgcagctg tacagtgcgc gccagcggcg gcggctgaac
      601 cggggcctgc ggcggaagca gcactccctg ctgaagcgcc tgcgcaaggc caagaaggag
      661 gcgccgccca tggagaagcc ggaagtggtg aagacgcacc tgcgggacat gatcatccta
      721 cccgagatgg tgggcagcat ggtgggcgtc tacaacggca agaccttcaa ccaggtggag
      781 atcaagcccg agatgatcgg ccactacctg ggcgagttct ccatcaccta caagcccgta
      841 aagcatggcc ggcccggcat cggggccacc cactcctccc gcttcatccc tctcaagtaa
      901 cagtaaaggt ggatacggat ccgaattcga gctccgtcga caagcttgcg gccgcactcg
      961 agcaccacca ccaccaccac tgagatccgg ctgctaacaa agcccgaaag gaagctgagt
     1021 tggctgctgc caccgctgag caataactag cataacccct tggggcctct aaacgggtct
     1081 tgaggggttt tttgctgaaa ggaggaacta tatccggatt ggcgaatggg acgcgccctg
     1141 tagcggcgca ttaagcgcgg cgggtgtggt ggttacgcgc agcgtgaccg ctacacttgc
     1201 cagcgcccta gcgcccgctc ctttcgcttt cttcccttcc tttctcgcca cgttcgccgg
     1261 ctttccccgt caagctctaa atcgggggct ccctttaggg ttccgattta gtgctttacg
     1321 gcacctcgac cccaaaaaac ttgattaggg tgatggttca cgtagtgggc catcgccctg
     1381 atagacggtt tttcgccctt tgacgttgga gtccacgttc tttaatagtg gactcttgtt
     1441 ccaaactgga acaacactca accctatctc ggtctattct tttgatttat aagggatttt
     1501 gccgatttcg gcctattggt taaaaaatga gctgatttaa caaaaattta acgcgaattt
     1561 taacaaaata ttaacgctta caatttaggt ggcacttttc ggggaaatgt gcgcggaacc
     1621 cctatttgtt tatttttcta aatacattca aatatgtatc cgctcatgaa ttaattctta
     1681 gaaaaactca tcgagcatca aatgaaactg caatttattc atatcaggat tatcaatacc
     1741 atatttttga aaaagccgtt tctgtaatga aggagaaaac tcaccgaggc agttccatag
     1801 gatggcaaga tcctggtatc ggtctgcgat tccgactcgt ccaacatcaa tacaacctat
     1861 taatttcccc tcgtcaaaaa taaggttatc aagtgagaaa tcaccatgag tgacgactga
     1921 atccggtgag aatggcaaaa gtttatgcat ttctttccag acttgttcaa caggccagcc
     1981 attacgctcg tcatcaaaat cactcgcatc aaccaaaccg ttattcattc gtgattgcgc
     2041 ctgagcgaga cgaaatacgc gatcgctgtt aaaaggacaa ttacaaacag gaatcgaatg
     2101 caaccggcgc aggaacactg ccagcgcatc aacaatattt tcacctgaat caggatattc
     2161 ttctaatacc tggaatgctg ttttcccggg gatcgcagtg gtgagtaacc atgcatcatc
     2221 aggagtacgg ataaaatgct tgatggtcgg aagaggcata aattccgtca gccagtttag
     2281 tctgaccatc tcatctgtaa catcattggc aacgctacct ttgccatgtt tcagaaacaa
     2341 ctctggcgca tcgggcttcc catacaatcg atagattgtc gcacctgatt gcccgacatt
     2401 atcgcgagcc catttatacc catataaatc agcatccatg ttggaattta atcgcggcct
     2461 agagcaagac gtttcccgtt gaatatggct cataacaccc cttgtattac tgtttatgta
     2521 agcagacagt tttattgttc atgaccaaaa tcccttaacg tgagttttcg ttccactgag
     2581 cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa
     2641 tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg ccggatcaag
     2701 agctaccaac tctttttccg aaggtaactg gcttcagcag agcgcagata ccaaatactg
     2761 tccttctagt gtagccgtag ttaggccacc acttcaagaa ctctgtagca ccgcctacat
     2821 acctcgctct gctaatcctg ttaccagtgg ctgctgccag tggcgataag tcgtgtctta
     2881 ccgggttgga ctcaagacga tagttaccgg ataaggcgca gcggtcgggc tgaacggggg
     2941 gttcgtgcac acagcccagc ttggagcgaa cgacctacac cgaactgaga tacctacagc
     3001 gtgagctatg agaaagcgcc acgcttcccg aagggagaaa ggcggacagg tatccggtaa
     3061 gcggcagggt cggaacagga gagcgcacga gggagcttcc agggggaaac gcctggtatc
     3121 tttatagtcc tgtcgggttt cgccacctct gacttgagcg tcgatttttg tgatgctcgt
     3181 caggggggcg gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct
     3241 tttgctggcc ttttgctcac atgttctttc ctgcgttatc ccctgattct gtggataacc
     3301 gtattaccgc ctttgagtga gctgataccg ctcgccgcag ccgaacgacc gagcgcagcg
     3361 agtcagtgag cgaggaagcg gaagagcgcc tgatgcggta ttttctcctt acgcatctgt
     3421 gcggtatttc acaccgcaat ggtgcactct cagtacaatc tgctctgatg ccgcatagtt
     3481 aagccagtat acactccgct atcgctacgt gactgggtca tggctgcgcc ccgacacccg
     3541 ccaacacccg ctgacgcgcc ctgacgggct tgtctgctcc cggcatccgc ttacagacaa
     3601 gctgtgaccg tctccgggag ctgcatgtgt cagaggtttt caccgtcatc accgaaacgc
     3661 gcgaggcagc tgcggtaaag ctcatcagcg tggtcgtgaa gcgattcaca gatgtctgcc
     3721 tgttcatccg cgtccagctc gttgagtttc tccagaagcg ttaatgtctg gcttctgata
     3781 aagcgggcca tgttaagggc ggttttttcc tgtttggtca ctgatgcctc cgtgtaaggg
     3841 ggatttctgt tcatgggggt aatgataccg atgaaacgag agaggatgct cacgatacgg
     3901 gttactgatg atgaacatgc ccggttactg gaacgttgtg agggtaaaca actggcggta
     3961 tggatgcggc gggaccagag aaaaatcact cagggtcaat gccagcgctt cgttaataca
     4021 gatgtaggtg ttccacaggg tagccagcag catcctgcga tgcagatccg gaacataatg
     4081 gtgcagggcg ctgacttccg cgtttccaga ctttacgaaa cacggaaacc gaagaccatt
     4141 catgttgttg ctcaggtcgc agacgttttg cagcagcagt cgcttcacgt tcgctcgcgt
     4201 atcggtgatt cattctgcta accagtaagg caaccccgcc agcctagccg ggtcctcaac
     4261 gacaggagca cgatcatgcg cacccgtggg gccgccatgc cggcgataat ggcctgcttc
     4321 tcgccgaaac gtttggtggc gggaccagtg acgaaggctt gagcgagggc gtgcaagatt
     4381 ccgaataccg caagcgacag gccgatcatc gtcgcgctcc agcgaaagcg gtcctcgccg
     4441 aaaatgaccc agagcgctgc cggcacctgt cctacgagtt gcatgataaa gaagacagtc
     4501 ataagtgcgg cgacgatagt catgccccgc gcccaccgga aggagctgac tgggttgaag
     4561 gctctcaagg gcatcggtcg agatcccggt gcctaatgag tgagctaact tacattaatt
     4621 gcgttgcgct cactgcccgc tttccagtcg ggaaacctgt cgtgccagct gcattaatga
     4681 atcggccaac gcgcggggag aggcggtttg cgtattgggc gccagggtgg tttttctttt
     4741 caccagtgag acgggcaaca gctgattgcc cttcaccgcc tggccctgag agagttgcag
     4801 caagcggtcc acgctggttt gccccagcag gcgaaaatcc tgtttgatgg tggttaacgg
     4861 cgggatataa catgagctgt cttcggtatc gtcgtatccc actaccgaga tatccgcacc
     4921 aacgcgcagc ccggactcgg taatggcgcg cattgcgccc agcgccatct gatcgttggc
     4981 aaccagcatc gcagtgggaa cgatgccctc attcagcatt tgcatggttt gttgaaaacc
     5041 ggacatggca ctccagtcgc cttcccgttc cgctatcggc tgaatttgat tgcgagtgag
     5101 atatttatgc cagccagcca gacgcagacg cgccgagaca gaacttaatg ggcccgctaa
     5161 cagcgcgatt tgctggtgac ccaatgcgac cagatgctcc acgcccagtc gcgtaccgtc
     5221 ttcatgggag aaaataatac tgttgatggg tgtctggtca gagacatcaa gaaataacgc
     5281 cggaacatta gtgcaggcag cttccacagc aatggcatcc tggtcatcca gcggatagtt
     5341 aatgatcagc ccactgacgc gttgcgcgag aagattgtgc accgccgctt tacaggcttc
     5401 gacgccgctt cgttctacca tcgacaccac cacgctggca cccagttgat cggcgcgaga
     5461 tttaatcgcc gcgacaattt gcgacggcgc gtgcagggcc agactggagg tggcaacgcc
     5521 aatcagcaac gactgtttgc ccgccagttg ttgtgccacg cggttgggaa tgtaattcag
     5581 ctccgccatc gccgcttcca ctttttcccg cgttttcgca gaaacgtggc tggcctggtt
     5641 caccacgcgg gaaacggtct gataagagac accggcatac tctgcgacat cgtataacgt
     5701 tactggtttc acattcacca ccctgaattg actctcttcc gggcgctatc atgccatacc
     5761 gcgaaaggtt ttgcgccatt cgatggtgtc cgggatctcg acgctctccc ttatgcgact
     5821 cctgcattag gaagcagccc agtagtaggt tgaggccgtt gagcaccgcc gccgcaagga
     5881 atggtgcatg caaggagatg gcgcccaaca gtcccccggc cacggggcct gccaccatac
     5941 ccacgccgaa acaagcgctc atgagcccga agtggcgagc ccgatcttcc ccatcggtga
     6001 tgtcggcgat ataggcgcca gcaaccgcac ctgtggcgcc ggtgatgccg gccacgatgc
     6061 gtccggcgta gaggatcgag atctcgatcc cgcgaaatta atacgactca ctatagg
//
