LOCUS       sequence-250144-        6216 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Production of recombinant human ribosomal protein L11.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6216)
  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 6216)
  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-L11
FEATURES             Location/Qualifiers
     source          1..6216
                     /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             119..445
                     /codon_start=1
                     /gene="trxA"
                     /product="E. coli thioredoxin"
                     /label=TrxA
                     /translation="MSDKIIHLTDDSFDTDVLKADGAILVDFWAEWCGPCKMIAPILDE
                     IADEYQGKLTVAKLNIDQNPGTAPKYGIRGIPTLLLFKNGEVAATKVGALSKGQLKEFL
                     DANLA"
     CDS             452..472
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQS"
     CDS             1081..1098
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(1151..1169)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      1165..1212
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      1249..1704
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1336..1355)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1546..1567
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(1796..2611)
                     /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(1796..2611)
                     /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     2519..2538
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     2603..2622
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     rep_origin      2733..3321
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3222..3241
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3475..3492
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    3507..3646
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(3572..3591)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3732..3754
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(3748..3939)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(4748..5830)
                     /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     5792..5811
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(5831..5908)
                     /gene="lacI"
                     /label=lacI promoter
     primer_bind     6114..6133
                     /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 cttctggtat
      121 gagcgataaa attattcacc tgactgacga cagttttgac acggatgtac tcaaagcgga
      181 cggggcgatc ctcgtcgatt tctgggcaga gtggtgcggt ccgtgcaaaa tgatcgcccc
      241 gattctggat gaaatcgctg acgaatatca gggcaaactg accgttgcaa aactgaacat
      301 cgatcaaaac cctggcactg cgccgaaata tggcatccgt ggtatcccga ctctgctgct
      361 gttcaaaaac ggtgaagtgg cggcaaccaa agtgggcgca ctgtctaaag gtcagttgaa
      421 agagttcctc gacgctaacc tggccggtac cgagaacttg tacttccaat ccggcggcgg
      481 catggcgcag gatcaaggtg aaaaggagaa ccccatgcgg gaacttcgca tccgcaaact
      541 ctgtctcaac atctgtgttg gggagagtgg agacagactg acgcgagcag ccaaggtgtt
      601 ggagcagctc acagggcaga cccctgtgtt ttccaaagct agatacactg tcagatcctt
      661 tggcatccgg agaaatgaaa agattgctgt ccactgcaca gttcgagggg ccaaggcaga
      721 agaaatcttg gagaagggtc taaaggtgcg ggagtatgag ttaagaaaaa acaacttctc
      781 agatactgga aactttggtt ttgggatcca ggaacacatc gatctgggta tcaaatatga
      841 cccaagcatt ggtatctacg gcctggactt ctatgtggtg ctgggtaggc caggtttcag
      901 catcgcagac aagaagcgca ggacaggctg cattggggcc aaacacagaa tcagcaaaga
      961 ggaggccatg cgctggttcc agcagaagta tgatgggatc atccttcctg gcaaataaca
     1021 gtaaaggtgg atacggatcc gaattcgagc tccgtcgaca agcttgcggc cgcactcgag
     1081 caccaccacc accaccactg agatccggct gctaacaaag cccgaaagga agctgagttg
     1141 gctgctgcca ccgctgagca ataactagca taaccccttg gggcctctaa acgggtcttg
     1201 aggggttttt tgctgaaagg aggaactata tccggattgg cgaatgggac gcgccctgta
     1261 gcggcgcatt aagcgcggcg ggtgtggtgg ttacgcgcag cgtgaccgct acacttgcca
     1321 gcgccctagc gcccgctcct ttcgctttct tcccttcctt tctcgccacg ttcgccggct
     1381 ttccccgtca agctctaaat cgggggctcc ctttagggtt ccgatttagt gctttacggc
     1441 acctcgaccc caaaaaactt gattagggtg atggttcacg tagtgggcca tcgccctgat
     1501 agacggtttt tcgccctttg acgttggagt ccacgttctt taatagtgga ctcttgttcc
     1561 aaactggaac aacactcaac cctatctcgg tctattcttt tgatttataa gggattttgc
     1621 cgatttcggc ctattggtta aaaaatgagc tgatttaaca aaaatttaac gcgaatttta
     1681 acaaaatatt aacgcttaca atttaggtgg cacttttcgg ggaaatgtgc gcggaacccc
     1741 tatttgttta tttttctaaa tacattcaaa tatgtatccg ctcatgaatt aattcttaga
     1801 aaaactcatc gagcatcaaa tgaaactgca atttattcat atcaggatta tcaataccat
     1861 atttttgaaa aagccgtttc tgtaatgaag gagaaaactc accgaggcag ttccatagga
     1921 tggcaagatc ctggtatcgg tctgcgattc cgactcgtcc aacatcaata caacctatta
     1981 atttcccctc gtcaaaaata aggttatcaa gtgagaaatc accatgagtg acgactgaat
     2041 ccggtgagaa tggcaaaagt ttatgcattt ctttccagac ttgttcaaca ggccagccat
     2101 tacgctcgtc atcaaaatca ctcgcatcaa ccaaaccgtt attcattcgt gattgcgcct
     2161 gagcgagacg aaatacgcga tcgctgttaa aaggacaatt acaaacagga atcgaatgca
     2221 accggcgcag gaacactgcc agcgcatcaa caatattttc acctgaatca ggatattctt
     2281 ctaatacctg gaatgctgtt ttcccgggga tcgcagtggt gagtaaccat gcatcatcag
     2341 gagtacggat aaaatgcttg atggtcggaa gaggcataaa ttccgtcagc cagtttagtc
     2401 tgaccatctc atctgtaaca tcattggcaa cgctaccttt gccatgtttc agaaacaact
     2461 ctggcgcatc gggcttccca tacaatcgat agattgtcgc acctgattgc ccgacattat
     2521 cgcgagccca tttataccca tataaatcag catccatgtt ggaatttaat cgcggcctag
     2581 agcaagacgt ttcccgttga atatggctca taacacccct tgtattactg tttatgtaag
     2641 cagacagttt tattgttcat gaccaaaatc ccttaacgtg agttttcgtt ccactgagcg
     2701 tcagaccccg tagaaaagat caaaggatct tcttgagatc ctttttttct gcgcgtaatc
     2761 tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc ggatcaagag
     2821 ctaccaactc tttttccgaa ggtaactggc ttcagcagag cgcagatacc aaatactgtc
     2881 cttctagtgt agccgtagtt aggccaccac ttcaagaact ctgtagcacc gcctacatac
     2941 ctcgctctgc taatcctgtt accagtggct gctgccagtg gcgataagtc gtgtcttacc
     3001 gggttggact caagacgata gttaccggat aaggcgcagc ggtcgggctg aacggggggt
     3061 tcgtgcacac agcccagctt ggagcgaacg acctacaccg aactgagata cctacagcgt
     3121 gagctatgag aaagcgccac gcttcccgaa gggagaaagg cggacaggta tccggtaagc
     3181 ggcagggtcg gaacaggaga gcgcacgagg gagcttccag ggggaaacgc ctggtatctt
     3241 tatagtcctg tcgggtttcg ccacctctga cttgagcgtc gatttttgtg atgctcgtca
     3301 ggggggcgga gcctatggaa aaacgccagc aacgcggcct ttttacggtt cctggccttt
     3361 tgctggcctt ttgctcacat gttctttcct gcgttatccc ctgattctgt ggataaccgt
     3421 attaccgcct ttgagtgagc tgataccgct cgccgcagcc gaacgaccga gcgcagcgag
     3481 tcagtgagcg aggaagcgga agagcgcctg atgcggtatt ttctccttac gcatctgtgc
     3541 ggtatttcac accgcaatgg tgcactctca gtacaatctg ctctgatgcc gcatagttaa
     3601 gccagtatac actccgctat cgctacgtga ctgggtcatg gctgcgcccc gacacccgcc
     3661 aacacccgct gacgcgccct gacgggcttg tctgctcccg gcatccgctt acagacaagc
     3721 tgtgaccgtc tccgggagct gcatgtgtca gaggttttca ccgtcatcac cgaaacgcgc
     3781 gaggcagctg cggtaaagct catcagcgtg gtcgtgaagc gattcacaga tgtctgcctg
     3841 ttcatccgcg tccagctcgt tgagtttctc cagaagcgtt aatgtctggc ttctgataaa
     3901 gcgggccatg ttaagggcgg ttttttcctg tttggtcact gatgcctccg tgtaaggggg
     3961 atttctgttc atgggggtaa tgataccgat gaaacgagag aggatgctca cgatacgggt
     4021 tactgatgat gaacatgccc ggttactgga acgttgtgag ggtaaacaac tggcggtatg
     4081 gatgcggcgg gaccagagaa aaatcactca gggtcaatgc cagcgcttcg ttaatacaga
     4141 tgtaggtgtt ccacagggta gccagcagca tcctgcgatg cagatccgga acataatggt
     4201 gcagggcgct gacttccgcg tttccagact ttacgaaaca cggaaaccga agaccattca
     4261 tgttgttgct caggtcgcag acgttttgca gcagcagtcg cttcacgttc gctcgcgtat
     4321 cggtgattca ttctgctaac cagtaaggca accccgccag cctagccggg tcctcaacga
     4381 caggagcacg atcatgcgca cccgtggggc cgccatgccg gcgataatgg cctgcttctc
     4441 gccgaaacgt ttggtggcgg gaccagtgac gaaggcttga gcgagggcgt gcaagattcc
     4501 gaataccgca agcgacaggc cgatcatcgt cgcgctccag cgaaagcggt cctcgccgaa
     4561 aatgacccag agcgctgccg gcacctgtcc tacgagttgc atgataaaga agacagtcat
     4621 aagtgcggcg acgatagtca tgccccgcgc ccaccggaag gagctgactg ggttgaaggc
     4681 tctcaagggc atcggtcgag atcccggtgc ctaatgagtg agctaactta cattaattgc
     4741 gttgcgctca ctgcccgctt tccagtcggg aaacctgtcg tgccagctgc attaatgaat
     4801 cggccaacgc gcggggagag gcggtttgcg tattgggcgc cagggtggtt tttcttttca
     4861 ccagtgagac gggcaacagc tgattgccct tcaccgcctg gccctgagag agttgcagca
     4921 agcggtccac gctggtttgc cccagcaggc gaaaatcctg tttgatggtg gttaacggcg
     4981 ggatataaca tgagctgtct tcggtatcgt cgtatcccac taccgagata tccgcaccaa
     5041 cgcgcagccc ggactcggta atggcgcgca ttgcgcccag cgccatctga tcgttggcaa
     5101 ccagcatcgc agtgggaacg atgccctcat tcagcatttg catggtttgt tgaaaaccgg
     5161 acatggcact ccagtcgcct tcccgttccg ctatcggctg aatttgattg cgagtgagat
     5221 atttatgcca gccagccaga cgcagacgcg ccgagacaga acttaatggg cccgctaaca
     5281 gcgcgatttg ctggtgaccc aatgcgacca gatgctccac gcccagtcgc gtaccgtctt
     5341 catgggagaa aataatactg ttgatgggtg tctggtcaga gacatcaaga aataacgccg
     5401 gaacattagt gcaggcagct tccacagcaa tggcatcctg gtcatccagc ggatagttaa
     5461 tgatcagccc actgacgcgt tgcgcgagaa gattgtgcac cgccgcttta caggcttcga
     5521 cgccgcttcg ttctaccatc gacaccacca cgctggcacc cagttgatcg gcgcgagatt
     5581 taatcgccgc gacaatttgc gacggcgcgt gcagggccag actggaggtg gcaacgccaa
     5641 tcagcaacga ctgtttgccc gccagttgtt gtgccacgcg gttgggaatg taattcagct
     5701 ccgccatcgc cgcttccact ttttcccgcg ttttcgcaga aacgtggctg gcctggttca
     5761 ccacgcggga aacggtctga taagagacac cggcatactc tgcgacatcg tataacgtta
     5821 ctggtttcac attcaccacc ctgaattgac tctcttccgg gcgctatcat gccataccgc
     5881 gaaaggtttt gcgccattcg atggtgtccg ggatctcgac gctctccctt atgcgactcc
     5941 tgcattagga agcagcccag tagtaggttg aggccgttga gcaccgccgc cgcaaggaat
     6001 ggtgcatgca aggagatggc gcccaacagt cccccggcca cggggcctgc caccataccc
     6061 acgccgaaac aagcgctcat gagcccgaag tggcgagccc gatcttcccc atcggtgatg
     6121 tcggcgatat aggcgccagc aaccgcacct gtggcgccgg tgatgccggc cacgatgcgt
     6181 ccggcgtaga ggatcgagat ctcgatcccg cgaaat
//
