LOCUS       sequence-250143-        5898 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 5898)
  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 5898)
  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-G-L11
FEATURES             Location/Qualifiers
     source          1..5898
                     /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             763..780
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(833..851)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      847..894
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      931..1386
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1018..1037)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1228..1249
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(1478..2293)
                     /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(1478..2293)
                     /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     2201..2220
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     2285..2304
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     rep_origin      2415..3003
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2904..2923
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3157..3174
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    3189..3328
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(3254..3273)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3414..3436
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(3430..3621)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(4430..5512)
                     /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     5474..5493
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(5513..5590)
                     /gene="lacI"
                     /label=lacI promoter
     primer_bind     5796..5815
                     /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 atccggcggc ggcatggcgc aggatcaagg
      181 tgaaaaggag aaccccatgc gggaacttcg catccgcaaa ctctgtctca acatctgtgt
      241 tggggagagt ggagacagac tgacgcgagc agccaaggtg ttggagcagc tcacagggca
      301 gacccctgtg ttttccaaag ctagatacac tgtcagatcc tttggcatcc ggagaaatga
      361 aaagattgct gtccactgca cagttcgagg ggccaaggca gaagaaatct tggagaaggg
      421 tctaaaggtg cgggagtatg agttaagaaa aaacaacttc tcagatactg gaaactttgg
      481 ttttgggatc caggaacaca tcgatctggg tatcaaatat gacccaagca ttggtatcta
      541 cggcctggac ttctatgtgg tgctgggtag gccaggtttc agcatcgcag acaagaagcg
      601 caggacaggc tgcattgggg ccaaacacag aatcagcaaa gaggaggcca tgcgctggtt
      661 ccagcagaag tatgatggga tcatccttcc tggcaaataa cagtaaaggt ggatacggat
      721 ccgaattcga gctccgtcga caagcttgcg gccgcactcg agcaccacca ccaccaccac
      781 tgagatccgg ctgctaacaa agcccgaaag gaagctgagt tggctgctgc caccgctgag
      841 caataactag cataacccct tggggcctct aaacgggtct tgaggggttt tttgctgaaa
      901 ggaggaacta tatccggatt ggcgaatggg acgcgccctg tagcggcgca ttaagcgcgg
      961 cgggtgtggt ggttacgcgc agcgtgaccg ctacacttgc cagcgcccta gcgcccgctc
     1021 ctttcgcttt cttcccttcc tttctcgcca cgttcgccgg ctttccccgt caagctctaa
     1081 atcgggggct ccctttaggg ttccgattta gtgctttacg gcacctcgac cccaaaaaac
     1141 ttgattaggg tgatggttca cgtagtgggc catcgccctg atagacggtt tttcgccctt
     1201 tgacgttgga gtccacgttc tttaatagtg gactcttgtt ccaaactgga acaacactca
     1261 accctatctc ggtctattct tttgatttat aagggatttt gccgatttcg gcctattggt
     1321 taaaaaatga gctgatttaa caaaaattta acgcgaattt taacaaaata ttaacgctta
     1381 caatttaggt ggcacttttc ggggaaatgt gcgcggaacc cctatttgtt tatttttcta
     1441 aatacattca aatatgtatc cgctcatgaa ttaattctta gaaaaactca tcgagcatca
     1501 aatgaaactg caatttattc atatcaggat tatcaatacc atatttttga aaaagccgtt
     1561 tctgtaatga aggagaaaac tcaccgaggc agttccatag gatggcaaga tcctggtatc
     1621 ggtctgcgat tccgactcgt ccaacatcaa tacaacctat taatttcccc tcgtcaaaaa
     1681 taaggttatc aagtgagaaa tcaccatgag tgacgactga atccggtgag aatggcaaaa
     1741 gtttatgcat ttctttccag acttgttcaa caggccagcc attacgctcg tcatcaaaat
     1801 cactcgcatc aaccaaaccg ttattcattc gtgattgcgc ctgagcgaga cgaaatacgc
     1861 gatcgctgtt aaaaggacaa ttacaaacag gaatcgaatg caaccggcgc aggaacactg
     1921 ccagcgcatc aacaatattt tcacctgaat caggatattc ttctaatacc tggaatgctg
     1981 ttttcccggg gatcgcagtg gtgagtaacc atgcatcatc aggagtacgg ataaaatgct
     2041 tgatggtcgg aagaggcata aattccgtca gccagtttag tctgaccatc tcatctgtaa
     2101 catcattggc aacgctacct ttgccatgtt tcagaaacaa ctctggcgca tcgggcttcc
     2161 catacaatcg atagattgtc gcacctgatt gcccgacatt atcgcgagcc catttatacc
     2221 catataaatc agcatccatg ttggaattta atcgcggcct agagcaagac gtttcccgtt
     2281 gaatatggct cataacaccc cttgtattac tgtttatgta agcagacagt tttattgttc
     2341 atgaccaaaa tcccttaacg tgagttttcg ttccactgag cgtcagaccc cgtagaaaag
     2401 atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa
     2461 aaaccaccgc taccagcggt ggtttgtttg ccggatcaag agctaccaac tctttttccg
     2521 aaggtaactg gcttcagcag agcgcagata ccaaatactg tccttctagt gtagccgtag
     2581 ttaggccacc acttcaagaa ctctgtagca ccgcctacat acctcgctct gctaatcctg
     2641 ttaccagtgg ctgctgccag tggcgataag tcgtgtctta ccgggttgga ctcaagacga
     2701 tagttaccgg ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac acagcccagc
     2761 ttggagcgaa cgacctacac cgaactgaga tacctacagc gtgagctatg agaaagcgcc
     2821 acgcttcccg aagggagaaa ggcggacagg tatccggtaa gcggcagggt cggaacagga
     2881 gagcgcacga gggagcttcc agggggaaac gcctggtatc tttatagtcc tgtcgggttt
     2941 cgccacctct gacttgagcg tcgatttttg tgatgctcgt caggggggcg gagcctatgg
     3001 aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct tttgctggcc ttttgctcac
     3061 atgttctttc ctgcgttatc ccctgattct gtggataacc gtattaccgc ctttgagtga
     3121 gctgataccg ctcgccgcag ccgaacgacc gagcgcagcg agtcagtgag cgaggaagcg
     3181 gaagagcgcc tgatgcggta ttttctcctt acgcatctgt gcggtatttc acaccgcaat
     3241 ggtgcactct cagtacaatc tgctctgatg ccgcatagtt aagccagtat acactccgct
     3301 atcgctacgt gactgggtca tggctgcgcc ccgacacccg ccaacacccg ctgacgcgcc
     3361 ctgacgggct tgtctgctcc cggcatccgc ttacagacaa gctgtgaccg tctccgggag
     3421 ctgcatgtgt cagaggtttt caccgtcatc accgaaacgc gcgaggcagc tgcggtaaag
     3481 ctcatcagcg tggtcgtgaa gcgattcaca gatgtctgcc tgttcatccg cgtccagctc
     3541 gttgagtttc tccagaagcg ttaatgtctg gcttctgata aagcgggcca tgttaagggc
     3601 ggttttttcc tgtttggtca ctgatgcctc cgtgtaaggg ggatttctgt tcatgggggt
     3661 aatgataccg atgaaacgag agaggatgct cacgatacgg gttactgatg atgaacatgc
     3721 ccggttactg gaacgttgtg agggtaaaca actggcggta tggatgcggc gggaccagag
     3781 aaaaatcact cagggtcaat gccagcgctt cgttaataca gatgtaggtg ttccacaggg
     3841 tagccagcag catcctgcga tgcagatccg gaacataatg gtgcagggcg ctgacttccg
     3901 cgtttccaga ctttacgaaa cacggaaacc gaagaccatt catgttgttg ctcaggtcgc
     3961 agacgttttg cagcagcagt cgcttcacgt tcgctcgcgt atcggtgatt cattctgcta
     4021 accagtaagg caaccccgcc agcctagccg ggtcctcaac gacaggagca cgatcatgcg
     4081 cacccgtggg gccgccatgc cggcgataat ggcctgcttc tcgccgaaac gtttggtggc
     4141 gggaccagtg acgaaggctt gagcgagggc gtgcaagatt ccgaataccg caagcgacag
     4201 gccgatcatc gtcgcgctcc agcgaaagcg gtcctcgccg aaaatgaccc agagcgctgc
     4261 cggcacctgt cctacgagtt gcatgataaa gaagacagtc ataagtgcgg cgacgatagt
     4321 catgccccgc gcccaccgga aggagctgac tgggttgaag gctctcaagg gcatcggtcg
     4381 agatcccggt gcctaatgag tgagctaact tacattaatt gcgttgcgct cactgcccgc
     4441 tttccagtcg ggaaacctgt cgtgccagct gcattaatga atcggccaac gcgcggggag
     4501 aggcggtttg cgtattgggc gccagggtgg tttttctttt caccagtgag acgggcaaca
     4561 gctgattgcc cttcaccgcc tggccctgag agagttgcag caagcggtcc acgctggttt
     4621 gccccagcag gcgaaaatcc tgtttgatgg tggttaacgg cgggatataa catgagctgt
     4681 cttcggtatc gtcgtatccc actaccgaga tatccgcacc aacgcgcagc ccggactcgg
     4741 taatggcgcg cattgcgccc agcgccatct gatcgttggc aaccagcatc gcagtgggaa
     4801 cgatgccctc attcagcatt tgcatggttt gttgaaaacc ggacatggca ctccagtcgc
     4861 cttcccgttc cgctatcggc tgaatttgat tgcgagtgag atatttatgc cagccagcca
     4921 gacgcagacg cgccgagaca gaacttaatg ggcccgctaa cagcgcgatt tgctggtgac
     4981 ccaatgcgac cagatgctcc acgcccagtc gcgtaccgtc ttcatgggag aaaataatac
     5041 tgttgatggg tgtctggtca gagacatcaa gaaataacgc cggaacatta gtgcaggcag
     5101 cttccacagc aatggcatcc tggtcatcca gcggatagtt aatgatcagc ccactgacgc
     5161 gttgcgcgag aagattgtgc accgccgctt tacaggcttc gacgccgctt cgttctacca
     5221 tcgacaccac cacgctggca cccagttgat cggcgcgaga tttaatcgcc gcgacaattt
     5281 gcgacggcgc gtgcagggcc agactggagg tggcaacgcc aatcagcaac gactgtttgc
     5341 ccgccagttg ttgtgccacg cggttgggaa tgtaattcag ctccgccatc gccgcttcca
     5401 ctttttcccg cgttttcgca gaaacgtggc tggcctggtt caccacgcgg gaaacggtct
     5461 gataagagac accggcatac tctgcgacat cgtataacgt tactggtttc acattcacca
     5521 ccctgaattg actctcttcc gggcgctatc atgccatacc gcgaaaggtt ttgcgccatt
     5581 cgatggtgtc cgggatctcg acgctctccc ttatgcgact cctgcattag gaagcagccc
     5641 agtagtaggt tgaggccgtt gagcaccgcc gccgcaagga atggtgcatg caaggagatg
     5701 gcgcccaaca gtcccccggc cacggggcct gccaccatac ccacgccgaa acaagcgctc
     5761 atgagcccga agtggcgagc ccgatcttcc ccatcggtga tgtcggcgat ataggcgcca
     5821 gcaaccgcac ctgtggcgcc ggtgatgccg gccacgatgc gtccggcgta gaggatcgag
     5881 atctcgatcc cgcgaaat
//
