LOCUS       sequence-222663-        6060 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Plasmid for highly efficient expression of engineered IL24 with 
            mutated binding sites.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6060)
  AUTHORS   Zahradnik J, Kolarova L, Peleg Y, Kolenko P, Svidenska S, Charnavets
            T, Unger T, Sussman JL, Schneider B
  TITLE     Flexible regions govern promiscuous binding of IL-24 to receptors 
            IL-20R1 and IL-22R1.
  JOURNAL   FEBS J. 2019 Oct;286(19):3858-3873. doi: 10.1111/febs.14945. Epub 
            2019 Jun 12.
  PUBMED    31152679
REFERENCE   2  (bases 1 to 6060)
  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: 
            "10.1111/febs.14945"; journalName: "FEBS J"; date: "2019-10"; 
            volume: "286"; issue: "19"; pages: "3858-3873"
COMMENT     Sequence Label: pSumo24P7
FEATURES             Location/Qualifiers
     source          1..6060
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     568..587
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(607..684)
                     /gene="lacI"
                     /label=lacI promoter
     primer_bind     890..909
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     993..1012
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        993..1011
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    1012..1036
                     /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             1051..1073
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             1917..1934
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(1987..2005)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      2001..2048
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      2085..2540
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2172..2191)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2382..2403
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(2632..3447)
                     /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(2632..3447)
                     /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     3355..3374
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     3439..3458
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     rep_origin      3569..4157
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4058..4077
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4311..4328
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    4343..4482
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(4408..4427)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4568..4590
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(4584..4775)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(join(5584..6060,1..606))
                     /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"
ORIGIN
        1 atgccagcca gccagacgca gacgcgccga gacagaactt aatgggcccg ctaacagcgc
       61 gatttgctgg tgacccaatg cgaccagatg ctccacgccc agtcgcgtac cgtcttcatg
      121 ggagaaaata atactgttga tgggtgtctg gtcagagaca tcaagaaata acgccggaac
      181 attagtgcag gcagcttcca cagcaatggc atcctggtca tccagcggat agttaatgat
      241 cagcccactg acgcgttgcg cgagaagatt gtgcaccgcc gctttacagg cttcgacgcc
      301 gcttcgttct accatcgaca ccaccacgct ggcacccagt tgatcggcgc gagatttaat
      361 cgccgcgaca atttgcgacg gcgcgtgcag ggccagactg gaggtggcaa cgccaatcag
      421 caacgactgt ttgcccgcca gttgttgtgc cacgcggttg ggaatgtaat tcagctccgc
      481 catcgccgct tccacttttt cccgcgtttt cgcagaaacg tggctggcct ggttcaccac
      541 gcgggaaacg gtctgataag agacaccggc atactctgcg acatcgtata acgttactgg
      601 tttcacattc accaccctga attgactctc ttccgggcgc tatcatgcca taccgcgaaa
      661 ggttttgcgc cattcgatgg tgtccgggat ctcgacgctc tcccttatgc gactcctgca
      721 ttaggaagca gcccagtagt aggttgaggc cgttgagcac cgccgccgca aggaatggtg
      781 catgcaagga gatggcgccc aacagtcccc cggccacggg gcctgccacc atacccacgc
      841 cgaaacaagc gctcatgagc ccgaagtggc gagcccgatc ttccccatcg gtgatgtcgg
      901 cgatataggc gccagcaacc gcacctgtgg cgccggtgat gccggccacg atgcgtccgg
      961 cgtagaggat cgagatctcg atcccgcgaa attaatacga ctcactatag gggaattgtg
     1021 agcggataac aattcccctc tagaaataat tttgtttaac tttaagaagg agatatacca
     1081 tgagcaagca tcaccatcat tcaggccatc accataccgg acaccaccat cattcaggca
     1141 gtcatcacca ttccggatct gctgcgggtg gcgaagaaga taagaaaccg gcaggtggcg
     1201 aaggtggcgg tgcccatatc aacctgaaag tgaaaggtca agacggcaac gaagtctttt
     1261 tccgcatcaa acgttctacc cagctgaaaa agctgatgaa cgcatactgt gaccgtcagt
     1321 ctgtagacat gaccgcaatt gctttcctgt ttgatggtcg tcgcctgcgt gcggaacaga
     1381 ccccggatga actggagatg gaagatggcg acgaaatcga cgcaatgctt catcagactg
     1441 gtggcgcgtt ccattttggt ccatgtcgtg tcgaaggtgt ggtgccgcaa gaactgtggg
     1501 aggccttctg ggcggttcgt gatacgttac aagctcagga caacatcacc gatgtacgtt
     1561 tacttcgtgc agaggtttta caaaatgttt cagatgcaga gagctgctat ttagtccatc
     1621 aactgctgcg cttctactta gacacagtct ttaaaaacta tcacaataag accgcagagc
     1681 ttcgtactct gaaaagtttt tcaacactgg cgaataattt cttacagatc cgcaaggatc
     1741 tgcaaccgtg tcaggaacaa aatatgtgct catcgcgtga ggaagcacac cgtcgctttt
     1801 tacaattcca acgtgcattt gagcaattgg atgtggaagc agcagcgaca aaggctctgg
     1861 gagagatcga catccttctt cgctggatgc aaaagtttta tcaactgtaa ctcgagcacc
     1921 accaccacca ccactgagat ccggctgcta acaaagcccg aaaggaagct gagttggctg
     1981 ctgccaccgc tgagcaataa ctagcataac cccttggggc ctctaaacgg gtcttgaggg
     2041 gttttttgct gaaaggagga actatatccg gattggcgaa tgggacgcgc cctgtagcgg
     2101 cgcattaagc gcggcgggtg tggtggttac gcgcagcgtg accgctacac ttgccagcgc
     2161 cctagcgccc gctcctttcg ctttcttccc ttcctttctc gccacgttcg ccggctttcc
     2221 ccgtcaagct ctaaatcggg ggctcccttt agggttccga tttagtgctt tacggcacct
     2281 cgaccccaaa aaacttgatt agggtgatgg ttcacgtagt gggccatcgc cctgatagac
     2341 ggtttttcgc cctttgacgt tggagtccac gttctttaat agtggactct tgttccaaac
     2401 tggaacaaca ctcaacccta tctcggtcta ttcttttgat ttataaggga ttttgccgat
     2461 ttcggcctat tggttaaaaa atgagctgat ttaacaaaaa tttaacgcga attttaacaa
     2521 aatattaacg cttacaattt aggtggcact tttcggggaa atgtgcgcgg aacccctatt
     2581 tgtttatttt tctaaataca ttcaaatatg tatccgctca tgaattaatt cttagaaaaa
     2641 ctcatcgagc atcaaatgaa actgcaattt attcatatca ggattatcaa taccatattt
     2701 ttgaaaaagc cgtttctgta atgaaggaga aaactcaccg aggcagttcc ataggatggc
     2761 aagatcctgg tatcggtctg cgattccgac tcgtccaaca tcaatacaac ctattaattt
     2821 cccctcgtca aaaataaggt tatcaagtga gaaatcacca tgagtgacga ctgaatccgg
     2881 tgagaatggc aaaagtttat gcatttcttt ccagacttgt tcaacaggcc agccattacg
     2941 ctcgtcatca aaatcactcg catcaaccaa accgttattc attcgtgatt gcgcctgagc
     3001 gagacgaaat acgcgatcgc tgttaaaagg acaattacaa acaggaatcg aatgcaaccg
     3061 gcgcaggaac actgccagcg catcaacaat attttcacct gaatcaggat attcttctaa
     3121 tacctggaat gctgttttcc cggggatcgc agtggtgagt aaccatgcat catcaggagt
     3181 acggataaaa tgcttgatgg tcggaagagg cataaattcc gtcagccagt ttagtctgac
     3241 catctcatct gtaacatcat tggcaacgct acctttgcca tgtttcagaa acaactctgg
     3301 cgcatcgggc ttcccataca atcgatagat tgtcgcacct gattgcccga cattatcgcg
     3361 agcccattta tacccatata aatcagcatc catgttggaa tttaatcgcg gcctagagca
     3421 agacgtttcc cgttgaatat ggctcataac accccttgta ttactgttta tgtaagcaga
     3481 cagttttatt gttcatgacc aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag
     3541 accccgtaga aaagatcaaa ggatcttctt gagatccttt ttttctgcgc gtaatctgct
     3601 gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg tttgccggat caagagctac
     3661 caactctttt tccgaaggta actggcttca gcagagcgca gataccaaat actgtccttc
     3721 tagtgtagcc gtagttaggc caccacttca agaactctgt agcaccgcct acatacctcg
     3781 ctctgctaat cctgttacca gtggctgctg ccagtggcga taagtcgtgt cttaccgggt
     3841 tggactcaag acgatagtta ccggataagg cgcagcggtc gggctgaacg gggggttcgt
     3901 gcacacagcc cagcttggag cgaacgacct acaccgaact gagataccta cagcgtgagc
     3961 tatgagaaag cgccacgctt cccgaaggga gaaaggcgga caggtatccg gtaagcggca
     4021 gggtcggaac aggagagcgc acgagggagc ttccaggggg aaacgcctgg tatctttata
     4081 gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg
     4141 ggcggagcct atggaaaaac gccagcaacg cggccttttt acggttcctg gccttttgct
     4201 ggccttttgc tcacatgttc tttcctgcgt tatcccctga ttctgtggat aaccgtatta
     4261 ccgcctttga gtgagctgat accgctcgcc gcagccgaac gaccgagcgc agcgagtcag
     4321 tgagcgagga agcggaagag cgcctgatgc ggtattttct ccttacgcat ctgtgcggta
     4381 tttcacaccg caatggtgca ctctcagtac aatctgctct gatgccgcat agttaagcca
     4441 gtatacactc cgctatcgct acgtgactgg gtcatggctg cgccccgaca cccgccaaca
     4501 cccgctgacg cgccctgacg ggcttgtctg ctcccggcat ccgcttacag acaagctgtg
     4561 accgtctccg ggagctgcat gtgtcagagg ttttcaccgt catcaccgaa acgcgcgagg
     4621 cagctgcggt aaagctcatc agcgtggtcg tgaagcgatt cacagatgtc tgcctgttca
     4681 tccgcgtcca gctcgttgag tttctccaga agcgttaatg tctggcttct gataaagcgg
     4741 gccatgttaa gggcggtttt ttcctgtttg gtcactgatg cctccgtgta agggggattt
     4801 ctgttcatgg gggtaatgat accgatgaaa cgagagagga tgctcacgat acgggttact
     4861 gatgatgaac atgcccggtt actggaacgt tgtgagggta aacaactggc ggtatggatg
     4921 cggcgggacc agagaaaaat cactcagggt caatgccagc gcttcgttaa tacagatgta
     4981 ggtgttccac agggtagcca gcagcatcct gcgatgcaga tccggaacat aatggtgcag
     5041 ggcgctgact tccgcgtttc cagactttac gaaacacgga aaccgaagac cattcatgtt
     5101 gttgctcagg tcgcagacgt tttgcagcag cagtcgcttc acgttcgctc gcgtatcggt
     5161 gattcattct gctaaccagt aaggcaaccc cgccagccta gccgggtcct caacgacagg
     5221 agcacgatca tgcgcacccg tggggccgcc atgccggcga taatggcctg cttctcgccg
     5281 aaacgtttgg tggcgggacc agtgacgaag gcttgagcga gggcgtgcaa gattccgaat
     5341 accgcaagcg acaggccgat catcgtcgcg ctccagcgaa agcggtcctc gccgaaaatg
     5401 acccagagcg ctgccggcac ctgtcctacg agttgcatga taaagaagac agtcataagt
     5461 gcggcgacga tagtcatgcc ccgcgcccac cggaaggagc tgactgggtt gaaggctctc
     5521 aagggcatcg gtcgagatcc cggtgcctaa tgagtgagct aacttacatt aattgcgttg
     5581 cgctcactgc ccgctttcca gtcgggaaac ctgtcgtgcc agctgcatta atgaatcggc
     5641 caacgcgcgg ggagaggcgg tttgcgtatt gggcgccagg gtggtttttc ttttcaccag
     5701 tgagacgggc aacagctgat tgcccttcac cgcctggccc tgagagagtt gcagcaagcg
     5761 gtccacgctg gtttgcccca gcaggcgaaa atcctgtttg atggtggtta acggcgggat
     5821 ataacatgag ctgtcttcgg tatcgtcgta tcccactacc gagatatccg caccaacgcg
     5881 cagcccggac tcggtaatgg cgcgcattgc gcccagcgcc atctgatcgt tggcaaccag
     5941 catcgcagtg ggaacgatgc cctcattcag catttgcatg gtttgttgaa aaccggacat
     6001 ggcactccag tcgccttccc gttccgctat cggctgaatt tgattgcgag tgagatattt
//
