LOCUS       sequence-415901-        4037 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Transient mammalian expression of IL-10.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4037)
  AUTHORS   Makri Pistikou AM, Cremers GAO, Nathalia BL, Meuleman TJ, Bogels 
            BWA, Eijkens BV, de Dreu A, Bezembinder MTH, Stassen OMJA, Bouten 
            CCV, Merkx M, Jerala R, de Greef TFA
  TITLE     Engineering a scalable and orthogonal platform for synthetic 
            communication in mammalian cells.
  JOURNAL   Nat Commun. 2023 Nov 2;14(1):7001. doi: 10.1038/s41467-023-42810-5.
  PUBMED    37919273
REFERENCE   2  (bases 1 to 4037)
  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: "Nat 
            Commun."; date: "2023-11-2"; volume: "14(1):7001. doi"; pages: " 
            10.1038/s41467-023-42810-5"
COMMENT     Sequence Label: pLS13-IL-10
FEATURES             Location/Qualifiers
     source          1..4037
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     polyA_signal    782..916
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(806..825)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     860..879
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(1059..1081)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     1222..1241
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     rep_origin      complement(1279..1867)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1359..1378)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2038..2829,2830..2898))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     2661..2680
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(2899..3003)
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3115..3153
                     /label=minimal CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     3150..3174
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
ORIGIN
        1 ctctcggact ccctacccca accccaggga ctgcaggttg tgccctgtgg ctgcctgcac
       61 cccaggaaag gagggggctc aggccatcca gccaccacct acagcccagt ggcctcgaga
      121 cctgcagccc aagcttaaaa cagctctggg gttgtaccca ccccagaggc ccacgtggcg
      181 gctagtactc cggtattgcg gtacccttgt acgcctgttt tatactccct tcccgtaact
      241 tagacgcaca aaaccaagtt caatagaagg gggtacaaac cagtaccacc acgaacaagc
      301 acttctgttt ccccggtgat gtcgtataga ctgcttgcgt ggttgaaagc gacggatccg
      361 ttatccgctt atgtacttcg agaagcccag taccacctcg gaatcttcga tgcgttgcgc
      421 tcagcactca accccagagt gtagcttagg ctgatgagtc tggacatccc tcaccggtga
      481 cggtggtcca ggctgcgttg gcggcctacc tatggctaac gccatgggac gctagttgtg
      541 aacaaggtgt gaagagccta ttgagctaca taagaatcct ccggcccctg aatgcggcta
      601 atcccaacct cggagcaggt ggtcacaaac cagtgattgg cctgtcgtaa cgcgcaagtc
      661 cgtggcggaa ccgactactt tgggtgtccg tgtttccttt tattttattg tggctgctta
      721 tggtgacaat cacagattgt tatcataaag cgaattggat tgcggccgca aagctagctt
      781 cgatccagac atgataagat acattgatga gtttggacaa accacaacta gaatgcagtg
      841 aaaaaaatgc tttatttgtg aaatttgtga tgctattgct ttatttgtaa ccattataag
      901 ctgcaataaa caagttaaca acaacaattg cattcatttt atgtttcagg ttcaggggga
      961 ggtgtgggag gttttttaaa gcaagtaaaa cctctacaaa tgtggtatgg ctgattatga
     1021 tccggctgcc tcgcgcgttt cggtgatgac ggtgaaaacc tctgacacat gcagctcccg
     1081 gagacggtca cagcttgtct gtaagcggat gccgggagca gacaagcccg tcagggcgcg
     1141 tcagcgggtg ttggcgggtg tcggggcgca gccatgaccc agtcacgtag cgatagcgga
     1201 gtgtatactg gcttaactat gcggcatcag agcagattgt actgagagtg caccatatcg
     1261 ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc acaaaaatcg
     1321 acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg cgtttccccc
     1381 tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat acctgtccgc
     1441 ctttctccct tcgggaagcg tggcgctttc tcatagctca cgctgtaggt atctcagttc
     1501 ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc agcccgaccg
     1561 ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg acttatcgcc
     1621 actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg gtgctacaga
     1681 gttcttgaag tggtggccta actacggcta cactagaagg acagtatttg gtatctgcgc
     1741 tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg gcaaacaaac
     1801 caccgctggt agcggtggtt tttttgtttg caagcagcag attacgcgca gaaaaaaagg
     1861 atctcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga acgaaaactc
     1921 acgttaaggg attttggtca tgagattatc aaaaaggatc ttcacctaga tccttttaaa
     1981 ttaaaaatga agttttaaat caatctaaag tatatatgag taaacttggt ctgacagtta
     2041 ccaatgctta atcagtgagg cacctatctc agcgatctgt ctatttcgtt catccatagt
     2101 tgcctgactc cccgtcgtgt agataactac gatacgggag ggcttaccat ctggccccag
     2161 tgctgcaatg ataccgcgag acccacgctc accggctcca gatttatcag caataaacca
     2221 gccagccgga agggccgagc gcagaagtgg tcctgcaact ttatccgcct ccatccagtc
     2281 tattaattgt tgccgggaag ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt
     2341 tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag
     2401 ctccggttcc caacgatcaa ggcgagttac atgatccccc atgttgtgca aaaaagcggt
     2461 tagctccttc ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt tatcactcat
     2521 ggttatggca gcactgcata attctcttac tgtcatgcca tccgtaagat gcttttctgt
     2581 gactggtgag tactcaacca agtcattctg agaatagtgt atgcggcgac cgagttgctc
     2641 ttgcccggcg tcaatacggg ataataccgc gccacatagc agaactttaa aagtgctcat
     2701 cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag
     2761 ttcgatgtaa cccactcgtg cacccaactg atcttcagca tcttttactt tcaccagcgt
     2821 ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg
     2881 gaaatgttga atactcatac tcttcctttt tcaatattat tgaagcattt atcagggtta
     2941 ttgtctcatg agcggataca tatttgaatg tatttagaaa aataaacaaa taggggttcc
     3001 gcgcacattt ccccgaaaag tgccacctga cgtcgtcgac atttcccgta aatcgtcgag
     3061 tcgacatttc ccgtaaatcg tcgacctgca ggtcgagctc ggtacccggg tcgaggtagg
     3121 cgtgtacggt gggaggccta tataagcaga gctcgtttag tgaaccgtca gatcgcctgg
     3181 agacgccatc cacgctgttt tgacctccat agaagacacc gggaccgatc cagcctccgc
     3241 ggaattcgag ctcgcccggg gatcctctag aatgcactca agcgctctcc tttgctgtct
     3301 tgttctcctc acaggagtac gcgcttcacc cggacaaggc acacagtctg aaaattcatg
     3361 cacacatttt cccggcaacc tcccaaatat gcttagggat ctccgcgacg cgtttagtag
     3421 agtaaagacg ttctttcaaa tgaaggacca attggacaat ttgctcttga aggagagcct
     3481 tctcgaagat ttcaagggtt accttggttg tcaagccctt tctgagatga tccaattcta
     3541 cttggaggaa gtaatgccac aagcggaaaa ccaagatcct gatattaaag ctcatgtaaa
     3601 ttctctggga gagaacctca agaccttgag gctccgcttg cgacggtgcc acagattctt
     3661 gccttgcgaa aataagagta aagccgtaga gcaagtcaaa aatgcgttta ataagttgca
     3721 agagaaggga atatataagg cgatgtccga gtttgacata ttcattaact acattgaggc
     3781 ttatatgaca atgaagatcc gcaactaagg gaccctgctg ctgctggaga cggccactgc
     3841 tccctgagtg tcccgtccct ggggctcctg cttccccatc ccggagttct cctgctcccc
     3901 acctcctgtc gtcctgcctg gcctccagcc cgagtcgtca tccccggagt ccctatacag
     3961 aggtcctgcc atggaacctt cccctccccg tgcgctctgg ggactgagcc catgacacca
     4021 aacctgcccc ttggctg
//
