LOCUS       sequence-487872-        4858 bp    DNA     circular SYN 02-OCT-2025
DEFINITION  Constitutive expression of human IL-10 (with its signal sequence).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4858)
  AUTHORS   Edelstein HI, Cosio A, Ezekiel ML, Corcoran WK, Morris AH, Leonard 
            JN
  TITLE     Conversion of natural cytokine receptors into orthogonal synthetic 
            biosensors.
  JOURNAL   Nat Chem Biol. 2025 Aug 22:10.1038/s41589-025-01986-1. doi: 
            10.1038/s41589-025-01986-1.
  PUBMED    40846994
REFERENCE   2  (bases 1 to 4858)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Oct 2, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Nat Chem 
            Biol. 2025 Aug 22:10.1038/s41589-025-01986-1. doi: 
            10.1038/s41589-025-01986-1."
COMMENT     Sequence Label: L4300 hIL10 in pcDNA backbone
FEATURES             Location/Qualifiers
     source          1..4858
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(44..63)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        235..614
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        615..818
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     769..789
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     863..882
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        863..881
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1488..1505)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    1494..1718
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      1764..2192
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1851..1870)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2061..2082
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        2215..2411
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      2262..2397
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     2324..2343
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     polyA_signal    2531..2652
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2568..2587)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2622..2641
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(2701..2717)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2701..2717)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2714..2736)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2725..2741
                     /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)."
     promoter        complement(join(2749..2755,2756..2773,2774..2779))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2794..2815
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2932..2949)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3103..3691)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3183..3202)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3862..4653,4654..4722))
                     /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     4485..4504
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4723..4827)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gacggatcgg gagatctccc gatcccctat ggtgcactct cagtacaatc tgctctgatg
       61 ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg
      121 cgagcaaaat ttaagctaca acaaggcaag gcttgaccga caattgcatg aagaatctgc
      181 ttagggttag gcgttttgcg ctgcttcgcg atgtacgggc cagatatacg cgttgacatt
      241 gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata
      301 tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc
      361 cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc
      421 attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt
      481 atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt
      541 atgcccagta catgacctta tgggactttc ctacttggca gtacatctac gtattagtca
      601 tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg
      661 actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc
      721 aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg
      781 gtaggcgtgt acggtgggag gtctatataa gcagagctct ctggctaact agagaaccca
      841 ctgcttactg gcttatcgaa attaatacga ctcactatag ggagtcccaa gctggctagc
      901 gccaccatgc atagctctgc cctgctgtgc tgtctggtcc tcttgacagg agtgagggct
      961 tcacctggac aaggaaccca gtctgaaaac agctgcactc actttcccgg taatctcccc
     1021 aatatgctgc gggatttgag ggacgccttt agtagggtga agacgttctt tcagatgaaa
     1081 gaccaactcg ataacttgct cctgaaagag tcactcttgg aggattttaa gggttatctt
     1141 ggttgccagg cattgagtga gatgattcag ttctatttgg aagaagtaat gcctcaggct
     1201 gagaaccagg atcccgacat taaagcgcac gtaaattccc tcggtgaaaa tttgaagaca
     1261 ctcaggctcc ggctgcgacg ctgccaccgc tttcttccgt gcgaaaataa gtctaaagcc
     1321 gtggagcagg tgaagaatgc tttcaataag ctccaagaga aaggcatata taaagcaatg
     1381 tcagagtttg acattttcat caactacatc gaagcctaca tgaccatgaa aatcagaaat
     1441 taggcggccg ctcgagtcta gagggcccgt ttaaacccgc tgatcagcct cgactgtgcc
     1501 ttctagttgc cagccatctg ttgtttgccc ctcccccgtg ccttccttga ccctggaagg
     1561 tgccactccc actgtccttt cctaataaaa tgaggaaatt gcatcgcatt gtctgagtag
     1621 gtgtcattct attctggggg gtggggtggg gcaggacagc aagggggagg attgggaagt
     1681 caatagcagg catgctgggg atgcggtggg ctctatggct tctgaggcgg aaagaaccag
     1741 ctggggctct agggggtatc cccacgcgcc ctgtagcggc gcattaagcg cggcgggtgt
     1801 ggtggttacg cgcagcgtga ccgctacact tgccagcgcc ctagcgcccg ctcctttcgc
     1861 tttcttccct tcctttctcg ccacgttcgc cggctttccc cgtcaagctc taaatcgggg
     1921 gctcccttta gggttccgat ttagtgcttt acggcacctc gaccccaaaa aacttgatta
     1981 gggtgatggt tcacgtagtg ggccatcgcc ctgatagacg gtttttcgcc ctttgacgtt
     2041 ggagtccacg ttctttaata gtggactctt gttccaaact ggaacaacac tcaaccctat
     2101 ctcggtctat tcttttgatt tataagggat tttgccgatt tcggcctatt ggttaaaaaa
     2161 tgagctgatt taacaaaaat ttaacgcgaa ttaattctgt ggaatgtgtg tcagttaggc
     2221 tcaattagtc agcaaccata gtcccgcccc taactccgcc catcccgccc ctaactccgc
     2281 ccagttccgc ccattctccg ccccatggct gactaatttt ttttatttat gcagaggccg
     2341 aggccgcctc tgcctctgag ctattccaga agtagtgagg aggctttttt ggaggcctag
     2401 gcttttgcaa aaagctcccg ggagcttgta tatccatttt cggaggttgg gcttcggaat
     2461 cgttttccgg gacgccggct ggatgatcct ccagcgcggg gatctcatgc tggagttctt
     2521 cgcccacccc aacttgttta ttgcagctta taatggttac aaataaagca atagcatcac
     2581 aaatttcaca aataaagcat ttttttcact gcattctagt tgtggtttgt ccaaactcat
     2641 caatgtatct tatcatgtct gtataccgtc gacctctagc tagagcttgg cgtaatcatg
     2701 gtcatagctg tttcctgtgt gaaattgtta tccgctcaca attccacaca acatacgagc
     2761 cggaagcata aagtgtaaag cctggggtgc ctaatgagtg agctaactca cattaattgc
     2821 gttgcgctca ctgcccgctt tccagtcggg aaacctgtcg tgccagctgc attaatgaat
     2881 cggccaacgc gcggggagag gcggtttgcg tattgggcgc tcttccgctt cctcgctcac
     2941 tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta tcagctcact caaaggcggt
     3001 aatacggtta tccacagaat caggggataa cgcaggaaag aacatgtgag caaaaggcca
     3061 gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc
     3121 ccctgacgag catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact
     3181 ataaagatac caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct
     3241 gccgcttacc ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcatag
     3301 ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca
     3361 cgaacccccc gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa
     3421 cccggtaaga cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc
     3481 gaggtatgta ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag
     3541 aagaacagta tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg
     3601 tagctcttga tccggcaaac aaaccaccgc tggtagcggt ggtttttttg tttgcaagca
     3661 gcagattacg cgcagaaaaa aaggatctca agaagatcct ttgatctttt ctacggggtc
     3721 tgacgctcag tggaacgaaa actcacgtta agggattttg gtcatgagat tatcaaaaag
     3781 gatcttcacc tagatccttt taaattaaaa atgaagtttt aaatcaatct aaagtatata
     3841 tgagtaaact tggtctgaca gttaccaatg cttaatcagt gaggcaccta tctcagcgat
     3901 ctgtctattt cgttcatcca tagttgcctg actccccgtc gtgtagataa ctacgatacg
     3961 ggagggctta ccatctggcc ccagtgctgc aatgataccg cgagatccac gctcaccggc
     4021 tccagattta tcagcaataa accagccagc cggaagggcc gagcgcagaa gtggtcctgc
     4081 aactttatcc gcctccatcc agtctattaa ttgttgccgg gaagctagag taagtagttc
     4141 gccagttaat agtttgcgca acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc
     4201 gtcgtttggt atggcttcat tcagctccgg ttcccaacga tcaaggcgag ttacatgatc
     4261 ccccatgttg tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg tcagaagtaa
     4321 gttggccgca gtgttatcac tcatggttat ggcagcactg cataattctc ttactgtcat
     4381 gccatccgta agatgctttt ctgtgactgg tgagtactca accaagtcat tctgagaata
     4441 gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata cgggataata ccgcgccaca
     4501 tagcagaact ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa aactctcaag
     4561 gatcttaccg ctgttgagat ccagttcgat gtaacccact cgtgcaccca actgatcttc
     4621 agcatctttt actttcacca gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc
     4681 aaaaaaggga ataagggcga cacggaaatg ttgaatactc atactcttcc tttttcaata
     4741 ttattgaagc atttatcagg gttattgtct catgagcgga tacatatttg aatgtattta
     4801 gaaaaataaa caaatagggg ttccgcgcac atttccccga aaagtgccac ctgacgtc
//
