LOCUS       sequence-489409-        8050 bp    DNA     circular SYN 15-OCT-2025
DEFINITION  Expression of a MESA CTEVp chain encoding (N to C): 
            IL10RaSS-3xFLAG-IL10RaECD-IL10RaTMD-CTEVp(190K)-PRS(M)-hcJun.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8050)
  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 8050)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Oct 15, 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: L4427 IL-10Ra CTEVp chain with cJun in TUPV3
FEATURES             Location/Qualifiers
     source          1..8050
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        350..1532
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation 
                     factor EF-1-alpha"
     intron          581..1523
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
     primer_bind     1480..1500
                     /label=EF1a-F
                     /note="Human elongation factor-1a promoter, forward primer"
     primer_bind     1534..1558
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     CDS             1708..1773
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             2878..2898
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQM"
     CDS             complement(2935..2946)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(3957..3974)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3963..4187
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      4956..5384
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5043..5062)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5253..5274
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        5407..5603
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      5454..5589
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     5516..5535
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     polyA_signal    5723..5844
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5760..5779)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5814..5833
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5893..5909)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5893..5909)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5906..5928)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5917..5933
                     /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(5941..5947,5948..5965,5966..5971))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5986..6007
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(6124..6141)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6295..6883)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6375..6394)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7054..7845,7846..7914))
                     /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     7677..7696
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7915..8019)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gacggatcgg gagatcgaag acggactaaa tagtagttca gacgccgtta agcgcaagat
       61 ctggggaggt cgctgagtag tgcgcgagca aaatttaagc tacaacaagg caaggcttga
      121 ccgacaattg catgaagaat ctgcttaggg ttaggcgttt tgcgctgctt cgcgatgtac
      181 gggccagata tacgcgtgag taattcatac aaaaggactc gcccctgcct tggggaatcc
      241 cagggaccgt cgttaaactc ccactaacgt agaacccaga gatcgctgcg ttcccgcccc
      301 ctcacccgcc cgctctcgtc atcactgagg tggagaagag catgcgtgag gctccggtgc
      361 ccgtcagtgg gcagagcgca catcgcccac agtccccgag aagttggggg gaggggtcgg
      421 caattgaacc ggtgcctaga gaaggtggcg cggggtaaac tgggaaagtg atgtcgtgta
      481 ctggctccgc ctttttcccg agggtggggg agaaccgtat ataagtgcag tagtcgccgt
      541 gaacgttctt tttcgcaacg ggtttgccgc cagaacacag gtaagtgccg tgtgtggttc
      601 ccgcgggcct ggcctcttta cgggttatgg cccttgcgtg ccttgaatta cttccacgcc
      661 cctggctgca gtacgtgatt cttgatcccg agcttcgggt tggaagtggg tgggagagtt
      721 cgaggccttg cgcttaagga gccccttcgc ctcgtgcttg agttgaggcc tggcttgggc
      781 gctggggccg ccgcgtgcga atctggtggc accttcgcgc ctgtctcgct gctttcgata
      841 agtctctagc catttaaaat ttttgatgac ctgctgcgac gctttttttc tggcaagata
      901 gtcttgtaaa tgcgggccaa gatctgcaca ctggtatttc ggtttttggg gccgcgggcg
      961 gcgacggggc ccgtgcgtcc cagcgcacat gttcggcgag gcggggcctg cgagcgcggc
     1021 caccgagaat cggacggggg tagtctcaag ctggccggcc tgctctggtg cctggcctcg
     1081 cgccgccgtg tatcgccccg ccctgggcgg caaggctggc ccggtcggca ccagttgcgt
     1141 gagcggaaag atggccgctt cccggccctg ctgcagggag ctcaaaatgg aggacgcggc
     1201 gctcgggaga gcgggcgggt gagtcaccca cacaaaggaa aagggccttt ccgtcctcag
     1261 ccgtcgcttc atgtgactcc acggagtacc gggcgccgtc caggcacctc gattagttct
     1321 cgagcttttg gagtacgtcg tctttaggtt ggggggaggg gttttatgcg atggagtttc
     1381 cccacactga gtgggtggag actgaagtta ggccagcttg gcacttgatg taattctcct
     1441 tggaatttgc cctttttgag tttggatctt ggttcattct caagcctcag acagtggttc
     1501 aaagtttttt tcttccattt caggtgtcgt gagagctcgt ttagtgaacc gtcagatcgc
     1561 ctggagacgc catccacgct gttttgacct ccatagaaga gaacgggacc gatccaaatt
     1621 ctcgaggcta gctgtacagc caccatgctg ccctgcctgg ttgttctgct tgccgctctg
     1681 ctgtctctga gactgggatc tgacgccgac tacaaggacc acgacggcga ttataaggat
     1741 catgacatcg attacaagga cgatgacgac aagcacggca ccgagctgcc ttctccacct
     1801 agcgtttggt tcgaggccga gttcttccac cacatcctgc actggacccc tattcctaac
     1861 cagagcgaga gcacctgtta cgaggtggcc ctgctgagat acggcatcga gagctggaac
     1921 agcatcagca actgcagcca gacactgagc tacgacctga ccgccgtgac actggatctg
     1981 taccacagca acggctaccg ggccagagtt agagccgtgg atggaagcag acactccaac
     2041 tggaccgtga ccaacaccag attcagcgtg gacgaagtga ccctgacagt gggcagcgtg
     2101 aacctggaaa tccacaacgg cttcatcctg ggcaagatcc agctgcctcg gcctaagatg
     2161 gcccctgcca atgataccta cgagagcatc ttcagccact tccgcgagta cgagatcgcc
     2221 atcagaaagg tgcccggcaa cttcaccttc acacacaaga aagtgaagca cgagaacttc
     2281 agcctgctga cctctggcga agtgggcgag ttctgcgtgc aagtgaaacc ctctgtggcc
     2341 agccggtcca acaaaggcat gtggtccaaa gaggaatgca tcagcctgac cagacagtac
     2401 ttcaccgtga caaacgtgat catcttcttc gccttcgtgc tgctgctgtc tggcgctctg
     2461 gcttattgtc tggccctgca gctgtacgtg cggcggagaa agaagggcgc caagagcatg
     2521 tctagcatgg tgtcagacac tagttgcaca ttcccttcat ctgatggcat attctggaag
     2581 cattggattc aaaccaagga tgggcagtgt ggcagtccat tagtatcaac tagagatggg
     2641 ttcattgttg gtatacactc agcatcgaat ttcaccaaca caaacaatta tttcacaagc
     2701 gtgccgaaaa acttcatgga attgaaaaca aatcaggagg cgcagcagtg ggttagtggt
     2761 tggcgattaa atgctgactc agtattgtgg gggggccata aagttttcat ggtgaaacct
     2821 gaagagcctt ttcagccagt taaggaagcg actcaactca tgaatcttaa gggctcggag
     2881 aacctgtatt ttcagatggg gggaggcagc gggggcatga ctgcaaagat ggaaacgacc
     2941 ttctatgacg atgccctcaa cgcctcgttc ctcccgtccg agagcggacc ttatggctac
     3001 agtaacccca agatcctgaa acagagcatg accctgaacc tggccgaccc agtggggagc
     3061 ctgaagccgc acctccgcgc caagaactcg gacctcctca cctcgcccga cgtggggctg
     3121 ctcaagctgg cgtcgcccga gctggagcgc ctgataatcc agtccagcaa cgggcacatc
     3181 accaccacgc cgacccccac ccagttcctg tgccccaaga acgtgacaga tgagcaggag
     3241 ggcttcgccg agggcttcgt gcgcgccctg gccgaactgc acagccagaa cacgctgccc
     3301 agcgtcacgt cggcggcgca gccggtcaac ggggcaggca tggtggctcc cgcggtagcc
     3361 tcggtggcag ggggcagcgg cagcggcggc ttcagcgcca gcctgcacag cgagccgccg
     3421 gtctacgcaa acctcagcaa cttcaaccca ggcgcgctga gcagcggcgg cggggcgccc
     3481 tcctacggcg cggccggcct ggcctttccc gcgcaacccc agcagcagca gcagccgccg
     3541 caccacctgc cccagcagat gcccgtgcag cacccgcggc tgcaggccct gaaggaggag
     3601 cctcagacag tgcccgagat gcccggcgag acaccgcccc tgtcccccat cgacatggag
     3661 tcccaggagc ggatcaaggc ggagaggaag cgcatgagga accgcatcgc tgcctccaag
     3721 tgccgaaaaa ggaagctgga gagaatcgcc cggctggagg aaaaagtgaa aaccttgaaa
     3781 gctcagaact cggagctggc gtccacggcc aacatgctca gggaacaggt ggcacagctt
     3841 aaacagaaag tcatgaacca cgttaacagt gggtgccaac tcatgctaac gcagcagttg
     3901 caaacatttt aagcggccgc tcgagtctag agggcccgtt taaacccgct gatcagcctc
     3961 gactgtgcct tctagttgcc agccatctgt tgtttgcccc tcccccgtgc cttccttgac
     4021 cctggaaggt gccactccca ctgtcctttc ctaataaaat gaggaaattg catcgcattg
     4081 tctgagtagg tgtcattcta ttctgggggg tggggtgggg caggacagca agggggagga
     4141 ttgggaagtc aatagcaggc atgctgggga tgcggtgggc tctatggctt ctgaggcagg
     4201 tcgaagagac tgcccgacac atcttagtga gcgcatcgcg ataagctcta gagggacagc
     4261 ccccccccaa agcccccagg gatgtaatta cgtccctccc ccgctagggg gcagcagcga
     4321 gccgcccggg gctccgctcc ggtccggcgc tccccccgca tccccgagcc ggcagcgtgc
     4381 ggggacagcc cgggcacggg gaaggtggca cgggatcgct ttcctctgaa cgcttctcgc
     4441 tgctctttga gcctgcagac acctgggggg atacggggaa aaagctttag gctgaaagag
     4501 agatttagaa tgacagaatc atagaacggc ctgcgtacgg atggggctct agctctagag
     4561 cttgatatcg aattcctgca gccccgctag agggacagcc cccccccaaa gcccccaggg
     4621 atgtaattac gtccctcccc cgctaggggg cagcagcgag ccgcccgggg ctccgctccg
     4681 gtccggcgct ccccccgcat ccccgagccg gcagcgtgcg gggacagccc gggcacgggg
     4741 aaggtggcac gggatcgctt tcctctgaac gcttctcgct gctctttgag cctgcagaca
     4801 cctgggggga tacggggaaa aagctttagg ctgaaagaga gatttagaat gacagaatca
     4861 tagaacggcc tgggcgcgcc tggtggcgaa cgatcaacag acgggagagg ttacaagtct
     4921 tcgctgacag cgaccggact ctagggggta tccccacgcg ccctgtagcg gcgcattaag
     4981 cgcggcgggt gtggtggtta cgcgcagcgt gaccgctaca cttgccagcg ccctagcgcc
     5041 cgctcctttc gctttcttcc cttcctttct cgccacgttc gccggctttc cccgtcaagc
     5101 tctaaatcgg gggctccctt tagggttccg atttagtgct ttacggcacc tcgaccccaa
     5161 aaaacttgat tagggtgatg gttcacgtag tgggccatcg ccctgataga cggtttttcg
     5221 ccctttgacg ttggagtcca cgttctttaa tagtggactc ttgttccaaa ctggaacaac
     5281 actcaaccct atctcggtct attcttttga tttataaggg attttgccga tttcggccta
     5341 ttggttaaaa aatgagctga tttaacaaaa atttaacgcg aattaattct gtggaatgtg
     5401 tgtcagttag gctcaattag tcagcaacca tagtcccgcc cctaactccg cccatcccgc
     5461 ccctaactcc gcccagttcc gcccattctc cgccccatgg ctgactaatt ttttttattt
     5521 atgcagaggc cgaggccgcc tctgcctctg agctattcca gaagtagtga ggaggctttt
     5581 ttggaggcct aggcttttgc aaaaagctcc cgggagcttg tatatccatt ttcggaggtt
     5641 gggcttcgga atcgttttcc gggacgccgg ctggatgatc ctccagcgcg gggatctcat
     5701 gctggagttc ttcgcccacc ccaacttgtt tattgcagct tataatggtt acaaataaag
     5761 caatagcatc acaaatttca caaataaagc atttttttca ctgcattcta gttgtggttt
     5821 gtccaaactc atcaatgtat cttatcatgt ctgtataccg tcgacctcta gctagagctt
     5881 ggcgtaatca tggtcatagc tgtttcctgt gtgaaattgt tatccgctca caattccaca
     5941 caacatacga gccggaagca taaagtgtaa agcctggggt gcctaatgag tgagctaact
     6001 cacattaatt gcgttgcgct cactgcccgc tttccagtcg ggaaacctgt cgtgccagct
     6061 gcattaatga atcggccaac gcgcggggag aggcggtttg cgtattgggc gctcttccgc
     6121 ttcctcgctc actgactcgc tgcgctcggt cgttcggctg cggcgagcgg tatcagctca
     6181 ctcaaaggcg gtaatacggt tatccacaga atcaggggat aacgcaggaa agaacatgtg
     6241 agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca
     6301 taggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa
     6361 cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg tgcgctctcc
     6421 tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg gaagcgtggc
     6481 gctttctcat agctcacgct gtaggtatct cagttcggtg taggtcgttc gctccaagct
     6541 gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg gtaactatcg
     6601 tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca ctggtaacag
     6661 gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt ggcctaacta
     6721 cggctacact agaagaacag tatttggtat ctgcgctctg ctgaagccag ttaccttcgg
     6781 aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg gtggtttttt
     6841 tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct caagaagatc ctttgatctt
     6901 ttctacgggg tctgacgctc agtggaacga aaactcacgt taagggattt tggtcatgag
     6961 attatcaaaa aggatcttca cctagatcct tttaaattaa aaatgaagtt ttaaatcaat
     7021 ctaaagtata tatgagtaaa cttggtctga cagttaccaa tgcttaatca gtgaggcacc
     7081 tatctcagcg atctgtctat ttcgttcatc catagttgcc tgactccccg tcgtgtagat
     7141 aactacgata cgggagggct taccatctgg ccccagtgct gcaatgatac cgcgagatcc
     7201 acgctcaccg gctccagatt tatcagcaat aaaccagcca gccggaaggg ccgagcgcag
     7261 aagtggtcct gcaactttat ccgcctccat ccagtctatt aattgttgcc gggaagctag
     7321 agtaagtagt tcgccagtta atagtttgcg caacgttgtt gccattgcta caggcatcgt
     7381 ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc ggttcccaac gatcaaggcg
     7441 agttacatga tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt
     7501 tgtcagaagt aagttggccg cagtgttatc actcatggtt atggcagcac tgcataattc
     7561 tcttactgtc atgccatccg taagatgctt ttctgtgact ggtgagtact caaccaagtc
     7621 attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa
     7681 taccgcgcca catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg
     7741 aaaactctca aggatcttac cgctgttgag atccagttcg atgtaaccca ctcgtgcacc
     7801 caactgatct tcagcatctt ttactttcac cagcgtttct gggtgagcaa aaacaggaag
     7861 gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa tgttgaatac tcatactctt
     7921 cctttttcaa tattattgaa gcatttatca gggttattgt ctcatgagcg gatacatatt
     7981 tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc
     8041 acctgacgtc
//
