LOCUS       sequence-489481-        5033 bp    DNA     circular SYN 16-OCT-2025
DEFINITION  Expression of a DsRed-Express2 reporter under a cJun-responsive 
            promoter (AP-1 binding sites).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5033)
  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 5033)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Oct 16, 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: L4437 x6TRE (cJun and AP-1) Ben DsRed-Express2...
FEATURES             Location/Qualifiers
     source          1..5033
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     regulatory      212..221
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             218..895
                     /codon_start=1
                     /product="noncytotoxic tetrameric variant of DsRed 
                     fluorescent protein (Strack et al., 2008)"
                     /label=DsRed-Express2
                     /note="mammalian codon-optimized"
                     /translation="MDSTENVIKPFMRFKVHMEGSVNGHEFEIEGEGEGKPYEGTQTAK
                     LQVTKGGPLPFAWDILSPQFQYGSKVYVKHPADIPDYKKLSFPEGFKWERVMNFEDGGV
                     VTVTQDSSLQDGTFIYHVKFIGVNFPSDGPVMQKKTLGWEPSTERLYPRDGVLKGEIHK
                     ALKLKGGGHYLVEFKSIYMAKKPVKLPGYYYVDSKLDITSHNEDYTVVEQYERAEARHH
                     LFQ"
     primer_bind     complement(398..418)
                     /label=DsRed1-N
                     /note="DsRed1, reverse primer"
     primer_bind     810..833
                     /label=DsRed1-C
                     /note="DsRed1, forward primer"
     primer_bind     complement(940..957)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    946..1170
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      1939..2367
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2026..2045)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2236..2257
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        2390..2586
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      2437..2572
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     2499..2518
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     polyA_signal    2706..2827
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2743..2762)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2797..2816
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(2876..2892)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2876..2892)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2889..2911)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2900..2916
                     /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(2924..2930,2931..2948,2949..2954))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2969..2990
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3107..3124)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3278..3866)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3358..3377)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4037..4828,4829..4897))
                     /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     4660..4679
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4898..5002)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gacggatcgg gagatcgaag acggtgccaa ggtcttgaca aacgagtgct agtacaagat
       61 ctgcgagtga gtcatcaagc actgagtcat caagcactga gtcatcaagc actgagtcat
      121 caagcactga gtcatcaagc actgagtcat caagcactct agagggtata taatgggggc
      181 cactagtcta ctaccagagc tcatcgctag cgccaccatg gatagcactg agaacgtcat
      241 caagcccttc atgcgcttca aggtgcacat ggagggctcc gtgaacggcc acgagttcga
      301 gatcgagggc gagggcgagg gcaagcccta cgagggcacc cagaccgcca agctccaggt
      361 gaccaagggc ggccccctgc ccttcgcctg ggacatcctg tccccccagt tccagtacgg
      421 ctccaaggtg tacgtgaagc accccgccga catccccgac tacaagaagc tgtccttccc
      481 cgagggcttc aagtgggagc gcgtgatgaa cttcgaggac ggcggcgtgg tgaccgtgac
      541 ccaggactcc tccctccagg acggcacctt catctaccac gtgaagttca tcggcgtgaa
      601 cttcccctcc gacggccccg taatgcagaa gaaaactctg ggctgggagc cctccaccga
      661 gcgcctgtac ccccgcgacg gcgtgctgaa gggcgagatc cacaaggcgc tgaagctgaa
      721 gggcggcggc cactacctgg tggagttcaa gtcaatctac atggccaaga agcccgtgaa
      781 gctgcccggc tactactacg tggactccaa gctggacatc acctcccaca acgaggacta
      841 caccgtggtg gagcagtacg agcgcgccga ggcccgccac cacctgttcc agtaggcggc
      901 cgctcgagtc tagagggccc gtttaaaccc gctgatcagc ctcgactgtg ccttctagtt
      961 gccagccatc tgttgtttgc ccctcccccg tgccttcctt gaccctggaa ggtgccactc
     1021 ccactgtcct ttcctaataa aatgaggaaa ttgcatcgca ttgtctgagt aggtgtcatt
     1081 ctattctggg gggtggggtg gggcaggaca gcaaggggga ggattgggaa gtcaatagca
     1141 ggcatgctgg ggatgcggtg ggctctatgg cttctgaggc aggtcgaggg tacatgcgcc
     1201 ttactccttg tgagcgcatc gcgataagct ctagagggac agcccccccc caaagccccc
     1261 agggatgtaa ttacgtccct cccccgctag ggggcagcag cgagccgccc ggggctccgc
     1321 tccggtccgg cgctcccccc gcatccccga gccggcagcg tgcggggaca gcccgggcac
     1381 ggggaaggtg gcacgggatc gctttcctct gaacgcttct cgctgctctt tgagcctgca
     1441 gacacctggg gggatacggg gaaaaagctt taggctgaaa gagagattta gaatgacaga
     1501 atcatagaac ggcctgcgta cggatggggc tctagctcta gagcttgata tcgaattcct
     1561 gcagccccgc tagagggaca gccccccccc aaagccccca gggatgtaat tacgtccctc
     1621 ccccgctagg gggcagcagc gagccgcccg gggctccgct ccggtccggc gctccccccg
     1681 catccccgag ccggcagcgt gcggggacag cccgggcacg gggaaggtgg cacgggatcg
     1741 ctttcctctg aacgcttctc gctgctcttt gagcctgcag acacctgggg ggatacgggg
     1801 aaaaagcttt aggctgaaag agagatttag aatgacagaa tcatagaacg gcctgggcgc
     1861 gcctggtggc gaacgatcaa cagacgggag agggcaaaag tcttcgctga cagcgaccgg
     1921 actctagggg gtatccccac gcgccctgta gcggcgcatt aagcgcggcg ggtgtggtgg
     1981 ttacgcgcag cgtgaccgct acacttgcca gcgccctagc gcccgctcct ttcgctttct
     2041 tcccttcctt tctcgccacg ttcgccggct ttccccgtca agctctaaat cgggggctcc
     2101 ctttagggtt ccgatttagt gctttacggc acctcgaccc caaaaaactt gattagggtg
     2161 atggttcacg tagtgggcca tcgccctgat agacggtttt tcgccctttg acgttggagt
     2221 ccacgttctt taatagtgga ctcttgttcc aaactggaac aacactcaac cctatctcgg
     2281 tctattcttt tgatttataa gggattttgc cgatttcggc ctattggtta aaaaatgagc
     2341 tgatttaaca aaaatttaac gcgaattaat tctgtggaat gtgtgtcagt taggctcaat
     2401 tagtcagcaa ccatagtccc gcccctaact ccgcccatcc cgcccctaac tccgcccagt
     2461 tccgcccatt ctccgcccca tggctgacta atttttttta tttatgcaga ggccgaggcc
     2521 gcctctgcct ctgagctatt ccagaagtag tgaggaggct tttttggagg cctaggcttt
     2581 tgcaaaaagc tcccgggagc ttgtatatcc attttcggag gttgggcttc ggaatcgttt
     2641 tccgggacgc cggctggatg atcctccagc gcggggatct catgctggag ttcttcgccc
     2701 accccaactt gtttattgca gcttataatg gttacaaata aagcaatagc atcacaaatt
     2761 tcacaaataa agcatttttt tcactgcatt ctagttgtgg tttgtccaaa ctcatcaatg
     2821 tatcttatca tgtctgtata ccgtcgacct ctagctagag cttggcgtaa tcatggtcat
     2881 agctgtttcc tgtgtgaaat tgttatccgc tcacaattcc acacaacata cgagccggaa
     2941 gcataaagtg taaagcctgg ggtgcctaat gagtgagcta actcacatta attgcgttgc
     3001 gctcactgcc cgctttccag tcgggaaacc tgtcgtgcca gctgcattaa tgaatcggcc
     3061 aacgcgcggg gagaggcggt ttgcgtattg ggcgctcttc cgcttcctcg ctcactgact
     3121 cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag gcggtaatac
     3181 ggttatccac agaatcaggg gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa
     3241 aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc cgcccccctg
     3301 acgagcatca caaaaatcga cgctcaagtc agaggtggcg aaacccgaca ggactataaa
     3361 gataccaggc gtttccccct ggaagctccc tcgtgcgctc tcctgttccg accctgccgc
     3421 ttaccggata cctgtccgcc tttctccctt cgggaagcgt ggcgctttct catagctcac
     3481 gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac
     3541 cccccgttca gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag tccaacccgg
     3601 taagacacga cttatcgcca ctggcagcag ccactggtaa caggattagc agagcgaggt
     3661 atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa ctacggctac actagaagaa
     3721 cagtatttgg tatctgcgct ctgctgaagc cagttacctt cggaaaaaga gttggtagct
     3781 cttgatccgg caaacaaacc accgctggta gcggtggttt ttttgtttgc aagcagcaga
     3841 ttacgcgcag aaaaaaagga tctcaagaag atcctttgat cttttctacg gggtctgacg
     3901 ctcagtggaa cgaaaactca cgttaaggga ttttggtcat gagattatca aaaaggatct
     3961 tcacctagat ccttttaaat taaaaatgaa gttttaaatc aatctaaagt atatatgagt
     4021 aaacttggtc tgacagttac caatgcttaa tcagtgaggc acctatctca gcgatctgtc
     4081 tatttcgttc atccatagtt gcctgactcc ccgtcgtgta gataactacg atacgggagg
     4141 gcttaccatc tggccccagt gctgcaatga taccgcgaga tccacgctca ccggctccag
     4201 atttatcagc aataaaccag ccagccggaa gggccgagcg cagaagtggt cctgcaactt
     4261 tatccgcctc catccagtct attaattgtt gccgggaagc tagagtaagt agttcgccag
     4321 ttaatagttt gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt
     4381 ttggtatggc ttcattcagc tccggttccc aacgatcaag gcgagttaca tgatccccca
     4441 tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat cgttgtcaga agtaagttgg
     4501 ccgcagtgtt atcactcatg gttatggcag cactgcataa ttctcttact gtcatgccat
     4561 ccgtaagatg cttttctgtg actggtgagt actcaaccaa gtcattctga gaatagtgta
     4621 tgcggcgacc gagttgctct tgcccggcgt caatacggga taataccgcg ccacatagca
     4681 gaactttaaa agtgctcatc attggaaaac gttcttcggg gcgaaaactc tcaaggatct
     4741 taccgctgtt gagatccagt tcgatgtaac ccactcgtgc acccaactga tcttcagcat
     4801 cttttacttt caccagcgtt tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa
     4861 agggaataag ggcgacacgg aaatgttgaa tactcatact cttccttttt caatattatt
     4921 gaagcattta tcagggttat tgtctcatga gcggatacat atttgaatgt atttagaaaa
     4981 ataaacaaat aggggttccg cgcacatttc cccgaaaagt gccacctgac gtc
//
