LOCUS       sequence-489441-        5034 bp    DNA     circular SYN 15-OCT-2025
DEFINITION  Expression of a DsRed-Express2 reporter under a cJun-responsive 
            promoter (cJun homodimer binding sites).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5034)
  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 5034)
  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: L4505 x6TRE2 (cJun2) Ben DsRed-Express2 in TUPV1
FEATURES             Location/Qualifiers
     source          1..5034
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     regulatory      213..222
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             219..896
                     /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(399..419)
                     /label=DsRed1-N
                     /note="DsRed1, reverse primer"
     primer_bind     811..834
                     /label=DsRed1-C
                     /note="DsRed1, forward primer"
     primer_bind     complement(941..958)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    947..1171
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      1940..2368
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2027..2046)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2237..2258
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        2391..2587
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      2438..2573
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     2500..2519
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     polyA_signal    2707..2828
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2744..2763)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2798..2817
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(2877..2893)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2877..2893)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2890..2912)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2901..2917
                     /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(2925..2931,2932..2949,2950..2955))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2970..2991
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3108..3125)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3279..3867)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3359..3378)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4038..4829,4830..4898))
                     /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     4661..4680
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4899..5003)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gacggatcgg gagatcgaag acggtgccaa ggtcttgaca aacgagtgct agtacaagat
       61 ctttacctca tcaagcactt acctcatcaa gcacttacct catcaagcac tgacatcatc
      121 aagcactgac atcatcaagc actgacatca tcaagcactc tagagggtat ataatggggg
      181 ccactagtct actaccagag ctcatcgcta gcgccaccat ggatagcact gagaacgtca
      241 tcaagccctt catgcgcttc aaggtgcaca tggagggctc cgtgaacggc cacgagttcg
      301 agatcgaggg cgagggcgag ggcaagccct acgagggcac ccagaccgcc aagctccagg
      361 tgaccaaggg cggccccctg cccttcgcct gggacatcct gtccccccag ttccagtacg
      421 gctccaaggt gtacgtgaag caccccgccg acatccccga ctacaagaag ctgtccttcc
      481 ccgagggctt caagtgggag cgcgtgatga acttcgagga cggcggcgtg gtgaccgtga
      541 cccaggactc ctccctccag gacggcacct tcatctacca cgtgaagttc atcggcgtga
      601 acttcccctc cgacggcccc gtaatgcaga agaaaactct gggctgggag ccctccaccg
      661 agcgcctgta cccccgcgac ggcgtgctga agggcgagat ccacaaggcg ctgaagctga
      721 agggcggcgg ccactacctg gtggagttca agtcaatcta catggccaag aagcccgtga
      781 agctgcccgg ctactactac gtggactcca agctggacat cacctcccac aacgaggact
      841 acaccgtggt ggagcagtac gagcgcgccg aggcccgcca ccacctgttc cagtaggcgg
      901 ccgctcgagt ctagagggcc cgtttaaacc cgctgatcag cctcgactgt gccttctagt
      961 tgccagccat ctgttgtttg cccctccccc gtgccttcct tgaccctgga aggtgccact
     1021 cccactgtcc tttcctaata aaatgaggaa attgcatcgc attgtctgag taggtgtcat
     1081 tctattctgg ggggtggggt ggggcaggac agcaaggggg aggattggga agtcaatagc
     1141 aggcatgctg gggatgcggt gggctctatg gcttctgagg caggtcgagg gtacatgcgc
     1201 cttactcctt gtgagcgcat cgcgataagc tctagaggga cagccccccc ccaaagcccc
     1261 cagggatgta attacgtccc tcccccgcta gggggcagca gcgagccgcc cggggctccg
     1321 ctccggtccg gcgctccccc cgcatccccg agccggcagc gtgcggggac agcccgggca
     1381 cggggaaggt ggcacgggat cgctttcctc tgaacgcttc tcgctgctct ttgagcctgc
     1441 agacacctgg ggggatacgg ggaaaaagct ttaggctgaa agagagattt agaatgacag
     1501 aatcatagaa cggcctgcgt acggatgggg ctctagctct agagcttgat atcgaattcc
     1561 tgcagccccg ctagagggac agcccccccc caaagccccc agggatgtaa ttacgtccct
     1621 cccccgctag ggggcagcag cgagccgccc ggggctccgc tccggtccgg cgctcccccc
     1681 gcatccccga gccggcagcg tgcggggaca gcccgggcac ggggaaggtg gcacgggatc
     1741 gctttcctct gaacgcttct cgctgctctt tgagcctgca gacacctggg gggatacggg
     1801 gaaaaagctt taggctgaaa gagagattta gaatgacaga atcatagaac ggcctgggcg
     1861 cgcctggtgg cgaacgatca acagacggga gagggcaaaa gtcttcgctg acagcgaccg
     1921 gactctaggg ggtatcccca cgcgccctgt agcggcgcat taagcgcggc gggtgtggtg
     1981 gttacgcgca gcgtgaccgc tacacttgcc agcgccctag cgcccgctcc tttcgctttc
     2041 ttcccttcct ttctcgccac gttcgccggc tttccccgtc aagctctaaa tcgggggctc
     2101 cctttagggt tccgatttag tgctttacgg cacctcgacc ccaaaaaact tgattagggt
     2161 gatggttcac gtagtgggcc atcgccctga tagacggttt ttcgcccttt gacgttggag
     2221 tccacgttct ttaatagtgg actcttgttc caaactggaa caacactcaa ccctatctcg
     2281 gtctattctt ttgatttata agggattttg ccgatttcgg cctattggtt aaaaaatgag
     2341 ctgatttaac aaaaatttaa cgcgaattaa ttctgtggaa tgtgtgtcag ttaggctcaa
     2401 ttagtcagca accatagtcc cgcccctaac tccgcccatc ccgcccctaa ctccgcccag
     2461 ttccgcccat tctccgcccc atggctgact aatttttttt atttatgcag aggccgaggc
     2521 cgcctctgcc tctgagctat tccagaagta gtgaggaggc ttttttggag gcctaggctt
     2581 ttgcaaaaag ctcccgggag cttgtatatc cattttcgga ggttgggctt cggaatcgtt
     2641 ttccgggacg ccggctggat gatcctccag cgcggggatc tcatgctgga gttcttcgcc
     2701 caccccaact tgtttattgc agcttataat ggttacaaat aaagcaatag catcacaaat
     2761 ttcacaaata aagcattttt ttcactgcat tctagttgtg gtttgtccaa actcatcaat
     2821 gtatcttatc atgtctgtat accgtcgacc tctagctaga gcttggcgta atcatggtca
     2881 tagctgtttc ctgtgtgaaa ttgttatccg ctcacaattc cacacaacat acgagccgga
     2941 agcataaagt gtaaagcctg gggtgcctaa tgagtgagct aactcacatt aattgcgttg
     3001 cgctcactgc ccgctttcca gtcgggaaac ctgtcgtgcc agctgcatta atgaatcggc
     3061 caacgcgcgg ggagaggcgg tttgcgtatt gggcgctctt ccgcttcctc gctcactgac
     3121 tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa ggcggtaata
     3181 cggttatcca cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa
     3241 aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt tccataggct ccgcccccct
     3301 gacgagcatc acaaaaatcg acgctcaagt cagaggtggc gaaacccgac aggactataa
     3361 agataccagg cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg
     3421 cttaccggat acctgtccgc ctttctccct tcgggaagcg tggcgctttc tcatagctca
     3481 cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa
     3541 ccccccgttc agcccgaccg ctgcgcctta tccggtaact atcgtcttga gtccaacccg
     3601 gtaagacacg acttatcgcc actggcagca gccactggta acaggattag cagagcgagg
     3661 tatgtaggcg gtgctacaga gttcttgaag tggtggccta actacggcta cactagaaga
     3721 acagtatttg gtatctgcgc tctgctgaag ccagttacct tcggaaaaag agttggtagc
     3781 tcttgatccg gcaaacaaac caccgctggt agcggtggtt tttttgtttg caagcagcag
     3841 attacgcgca gaaaaaaagg atctcaagaa gatcctttga tcttttctac ggggtctgac
     3901 gctcagtgga acgaaaactc acgttaaggg attttggtca tgagattatc aaaaaggatc
     3961 ttcacctaga tccttttaaa ttaaaaatga agttttaaat caatctaaag tatatatgag
     4021 taaacttggt ctgacagtta ccaatgctta atcagtgagg cacctatctc agcgatctgt
     4081 ctatttcgtt catccatagt tgcctgactc cccgtcgtgt agataactac gatacgggag
     4141 ggcttaccat ctggccccag tgctgcaatg ataccgcgag atccacgctc accggctcca
     4201 gatttatcag caataaacca gccagccgga agggccgagc gcagaagtgg tcctgcaact
     4261 ttatccgcct ccatccagtc tattaattgt tgccgggaag ctagagtaag tagttcgcca
     4321 gttaatagtt tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg
     4381 tttggtatgg cttcattcag ctccggttcc caacgatcaa ggcgagttac atgatccccc
     4441 atgttgtgca aaaaagcggt tagctccttc ggtcctccga tcgttgtcag aagtaagttg
     4501 gccgcagtgt tatcactcat ggttatggca gcactgcata attctcttac tgtcatgcca
     4561 tccgtaagat gcttttctgt gactggtgag tactcaacca agtcattctg agaatagtgt
     4621 atgcggcgac cgagttgctc ttgcccggcg tcaatacggg ataataccgc gccacatagc
     4681 agaactttaa aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc
     4741 ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg cacccaactg atcttcagca
     4801 tcttttactt tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa
     4861 aagggaataa gggcgacacg gaaatgttga atactcatac tcttcctttt tcaatattat
     4921 tgaagcattt atcagggtta ttgtctcatg agcggataca tatttgaatg tatttagaaa
     4981 aataaacaaa taggggttcc gcgcacattt ccccgaaaag tgccacctga cgtc
//
