LOCUS       sequence-489411-        5035 bp    DNA     circular SYN 15-OCT-2025
DEFINITION  Expression of a DsRed-Express2 reporter under a hybrid 
            synTF-responsive promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5035)
  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 5035)
  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: L4832 ZF1+ZF6 x6c interspersed Ben DsRed-Expre...
FEATURES             Location/Qualifiers
     source          1..5035
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     regulatory      214..223
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             220..897
                     /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(400..420)
                     /label=DsRed1-N
                     /note="DsRed1, reverse primer"
     primer_bind     812..835
                     /label=DsRed1-C
                     /note="DsRed1, forward primer"
     primer_bind     complement(942..959)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    948..1172
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      1941..2369
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2028..2047)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2238..2259
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        2392..2588
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      2439..2574
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     2501..2520
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     polyA_signal    2708..2829
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2745..2764)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2799..2818
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(2878..2894)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2878..2894)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2891..2913)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2902..2918
                     /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(2926..2932,2933..2950,2951..2956))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2971..2992
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3109..3126)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3280..3868)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3360..3379)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4039..4830,4831..4899))
                     /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     4662..4681
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4900..5004)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gacggatcgg gagatcgaag acggtgccaa ggtcttgaca aacgagtgct agtacaagat
       61 ctgcgagaga gtgaggactc gtggtgtagg ggttataaga gtgaggacaa ttggtgtagg
      121 ggtatgtaga gtgaggacaa cgggtgtagg ggtacgctct ctagagggta tataatgggg
      181 gccactagtc tactaccaga gctcatcgct agcgccacca tggatagcac tgagaacgtc
      241 atcaagccct tcatgcgctt caaggtgcac atggagggct ccgtgaacgg ccacgagttc
      301 gagatcgagg gcgagggcga gggcaagccc tacgagggca cccagaccgc caagctccag
      361 gtgaccaagg gcggccccct gcccttcgcc tgggacatcc tgtcccccca gttccagtac
      421 ggctccaagg tgtacgtgaa gcaccccgcc gacatccccg actacaagaa gctgtccttc
      481 cccgagggct tcaagtggga gcgcgtgatg aacttcgagg acggcggcgt ggtgaccgtg
      541 acccaggact cctccctcca ggacggcacc ttcatctacc acgtgaagtt catcggcgtg
      601 aacttcccct ccgacggccc cgtaatgcag aagaaaactc tgggctggga gccctccacc
      661 gagcgcctgt acccccgcga cggcgtgctg aagggcgaga tccacaaggc gctgaagctg
      721 aagggcggcg gccactacct ggtggagttc aagtcaatct acatggccaa gaagcccgtg
      781 aagctgcccg gctactacta cgtggactcc aagctggaca tcacctccca caacgaggac
      841 tacaccgtgg tggagcagta cgagcgcgcc gaggcccgcc accacctgtt ccagtaggcg
      901 gccgctcgag tctagagggc ccgtttaaac ccgctgatca gcctcgactg tgccttctag
      961 ttgccagcca tctgttgttt gcccctcccc cgtgccttcc ttgaccctgg aaggtgccac
     1021 tcccactgtc ctttcctaat aaaatgagga aattgcatcg cattgtctga gtaggtgtca
     1081 ttctattctg gggggtgggg tggggcagga cagcaagggg gaggattggg aagtcaatag
     1141 caggcatgct ggggatgcgg tgggctctat ggcttctgag gcaggtcgag ggtacatgcg
     1201 ccttactcct tgtgagcgca tcgcgataag ctctagaggg acagcccccc cccaaagccc
     1261 ccagggatgt aattacgtcc ctcccccgct agggggcagc agcgagccgc ccggggctcc
     1321 gctccggtcc ggcgctcccc ccgcatcccc gagccggcag cgtgcgggga cagcccgggc
     1381 acggggaagg tggcacggga tcgctttcct ctgaacgctt ctcgctgctc tttgagcctg
     1441 cagacacctg gggggatacg gggaaaaagc tttaggctga aagagagatt tagaatgaca
     1501 gaatcataga acggcctgcg tacggatggg gctctagctc tagagcttga tatcgaattc
     1561 ctgcagcccc gctagaggga cagccccccc ccaaagcccc cagggatgta attacgtccc
     1621 tcccccgcta gggggcagca gcgagccgcc cggggctccg ctccggtccg gcgctccccc
     1681 cgcatccccg agccggcagc gtgcggggac agcccgggca cggggaaggt ggcacgggat
     1741 cgctttcctc tgaacgcttc tcgctgctct ttgagcctgc agacacctgg ggggatacgg
     1801 ggaaaaagct ttaggctgaa agagagattt agaatgacag aatcatagaa cggcctgggc
     1861 gcgcctggtg gcgaacgatc aacagacggg agagggcaaa agtcttcgct gacagcgacc
     1921 ggactctagg gggtatcccc acgcgccctg tagcggcgca ttaagcgcgg cgggtgtggt
     1981 ggttacgcgc agcgtgaccg ctacacttgc cagcgcccta gcgcccgctc ctttcgcttt
     2041 cttcccttcc tttctcgcca cgttcgccgg ctttccccgt caagctctaa atcgggggct
     2101 ccctttaggg ttccgattta gtgctttacg gcacctcgac cccaaaaaac ttgattaggg
     2161 tgatggttca cgtagtgggc catcgccctg atagacggtt tttcgccctt tgacgttgga
     2221 gtccacgttc tttaatagtg gactcttgtt ccaaactgga acaacactca accctatctc
     2281 ggtctattct tttgatttat aagggatttt gccgatttcg gcctattggt taaaaaatga
     2341 gctgatttaa caaaaattta acgcgaatta attctgtgga atgtgtgtca gttaggctca
     2401 attagtcagc aaccatagtc ccgcccctaa ctccgcccat cccgccccta actccgccca
     2461 gttccgccca ttctccgccc catggctgac taattttttt tatttatgca gaggccgagg
     2521 ccgcctctgc ctctgagcta ttccagaagt agtgaggagg cttttttgga ggcctaggct
     2581 tttgcaaaaa gctcccggga gcttgtatat ccattttcgg aggttgggct tcggaatcgt
     2641 tttccgggac gccggctgga tgatcctcca gcgcggggat ctcatgctgg agttcttcgc
     2701 ccaccccaac ttgtttattg cagcttataa tggttacaaa taaagcaata gcatcacaaa
     2761 tttcacaaat aaagcatttt tttcactgca ttctagttgt ggtttgtcca aactcatcaa
     2821 tgtatcttat catgtctgta taccgtcgac ctctagctag agcttggcgt aatcatggtc
     2881 atagctgttt cctgtgtgaa attgttatcc gctcacaatt ccacacaaca tacgagccgg
     2941 aagcataaag tgtaaagcct ggggtgccta atgagtgagc taactcacat taattgcgtt
     3001 gcgctcactg cccgctttcc agtcgggaaa cctgtcgtgc cagctgcatt aatgaatcgg
     3061 ccaacgcgcg gggagaggcg gtttgcgtat tgggcgctct tccgcttcct cgctcactga
     3121 ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca gctcactcaa aggcggtaat
     3181 acggttatcc acagaatcag gggataacgc aggaaagaac atgtgagcaa aaggccagca
     3241 aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc tccgcccccc
     3301 tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga caggactata
     3361 aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc
     3421 gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt ctcatagctc
     3481 acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga
     3541 accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc
     3601 ggtaagacac gacttatcgc cactggcagc agccactggt aacaggatta gcagagcgag
     3661 gtatgtaggc ggtgctacag agttcttgaa gtggtggcct aactacggct acactagaag
     3721 aacagtattt ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa gagttggtag
     3781 ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt gcaagcagca
     3841 gattacgcgc agaaaaaaag gatctcaaga agatcctttg atcttttcta cggggtctga
     3901 cgctcagtgg aacgaaaact cacgttaagg gattttggtc atgagattat caaaaaggat
     3961 cttcacctag atccttttaa attaaaaatg aagttttaaa tcaatctaaa gtatatatga
     4021 gtaaacttgg tctgacagtt accaatgctt aatcagtgag gcacctatct cagcgatctg
     4081 tctatttcgt tcatccatag ttgcctgact ccccgtcgtg tagataacta cgatacggga
     4141 gggcttacca tctggcccca gtgctgcaat gataccgcga gatccacgct caccggctcc
     4201 agatttatca gcaataaacc agccagccgg aagggccgag cgcagaagtg gtcctgcaac
     4261 tttatccgcc tccatccagt ctattaattg ttgccgggaa gctagagtaa gtagttcgcc
     4321 agttaatagt ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc
     4381 gtttggtatg gcttcattca gctccggttc ccaacgatca aggcgagtta catgatcccc
     4441 catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca gaagtaagtt
     4501 ggccgcagtg ttatcactca tggttatggc agcactgcat aattctctta ctgtcatgcc
     4561 atccgtaaga tgcttttctg tgactggtga gtactcaacc aagtcattct gagaatagtg
     4621 tatgcggcga ccgagttgct cttgcccggc gtcaatacgg gataataccg cgccacatag
     4681 cagaacttta aaagtgctca tcattggaaa acgttcttcg gggcgaaaac tctcaaggat
     4741 cttaccgctg ttgagatcca gttcgatgta acccactcgt gcacccaact gatcttcagc
     4801 atcttttact ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa
     4861 aaagggaata agggcgacac ggaaatgttg aatactcata ctcttccttt ttcaatatta
     4921 ttgaagcatt tatcagggtt attgtctcat gagcggatac atatttgaat gtatttagaa
     4981 aaataaacaa ataggggttc cgcgcacatt tccccgaaaa gtgccacctg acgtc
//
