LOCUS       sequence-489483-        5057 bp    DNA     circular SYN 16-OCT-2025
DEFINITION  Expression of a DsRed-Express2 reporter under a BATF-responsive 
            promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5057)
  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 5057)
  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: L4459 x6BRE (BATF) Ben DsRed-Express2 in TUPV1
FEATURES             Location/Qualifiers
     source          1..5057
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     regulatory      236..245
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             242..919
                     /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(422..442)
                     /label=DsRed1-N
                     /note="DsRed1, reverse primer"
     primer_bind     834..857
                     /label=DsRed1-C
                     /note="DsRed1, forward primer"
     primer_bind     complement(964..981)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    970..1194
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      1963..2391
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2050..2069)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2260..2281
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        2414..2610
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      2461..2596
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     2523..2542
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     polyA_signal    2730..2851
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2767..2786)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2821..2840
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(2900..2916)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2900..2916)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2913..2935)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2924..2940
                     /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(2948..2954,2955..2972,2973..2978))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2993..3014
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3131..3148)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3302..3890)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3382..3401)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4061..4852,4853..4921))
                     /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     4684..4703
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4922..5026)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gacggatcgg gagatcgaag acggtgccaa ggtcttgaca aacgagtgct agtacaagat
       61 ctgcgaggaa atgagtcatc aagcacgaaa tgagtcatca agcacgaaat gagtcatcaa
      121 gcacgaaatg agtcatcaag cacgaaatga gtcatcaagc acgaaatgag tcatcaagca
      181 ctctagaggg tatataatgg gggccactag tctactacca gagctcatcg ctagcgccac
      241 catggatagc actgagaacg tcatcaagcc cttcatgcgc ttcaaggtgc acatggaggg
      301 ctccgtgaac ggccacgagt tcgagatcga gggcgagggc gagggcaagc cctacgaggg
      361 cacccagacc gccaagctcc aggtgaccaa gggcggcccc ctgcccttcg cctgggacat
      421 cctgtccccc cagttccagt acggctccaa ggtgtacgtg aagcaccccg ccgacatccc
      481 cgactacaag aagctgtcct tccccgaggg cttcaagtgg gagcgcgtga tgaacttcga
      541 ggacggcggc gtggtgaccg tgacccagga ctcctccctc caggacggca ccttcatcta
      601 ccacgtgaag ttcatcggcg tgaacttccc ctccgacggc cccgtaatgc agaagaaaac
      661 tctgggctgg gagccctcca ccgagcgcct gtacccccgc gacggcgtgc tgaagggcga
      721 gatccacaag gcgctgaagc tgaagggcgg cggccactac ctggtggagt tcaagtcaat
      781 ctacatggcc aagaagcccg tgaagctgcc cggctactac tacgtggact ccaagctgga
      841 catcacctcc cacaacgagg actacaccgt ggtggagcag tacgagcgcg ccgaggcccg
      901 ccaccacctg ttccagtagg cggccgctcg agtctagagg gcccgtttaa acccgctgat
      961 cagcctcgac tgtgccttct agttgccagc catctgttgt ttgcccctcc cccgtgcctt
     1021 ccttgaccct ggaaggtgcc actcccactg tcctttccta ataaaatgag gaaattgcat
     1081 cgcattgtct gagtaggtgt cattctattc tggggggtgg ggtggggcag gacagcaagg
     1141 gggaggattg ggaagtcaat agcaggcatg ctggggatgc ggtgggctct atggcttctg
     1201 aggcaggtcg agggtacatg cgccttactc cttgtgagcg catcgcgata agctctagag
     1261 ggacagcccc cccccaaagc ccccagggat gtaattacgt ccctcccccg ctagggggca
     1321 gcagcgagcc gcccggggct ccgctccggt ccggcgctcc ccccgcatcc ccgagccggc
     1381 agcgtgcggg gacagcccgg gcacggggaa ggtggcacgg gatcgctttc ctctgaacgc
     1441 ttctcgctgc tctttgagcc tgcagacacc tggggggata cggggaaaaa gctttaggct
     1501 gaaagagaga tttagaatga cagaatcata gaacggcctg cgtacggatg gggctctagc
     1561 tctagagctt gatatcgaat tcctgcagcc ccgctagagg gacagccccc ccccaaagcc
     1621 cccagggatg taattacgtc cctcccccgc tagggggcag cagcgagccg cccggggctc
     1681 cgctccggtc cggcgctccc cccgcatccc cgagccggca gcgtgcgggg acagcccggg
     1741 cacggggaag gtggcacggg atcgctttcc tctgaacgct tctcgctgct ctttgagcct
     1801 gcagacacct ggggggatac ggggaaaaag ctttaggctg aaagagagat ttagaatgac
     1861 agaatcatag aacggcctgg gcgcgcctgg tggcgaacga tcaacagacg ggagagggca
     1921 aaagtcttcg ctgacagcga ccggactcta gggggtatcc ccacgcgccc tgtagcggcg
     1981 cattaagcgc ggcgggtgtg gtggttacgc gcagcgtgac cgctacactt gccagcgccc
     2041 tagcgcccgc tcctttcgct ttcttccctt cctttctcgc cacgttcgcc ggctttcccc
     2101 gtcaagctct aaatcggggg ctccctttag ggttccgatt tagtgcttta cggcacctcg
     2161 accccaaaaa acttgattag ggtgatggtt cacgtagtgg gccatcgccc tgatagacgg
     2221 tttttcgccc tttgacgttg gagtccacgt tctttaatag tggactcttg ttccaaactg
     2281 gaacaacact caaccctatc tcggtctatt cttttgattt ataagggatt ttgccgattt
     2341 cggcctattg gttaaaaaat gagctgattt aacaaaaatt taacgcgaat taattctgtg
     2401 gaatgtgtgt cagttaggct caattagtca gcaaccatag tcccgcccct aactccgccc
     2461 atcccgcccc taactccgcc cagttccgcc cattctccgc cccatggctg actaattttt
     2521 tttatttatg cagaggccga ggccgcctct gcctctgagc tattccagaa gtagtgagga
     2581 ggcttttttg gaggcctagg cttttgcaaa aagctcccgg gagcttgtat atccattttc
     2641 ggaggttggg cttcggaatc gttttccggg acgccggctg gatgatcctc cagcgcgggg
     2701 atctcatgct ggagttcttc gcccacccca acttgtttat tgcagcttat aatggttaca
     2761 aataaagcaa tagcatcaca aatttcacaa ataaagcatt tttttcactg cattctagtt
     2821 gtggtttgtc caaactcatc aatgtatctt atcatgtctg tataccgtcg acctctagct
     2881 agagcttggc gtaatcatgg tcatagctgt ttcctgtgtg aaattgttat ccgctcacaa
     2941 ttccacacaa catacgagcc ggaagcataa agtgtaaagc ctggggtgcc taatgagtga
     3001 gctaactcac attaattgcg ttgcgctcac tgcccgcttt ccagtcggga aacctgtcgt
     3061 gccagctgca ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt attgggcgct
     3121 cttccgcttc ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat
     3181 cagctcactc aaaggcggta atacggttat ccacagaatc aggggataac gcaggaaaga
     3241 acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt
     3301 ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt
     3361 ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc
     3421 gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa
     3481 gcgtggcgct ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct
     3541 ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta
     3601 actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg
     3661 gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc
     3721 ctaactacgg ctacactaga agaacagtat ttggtatctg cgctctgctg aagccagtta
     3781 ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg
     3841 gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt
     3901 tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg
     3961 tcatgagatt atcaaaaagg atcttcacct agatcctttt aaattaaaaa tgaagtttta
     4021 aatcaatcta aagtatatat gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg
     4081 aggcacctat ctcagcgatc tgtctatttc gttcatccat agttgcctga ctccccgtcg
     4141 tgtagataac tacgatacgg gagggcttac catctggccc cagtgctgca atgataccgc
     4201 gagatccacg ctcaccggct ccagatttat cagcaataaa ccagccagcc ggaagggccg
     4261 agcgcagaag tggtcctgca actttatccg cctccatcca gtctattaat tgttgccggg
     4321 aagctagagt aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc attgctacag
     4381 gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt cagctccggt tcccaacgat
     4441 caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc
     4501 cgatcgttgt cagaagtaag ttggccgcag tgttatcact catggttatg gcagcactgc
     4561 ataattctct tactgtcatg ccatccgtaa gatgcttttc tgtgactggt gagtactcaa
     4621 ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac
     4681 gggataatac cgcgccacat agcagaactt taaaagtgct catcattgga aaacgttctt
     4741 cggggcgaaa actctcaagg atcttaccgc tgttgagatc cagttcgatg taacccactc
     4801 gtgcacccaa ctgatcttca gcatctttta ctttcaccag cgtttctggg tgagcaaaaa
     4861 caggaaggca aaatgccgca aaaaagggaa taagggcgac acggaaatgt tgaatactca
     4921 tactcttcct ttttcaatat tattgaagca tttatcaggg ttattgtctc atgagcggat
     4981 acatatttga atgtatttag aaaaataaac aaataggggt tccgcgcaca tttccccgaa
     5041 aagtgccacc tgacgtc
//
