LOCUS       sequence-466377-        7691 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Optimized IL-2-sensing eLIDAR with IL-2 beta chain ecto+TM domains 
            fused with ADAR2dd, with Golgi traffic sequence and ER export 
            sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7691)
  AUTHORS   Zhang X, Mille-Fragoso LS, Kaseniit KE, Lee AP, Zhang M, Call CC, Hu
            Y, Xie Y, Gao XJ
  TITLE     Post-transcriptional modular synthetic receptors.
  JOURNAL   Nat Chem Biol. 2025 Mar 28. doi: 10.1038/s41589-025-01872-w.
  PUBMED    40155716
REFERENCE   2  (bases 1 to 7691)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Nat Chem 
            Biol. 2025 Mar 28. doi: 10.1038/s41589-025-01872-w."
COMMENT     Sequence Label: XZ375
            EF1a-IL2Rbsp-3xFLAG-IL2Rb(ECD+TM)-TS-ADA...
FEATURES             Location/Qualifiers
     source          1..7691
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(300..323)
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     regulatory      394..403
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     regulatory      398..407
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     intron          429..1367
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
     primer_bind     complement(452..472)
                     /label=EF1a-F
                     /note="Human elongation factor-1a promoter, forward primer"
     promoter        complement(1368..1579)
                     /label=EF-1-alpha core promoter
                     /note="core promoter for human elongation factor 
                     EF-1-alpha"
     primer_bind     1706..1725
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        1800..1904
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1905..1973,1974..2765)
                     /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     complement(2123..2142)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2936..3524
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3425..3444
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3678..3695
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    3812..3833
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(3848..3853,3854..3871,3872..3878)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3886..3902
                     /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)."
     primer_bind     3891..3913
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     3910..3926
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     3910..3926
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     polyA_signal    3975..4096
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3986..4005)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     4040..4059
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     CDS             complement(4226..5248)
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="KKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRGY
                     VLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLPE
                     TELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQT
                     VMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMFG
                     DSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDGN
                     FDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAKE
                     "
     CDS             complement(4226..5248)
                     /codon_start=1
                     /gene="aph(4)-Ia"
                     /product="aminoglycoside phosphotransferase from E. coli"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="KKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRGY
                     VLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLPE
                     TELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQT
                     VMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMFG
                     DSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDGN
                     FDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAKE
                     "
     protein_bind    complement(5256..5303)
                     /label=FRT
                     /bound_moiety="FLP recombinase from the Saccharomyces 
                     cerevisiae 2u plasmid"
                     /note="FLP-mediated recombination occurs in the 8-bp core 
                     sequence TCTAGAAA (Turan and Bode, 2011)."
     primer_bind     5435..5454
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     polyA_signal    5626..5850
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     5839..5856
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     CDS             complement(5912..5941)
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
ORIGIN
        1 aagtcctgga tggccatcac cctccatcgc accccctcac ggcacagcac cctcagggtg
       61 acgatgtcaa ctgtggtcag tttctgagaa tctggggctc cgaggatcag gttgcaggcc
      121 caggatgctt gactcacggg gagcagctca caggtttggt tccaccgccg tctgtccggc
      181 caggcatgga cttggcagga agtgtcctgc agagccccat cttggctcca gacacaggag
      241 atgttggctc tcgaattgta gaagcatgtg aactgggaag tgccattcac cgcggtaccc
      301 ttatcatcgt catccttgta gtctgcagat gcccaagagg tagccagggg caggaggagg
      361 atgaggaggg gcagacgcca ggacagagca ggggccgcca tggtggcgaa ctagtagctt
      421 cacgacacct gaaatggaag aaaaaaactt tgaaccactg tctgaggctt gagaatgaac
      481 caagatccaa actcaaaaag ggcaaattcc aaggagaatt acatcaagtg ccaagctggc
      541 ctaacttcag tctccaccca ctcagtgtgg ggaaactcca tcgcataaaa cccctccccc
      601 caacctaaag acgacgtact ccaaaagcgc gagaactaat cgaggtgcct ggacggcgcc
      661 cggtactccg tggagtcaca tgaagcgacg gctgaggacg gaaaggccct tttcctttgt
      721 gtgggtgact cacccgcccg ctctcccgag cgccgcgtcc tccattttga gctccctgca
      781 gcagggccgg gaagcggcca tctttccgct cacgcaactg gtgccgaccg ggccagcctt
      841 gccgcccagg gcggggcgat acacggcggc gcgaggccag gcaccagagc aggccggcca
      901 gcttgagact acccccgtcc gattctcggt ggccgcgctc gcaggccccg cctcgccgaa
      961 catgtgcgct gggacgcacg ggccccgtcg ccgcccgcgg ccccaaaaac cgaaatacca
     1021 gtgtgcagat cttggcccgc atttacaaga ctatcttgcc agaaaaaaag cgtcgcagca
     1081 ggtcatcaaa aattttaaat ggctagagac ttatcgaaag cagcgagaca ggcgcgaagg
     1141 tgccaccaga ttcgcacgcg gcggccccag cgcccaggcc aggcctcaac tcaagcacga
     1201 ggcgaagggg ctccttaagc gcaaggcctc gaactctccc acccacttcc aacccgaagc
     1261 tcgggatcaa gaatcacgta ctgcagccag gtggaagtaa ttcaaggcac gcaagggcca
     1321 taacccgtaa agaggccagg cccgcgggaa ccacacacgg cacttacctg tgttctggcg
     1381 gcaaacccgt tgcgaaaaag aacgttcacg gcgactactg cacttatata cggttctccc
     1441 ccaccctcgg gaaaaaggcg gagccagtac acgacatcac tttcccagtt taccccgcgc
     1501 caccttctct aggcaccggt tcaattgccg acccctcccc ccaacttctc ggggactgtg
     1561 ggcgatgtgc gctctgccca ctgacgggca cgattcttca tgcaattgtc ggtcaagcct
     1621 tgccttgttg tagcttaaat tttgctcgcg cactactcag cgacctccaa cacacaagca
     1681 gggagcagat actggcttaa ctatgcggca tcagagcaga ttgtactgag agtgcaccat
     1741 aggggatcgg gagatctccc gatccgtcga cgtcaggtgg cacttttcgg ggaaatgtgc
     1801 gcggaacccc tatttgttta tttttctaaa tacattcaaa tatgtatccg ctcatgagac
     1861 aataaccctg ataaatgctt caataatatt gaaaaaggaa gagtatgagt attcaacatt
     1921 tccgtgtcgc ccttattccc ttttttgcgg cattttgcct tcctgttttt gctcacccag
     1981 aaacgctggt gaaagtaaaa gatgctgaag atcagttggg tgcacgagtg ggttacatcg
     2041 aactggatct caacagcggt aagatccttg agagttttcg ccccgaagaa cgttttccaa
     2101 tgatgagcac ttttaaagtt ctgctatgtg gcgcggtatt atcccgtatt gacgccgggc
     2161 aagagcaact cggtcgccgc atacactatt ctcagaatga cttggttgag tactcaccag
     2221 tcacagaaaa gcatcttacg gatggcatga cagtaagaga attatgcagt gctgccataa
     2281 ccatgagtga taacactgcg gccaacttac ttctgacaac gatcggagga ccgaaggagc
     2341 taaccgcttt tttgcacaac atgggggatc atgtaactcg ccttgatcgt tgggaaccgg
     2401 agctgaatga agccatacca aacgacgagc gtgacaccac gatgcctgta gcaatggcaa
     2461 caacgttgcg caaactatta actggcgaac tacttactct agcttcccgg caacaattaa
     2521 tagactggat ggaggcggat aaagttgcag gaccacttct gcgctcggcc cttccggctg
     2581 gctggtttat tgctgataaa tctggagccg gtgagcgtgg gtctcgcggt atcattgcag
     2641 cactggggcc agatggtaag ccctcccgta tcgtagttat ctacacgacg gggagtcagg
     2701 caactatgga tgaacgaaat agacagatcg ctgagatagg tgcctcactg attaagcatt
     2761 ggtaactgtc agaccaagtt tactcatata tactttagat tgatttaaaa cttcattttt
     2821 aatttaaaag gatctaggtg aagatccttt ttgataatct catgaccaaa atcccttaac
     2881 gtgagttttc gttccactga gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag
     2941 atcctttttt tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg
     3001 tggtttgttt gccggatcaa gagctaccaa ctctttttcc gaaggtaact ggcttcagca
     3061 gagcgcagat accaaatact gttcttctag tgtagccgta gttaggccac cacttcaaga
     3121 actctgtagc accgcctaca tacctcgctc tgctaatcct gttaccagtg gctgctgcca
     3181 gtggcgataa gtcgtgtctt accgggttgg actcaagacg atagttaccg gataaggcgc
     3241 agcggtcggg ctgaacgggg ggttcgtgca cacagcccag cttggagcga acgacctaca
     3301 ccgaactgag atacctacag cgtgagctat gagaaagcgc cacgcttccc gaagggagaa
     3361 aggcggacag gtatccggta agcggcaggg tcggaacagg agagcgcacg agggagcttc
     3421 cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc
     3481 gtcgattttt gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg
     3541 cctttttacg gttcctggcc ttttgctggc cttttgctca catgttcttt cctgcgttat
     3601 cccctgattc tgtggataac cgtattaccg cctttgagtg agctgatacc gctcgccgca
     3661 gccgaacgac cgagcgcagc gagtcagtga gcgaggaagc ggaagagcgc ccaatacgca
     3721 aaccgcctct ccccgcgcgt tggccgattc attaatgcag ctggcacgac aggtttcccg
     3781 actggaaagc gggcagtgag cgcaacgcaa ttaatgtgag ttagctcact cattaggcac
     3841 cccaggcttt acactttatg cttccggctc gtatgttgtg tggaattgtg agcggataac
     3901 aatttcacac aggaaacagc tatgaccatg attacgccaa gctctagcta gaggtcgacg
     3961 gtatacagac atgataagat acattgatga gtttggacaa accacaacta gaatgcagtg
     4021 aaaaaaatgc tttatttgtg aaatttgtga tgctattgct ttatttgtaa ccattataag
     4081 ctgcaataaa caagttgggg tgggcgaaga actccagcat gagatccccg cgctggagga
     4141 tcatccagcc ggcgtcccgg aaaacgattc cgaagcccaa cctttcatag aaggcggcgg
     4201 tggaatcgaa atctcgtagt acgtgctatt cctttgccct cggacgagtg ctggggcgtc
     4261 ggtttccact atcggcgagt acttctacac agccatcggt ccagacggcc gcgcttctgc
     4321 gggcgatttg tgtacgcccg acagtcccgg ctccggatcg gacgattgcg tcgcatcgac
     4381 cctgcgccca agctgcatca tcgaaattgc cgtcaaccaa gctctgatag agttggtcaa
     4441 gaccaatgcg gagcatatac gcccggagcc gcggcgatcc tgcaagctcc ggatgcctcc
     4501 gctcgaagta gcgcgtctgc tgctccatac aagccaacca cggcctccag aagaagatgt
     4561 tggcgacctc gtattgggaa tccccgaaca tcgcctcgct ccagtcaatg accgctgtta
     4621 tgcggccatt gtccgtcagg acattgttgg agccgaaatc cgcgtgcacg aggtgccgga
     4681 cttcggggca gtcctcggcc caaagcatca gctcatcgag agcctgcgcg acggacgcac
     4741 tgacggtgtc gtccatcaca gtttgccagt gatacacatg gggatcagca atcgcgcata
     4801 tgaaatcacg ccatgtagtg tattgaccga ttccttgcgg tccgaatggg ccgaacccgc
     4861 tcgtctggct aagatcggcc gcagcgatcg catccatggc ctccgcgacc ggctgcagaa
     4921 cagcgggcag ttcggtttca ggcaggtctt gcaacgtgac accctgtgca cggcgggaga
     4981 tgcaataggt caggctctcg ctgaattccc caatgtcaag cacttccgga atcgggagcg
     5041 cggccgatgc aaagtgccga taaacataac gatctttgta gaaaccatcg gcgcagctat
     5101 ttacccgcag gacatatcca cgccctccta catcgaagct gaaagcacga gattcttcgc
     5161 cctccgagag ctgcatcagg tcggagacgc tgtcgaactt ttcgatcaga aacttctcga
     5221 cagacgtcgc ggtgagttca ggctttttgg ccaaggaagt tcctatactt tctagagaat
     5281 aggaacttcg gaataggaac ttctaggtac gtgaaccatc accctaatca agttttttgg
     5341 ggtcgaggtg ccgtaaagca ctaaatcgga accctaaagg gagcccccga tttagagctt
     5401 gacggggaaa gccggcgaac gtggcgagaa aggaagggaa gaaagcgaaa ggagcgggcg
     5461 ctagggcgct ggcaagtgta gcggtcacgc tgcgcgtaac caccacaccc gccgcgctta
     5521 atgcgccgct acagggcgcg tggggatacc ccctagagcc ccagctggtt ctttccgcct
     5581 cagaagccat agagcccacc gcatccccag aacatcgata gatctccata gagcccaccg
     5641 catccccagc atgcctgcta ttgtcttccc aatcctcccc cttgctgtcc tgccccaccc
     5701 caccccccag aatagaatga cacctactca gacaatgcga tgcaatttcc tcattttatt
     5761 aggaaaggac agtgggagtg gcaccttcca gggtcaagga aggcacgggg gaggggcaaa
     5821 caacagatgg ctggcaacta gaaggcacag tcgaggctga tcagcgggtt taaactcaat
     5881 gatgattatg atgatggtcg acggcgctat tcagatcctc ttctgagatg agtttttgtt
     5941 cgggccctcc tcgagcggct cacacctcgt tctcgtagca gaagggcgtg agtgagaact
     6001 ggtcctgctc ggtgggcttc tccacccagg cccccagccc cgccttgatg aaggctgtga
     6061 acagacgcgc cttggcggcc tggtactcct ttgccgccag cttggactca tggtacacgt
     6121 tgggtttggt aatcttggag cgtagtaagt gggagggaac cttgccgtgc acacgcatcc
     6181 agcgacagta caacgcgtgc ttacacaggc gggacgcgcg gcccagctca tccttcccag
     6241 tcgtggcgtt gatgacctca atagcggagt cgcctaccgt ccagttgaca ctgaagttgg
     6301 gggccttccc tggctgccgt gcttctgcat tgctgatgcc actgagcaaa ggcttgttga
     6361 gggtgtagag aggtggcagg tcctctatgt tggagatccg ctggtacatg gccctggaaa
     6421 ggtggtcccc gtggtaaagg ctgcccagga tgatgctcga gaagtaaatg ggctccacga
     6481 aaatgctgag cagggatccc tggatgccca ccacgttcca gcgtgcaatc ttgtcactgc
     6541 aggacatggt gagcagccgc tccccttgca gcaccccgtc ccacgcttgg atgctcgcat
     6601 tggagcgcac tggaatcgtc ccctgaccag actctatttt ggtccgtagc tgtcctcttg
     6661 ctttacgatt tgggtgtcta tctgctggtt cttccaggat tggctcatgt ggtgagaaga
     6721 ttctggcatc tccacaggga gaggtgctga tgtacagatg aaactggaca ttctccttca
     6781 gcctaaaccc ccctcgctct gatttctgaa agatggatct tttttgatca tctttgttat
     6841 ttaagtaaag ctcaagttgt gtataaagaa atctgagcaa ggatctccga gatattattt
     6901 ctgcatggca gtcatttaat gcaaggccac gatcactcat gtattcacca ttaatacatt
     6961 ttgttcctgt agaaacactt atcaccttgg catctttaac atctgtgcct gttgtcatga
     7021 cgactccagc cagcactttt ctgcgagcgt gaggggagga gaagttgtcg gtcaggtcac
     7081 caaacttacc caggaccagg cgtgagacag cgtcagctaa aacctgcggt aaatgcagct
     7141 gaagaccctc actgggaata ggctggcgag atggcgtttg atccaagtgc aacacgttga
     7201 tgtcgatctg gtccaggggg atgtactcgc cctcgctggt gatcctgctc ttggcggccg
     7261 cggtaccgat gagcaagtac actaagatga tgaagccaaa agccccgctg aggcccacga
     7321 ggaggtggcc gagccacgga atggtgtcct tcccaagggc tgcaggcttt gtcctgaagg
     7381 ccaggggctg gctccagggg ctccaggtcg tgaactcgcc ttgcagaggc ttgacccgca
     7441 cctgaaactc atactgggtg tctggggtga gcgtctccag gcagatccat tcctgcttct
     7501 gcttgagagt cagcaggggg gcctcctccc aggtgtggcc tggggacagc gtccgggcct
     7561 cgaactccag gtgtctttca aagtagtggg aggcttggga gatttcccag cttatgttgc
     7621 atctgtgggt ctccacgtgg acaacttgga gggagatggg ggccatcagg cgaaggttct
     7681 caaagggctt g
//
