LOCUS       sequence-466382-        7346 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Optimized IL-2-sensing eLIDAR with IL-2 gamma chain ecto+TM domains 
            fused with tdMCP, with Golgi traffic sequence and ER export 
            sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7346)
  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 7346)
  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: XZ374
            EF1a-IL2Rgsp-3xHA-IL2Rg(ECD+TM)-TS-tdMCP...
FEATURES             Location/Qualifiers
     source          1..7346
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             17..1039
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="KKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRGY
                     VLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLPE
                     TELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQT
                     VMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMFG
                     DSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDGN
                     FDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAKE
                     "
     CDS             17..1039
                     /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
                     "
     polyA_signal    1169..1290
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1206..1225)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     1260..1279
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(1339..1355)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1339..1355)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1352..1374)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1363..1379
                     /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(1387..1393,1394..1411,1412..1417))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1432..1453
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(1570..1587)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1741..2329)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1821..1840)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2500..3291,3292..3360))
                     /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     3123..3142
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3361..3465)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(3540..3559)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        3686..3897
                     /label=EF-1-alpha core promoter
                     /note="core promoter for human elongation factor 
                     EF-1-alpha"
     intron          3898..4836
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
     primer_bind     4793..4813
                     /label=EF1a-F
                     /note="Human elongation factor-1a promoter, forward primer"
     CDS             4939..5028
                     /codon_start=1
                     /product="three tandem HA epitope tags"
                     /label=3xHA
                     /translation="YPYDVPDYAGYPYDVPDYAGSYPYDVPDYA"
     CDS             6670..6699
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     primer_bind     complement(6755..6772)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    6761..6985
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(7157..7176)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     protein_bind    join(7308..7346,1..9)
                     /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)."
ORIGIN
        1 aggaacttcc ttggccaaaa agcctgaact caccgcgacg tctgtcgaga agtttctgat
       61 cgaaaagttc gacagcgtct ccgacctgat gcagctctcg gagggcgaag aatctcgtgc
      121 tttcagcttc gatgtaggag ggcgtggata tgtcctgcgg gtaaatagct gcgccgatgg
      181 tttctacaaa gatcgttatg tttatcggca ctttgcatcg gccgcgctcc cgattccgga
      241 agtgcttgac attggggaat tcagcgagag cctgacctat tgcatctccc gccgtgcaca
      301 gggtgtcacg ttgcaagacc tgcctgaaac cgaactgccc gctgttctgc agccggtcgc
      361 ggaggccatg gatgcgatcg ctgcggccga tcttagccag acgagcgggt tcggcccatt
      421 cggaccgcaa ggaatcggtc aatacactac atggcgtgat ttcatatgcg cgattgctga
      481 tccccatgtg tatcactggc aaactgtgat ggacgacacc gtcagtgcgt ccgtcgcgca
      541 ggctctcgat gagctgatgc tttgggccga ggactgcccc gaagtccggc acctcgtgca
      601 cgcggatttc ggctccaaca atgtcctgac ggacaatggc cgcataacag cggtcattga
      661 ctggagcgag gcgatgttcg gggattccca atacgaggtc gccaacatct tcttctggag
      721 gccgtggttg gcttgtatgg agcagcagac gcgctacttc gagcggaggc atccggagct
      781 tgcaggatcg ccgcggctcc gggcgtatat gctccgcatt ggtcttgacc aactctatca
      841 gagcttggtt gacggcaatt tcgatgatgc agcttgggcg cagggtcgat gcgacgcaat
      901 cgtccgatcc ggagccggga ctgtcgggcg tacacaaatc gcccgcagaa gcgcggccgt
      961 ctggaccgat ggctgtgtag aagtactcgc cgatagtgga aaccgacgcc ccagcactcg
     1021 tccgagggca aaggaatagc acgtactacg agatttcgat tccaccgccg ccttctatga
     1081 aaggttgggc ttcggaatcg ttttccggga cgccggctgg atgatcctcc agcgcgggga
     1141 tctcatgctg gagttcttcg cccaccccaa cttgtttatt gcagcttata atggttacaa
     1201 ataaagcaat agcatcacaa atttcacaaa taaagcattt ttttcactgc attctagttg
     1261 tggtttgtcc aaactcatca atgtatctta tcatgtctgt ataccgtcga cctctagcta
     1321 gagcttggcg taatcatggt catagctgtt tcctgtgtga aattgttatc cgctcacaat
     1381 tccacacaac atacgagccg gaagcataaa gtgtaaagcc tggggtgcct aatgagtgag
     1441 ctaactcaca ttaattgcgt tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg
     1501 ccagctgcat taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta ttgggcgctc
     1561 ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc
     1621 agctcactca aaggcggtaa tacggttatc cacagaatca ggggataacg caggaaagaa
     1681 catgtgagca aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt
     1741 tttccatagg ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg
     1801 gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggaagct ccctcgtgcg
     1861 ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag
     1921 cgtggcgctt tctcatagct cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc
     1981 caagctgggc tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa
     2041 ctatcgtctt gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccactgg
     2101 taacaggatt agcagagcga ggtatgtagg cggtgctaca gagttcttga agtggtggcc
     2161 taactacggc tacactagaa gaacagtatt tggtatctgc gctctgctga agccagttac
     2221 cttcggaaaa agagttggta gctcttgatc cggcaaacaa accaccgctg gtagcggtgg
     2281 tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag aagatccttt
     2341 gatcttttct acggggtctg acgctcagtg gaacgaaaac tcacgttaag ggattttggt
     2401 catgagatta tcaaaaagga tcttcaccta gatcctttta aattaaaaat gaagttttaa
     2461 atcaatctaa agtatatatg agtaaacttg gtctgacagt taccaatgct taatcagtga
     2521 ggcacctatc tcagcgatct gtctatttcg ttcatccata gttgcctgac tccccgtcgt
     2581 gtagataact acgatacggg agggcttacc atctggcccc agtgctgcaa tgataccgcg
     2641 agacccacgc tcaccggctc cagatttatc agcaataaac cagccagccg gaagggccga
     2701 gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag tctattaatt gttgccggga
     2761 agctagagta agtagttcgc cagttaatag tttgcgcaac gttgttgcca ttgctacagg
     2821 catcgtggtg tcacgctcgt cgtttggtat ggcttcattc agctccggtt cccaacgatc
     2881 aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc
     2941 gatcgttgtc agaagtaagt tggccgcagt gttatcactc atggttatgg cagcactgca
     3001 taattctctt actgtcatgc catccgtaag atgcttttct gtgactggtg agtactcaac
     3061 caagtcattc tgagaatagt gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg
     3121 ggataatacc gcgccacata gcagaacttt aaaagtgctc atcattggaa aacgttcttc
     3181 ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc agttcgatgt aacccactcg
     3241 tgcacccaac tgatcttcag catcttttac tttcaccagc gtttctgggt gagcaaaaac
     3301 aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt gaatactcat
     3361 actcttcctt tttcaatatt attgaagcat ttatcagggt tattgtctca tgagcggata
     3421 catatttgaa tgtatttaga aaaataaaca aataggggtt ccgcgcacat ttccccgaaa
     3481 agtgccacct gacgtcgacg gatcgggaga tctcccgatc ccctatggtg cactctcagt
     3541 acaatctgct ctgatgccgc atagttaagc cagtatctgc tccctgcttg tgtgttggag
     3601 gtcgctgagt agtgcgcgag caaaatttaa gctacaacaa ggcaaggctt gaccgacaat
     3661 tgcatgaaga atcgtgcccg tcagtgggca gagcgcacat cgcccacagt ccccgagaag
     3721 ttggggggag gggtcggcaa ttgaaccggt gcctagagaa ggtggcgcgg ggtaaactgg
     3781 gaaagtgatg tcgtgtactg gctccgcctt tttcccgagg gtgggggaga accgtatata
     3841 agtgcagtag tcgccgtgaa cgttcttttt cgcaacgggt ttgccgccag aacacaggta
     3901 agtgccgtgt gtggttcccg cgggcctggc ctctttacgg gttatggccc ttgcgtgcct
     3961 tgaattactt ccacctggct gcagtacgtg attcttgatc ccgagcttcg ggttggaagt
     4021 gggtgggaga gttcgaggcc ttgcgcttaa ggagcccctt cgcctcgtgc ttgagttgag
     4081 gcctggcctg ggcgctgggg ccgccgcgtg cgaatctggt ggcaccttcg cgcctgtctc
     4141 gctgctttcg ataagtctct agccatttaa aatttttgat gacctgctgc gacgcttttt
     4201 ttctggcaag atagtcttgt aaatgcgggc caagatctgc acactggtat ttcggttttt
     4261 ggggccgcgg gcggcgacgg ggcccgtgcg tcccagcgca catgttcggc gaggcggggc
     4321 ctgcgagcgc ggccaccgag aatcggacgg gggtagtctc aagctggccg gcctgctctg
     4381 gtgcctggcc tcgcgccgcc gtgtatcgcc ccgccctggg cggcaaggct ggcccggtcg
     4441 gcaccagttg cgtgagcgga aagatggccg cttcccggcc ctgctgcagg gagctcaaaa
     4501 tggaggacgc ggcgctcggg agagcgggcg ggtgagtcac ccacacaaag gaaaagggcc
     4561 tttccgtcct cagccgtcgc ttcatgtgac tccacggagt accgggcgcc gtccaggcac
     4621 ctcgattagt tctcgcgctt ttggagtacg tcgtctttag gttgggggga ggggttttat
     4681 gcgatggagt ttccccacac tgagtgggtg gagactgaag ttaggccagc ttggcacttg
     4741 atgtaattct ccttggaatt tgcccttttt gagtttggat cttggttcat tctcaagcct
     4801 cagacagtgg ttcaaagttt ttttcttcca tttcaggtgt cgtgaagcta ctagttcgcc
     4861 accatgttga agccatcatt accattcaca tccctcttat tcctgcagct gcccctgctg
     4921 ggagtggggg cggccgttta cccatacgat gttcctgact atgcgggcta tccctatgac
     4981 gtcccggact atgcaggatc ctatccatat gacgttccag attacgctcc ggccgccggt
     5041 accctgaaca cgacaattct gacgcccaat gggaatgaag acaccacagc tgatttcttc
     5101 ctgaccacta tgcccactga ctccctcagt gtttccactc tgcccctccc agaggttcag
     5161 tgttttgtgt tcaatgtcga gtacatgaat tgcacttgga acagcagctc tgagccccag
     5221 cctaccaacc tcactctgca ttattggtac aagaactcgg ataatgataa agtccagaag
     5281 tgcagccact atctattctc tgaagaaatc acttctggct gtcagttgca aaaaaaggag
     5341 atccacctct accaaacatt tgttgttcag ctccaggacc cacgggaacc caggagacag
     5401 gccacacaga tgctaaaact gcagaatctg gtgatcccct gggctccaga gaacctaaca
     5461 cttcacaaac tgagtgaatc ccagctagaa ctgaactgga acaacagatt cttgaaccac
     5521 tgtttggagc acttggtgca gtaccggact gactgggacc acagctggac tgaacaatca
     5581 gtggattata gacataagtt ctccttgcct agtgtggatg ggcagaaacg ctacacgttt
     5641 cgtgttcgga gccgctttaa cccactctgt ggaagtgctc agcattggag tgaatggagc
     5701 cacccaatcc actgggggag caatacttca aaagagaatc ctttcctgtt tgcattggaa
     5761 gccgtggtta tctctgttgg ctccatggga ttgattatca gccttctctg tgtgtatttc
     5821 tggctgggta ccgcggccgc caagagcagg atcaccagcg agggcgagta catccccctg
     5881 gaccagatcg acatcaacgt gatggcgagc aactttacgc aatttgtgtt ggttgacaat
     5941 ggtggcaccg gagacgtaac tgttgcgcca agcaattttg cgaacgggat tgcagaatgg
     6001 ataagtagta acagccggag tcaggcatat aaagtcacat gctcagttcg ccaaagttcc
     6061 gcacagaacc gaaaatatac cataaaggta gaagtgccaa aaggggcgtg gcgatcttat
     6121 cttaatatgg agcttacgat cccaatcttt gcgactaatt cagactgcga gctgattgtc
     6181 aaggccatgc aaggtctttt gaaagacggt aacccgattc cctctgcgat cgccgccaac
     6241 tcaggaatat atggtggcgg aaccggtggc gggtccatgg cctccaattt tacacagttt
     6301 gtgctcgtgg acaacggcgg cactggagac gtcacagtag ctccaagtaa ctttgcgaat
     6361 gggatagccg agtggatctc ttccaattca agatcacaag cctataaggt gacttgtagt
     6421 gtacgccaat ctagtgccca gaaccggaag tataccatta aagtggaggt ccccaaaggt
     6481 gcatggcgct catacttgaa tatggaattg acgattccga tatttgcgac caatagcgac
     6541 tgcgagctta ttgtaaaagc catgcaagga cttctgaaag atggcaaccc gataccttcc
     6601 gcaatcgctg cgaatagcgg aatttacttc tgctacgaga acgaggtgtg agccgctcga
     6661 ggagggcccg aacaaaaact catctcagaa gaggatctga atagcgccgt cgaccatcat
     6721 cataatcatc attgagttta aacccgctga tcagcctcga ctgtgccttc tagttgccag
     6781 ccatctgttg tttgcccctc ccccgtgcct tccttgaccc tggaaggtgc cactcccact
     6841 gtcctttcct aataaaatga ggaaattgca tcgcattgtc tgagtaggtg tcattctatt
     6901 ctggggggtg gggtggggca ggacagcaag ggggaggatt gggaagacaa tagcaggcat
     6961 gctggggatg cggtgggctc tatggagatc tatcgatgtt ctggggatgc ggtgggctct
     7021 atggcttctg aggcggaaag aaccagctgg ggctctaggg ggtatcccca cgcgccctgt
     7081 agcggcgcat taagcgcggc gggtgtggtg gttacgcgca gcgtgaccgc tacacttgcc
     7141 agcgccctag cgcccgctcc tttcgctttc ttcccttcct ttctcgccac gttcgccggc
     7201 tttccccgtc aagctctaaa tcgggggctc cctttagggt tccgatttag tgctttacgg
     7261 cacctcgacc ccaaaaaact tgattagggt gatggttcac gtacctagaa gttcctattc
     7321 cgaagttcct attctctaga aagtat
//
