LOCUS       sequence-466376-        5628 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  secreted human IL-2.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5628)
  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 5628)
  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: CMVTO-hIL2
FEATURES             Location/Qualifiers
     source          1..5628
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     polyA_signal    492..613
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(529..548)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     583..602
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(662..678)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(662..678)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(675..697)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    686..702
                     /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(710..716,717..734,735..740))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    755..776
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(893..910)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1064..1652)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1144..1163)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(1823..2614,2615..2683))
                     /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     2446..2465
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(2684..2788)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(2863..2882)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        3049..3428
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        3429..3632
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     3583..3603
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     protein_bind    3634..3652
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    3655..3673
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     primer_bind     3683..3707
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     CDS             4275..4304
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     primer_bind     complement(4360..4377)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    4366..4590
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(4762..4781)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     protein_bind    4913..4960
                     /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)."
     CDS             join(4968..5628,1..362)
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="KKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRGY
                     VLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLPE
                     TELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQT
                     VMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMFG
                     DSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDGN
                     FDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAKE
                     "
     CDS             join(4968..5628,1..362)
                     /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
                     "
ORIGIN
        1 tcggggattc ccaatacgag gtcgccaaca tcttcttctg gaggccgtgg ttggcttgta
       61 tggagcagca gacgcgctac ttcgagcgga ggcatccgga gcttgcagga tcgccgcggc
      121 tccgggcgta tatgctccgc attggtcttg accaactcta tcagagcttg gttgacggca
      181 atttcgatga tgcagcttgg gcgcagggtc gatgcgacgc aatcgtccga tccggagccg
      241 ggactgtcgg gcgtacacaa atcgcccgca gaagcgcggc cgtctggacc gatggctgtg
      301 tagaagtact cgccgatagt ggaaaccgac gccccagcac tcgtccgagg gcaaaggaat
      361 agcacgtact acgagatttc gattccaccg ccgccttcta tgaaaggttg ggcttcggaa
      421 tcgttttccg ggacgccggc tggatgatcc tccagcgcgg ggatctcatg ctggagttct
      481 tcgcccaccc caacttgttt attgcagctt ataatggtta caaataaagc aatagcatca
      541 caaatttcac aaataaagca tttttttcac tgcattctag ttgtggtttg tccaaactca
      601 tcaatgtatc ttatcatgtc tgtataccgt cgacctctag ctagagcttg gcgtaatcat
      661 ggtcatagct gtttcctgtg tgaaattgtt atccgctcac aattccacac aacatacgag
      721 ccggaagcat aaagtgtaaa gcctggggtg cctaatgagt gagctaactc acattaattg
      781 cgttgcgctc actgcccgct ttccagtcgg gaaacctgtc gtgccagctg cattaatgaa
      841 tcggccaacg cgcggggaga ggcggtttgc gtattgggcg ctcttccgct tcctcgctca
      901 ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg
      961 taatacggtt atccacagaa tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc
     1021 agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc gtttttccat aggctccgcc
     1081 cccctgacga gcatcacaaa aatcgacgct caagtcagag gtggcgaaac ccgacaggac
     1141 tataaagata ccaggcgttt ccccctggaa gctccctcgt gcgctctcct gttccgaccc
     1201 tgccgcttac cggatacctg tccgcctttc tcccttcggg aagcgtggcg ctttctcata
     1261 gctcacgctg taggtatctc agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc
     1321 acgaaccccc cgttcagccc gaccgctgcg ccttatccgg taactatcgt cttgagtcca
     1381 acccggtaag acacgactta tcgccactgg cagcagccac tggtaacagg attagcagag
     1441 cgaggtatgt aggcggtgct acagagttct tgaagtggtg gcctaactac ggctacacta
     1501 gaagaacagt atttggtatc tgcgctctgc tgaagccagt taccttcgga aaaagagttg
     1561 gtagctcttg atccggcaaa caaaccaccg ctggtagcgg tggttttttt gtttgcaagc
     1621 agcagattac gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt tctacggggt
     1681 ctgacgctca gtggaacgaa aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa
     1741 ggatcttcac ctagatcctt ttaaattaaa aatgaagttt taaatcaatc taaagtatat
     1801 atgagtaaac ttggtctgac agttaccaat gcttaatcag tgaggcacct atctcagcga
     1861 tctgtctatt tcgttcatcc atagttgcct gactccccgt cgtgtagata actacgatac
     1921 gggagggctt accatctggc cccagtgctg caatgatacc gcgagaccca cgctcaccgg
     1981 ctccagattt atcagcaata aaccagccag ccggaagggc cgagcgcaga agtggtcctg
     2041 caactttatc cgcctccatc cagtctatta attgttgccg ggaagctaga gtaagtagtt
     2101 cgccagttaa tagtttgcgc aacgttgttg ccattgctac aggcatcgtg gtgtcacgct
     2161 cgtcgtttgg tatggcttca ttcagctccg gttcccaacg atcaaggcga gttacatgat
     2221 cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt gtcagaagta
     2281 agttggccgc agtgttatca ctcatggtta tggcagcact gcataattct cttactgtca
     2341 tgccatccgt aagatgcttt tctgtgactg gtgagtactc aaccaagtca ttctgagaat
     2401 agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat acgggataat accgcgccac
     2461 atagcagaac tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga aaactctcaa
     2521 ggatcttacc gctgttgaga tccagttcga tgtaacccac tcgtgcaccc aactgatctt
     2581 cagcatcttt tactttcacc agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg
     2641 caaaaaaggg aataagggcg acacggaaat gttgaatact catactcttc ctttttcaat
     2701 attattgaag catttatcag ggttattgtc tcatgagcgg atacatattt gaatgtattt
     2761 agaaaaataa acaaataggg gttccgcgca catttccccg aaaagtgcca cctgacgtcg
     2821 acggatcggg agatctcccg atcccctatg gtgcactctc agtacaatct gctctgatgc
     2881 cgcatagtta agccagtatc tgctccctgc ttgtgtgttg gaggtcgctg agtagtgcgc
     2941 gagcaaaatt taagctacaa caaggcaagg cttgaccgac aattgcatga agaatctgct
     3001 tagggttagg cgttttgcgc tgcttcgtac gggccagata tacgcgttga cattgattat
     3061 tgactagtta ttaatagtaa tcaattacgg ggtcattagt tcatagccca tatatggagt
     3121 tccgcgttac ataacttacg gtaaatggcc cgcctggctg accgcccaac gacccccgcc
     3181 cattgacgtc aataatgacg tatgttccca tagtaacgcc aatagggact ttccattgac
     3241 gtcaatgggt ggagtattta cggtaaactg cccacttggc agtacatcaa gtgtatcata
     3301 tgccaagtac gccccctatt gacgtcaatg acggtaaatg gcccgcctgg cattatgccc
     3361 agtacatgac cttatgggac tttcctactt ggcagtacat ctacgtatta gtcatcgcta
     3421 ttaccatggt gatgcggttt tggcagtaca tcaatgggcg tggatagcgg tttgactcac
     3481 ggggatttcc aagtctccac cccattgacg tcaatgggag tttgttttgg caccaaaatc
     3541 aacgggactt tccaaaatgt cgtaacaact ccgccccatt gacgcaaatg ggcggtaggc
     3601 gtgtacggtg ggaggtctat ataagcagag ctctccctat cagtgataga gatctcccta
     3661 tcagtgatag agatcgtcga cgagctcgtt tagtgaaccg tcagatcgcc tggagacgcc
     3721 atccacgctg ttttgacctc catagaagac accgggaccg atccagcctc cggactctag
     3781 cgtttaaact taagcttcgc caccatgtac aggatgcaac tcctgtcttg cattgcacta
     3841 agtcttgcac ttgtcacaaa cgctagcgca cctacttcaa gttctacaaa gaaaacacag
     3901 ctacaactgg agcatttact gctggattta cagatgattt tgaatggaat taataattac
     3961 aagaatccca aactcaccag gatgctcaca tttaagtttt acatgcccaa gaaggccaca
     4021 gaactgaaac atcttcagtg tctagaagaa gaactcaaac ctctggagga agtgctaaat
     4081 ttagctcaaa gcaaaaactt tcacttaaga cccagggact taatcagcaa tatcaacgta
     4141 atagttctgg aactaaaggg atctgaaaca acattcatgt gtgaatatgc tgatgagaca
     4201 gcaaccattg tagaatttct gaacagatgg attacctttt gtcaaagcat catctcaaca
     4261 ctgacttagg gcccgaacaa aaactcatct cagaagagga tctgaatagc gccgtcgacc
     4321 atcatcataa tcatcattga gtttaaaccc gctgatcagc ctcgactgtg ccttctagtt
     4381 gccagccatc tgttgtttgc ccctcccccg tgccttcctt gaccctggaa ggtgccactc
     4441 ccactgtcct ttcctaataa aatgaggaaa ttgcatcgca ttgtctgagt aggtgtcatt
     4501 ctattctggg gggtggggtg gggcaggaca gcaaggggga ggattgggaa gacaatagca
     4561 ggcatgctgg ggatgcggtg ggctctatgg agatctatcg atgttctggg gatgcggtgg
     4621 gctctatggc ttctgaggcg gaaagaacca gctggggctc tagggggtat ccccacgcgc
     4681 cctgtagcgg cgcattaagc gcggcgggtg tggtggttac gcgcagcgtg accgctacac
     4741 ttgccagcgc cctagcgccc gctcctttcg ctttcttccc ttcctttctc gccacgttcg
     4801 ccggctttcc ccgtcaagct ctaaatcggg ggctcccttt agggttccga tttagtgctt
     4861 tacggcacct cgaccccaaa aaacttgatt agggtgatgg ttcacgtacc tagaagttcc
     4921 tattccgaag ttcctattct ctagaaagta taggaacttc cttggccaaa aagcctgaac
     4981 tcaccgcgac gtctgtcgag aagtttctga tcgaaaagtt cgacagcgtc tccgacctga
     5041 tgcagctctc ggagggcgaa gaatctcgtg ctttcagctt cgatgtagga gggcgtggat
     5101 atgtcctgcg ggtaaatagc tgcgccgatg gtttctacaa agatcgttat gtttatcggc
     5161 actttgcatc ggccgcgctc ccgattccgg aagtgcttga cattggggaa ttcagcgaga
     5221 gcctgaccta ttgcatctcc cgccgtgcac agggtgtcac gttgcaagac ctgcctgaaa
     5281 ccgaactgcc cgctgttctg cagccggtcg cggaggccat ggatgcgatc gctgcggccg
     5341 atcttagcca gacgagcggg ttcggcccat tcggaccgca aggaatcggt caatacacta
     5401 catggcgtga tttcatatgc gcgattgctg atccccatgt gtatcactgg caaactgtga
     5461 tggacgacac cgtcagtgcg tccgtcgcgc aggctctcga tgagctgatg ctttgggccg
     5521 aggactgccc cgaagtccgg cacctcgtgc acgcggattt cggctccaac aatgtcctga
     5581 cggacaatgg ccgcataaca gcggtcattg actggagcga ggcgatgt
//
