LOCUS       sequence-466402-        8159 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  IVT template for AVPR2-V2-ADAR2dd(E488Q, T501A).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8159)
  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 8159)
  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: XZ387 (IVT) CMV-TO-T7-AVPR2-V2-ADAR2dd-hybridUTR3
FEATURES             Location/Qualifiers
     source          1..8159
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      731..1319
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1220..1239
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     1473..1490
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    1607..1628
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(1643..1648,1649..1666,1667..1673)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1681..1697
                     /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     1686..1708
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     1705..1721
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     1705..1721
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     polyA_signal    1770..1891
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1781..1800)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     1835..1854
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     CDS             complement(2021..3043)
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="KKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRGY
                     VLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLPE
                     TELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQT
                     VMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMFG
                     DSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDGN
                     FDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAKE
                     "
     CDS             complement(2021..3043)
                     /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(3051..3098)
                     /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     3230..3249
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     polyA_signal    3421..3645
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     3634..3651
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     CDS             complement(3707..3736)
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     primer_bind     complement(6706..6725)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(6707..6725)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(6835..6859)
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     protein_bind    6869..6887
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    6890..6908
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     promoter        complement(6910..7113)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(6939..6959)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        7114..7493
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     primer_bind     7660..7679
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        7754..7858
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(7859..7927,7928..8159,1..560)
                     /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(8077..8096)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
ORIGIN
        1 ttgagtactc accagtcaca gaaaagcatc ttacggatgg catgacagta agagaattat
       61 gcagtgctgc cataaccatg agtgataaca ctgcggccaa cttacttctg acaacgatcg
      121 gaggaccgaa ggagctaacc gcttttttgc acaacatggg ggatcatgta actcgccttg
      181 atcgttggga accggagctg aatgaagcca taccaaacga cgagcgtgac accacgatgc
      241 ctgtagcaat ggcaacaacg ttgcgcaaac tattaactgg cgaactactt actctagctt
      301 cccggcaaca attaatagac tggatggagg cggataaagt tgcaggacca cttctgcgct
      361 cggcccttcc ggctggctgg tttattgctg ataaatctgg agccggtgag cgtgggtctc
      421 gcggtatcat tgcagcactg gggccagatg gtaagccctc ccgtatcgta gttatctaca
      481 cgacggggag tcaggcaact atggatgaac gaaatagaca gatcgctgag ataggtgcct
      541 cactgattaa gcattggtaa ctgtcagacc aagtttactc atatatactt tagattgatt
      601 taaaacttca tttttaattt aaaaggatct aggtgaagat cctttttgat aatctcatga
      661 ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc agaccccgta gaaaagatca
      721 aaggatcttc ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac
      781 caccgctacc agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg
      841 taactggctt cagcagagcg cagataccaa atactgttct tctagtgtag ccgtagttag
      901 gccaccactt caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac
      961 cagtggctgc tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt
     1021 taccggataa ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg
     1081 agcgaacgac ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc
     1141 ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc
     1201 gcacgaggga gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc
     1261 acctctgact tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa
     1321 acgccagcaa cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgt
     1381 tctttcctgc gttatcccct gattctgtgg ataaccgtat taccgccttt gagtgagctg
     1441 ataccgctcg ccgcagccga acgaccgagc gcagcgagtc agtgagcgag gaagcggaag
     1501 agcgcccaat acgcaaaccg cctctccccg cgcgttggcc gattcattaa tgcagctggc
     1561 acgacaggtt tcccgactgg aaagcgggca gtgagcgcaa cgcaattaat gtgagttagc
     1621 tcactcatta ggcaccccag gctttacact ttatgcttcc ggctcgtatg ttgtgtggaa
     1681 ttgtgagcgg ataacaattt cacacaggaa acagctatga ccatgattac gccaagctct
     1741 agctagaggt cgacggtata cagacatgat aagatacatt gatgagtttg gacaaaccac
     1801 aactagaatg cagtgaaaaa aatgctttat ttgtgaaatt tgtgatgcta ttgctttatt
     1861 tgtaaccatt ataagctgca ataaacaagt tggggtgggc gaagaactcc agcatgagat
     1921 ccccgcgctg gaggatcatc cagccggcgt cccggaaaac gattccgaag cccaaccttt
     1981 catagaaggc ggcggtggaa tcgaaatctc gtagtacgtg ctattccttt gccctcggac
     2041 gagtgctggg gcgtcggttt ccactatcgg cgagtacttc tacacagcca tcggtccaga
     2101 cggccgcgct tctgcgggcg atttgtgtac gcccgacagt cccggctccg gatcggacga
     2161 ttgcgtcgca tcgaccctgc gcccaagctg catcatcgaa attgccgtca accaagctct
     2221 gatagagttg gtcaagacca atgcggagca tatacgcccg gagccgcggc gatcctgcaa
     2281 gctccggatg cctccgctcg aagtagcgcg tctgctgctc catacaagcc aaccacggcc
     2341 tccagaagaa gatgttggcg acctcgtatt gggaatcccc gaacatcgcc tcgctccagt
     2401 caatgaccgc tgttatgcgg ccattgtccg tcaggacatt gttggagccg aaatccgcgt
     2461 gcacgaggtg ccggacttcg gggcagtcct cggcccaaag catcagctca tcgagagcct
     2521 gcgcgacgga cgcactgacg gtgtcgtcca tcacagtttg ccagtgatac acatggggat
     2581 cagcaatcgc gcatatgaaa tcacgccatg tagtgtattg accgattcct tgcggtccga
     2641 atgggccgaa cccgctcgtc tggctaagat cggccgcagc gatcgcatcc atggcctccg
     2701 cgaccggctg cagaacagcg ggcagttcgg tttcaggcag gtcttgcaac gtgacaccct
     2761 gtgcacggcg ggagatgcaa taggtcaggc tctcgctgaa ttccccaatg tcaagcactt
     2821 ccggaatcgg gagcgcggcc gatgcaaagt gccgataaac ataacgatct ttgtagaaac
     2881 catcggcgca gctatttacc cgcaggacat atccacgccc tcctacatcg aagctgaaag
     2941 cacgagattc ttcgccctcc gagagctgca tcaggtcgga gacgctgtcg aacttttcga
     3001 tcagaaactt ctcgacagac gtcgcggtga gttcaggctt tttggccaag gaagttccta
     3061 tactttctag agaataggaa cttcggaata ggaacttcta ggtacgtgaa ccatcaccct
     3121 aatcaagttt tttggggtcg aggtgccgta aagcactaaa tcggaaccct aaagggagcc
     3181 cccgatttag agcttgacgg ggaaagccgg cgaacgtggc gagaaaggaa gggaagaaag
     3241 cgaaaggagc gggcgctagg gcgctggcaa gtgtagcggt cacgctgcgc gtaaccacca
     3301 cacccgccgc gcttaatgcg ccgctacagg gcgcgtgggg atacccccta gagccccagc
     3361 tggttctttc cgcctcagaa gccatagagc ccaccgcatc cccagaacat cgatagatct
     3421 ccatagagcc caccgcatcc ccagcatgcc tgctattgtc ttcccaatcc tcccccttgc
     3481 tgtcctgccc caccccaccc cccagaatag aatgacacct actcagacaa tgcgatgcaa
     3541 tttcctcatt ttattaggaa aggacagtgg gagtggcacc ttccagggtc aaggaaggca
     3601 cgggggaggg gcaaacaaca gatggctggc aactagaagg cacagtcgag gctgatcagc
     3661 gggtttaaac tcaatgatga ttatgatgat ggtcgacggc gctattcaga tcctcttctg
     3721 agatgagttt ttgttcgggc ccgctagctc cagggtgtgg ctggcacgaa attgaccaac
     3781 cctggggtta gtatagctta gttaaacttt cgtttattgc taaaggttaa tcactgctgt
     3841 ttcccgtggg ggtgtggcta ggctaagcgt tttgagctgc attgctgcgt gcttgggagg
     3901 tgtctggaac tagcagaggt ggtgagtggg gcaggtggag gtgggagcat acctgggacc
     3961 cgaggtcggg ggagactcgg ggtacccagg acgggaaagg ggcagctagc attgcgtgca
     4021 tgcagtacca gctcgagtca gggcgtgagt gagaactggt cctgctcggt gggcttctcc
     4081 acccaggccc ccagccccgc cttgatgaag gctgtgaaca gacgcgcctt ggcggcctgg
     4141 tactcctttg ccgccagctt ggactcatgg tacacgttgg gtttggtaat cttggagcgt
     4201 agtaagtggg agggaacctt gccgtgcaca cgcatccagc gacagtacaa cgcgtgctta
     4261 cacaggcggg acgcgcggcc cagctcatcc ttcccagtcg tggcgttgat gacctcaata
     4321 gcggagtcgc ctaccgtcca gttgacactg aagttggggg ccttccctgg ctgccgtgct
     4381 tctgcattgc tgatgccact gagcaaaggc ttgttgaggg tgtagagagg tggcaggtcc
     4441 tctatgttgg agatccgctg gtacatggcc ctggaaaggt ggtccccgtg gtaaaggctg
     4501 cccaggatga tgctcgagaa gtaaatgggc tccacgaaaa tgctgagcag ggatccctgg
     4561 atgcccacca cgttccagcg tgcaatcttg tcactgcagg acatggtgag cagccgctcc
     4621 ccttgcagca ccccgtccca cgcttggatg ctcgcattgg agcgcactgg aatcgtcccc
     4681 tgaccagact ctattttggt ccgtagctgt cctcttgctt tacgatttgg gtgtctatct
     4741 gctggttctt ccaggattgg ctcatgtggt gagaagattc tggcatctcc acagggagag
     4801 gtgctgatgt acagatgaaa ctggacattc tccttcagcc taaacccccc tcgctctgat
     4861 ttctgaaaga tggatctttt ttgatcatct ttgttattta agtaaagctc aagttgtgta
     4921 taaagaaatc tgagcaagga tctccgagat attatttctg catggcagtc atttaatgca
     4981 aggccacgat cactcatgta ttcaccatta atacattttg ttcctgtaga aacacttatc
     5041 accttggcat ctttaacatc tgtgcctgtt gtcatgacga ctccagccag cacttttctg
     5101 cgagcgtgag gggaggagaa gttgtcggtc aggtcaccaa acttacccag gaccaggcgt
     5161 gagacagcgt cagctaaaac ctgcggtaaa tgcagctgaa gaccctcact gggaataggc
     5221 tggcgagatg gcgtttgatc caagtgcaaa gcaccggtac ccgatgaagt gtccttggcc
     5281 agggaggagc tggcggtggt gcaggactca tcttggggac ccaggctggg tggggtgcgt
     5341 ccaccggtat cgatagaaga ggtgtctttg gccagggaac tggaggcggt agtgcaggat
     5401 tcgtcctgtg gacccaggct tgggggggtc ctgccccggg cgcaacagag aagactccgc
     5461 agctcggagc taacactgga ggagaaagag gcgtaaatcc aaggattggt acagctattc
     5521 agtgaagcga gaagcatgag cagaacgaat ggagcacctt ccaggggtgc ttcggggtcc
     5581 caggccgccc agagctgcac caaaaaaaac ggggcccagc aaagaacgta aacgacgacg
     5641 atgaccagtg tcatgcgtac ggtcttagct accgcagctg atacgtgggc gccttcgcct
     5701 ggagagccag tccttcgacc ccgtcttctt ccaccagggc gctcgcttgg cccaggaacc
     5761 aaggaggcgt gtatttctct gaatatcagg acctggcatg cggcgatccc cagggtggga
     5821 gcaacgaaca ccatgagagc aatccacgtc acgtaggtcc ttctgcccca tggctcagcg
     5881 aagcaagccc agcaatcggt cacgccagat ccgccctcca catttctctg tgcgaaaata
     5941 aacaactggg gcaagctcag aagcagggaa aaagcccacg caaccagaac tggcctgttc
     6001 caatgggctc cggaaccgtg cctatacgcc agcatggggc gacagatagc cctgtgtctg
     6061 tccaaggtca tagcaagtat catatatgaa ctggcataca ttcccaccat ctgcaggtat
     6121 ttcacggccc gacacagtgc gtctggcccc cggaatctat cggtggcttt ccaagccagt
     6181 tgagggagca cctgaaacag cgccacggcc aggtctgcaa gacagagatg gccgataaag
     6241 acatgaatag gcgcccagtg gcctcggcgg cctcgtcgcg ccagggcggc cagcaccaga
     6301 ccattgctca acgcgacggc aacgaaaacg atggagagca aggcgagctc cgctcttgca
     6361 agaagtgggt cccgggtatc cagaggcctc tcctgtgagc tgttggaggg aagacttgga
     6421 agagatggat gtccgggcac ggcgcttgtg gtagaggcca tgagcatatc gatgctggcg
     6481 tcatcatcgt ccttgtagtc ggcgaatacc aggcagaaga tgtagctcag ggcgatgatc
     6541 gtcttcatgg tggcgaagct ttatcgttcg taaatggtga aaattttcag aaaattgctt
     6601 ttgctttaaa agaaatgatt taaattgctg caatagaagt agaatgcttg attgcttgag
     6661 attcgtttgt tttgtatatg ttgtgttgag atttgttatt tttctcccta tagtgagtcg
     6721 tattagcggc cgcggatctc tagcgagctt aagtttaaac gctagagtcc ggaggctgga
     6781 tcggtcccgg tgtcttctat ggaggtcaaa acagcgtgga tggcgtctcc aggcgatctg
     6841 acggttcact aaacgagctc gtcgacgatc tctatcactg atagggagat ctctatcact
     6901 gatagggaga gctctgctta tatagacctc ccaccgtaca cgcctaccgc ccatttgcgt
     6961 caatggggcg gagttgttac gacattttgg aaagtcccgt tgattttggt gccaaaacaa
     7021 actcccattg acgtcaatgg ggtggagact tggaaatccc cgtgagtcaa accgctatcc
     7081 acgcccattg atgtactgcc aaaaccgcat caccatggta atagcgatga ctaatacgta
     7141 gatgtactgc caagtaggaa agtcccataa ggtcatgtac tgggcataat gccaggcggg
     7201 ccatttaccg tcattgacgt caataggggg cgtacttggc atatgataca cttgatgtac
     7261 tgccaagtgg gcagtttacc gtaaatactc cacccattga cgtcaatgga aagtccctat
     7321 tggcgttact atgggaacat acgtcattat tgacgtcaat gggcgggggt cgttgggcgg
     7381 tcagccaggc gggccattta ccgtaagtta tgtaacgcgg aactccatat atgggctatg
     7441 aactaatgac cccgtaattg attactatta ataactagtc aataatcaat gtcaacgcgt
     7501 atatctggcc cgtacgaagc agcgcaaaac gcctaaccct aagcagattc ttcatgcaat
     7561 tgtcggtcaa gccttgcctt gttgtagctt aaattttgct cgcgcactac tcagcgacct
     7621 ccaacacaca agcagggagc agatactggc ttaactatgc ggcatcagag cagattgtac
     7681 tgagagtgca ccatagggga tcgggagatc tcccgatccg tcgacgtcag gtggcacttt
     7741 tcggggaaat gtgcgcggaa cccctatttg tttatttttc taaatacatt caaatatgta
     7801 tccgctcatg agacaataac cctgataaat gcttcaataa tattgaaaaa ggaagagtat
     7861 gagtattcaa catttccgtg tcgcccttat tccctttttt gcggcatttt gccttcctgt
     7921 ttttgctcac ccagaaacgc tggtgaaagt aaaagatgct gaagatcagt tgggtgcacg
     7981 agtgggttac atcgaactgg atctcaacag cggtaagatc cttgagagtt ttcgccccga
     8041 agaacgtttt ccaatgatga gcacttttaa agttctgcta tgtggcgcgg tattatcccg
     8101 tattgacgcc gggcaagagc aactcggtcg ccgcatacac tattctcaga atgacttgg
//
