LOCUS       sequence-466391-        6733 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  ER-retained catalytically inactive ADAR2dd (E396A mutation).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6733)
  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 6733)
  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: YH001 CMV-TO-tm-GS+XTEN-ADAR2(DD-E396A)-RXR (F...
FEATURES             Location/Qualifiers
     source          1..6733
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     563..582
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(801..905)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(980..999)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        1166..1545
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        1546..1749
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     1700..1720
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     protein_bind    1751..1769
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    1772..1790
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     primer_bind     1800..1824
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     regulatory      1916..1925
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             3497..3526
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     primer_bind     complement(3582..3599)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3588..3812
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(3984..4003)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     protein_bind    4135..4182
                     /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             4190..5212
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="KKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRGY
                     VLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLPE
                     TELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQT
                     VMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMFG
                     DSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDGN
                     FDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAKE
                     "
     CDS             4190..5212
                     /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    5342..5463
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5379..5398)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5433..5452
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5512..5528)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5512..5528)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5525..5547)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5536..5552
                     /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(5560..5566,5567..5584,5585..5590))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5605..5626
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5743..5760)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5914..6502)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5994..6013)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6673..6733,1..731,732..800))
                     /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"
ORIGIN
        1 gttgcctgac tccccgtcgt gtagataact acgatacggg agggcttacc atctggcccc
       61 agtgctgcaa tgataccgcg agacccacgc tcaccggctc cagatttatc agcaataaac
      121 cagccagccg gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag
      181 tctattaatt gttgccggga agctagagta agtagttcgc cagttaatag tttgcgcaac
      241 gttgttgcca ttgctacagg catcgtggtg tcacgctcgt cgtttggtat ggcttcattc
      301 agctccggtt cccaacgatc aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg
      361 gttagctcct tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt gttatcactc
      421 atggttatgg cagcactgca taattctctt actgtcatgc catccgtaag atgcttttct
      481 gtgactggtg agtactcaac caagtcattc tgagaatagt gtatgcggcg accgagttgc
      541 tcttgcccgg cgtcaatacg ggataatacc gcgccacata gcagaacttt aaaagtgctc
      601 atcattggaa aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc
      661 agttcgatgt aacccactcg tgcacccaac tgatcttcag catcttttac tttcaccagc
      721 gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca
      781 cggaaatgtt gaatactcat actcttcctt tttcaatatt attgaagcat ttatcagggt
      841 tattgtctca tgagcggata catatttgaa tgtatttaga aaaataaaca aataggggtt
      901 ccgcgcacat ttccccgaaa agtgccacct gacgtcgacg gatcgggaga tctcccgatc
      961 ccctatggtg cactctcagt acaatctgct ctgatgccgc atagttaagc cagtatctgc
     1021 tccctgcttg tgtgttggag gtcgctgagt agtgcgcgag caaaatttaa gctacaacaa
     1081 ggcaaggctt gaccgacaat tgcatgaaga atctgcttag ggttaggcgt tttgcgctgc
     1141 ttcgtacggg ccagatatac gcgttgacat tgattattga ctagttatta atagtaatca
     1201 attacggggt cattagttca tagcccatat atggagttcc gcgttacata acttacggta
     1261 aatggcccgc ctggctgacc gcccaacgac ccccgcccat tgacgtcaat aatgacgtat
     1321 gttcccatag taacgccaat agggactttc cattgacgtc aatgggtgga gtatttacgg
     1381 taaactgccc acttggcagt acatcaagtg tatcatatgc caagtacgcc ccctattgac
     1441 gtcaatgacg gtaaatggcc cgcctggcat tatgcccagt acatgacctt atgggacttt
     1501 cctacttggc agtacatcta cgtattagtc atcgctatta ccatggtgat gcggttttgg
     1561 cagtacatca atgggcgtgg atagcggttt gactcacggg gatttccaag tctccacccc
     1621 attgacgtca atgggagttt gttttggcac caaaatcaac gggactttcc aaaatgtcgt
     1681 aacaactccg ccccattgac gcaaatgggc ggtaggcgtg tacggtggga ggtctatata
     1741 agcagagctc tccctatcag tgatagagat ctccctatca gtgatagaga tcgtcgacga
     1801 gctcgtttag tgaaccgtca gatcgcctgg agacgccatc cacgctgttt tgacctccat
     1861 agaagacacc gggaccgatc cagcctccgg actctagcgt ttaaacttaa gcttcgccac
     1921 catggcgctc cctgtcaccg cactgcttct tccgctggca ctgctgctgc acgctgcacg
     1981 gcctggatcc ctggtcgtgg ttgctggagt cctgttttgt tatggcttgc tagtgacagt
     2041 ggctctttgt gttgaaaatt ccgggtcagg cggttccggc agcggggcgg gtagcgggtc
     2101 cagtgggtca gaaacacccg gcacgagtga gtcagcaacg ccagagagtg ctagcgcttt
     2161 gcacttggat caaacgccat ctcgccagcc tattcccagt gagggtcttc agctgcattt
     2221 accgcaggtt ttagctgacg ctgtctcacg cctggtcctg ggtaagtttg gtgacctgac
     2281 cgacaacttc tcctcccctc acgctcgcag aaaagtgctg gctggagtcg tcatgacaac
     2341 aggcacagat gttaaagatg ccaaggtgat aagtgtttct acaggaacaa aatgtattaa
     2401 tggtgaatac atgagtgatc gtggccttgc attaaatgac tgccatgcag ccataatatc
     2461 tcggagatcc ttgctcagat ttctttatac acaacttgag ctttacttaa ataacaaaga
     2521 tgatcaaaaa agatccatct ttcagaaatc agagcgaggg gggtttaggc tgaaggagaa
     2581 tgtccagttt catctgtaca tcagcacctc tccctgtgga gatgccagaa tcttctcacc
     2641 acatgagcca atcctggaag aaccagcaga tagacaccca aatcgtaaag caagaggaca
     2701 gctacggacc aaaatagagt ctggtgaggg gacgattcca gtgcgctcca atgcgagcat
     2761 ccaaacgtgg gacggggtgc tgcaagggga gcggctgctc accatgtcct gcagtgacaa
     2821 gattgcacgc tggaacgtgg tgggcatcca gggatccctg ctcagcattt tcgtggagcc
     2881 catttacttc tcgagcatca tcctgggcag cctttaccac ggggaccacc tttccagggc
     2941 catgtaccag cggatctcca acatagagga cctgccacct ctctacaccc tcaacaagcc
     3001 tttgctcagt ggcatcagca atgcagaagc acggcagcca gggaaggccc ccaacttcag
     3061 tgtcaactgg acggtaggcg actccgctat tgaggtcatc aacgccacga ctgggaagga
     3121 tgagctgggc cgcgcgtccc gcctgtgtaa gcacgcgttg tactgtcgct ggatgcgtgt
     3181 gcacggcaag gttccctccc acttactacg ctccaagatt accaaaccca acgtgtacca
     3241 tgagtccaag ctggcggcaa aggagtacca ggccgccaag gcgcgtctgt tcacagcctt
     3301 catcaaggcg gggctggggg cctgggtgga gaagcccacc gagcaggacc agttctcact
     3361 cacgcccctt ctcgaagccc tgaccttagc cagtgcacgg ggacccctga ggaagagatc
     3421 tgtgccaatg gctaaggcga aaccgaaatt ctccatctca cctgacagct tgagctgagc
     3481 cgctcgagga gggcccgaac aaaaactcat ctcagaagag gatctgaata gcgccgtcga
     3541 ccatcatcat aatcatcatt gagtttaaac ccgctgatca gcctcgactg tgccttctag
     3601 ttgccagcca tctgttgttt gcccctcccc cgtgccttcc ttgaccctgg aaggtgccac
     3661 tcccactgtc ctttcctaat aaaatgagga aattgcatcg cattgtctga gtaggtgtca
     3721 ttctattctg gggggtgggg tggggcagga cagcaagggg gaggattggg aagacaatag
     3781 caggcatgct ggggatgcgg tgggctctat ggagatctat cgatgttctg gggatgcggt
     3841 gggctctatg gcttctgagg cggaaagaac cagctggggc tctagggggt atccccacgc
     3901 gccctgtagc ggcgcattaa gcgcggcggg tgtggtggtt acgcgcagcg tgaccgctac
     3961 acttgccagc gccctagcgc ccgctccttt cgctttcttc ccttcctttc tcgccacgtt
     4021 cgccggcttt ccccgtcaag ctctaaatcg ggggctccct ttagggttcc gatttagtgc
     4081 tttacggcac ctcgacccca aaaaacttga ttagggtgat ggttcacgta cctagaagtt
     4141 cctattccga agttcctatt ctctagaaag tataggaact tccttggcca aaaagcctga
     4201 actcaccgcg acgtctgtcg agaagtttct gatcgaaaag ttcgacagcg tctccgacct
     4261 gatgcagctc tcggagggcg aagaatctcg tgctttcagc ttcgatgtag gagggcgtgg
     4321 atatgtcctg cgggtaaata gctgcgccga tggtttctac aaagatcgtt atgtttatcg
     4381 gcactttgca tcggccgcgc tcccgattcc ggaagtgctt gacattgggg aattcagcga
     4441 gagcctgacc tattgcatct cccgccgtgc acagggtgtc acgttgcaag acctgcctga
     4501 aaccgaactg cccgctgttc tgcagccggt cgcggaggcc atggatgcga tcgctgcggc
     4561 cgatcttagc cagacgagcg ggttcggccc attcggaccg caaggaatcg gtcaatacac
     4621 tacatggcgt gatttcatat gcgcgattgc tgatccccat gtgtatcact ggcaaactgt
     4681 gatggacgac accgtcagtg cgtccgtcgc gcaggctctc gatgagctga tgctttgggc
     4741 cgaggactgc cccgaagtcc ggcacctcgt gcacgcggat ttcggctcca acaatgtcct
     4801 gacggacaat ggccgcataa cagcggtcat tgactggagc gaggcgatgt tcggggattc
     4861 ccaatacgag gtcgccaaca tcttcttctg gaggccgtgg ttggcttgta tggagcagca
     4921 gacgcgctac ttcgagcgga ggcatccgga gcttgcagga tcgccgcggc tccgggcgta
     4981 tatgctccgc attggtcttg accaactcta tcagagcttg gttgacggca atttcgatga
     5041 tgcagcttgg gcgcagggtc gatgcgacgc aatcgtccga tccggagccg ggactgtcgg
     5101 gcgtacacaa atcgcccgca gaagcgcggc cgtctggacc gatggctgtg tagaagtact
     5161 cgccgatagt ggaaaccgac gccccagcac tcgtccgagg gcaaaggaat agcacgtact
     5221 acgagatttc gattccaccg ccgccttcta tgaaaggttg ggcttcggaa tcgttttccg
     5281 ggacgccggc tggatgatcc tccagcgcgg ggatctcatg ctggagttct tcgcccaccc
     5341 caacttgttt attgcagctt ataatggtta caaataaagc aatagcatca caaatttcac
     5401 aaataaagca tttttttcac tgcattctag ttgtggtttg tccaaactca tcaatgtatc
     5461 ttatcatgtc tgtataccgt cgacctctag ctagagcttg gcgtaatcat ggtcatagct
     5521 gtttcctgtg tgaaattgtt atccgctcac aattccacac aacatacgag ccggaagcat
     5581 aaagtgtaaa gcctggggtg cctaatgagt gagctaactc acattaattg cgttgcgctc
     5641 actgcccgct ttccagtcgg gaaacctgtc gtgccagctg cattaatgaa tcggccaacg
     5701 cgcggggaga ggcggtttgc gtattgggcg ctcttccgct tcctcgctca ctgactcgct
     5761 gcgctcggtc gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg taatacggtt
     5821 atccacagaa tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc
     5881 caggaaccgt aaaaaggccg cgttgctggc gtttttccat aggctccgcc cccctgacga
     5941 gcatcacaaa aatcgacgct caagtcagag gtggcgaaac ccgacaggac tataaagata
     6001 ccaggcgttt ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac
     6061 cggatacctg tccgcctttc tcccttcggg aagcgtggcg ctttctcata gctcacgctg
     6121 taggtatctc agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc
     6181 cgttcagccc gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag
     6241 acacgactta tcgccactgg cagcagccac tggtaacagg attagcagag cgaggtatgt
     6301 aggcggtgct acagagttct tgaagtggtg gcctaactac ggctacacta gaagaacagt
     6361 atttggtatc tgcgctctgc tgaagccagt taccttcgga aaaagagttg gtagctcttg
     6421 atccggcaaa caaaccaccg ctggtagcgg tggttttttt gtttgcaagc agcagattac
     6481 gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt tctacggggt ctgacgctca
     6541 gtggaacgaa aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac
     6601 ctagatcctt ttaaattaaa aatgaagttt taaatcaatc taaagtatat atgagtaaac
     6661 ttggtctgac agttaccaat gcttaatcag tgaggcacct atctcagcga tctgtctatt
     6721 tcgttcatcc ata
//
