LOCUS       sequence-466410-        6844 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  FKBP fused with ADAR2dd, with E488Q and T501A double mutation.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6844)
  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 6844)
  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: XZ135 CMV-TO-FKBP-ADARdd(QA) (FLP-IN)
FEATURES             Location/Qualifiers
     source          1..6844
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     polyA_signal    928..1049
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(965..984)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     1019..1038
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(1098..1114)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1098..1114)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1111..1133)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1122..1138
                     /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(1146..1152,1153..1170,1171..1176))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1191..1212
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(1329..1346)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1500..2088)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1580..1599)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2259..3050,3051..3119))
                     /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     2882..2901
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3120..3224)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(3299..3318)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        3485..3864
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        3865..4068
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     4019..4039
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     protein_bind    4070..4088
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    4091..4109
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     primer_bind     4119..4143
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     regulatory      4235..4244
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(4250..4252,4253..4573)
                     /codon_start=1
                     /product="human FK506-binding protein FKBP12"
                     /label=FKBP
                     /translation="MGVQVETISPGDGRTFPKRGQTCVVHYTGMLEDGKKFDSSRDRNK
                     PFKFMLGKQEVIRGWEEGVAQMSVGQRAKLTISPDYAYGATGHPGIIPPHATLVFDVEL
                     LKLE"
     CDS             5927..5956
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     primer_bind     complement(6012..6029)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    6018..6242
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(6414..6433)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     protein_bind    6565..6612
                     /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(6620..6844,1..798)
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="KKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRGY
                     VLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLPE
                     TELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQT
                     VMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMFG
                     DSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDGN
                     FDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAKE
                     "
     CDS             join(6620..6844,1..798)
                     /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 gtgcttgaca ttggggaatt cagcgagagc ctgacctatt gcatctcccg ccgtgcacag
       61 ggtgtcacgt tgcaagacct gcctgaaacc gaactgcccg ctgttctgca gccggtcgcg
      121 gaggccatgg atgcgatcgc tgcggccgat cttagccaga cgagcgggtt cggcccattc
      181 ggaccgcaag gaatcggtca atacactaca tggcgtgatt tcatatgcgc gattgctgat
      241 ccccatgtgt atcactggca aactgtgatg gacgacaccg tcagtgcgtc cgtcgcgcag
      301 gctctcgatg agctgatgct ttgggccgag gactgccccg aagtccggca cctcgtgcac
      361 gcggatttcg gctccaacaa tgtcctgacg gacaatggcc gcataacagc ggtcattgac
      421 tggagcgagg cgatgttcgg ggattcccaa tacgaggtcg ccaacatctt cttctggagg
      481 ccgtggttgg cttgtatgga gcagcagacg cgctacttcg agcggaggca tccggagctt
      541 gcaggatcgc cgcggctccg ggcgtatatg ctccgcattg gtcttgacca actctatcag
      601 agcttggttg acggcaattt cgatgatgca gcttgggcgc agggtcgatg cgacgcaatc
      661 gtccgatccg gagccgggac tgtcgggcgt acacaaatcg cccgcagaag cgcggccgtc
      721 tggaccgatg gctgtgtaga agtactcgcc gatagtggaa accgacgccc cagcactcgt
      781 ccgagggcaa aggaatagca cgtactacga gatttcgatt ccaccgccgc cttctatgaa
      841 aggttgggct tcggaatcgt tttccgggac gccggctgga tgatcctcca gcgcggggat
      901 ctcatgctgg agttcttcgc ccaccccaac ttgtttattg cagcttataa tggttacaaa
      961 taaagcaata gcatcacaaa tttcacaaat aaagcatttt tttcactgca ttctagttgt
     1021 ggtttgtcca aactcatcaa tgtatcttat catgtctgta taccgtcgac ctctagctag
     1081 agcttggcgt aatcatggtc atagctgttt cctgtgtgaa attgttatcc gctcacaatt
     1141 ccacacaaca tacgagccgg aagcataaag tgtaaagcct ggggtgccta atgagtgagc
     1201 taactcacat taattgcgtt gcgctcactg cccgctttcc agtcgggaaa cctgtcgtgc
     1261 cagctgcatt aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat tgggcgctct
     1321 tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca
     1381 gctcactcaa aggcggtaat acggttatcc acagaatcag gggataacgc aggaaagaac
     1441 atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt
     1501 ttccataggc tccgcccccc tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg
     1561 cgaaacccga caggactata aagataccag gcgtttcccc ctggaagctc cctcgtgcgc
     1621 tctcctgttc cgaccctgcc gcttaccgga tacctgtccg cctttctccc ttcgggaagc
     1681 gtggcgcttt ctcatagctc acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc
     1741 aagctgggct gtgtgcacga accccccgtt cagcccgacc gctgcgcctt atccggtaac
     1801 tatcgtcttg agtccaaccc ggtaagacac gacttatcgc cactggcagc agccactggt
     1861 aacaggatta gcagagcgag gtatgtaggc ggtgctacag agttcttgaa gtggtggcct
     1921 aactacggct acactagaag aacagtattt ggtatctgcg ctctgctgaa gccagttacc
     1981 ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt
     2041 ttttttgttt gcaagcagca gattacgcgc agaaaaaaag gatctcaaga agatcctttg
     2101 atcttttcta cggggtctga cgctcagtgg aacgaaaact cacgttaagg gattttggtc
     2161 atgagattat caaaaaggat cttcacctag atccttttaa attaaaaatg aagttttaaa
     2221 tcaatctaaa gtatatatga gtaaacttgg tctgacagtt accaatgctt aatcagtgag
     2281 gcacctatct cagcgatctg tctatttcgt tcatccatag ttgcctgact ccccgtcgtg
     2341 tagataacta cgatacggga gggcttacca tctggcccca gtgctgcaat gataccgcga
     2401 gacccacgct caccggctcc agatttatca gcaataaacc agccagccgg aagggccgag
     2461 cgcagaagtg gtcctgcaac tttatccgcc tccatccagt ctattaattg ttgccgggaa
     2521 gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat tgctacaggc
     2581 atcgtggtgt cacgctcgtc gtttggtatg gcttcattca gctccggttc ccaacgatca
     2641 aggcgagtta catgatcccc catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg
     2701 atcgttgtca gaagtaagtt ggccgcagtg ttatcactca tggttatggc agcactgcat
     2761 aattctctta ctgtcatgcc atccgtaaga tgcttttctg tgactggtga gtactcaacc
     2821 aagtcattct gagaatagtg tatgcggcga ccgagttgct cttgcccggc gtcaatacgg
     2881 gataataccg cgccacatag cagaacttta aaagtgctca tcattggaaa acgttcttcg
     2941 gggcgaaaac tctcaaggat cttaccgctg ttgagatcca gttcgatgta acccactcgt
     3001 gcacccaact gatcttcagc atcttttact ttcaccagcg tttctgggtg agcaaaaaca
     3061 ggaaggcaaa atgccgcaaa aaagggaata agggcgacac ggaaatgttg aatactcata
     3121 ctcttccttt ttcaatatta ttgaagcatt tatcagggtt attgtctcat gagcggatac
     3181 atatttgaat gtatttagaa aaataaacaa ataggggttc cgcgcacatt tccccgaaaa
     3241 gtgccacctg acgtcgacgg atcgggagat ctcccgatcc cctatggtgc actctcagta
     3301 caatctgctc tgatgccgca tagttaagcc agtatctgct ccctgcttgt gtgttggagg
     3361 tcgctgagta gtgcgcgagc aaaatttaag ctacaacaag gcaaggcttg accgacaatt
     3421 gcatgaagaa tctgcttagg gttaggcgtt ttgcgctgct tcgtacgggc cagatatacg
     3481 cgttgacatt gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat
     3541 agcccatata tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg
     3601 cccaacgacc cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata
     3661 gggactttcc attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta
     3721 catcaagtgt atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc
     3781 gcctggcatt atgcccagta catgacctta tgggactttc ctacttggca gtacatctac
     3841 gtattagtca tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga
     3901 tagcggtttg actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg
     3961 ttttggcacc aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg
     4021 caaatgggcg gtaggcgtgt acggtgggag gtctatataa gcagagctct ccctatcagt
     4081 gatagagatc tccctatcag tgatagagat cgtcgacgag ctcgtttagt gaaccgtcag
     4141 atcgcctgga gacgccatcc acgctgtttt gacctccata gaagacaccg ggaccgatcc
     4201 agcctccgga ctctagcgtt taaacttaag cttcgccacc atggctagca tgggagtgca
     4261 ggtggaaacc atctccccag gagacgggcg caccttcccc aagcgcggcc agacctgcgt
     4321 ggtgcactac accgggatgc ttgaagatgg aaagaaattt gattcctccc gggacagaaa
     4381 caagcccttt aagtttatgc taggcaagca ggaggtgatc cgaggctggg aagaaggggt
     4441 tgcccagatg agtgtgggtc agagagccaa actgactata tctccagatt atgcctatgg
     4501 tgccactggg cacccaggca tcatcccacc acatgccact ctcgtcttcg atgtggagct
     4561 tctaaaactg gaaggcgccg ggacaggagg gtctgggggt ggcactggcg gatctggagg
     4621 agactacaag gacgatgacg acaacggcgg atcggggggg tctggtggta gtggcggttc
     4681 ggggggaggt accggtttgc acttggatca aacgccatct cgccagccta ttcccagtga
     4741 gggtcttcag ctgcatttac cgcaggtttt agctgacgct gtctcacgcc tggtcctggg
     4801 taagtttggt gacctgaccg acaacttctc ctcccctcac gctcgcagaa aagtgctggc
     4861 tggagtcgtc atgacaacag gcacagatgt taaagatgcc aaggtgataa gtgtttctac
     4921 aggaacaaaa tgtattaatg gtgaatacat gagtgatcgt ggccttgcat taaatgactg
     4981 ccatgcagaa ataatatctc ggagatcctt gctcagattt ctttatacac aacttgagct
     5041 ttacttaaat aacaaagatg atcaaaaaag atccatcttt cagaaatcag agcgaggggg
     5101 gtttaggctg aaggagaatg tccagtttca tctgtacatc agcacctctc cctgtggaga
     5161 tgccagaatc ttctcaccac atgagccaat cctggaagaa ccagcagata gacacccaaa
     5221 tcgtaaagca agaggacagc tacggaccaa aatagagtct ggtcagggga cgattccagt
     5281 gcgctccaat gcgagcatcc aagcgtggga cggggtgctg caaggggagc ggctgctcac
     5341 catgtcctgc agtgacaaga ttgcacgctg gaacgtggtg ggcatccagg gatccctgct
     5401 cagcattttc gtggagccca tttacttctc gagcatcatc ctgggcagcc tttaccacgg
     5461 ggaccacctt tccagggcca tgtaccagcg gatctccaac atagaggacc tgccacctct
     5521 ctacaccctc aacaagcctt tgctcagtgg catcagcaat gcagaagcac ggcagccagg
     5581 gaaggccccc aacttcagtg tcaactggac ggtaggcgac tccgctattg aggtcatcaa
     5641 cgccacgact gggaaggatg agctgggccg cgcgtcccgc ctgtgtaagc acgcgttgta
     5701 ctgtcgctgg atgcgtgtgc acggcaaggt tccctcccac ttactacgct ccaagattac
     5761 caaacccaac gtgtaccatg agtccaagct ggcggcaaag gagtaccagg ccgccaaggc
     5821 gcgtctgttc acagccttca tcaaggcggg gctgggggcc tgggtggaga agcccaccga
     5881 gcaggaccag ttctcactca cgccctgagc cgctcgagga gggcccgaac aaaaactcat
     5941 ctcagaagag gatctgaata gcgccgtcga ccatcatcat aatcatcatt gagtttaaac
     6001 ccgctgatca gcctcgactg tgccttctag ttgccagcca tctgttgttt gcccctcccc
     6061 cgtgccttcc ttgaccctgg aaggtgccac tcccactgtc ctttcctaat aaaatgagga
     6121 aattgcatcg cattgtctga gtaggtgtca ttctattctg gggggtgggg tggggcagga
     6181 cagcaagggg gaggattggg aagacaatag caggcatgct ggggatgcgg tgggctctat
     6241 ggagatctat cgatgttctg gggatgcggt gggctctatg gcttctgagg cggaaagaac
     6301 cagctggggc tctagggggt atccccacgc gccctgtagc ggcgcattaa gcgcggcggg
     6361 tgtggtggtt acgcgcagcg tgaccgctac acttgccagc gccctagcgc ccgctccttt
     6421 cgctttcttc ccttcctttc tcgccacgtt cgccggcttt ccccgtcaag ctctaaatcg
     6481 ggggctccct ttagggttcc gatttagtgc tttacggcac ctcgacccca aaaaacttga
     6541 ttagggtgat ggttcacgta cctagaagtt cctattccga agttcctatt ctctagaaag
     6601 tataggaact tccttggcca aaaagcctga actcaccgcg acgtctgtcg agaagtttct
     6661 gatcgaaaag ttcgacagcg tctccgacct gatgcagctc tcggagggcg aagaatctcg
     6721 tgctttcagc ttcgatgtag gagggcgtgg atatgtcctg cgggtaaata gctgcgccga
     6781 tggtttctac aaagatcgtt atgtttatcg gcactttgca tcggccgcgc tcccgattcc
     6841 ggaa
//
