LOCUS       sequence-466384-        7687 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AVPR2-V2-ADAR2dd(E488Q, T501A) with a short 4 amino acid linker and 
            extra V2 tail.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7687)
  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 7687)
  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: XZ030 CMV-TO-AVPR2-ADAR2(DD-E488Q-T501A) (FLP-IN)
FEATURES             Location/Qualifiers
     source          1..7687
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     1..20
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        95..199
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(200..268,269..1060)
                     /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(418..437)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      1231..1819
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1720..1739
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     1973..1990
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    2107..2128
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(2143..2148,2149..2166,2167..2173)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2181..2197
                     /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     2186..2208
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     2205..2221
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     2205..2221
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     polyA_signal    2270..2391
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2281..2300)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2335..2354
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     CDS             complement(2521..3543)
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="KKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRGY
                     VLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLPE
                     TELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQT
                     VMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMFG
                     DSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDGN
                     FDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAKE
                     "
     CDS             complement(2521..3543)
                     /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(3551..3598)
                     /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     3730..3749
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     polyA_signal    3921..4145
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     4134..4151
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     CDS             complement(4207..4236)
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     primer_bind     complement(6863..6887)
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     protein_bind    6897..6915
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    6918..6936
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     promoter        complement(6938..7141)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(6967..6987)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        7142..7521
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
ORIGIN
        1 cggcatcaga gcagattgta ctgagagtgc accatagggg atcgggagat ctcccgatcc
       61 gtcgacgtca ggtggcactt ttcggggaaa tgtgcgcgga acccctattt gtttattttt
      121 ctaaatacat tcaaatatgt atccgctcat gagacaataa ccctgataaa tgcttcaata
      181 atattgaaaa aggaagagta tgagtattca acatttccgt gtcgccctta ttcccttttt
      241 tgcggcattt tgccttcctg tttttgctca cccagaaacg ctggtgaaag taaaagatgc
      301 tgaagatcag ttgggtgcac gagtgggtta catcgaactg gatctcaaca gcggtaagat
      361 ccttgagagt tttcgccccg aagaacgttt tccaatgatg agcactttta aagttctgct
      421 atgtggcgcg gtattatccc gtattgacgc cgggcaagag caactcggtc gccgcataca
      481 ctattctcag aatgacttgg ttgagtactc accagtcaca gaaaagcatc ttacggatgg
      541 catgacagta agagaattat gcagtgctgc cataaccatg agtgataaca ctgcggccaa
      601 cttacttctg acaacgatcg gaggaccgaa ggagctaacc gcttttttgc acaacatggg
      661 ggatcatgta actcgccttg atcgttggga accggagctg aatgaagcca taccaaacga
      721 cgagcgtgac accacgatgc ctgtagcaat ggcaacaacg ttgcgcaaac tattaactgg
      781 cgaactactt actctagctt cccggcaaca attaatagac tggatggagg cggataaagt
      841 tgcaggacca cttctgcgct cggcccttcc ggctggctgg tttattgctg ataaatctgg
      901 agccggtgag cgtgggtctc gcggtatcat tgcagcactg gggccagatg gtaagccctc
      961 ccgtatcgta gttatctaca cgacggggag tcaggcaact atggatgaac gaaatagaca
     1021 gatcgctgag ataggtgcct cactgattaa gcattggtaa ctgtcagacc aagtttactc
     1081 atatatactt tagattgatt taaaacttca tttttaattt aaaaggatct aggtgaagat
     1141 cctttttgat aatctcatga ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc
     1201 agaccccgta gaaaagatca aaggatcttc ttgagatcct ttttttctgc gcgtaatctg
     1261 ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt tgtttgccgg atcaagagct
     1321 accaactctt tttccgaagg taactggctt cagcagagcg cagataccaa atactgttct
     1381 tctagtgtag ccgtagttag gccaccactt caagaactct gtagcaccgc ctacatacct
     1441 cgctctgcta atcctgttac cagtggctgc tgccagtggc gataagtcgt gtcttaccgg
     1501 gttggactca agacgatagt taccggataa ggcgcagcgg tcgggctgaa cggggggttc
     1561 gtgcacacag cccagcttgg agcgaacgac ctacaccgaa ctgagatacc tacagcgtga
     1621 gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg
     1681 cagggtcgga acaggagagc gcacgaggga gcttccaggg ggaaacgcct ggtatcttta
     1741 tagtcctgtc gggtttcgcc acctctgact tgagcgtcga tttttgtgat gctcgtcagg
     1801 ggggcggagc ctatggaaaa acgccagcaa cgcggccttt ttacggttcc tggccttttg
     1861 ctggcctttt gctcacatgt tctttcctgc gttatcccct gattctgtgg ataaccgtat
     1921 taccgccttt gagtgagctg ataccgctcg ccgcagccga acgaccgagc gcagcgagtc
     1981 agtgagcgag gaagcggaag agcgcccaat acgcaaaccg cctctccccg cgcgttggcc
     2041 gattcattaa tgcagctggc acgacaggtt tcccgactgg aaagcgggca gtgagcgcaa
     2101 cgcaattaat gtgagttagc tcactcatta ggcaccccag gctttacact ttatgcttcc
     2161 ggctcgtatg ttgtgtggaa ttgtgagcgg ataacaattt cacacaggaa acagctatga
     2221 ccatgattac gccaagctct agctagaggt cgacggtata cagacatgat aagatacatt
     2281 gatgagtttg gacaaaccac aactagaatg cagtgaaaaa aatgctttat ttgtgaaatt
     2341 tgtgatgcta ttgctttatt tgtaaccatt ataagctgca ataaacaagt tggggtgggc
     2401 gaagaactcc agcatgagat ccccgcgctg gaggatcatc cagccggcgt cccggaaaac
     2461 gattccgaag cccaaccttt catagaaggc ggcggtggaa tcgaaatctc gtagtacgtg
     2521 ctattccttt gccctcggac gagtgctggg gcgtcggttt ccactatcgg cgagtacttc
     2581 tacacagcca tcggtccaga cggccgcgct tctgcgggcg atttgtgtac gcccgacagt
     2641 cccggctccg gatcggacga ttgcgtcgca tcgaccctgc gcccaagctg catcatcgaa
     2701 attgccgtca accaagctct gatagagttg gtcaagacca atgcggagca tatacgcccg
     2761 gagccgcggc gatcctgcaa gctccggatg cctccgctcg aagtagcgcg tctgctgctc
     2821 catacaagcc aaccacggcc tccagaagaa gatgttggcg acctcgtatt gggaatcccc
     2881 gaacatcgcc tcgctccagt caatgaccgc tgttatgcgg ccattgtccg tcaggacatt
     2941 gttggagccg aaatccgcgt gcacgaggtg ccggacttcg gggcagtcct cggcccaaag
     3001 catcagctca tcgagagcct gcgcgacgga cgcactgacg gtgtcgtcca tcacagtttg
     3061 ccagtgatac acatggggat cagcaatcgc gcatatgaaa tcacgccatg tagtgtattg
     3121 accgattcct tgcggtccga atgggccgaa cccgctcgtc tggctaagat cggccgcagc
     3181 gatcgcatcc atggcctccg cgaccggctg cagaacagcg ggcagttcgg tttcaggcag
     3241 gtcttgcaac gtgacaccct gtgcacggcg ggagatgcaa taggtcaggc tctcgctgaa
     3301 ttccccaatg tcaagcactt ccggaatcgg gagcgcggcc gatgcaaagt gccgataaac
     3361 ataacgatct ttgtagaaac catcggcgca gctatttacc cgcaggacat atccacgccc
     3421 tcctacatcg aagctgaaag cacgagattc ttcgccctcc gagagctgca tcaggtcgga
     3481 gacgctgtcg aacttttcga tcagaaactt ctcgacagac gtcgcggtga gttcaggctt
     3541 tttggccaag gaagttccta tactttctag agaataggaa cttcggaata ggaacttcta
     3601 ggtacgtgaa ccatcaccct aatcaagttt tttggggtcg aggtgccgta aagcactaaa
     3661 tcggaaccct aaagggagcc cccgatttag agcttgacgg ggaaagccgg cgaacgtggc
     3721 gagaaaggaa gggaagaaag cgaaaggagc gggcgctagg gcgctggcaa gtgtagcggt
     3781 cacgctgcgc gtaaccacca cacccgccgc gcttaatgcg ccgctacagg gcgcgtgggg
     3841 atacccccta gagccccagc tggttctttc cgcctcagaa gccatagagc ccaccgcatc
     3901 cccagaacat cgatagatct ccatagagcc caccgcatcc ccagcatgcc tgctattgtc
     3961 ttcccaatcc tcccccttgc tgtcctgccc caccccaccc cccagaatag aatgacacct
     4021 actcagacaa tgcgatgcaa tttcctcatt ttattaggaa aggacagtgg gagtggcacc
     4081 ttccagggtc aaggaaggca cgggggaggg gcaaacaaca gatggctggc aactagaagg
     4141 cacagtcgag gctgatcagc gggtttaaac tcaatgatga ttatgatgat ggtcgacggc
     4201 gctattcaga tcctcttctg agatgagttt ttgttcgggc cctcctcgag cggctcaggg
     4261 cgtgagtgag aactggtcct gctcggtggg cttctccacc caggccccca gccccgcctt
     4321 gatgaaggct gtgaacagac gcgccttggc ggcctggtac tcctttgccg ccagcttgga
     4381 ctcatggtac acgttgggtt tggtaatctt ggagcgtagt aagtgggagg gaaccttgcc
     4441 gtgcacacgc atccagcgac agtacaacgc gtgcttacac aggcgggacg cgcggcccag
     4501 ctcatccttc ccagtcgtgg cgttgatgac ctcaatagcg gagtcgccta ccgtccagtt
     4561 gacactgaag ttgggggcct tccctggctg ccgtgcttct gcattgctga tgccactgag
     4621 caaaggcttg ttgagggtgt agagaggtgg caggtcctct atgttggaga tccgctggta
     4681 catggccctg gaaaggtggt ccccgtggta aaggctgccc aggatgatgc tcgagaagta
     4741 aatgggctcc acgaaaatgc tgagcaggga tccctggatg cccaccacgt tccagcgtgc
     4801 aatcttgtca ctgcaggaca tggtgagcag ccgctcccct tgcagcaccc cgtcccacgc
     4861 ttggatgctc gcattggagc gcactggaat cgtcccctga ccagactcta ttttggtccg
     4921 tagctgtcct cttgctttac gatttgggtg tctatctgct ggttcttcca ggattggctc
     4981 atgtggtgag aagattctgg catctccaca gggagaggtg ctgatgtaca gatgaaactg
     5041 gacattctcc ttcagcctaa acccccctcg ctctgatttc tgaaagatgg atcttttttg
     5101 atcatctttg ttatttaagt aaagctcaag ttgtgtataa agaaatctga gcaaggatct
     5161 ccgagatatt atttctgcat ggcagtcatt taatgcaagg ccacgatcac tcatgtattc
     5221 accattaata cattttgttc ctgtagaaac acttatcacc ttggcatctt taacatctgt
     5281 gcctgttgtc atgacgactc cagccagcac ttttctgcga gcgtgagggg aggagaagtt
     5341 gtcggtcagg tcaccaaact tacccaggac caggcgtgag acagcgtcag ctaaaacctg
     5401 cggtaaatgc agctgaagac cctcactggg aataggctgg cgagatggcg tttgatccaa
     5461 gtgcaaagca ccggtacccg atgaagtgtc cttggccagg gaggagctgg cggtggtgca
     5521 ggactcatct tggggaccca ggctgggtgg ggtgcgtcca ccggtatcga tagaagaggt
     5581 gtctttggcc agggaactgg aggcggtagt gcaggattcg tcctgtggac ccaggcttgg
     5641 gggggtcctg ccccgggcgc aacagagaag actccgcagc tcggagctaa cactggagga
     5701 gaaagaggcg taaatccaag gattggtaca gctattcagt gaagcgagaa gcatgagcag
     5761 aacgaatgga gcaccttcca ggggtgcttc ggggtcccag gccgcccaga gctgcaccaa
     5821 aaaaaacggg gcccagcaaa gaacgtaaac gacgacgatg accagtgtca tgcgtacggt
     5881 cttagctacc gcagctgata cgtgggcgcc ttcgcctgga gagccagtcc ttcgaccccg
     5941 tcttcttcca ccagggcgct cgcttggccc aggaaccaag gaggcgtgta tttctctgaa
     6001 tatcaggacc tggcatgcgg cgatccccag ggtgggagca acgaacacca tgagagcaat
     6061 ccacgtcacg taggtccttc tgccccatgg ctcagcgaag caagcccagc aatcggtcac
     6121 gccagatccg ccctccacat ttctctgtgc gaaaataaac aactggggca agctcagaag
     6181 cagggaaaaa gcccacgcaa ccagaactgg cctgttccaa tgggctccgg aaccgtgcct
     6241 atacgccagc atggggcgac agatagccct gtgtctgtcc aaggtcatag caagtatcat
     6301 atatgaactg gcatacattc ccaccatctg caggtatttc acggcccgac acagtgcgtc
     6361 tggcccccgg aatctatcgg tggctttcca agccagttga gggagcacct gaaacagcgc
     6421 cacggccagg tctgcaagac agagatggcc gataaagaca tgaataggcg cccagtggcc
     6481 tcggcggcct cgtcgcgcca gggcggccag caccagacca ttgctcaacg cgacggcaac
     6541 gaaaacgatg gagagcaagg cgagctccgc tcttgcaaga agtgggtccc gggtatccag
     6601 aggcctctcc tgtgagctgt tggagggaag acttggaaga gatggatgtc cgggcacggc
     6661 gcttgtggta gaggccatga gcatatcgat gctggcgtca tcatcgtcct tgtagtcggc
     6721 gaataccagg cagaagatgt agctcagggc gatgatcgtc ttcatggtgg cgaagcttaa
     6781 gtttaaacgc tagagtccgg aggctggatc ggtcccggtg tcttctatgg aggtcaaaac
     6841 agcgtggatg gcgtctccag gcgatctgac ggttcactaa acgagctcgt cgacgatctc
     6901 tatcactgat agggagatct ctatcactga tagggagagc tctgcttata tagacctccc
     6961 accgtacacg cctaccgccc atttgcgtca atggggcgga gttgttacga cattttggaa
     7021 agtcccgttg attttggtgc caaaacaaac tcccattgac gtcaatgggg tggagacttg
     7081 gaaatccccg tgagtcaaac cgctatccac gcccattgat gtactgccaa aaccgcatca
     7141 ccatggtaat agcgatgact aatacgtaga tgtactgcca agtaggaaag tcccataagg
     7201 tcatgtactg ggcataatgc caggcgggcc atttaccgtc attgacgtca atagggggcg
     7261 tacttggcat atgatacact tgatgtactg ccaagtgggc agtttaccgt aaatactcca
     7321 cccattgacg tcaatggaaa gtccctattg gcgttactat gggaacatac gtcattattg
     7381 acgtcaatgg gcgggggtcg ttgggcggtc agccaggcgg gccatttacc gtaagttatg
     7441 taacgcggaa ctccatatat gggctatgaa ctaatgaccc cgtaattgat tactattaat
     7501 aactagtcaa taatcaatgt caacgcgtat atctggcccg tacgaagcag cgcaaaacgc
     7561 ctaaccctaa gcagattctt catgcaattg tcggtcaagc cttgccttgt tgtagcttaa
     7621 attttgctcg cgcactactc agcgacctcc aacacacaag cagggagcag atactggctt
     7681 aactatg
//
