LOCUS       sequence-466393-        7714 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  _-2 adrenergic receptor-ADAR2dd(E488Q, T501A), no V2 tail.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7714)
  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 7714)
  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: XZ142 CMV-TO-signal-B2AR-ADAR2dd(DD-E488Q-T501...
FEATURES             Location/Qualifiers
     source          1..7714
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      791..1379
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1280..1299
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     1533..1550
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    1667..1688
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(1703..1708,1709..1726,1727..1733)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1741..1757
                     /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     1746..1768
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     1765..1781
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     1765..1781
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     polyA_signal    1830..1951
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1841..1860)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     1895..1914
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     CDS             complement(2081..3103)
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="KKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRGY
                     VLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLPE
                     TELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQT
                     VMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMFG
                     DSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDGN
                     FDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAKE
                     "
     CDS             complement(2081..3103)
                     /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(3111..3158)
                     /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     3290..3309
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     polyA_signal    3481..3705
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     3694..3711
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     CDS             complement(3767..3796)
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     primer_bind     complement(6450..6474)
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     protein_bind    6484..6502
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    6505..6523
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     promoter        complement(6525..6728)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(6554..6574)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        6729..7108
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     primer_bind     7275..7294
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        7369..7473
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(7474..7542,7543..7714,1..620)
                     /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(7692..7711)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
ORIGIN
        1 gtattgacgc cgggcaagag caactcggtc gccgcataca ctattctcag aatgacttgg
       61 ttgagtactc accagtcaca gaaaagcatc ttacggatgg catgacagta agagaattat
      121 gcagtgctgc cataaccatg agtgataaca ctgcggccaa cttacttctg acaacgatcg
      181 gaggaccgaa ggagctaacc gcttttttgc acaacatggg ggatcatgta actcgccttg
      241 atcgttggga accggagctg aatgaagcca taccaaacga cgagcgtgac accacgatgc
      301 ctgtagcaat ggcaacaacg ttgcgcaaac tattaactgg cgaactactt actctagctt
      361 cccggcaaca attaatagac tggatggagg cggataaagt tgcaggacca cttctgcgct
      421 cggcccttcc ggctggctgg tttattgctg ataaatctgg agccggtgag cgtgggtctc
      481 gcggtatcat tgcagcactg gggccagatg gtaagccctc ccgtatcgta gttatctaca
      541 cgacggggag tcaggcaact atggatgaac gaaatagaca gatcgctgag ataggtgcct
      601 cactgattaa gcattggtaa ctgtcagacc aagtttactc atatatactt tagattgatt
      661 taaaacttca tttttaattt aaaaggatct aggtgaagat cctttttgat aatctcatga
      721 ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc agaccccgta gaaaagatca
      781 aaggatcttc ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac
      841 caccgctacc agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg
      901 taactggctt cagcagagcg cagataccaa atactgttct tctagtgtag ccgtagttag
      961 gccaccactt caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac
     1021 cagtggctgc tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt
     1081 taccggataa ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg
     1141 agcgaacgac ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc
     1201 ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc
     1261 gcacgaggga gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc
     1321 acctctgact tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa
     1381 acgccagcaa cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgt
     1441 tctttcctgc gttatcccct gattctgtgg ataaccgtat taccgccttt gagtgagctg
     1501 ataccgctcg ccgcagccga acgaccgagc gcagcgagtc agtgagcgag gaagcggaag
     1561 agcgcccaat acgcaaaccg cctctccccg cgcgttggcc gattcattaa tgcagctggc
     1621 acgacaggtt tcccgactgg aaagcgggca gtgagcgcaa cgcaattaat gtgagttagc
     1681 tcactcatta ggcaccccag gctttacact ttatgcttcc ggctcgtatg ttgtgtggaa
     1741 ttgtgagcgg ataacaattt cacacaggaa acagctatga ccatgattac gccaagctct
     1801 agctagaggt cgacggtata cagacatgat aagatacatt gatgagtttg gacaaaccac
     1861 aactagaatg cagtgaaaaa aatgctttat ttgtgaaatt tgtgatgcta ttgctttatt
     1921 tgtaaccatt ataagctgca ataaacaagt tggggtgggc gaagaactcc agcatgagat
     1981 ccccgcgctg gaggatcatc cagccggcgt cccggaaaac gattccgaag cccaaccttt
     2041 catagaaggc ggcggtggaa tcgaaatctc gtagtacgtg ctattccttt gccctcggac
     2101 gagtgctggg gcgtcggttt ccactatcgg cgagtacttc tacacagcca tcggtccaga
     2161 cggccgcgct tctgcgggcg atttgtgtac gcccgacagt cccggctccg gatcggacga
     2221 ttgcgtcgca tcgaccctgc gcccaagctg catcatcgaa attgccgtca accaagctct
     2281 gatagagttg gtcaagacca atgcggagca tatacgcccg gagccgcggc gatcctgcaa
     2341 gctccggatg cctccgctcg aagtagcgcg tctgctgctc catacaagcc aaccacggcc
     2401 tccagaagaa gatgttggcg acctcgtatt gggaatcccc gaacatcgcc tcgctccagt
     2461 caatgaccgc tgttatgcgg ccattgtccg tcaggacatt gttggagccg aaatccgcgt
     2521 gcacgaggtg ccggacttcg gggcagtcct cggcccaaag catcagctca tcgagagcct
     2581 gcgcgacgga cgcactgacg gtgtcgtcca tcacagtttg ccagtgatac acatggggat
     2641 cagcaatcgc gcatatgaaa tcacgccatg tagtgtattg accgattcct tgcggtccga
     2701 atgggccgaa cccgctcgtc tggctaagat cggccgcagc gatcgcatcc atggcctccg
     2761 cgaccggctg cagaacagcg ggcagttcgg tttcaggcag gtcttgcaac gtgacaccct
     2821 gtgcacggcg ggagatgcaa taggtcaggc tctcgctgaa ttccccaatg tcaagcactt
     2881 ccggaatcgg gagcgcggcc gatgcaaagt gccgataaac ataacgatct ttgtagaaac
     2941 catcggcgca gctatttacc cgcaggacat atccacgccc tcctacatcg aagctgaaag
     3001 cacgagattc ttcgccctcc gagagctgca tcaggtcgga gacgctgtcg aacttttcga
     3061 tcagaaactt ctcgacagac gtcgcggtga gttcaggctt tttggccaag gaagttccta
     3121 tactttctag agaataggaa cttcggaata ggaacttcta ggtacgtgaa ccatcaccct
     3181 aatcaagttt tttggggtcg aggtgccgta aagcactaaa tcggaaccct aaagggagcc
     3241 cccgatttag agcttgacgg ggaaagccgg cgaacgtggc gagaaaggaa gggaagaaag
     3301 cgaaaggagc gggcgctagg gcgctggcaa gtgtagcggt cacgctgcgc gtaaccacca
     3361 cacccgccgc gcttaatgcg ccgctacagg gcgcgtgggg atacccccta gagccccagc
     3421 tggttctttc cgcctcagaa gccatagagc ccaccgcatc cccagaacat cgatagatct
     3481 ccatagagcc caccgcatcc ccagcatgcc tgctattgtc ttcccaatcc tcccccttgc
     3541 tgtcctgccc caccccaccc cccagaatag aatgacacct actcagacaa tgcgatgcaa
     3601 tttcctcatt ttattaggaa aggacagtgg gagtggcacc ttccagggtc aaggaaggca
     3661 cgggggaggg gcaaacaaca gatggctggc aactagaagg cacagtcgag gctgatcagc
     3721 gggtttaaac tcaatgatga ttatgatgat ggtcgacggc gctattcaga tcctcttctg
     3781 agatgagttt ttgttcgggc cctcctcgag cggctcaggg cgtgagtgag aactggtcct
     3841 gctcggtggg cttctccacc caggccccca gccccgcctt gatgaaggct gtgaacagac
     3901 gcgccttggc ggcctggtac tcctttgccg ccagcttgga ctcatggtac acgttgggtt
     3961 tggtaatctt ggagcgtagt aagtgggagg gaaccttgcc gtgcacacgc atccagcgac
     4021 agtacaacgc gtgcttacac aggcgggacg cgcggcccag ctcatccttc ccagtcgtgg
     4081 cgttgatgac ctcaatagcg gagtcgccta ccgtccagtt gacactgaag ttgggggcct
     4141 tccctggctg ccgtgcttct gcattgctga tgccactgag caaaggcttg ttgagggtgt
     4201 agagaggtgg caggtcctct atgttggaga tccgctggta catggccctg gaaaggtggt
     4261 ccccgtggta aaggctgccc aggatgatgc tcgagaagta aatgggctcc acgaaaatgc
     4321 tgagcaggga tccctggatg cccaccacgt tccagcgtgc aatcttgtca ctgcaggaca
     4381 tggtgagcag ccgctcccct tgcagcaccc cgtcccacgc ttggatgctc gcattggagc
     4441 gcactggaat cgtcccctga ccagactcta ttttggtccg tagctgtcct cttgctttac
     4501 gatttgggtg tctatctgct ggttcttcca ggattggctc atgtggtgag aagattctgg
     4561 catctccaca gggagaggtg ctgatgtaca gatgaaactg gacattctcc ttcagcctaa
     4621 acccccctcg ctctgatttc tgaaagatgg atcttttttg atcatctttg ttatttaagt
     4681 aaagctcaag ttgtgtataa agaaatctga gcaaggatct ccgagatatt atttctgcat
     4741 ggcagtcatt taatgcaagg ccacgatcac tcatgtattc accattaata cattttgttc
     4801 ctgtagaaac acttatcacc ttggcatctt taacatctgt gcctgttgtc atgacgactc
     4861 cagccagcac ttttctgcga gcgtgagggg aggagaagtt gtcggtcagg tcaccaaact
     4921 tacccaggac caggcgtgag acagcgtcag ctaaaacctg cggtaaatgc agctgaagac
     4981 cctcactggg aataggctgg cgagatggcg tttgatccaa gtgcaaagca ccggtaccca
     5041 gcagtgagtc atttgtacta caattcctcc cttgtgaatc aatgttatcg ctaggcacag
     5101 taccttgatg gcccacaaag tcttccgtgc ctgggaggtc ttcacacagc agtttatttt
     5161 ctttctcctg ttccacgtga tatccactct gctcccctgt gttgccgttg ctggagtagc
     5221 cattcccata ggccttcaaa gaagacctgc gcaggcacag aagctcctgg aaggcaatcc
     5281 tgaaatctgg gctccggcag tagataaggg gattgaaacc agaattgaca tagcctatcc
     5341 aatttaggag gatgtaaact tccttacgga tgaggttatc ctggatcaca tgcacaatgt
     5401 taacgatgaa gaagggcagc cagcagaggg tgaaagtgcc catgatgatg cctaacgtct
     5461 tgagggcttt gtgctccttc aagcagaact tggaagatct gcggagtcca tgccccgtcc
     5521 gcccatcctg ctccacctgg ctaaggttct ggacatggaa gcggccctca gatttgtcaa
     5581 tcttctggag ctgccttttg gcctcctgaa agaccctgga gtagacgaag accatgatca
     5641 ccaggggaac gtagaaggac acgatggaag aggcaatggc ataggcttgg ttcgtgaaga
     5701 agtcacagca ggtctcattg gcatagcagt tgatggcttc ctggtgggtg gcccgatacc
     5761 agtgcatctg aatgggcaag aaggaggtaa ggcctgacac aatccacacc atcagaatga
     5821 tcacccgggc cttattcttg gtcagcaggc tctggtactt gaaaggtgaa gtaatggcaa
     5881 agtagcgatc cactgcgatc acgcacaggg tctcaatgct ggccgtgacg cacagcacat
     5941 caatggaagt ccaaaactcg caccagaagt tgccaaaagt ccacattttc ataagaatat
     6001 gggcggcccc aaagggcacc actgccaggc ccatgaccag atcagcacag gccagtgaag
     6061 tgatgaagta gttggtgacc gtctgcagac gctcgaactt ggcaatggct gtgatgacca
     6121 gcacattgcc aaacacgatg gccaggacga tgagagacat gacgatgccc atgcccacca
     6181 cccacacctc gtccctttgc tgcgtgacgt cgtggtccgg cgcatggctt ctattgggtg
     6241 ccagcaagaa ggcgctgccg ttcccgggtt gcccgccgct ggcgtcatca tcgtccttgt
     6301 agtcggcgaa taccaggcag aagatgtagc tcagggcgat gatcgtcttc atggtggcga
     6361 agcttaagtt taaacgctag agtccggagg ctggatcggt cccggtgtct tctatggagg
     6421 tcaaaacagc gtggatggcg tctccaggcg atctgacggt tcactaaacg agctcgtcga
     6481 cgatctctat cactgatagg gagatctcta tcactgatag ggagagctct gcttatatag
     6541 acctcccacc gtacacgcct accgcccatt tgcgtcaatg gggcggagtt gttacgacat
     6601 tttggaaagt cccgttgatt ttggtgccaa aacaaactcc cattgacgtc aatggggtgg
     6661 agacttggaa atccccgtga gtcaaaccgc tatccacgcc cattgatgta ctgccaaaac
     6721 cgcatcacca tggtaatagc gatgactaat acgtagatgt actgccaagt aggaaagtcc
     6781 cataaggtca tgtactgggc ataatgccag gcgggccatt taccgtcatt gacgtcaata
     6841 gggggcgtac ttggcatatg atacacttga tgtactgcca agtgggcagt ttaccgtaaa
     6901 tactccaccc attgacgtca atggaaagtc cctattggcg ttactatggg aacatacgtc
     6961 attattgacg tcaatgggcg ggggtcgttg ggcggtcagc caggcgggcc atttaccgta
     7021 agttatgtaa cgcggaactc catatatggg ctatgaacta atgaccccgt aattgattac
     7081 tattaataac tagtcaataa tcaatgtcaa cgcgtatatc tggcccgtac gaagcagcgc
     7141 aaaacgccta accctaagca gattcttcat gcaattgtcg gtcaagcctt gccttgttgt
     7201 agcttaaatt ttgctcgcgc actactcagc gacctccaac acacaagcag ggagcagata
     7261 ctggcttaac tatgcggcat cagagcagat tgtactgaga gtgcaccata ggggatcggg
     7321 agatctcccg atccgtcgac gtcaggtggc acttttcggg gaaatgtgcg cggaacccct
     7381 atttgtttat ttttctaaat acattcaaat atgtatccgc tcatgagaca ataaccctga
     7441 taaatgcttc aataatattg aaaaaggaag agtatgagta ttcaacattt ccgtgtcgcc
     7501 cttattccct tttttgcggc attttgcctt cctgtttttg ctcacccaga aacgctggtg
     7561 aaagtaaaag atgctgaaga tcagttgggt gcacgagtgg gttacatcga actggatctc
     7621 aacagcggta agatccttga gagttttcgc cccgaagaac gttttccaat gatgagcact
     7681 tttaaagttc tgctatgtgg cgcggtatta tccc
//
