LOCUS       sequence-466389-        7147 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  ADAR2dd(E488Q,T501A)-MCP (positive control that fuses ADAR2dd and 
            MCP on the same protein).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7147)
  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 7147)
  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: LSM101 CMV-TO-ADAR2dd(E488Q,T501A)-MCP
FEATURES             Location/Qualifiers
     source          1..7147
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      547..1135
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1036..1055
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     1289..1306
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    1423..1444
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(1459..1464,1465..1482,1483..1489)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1497..1513
                     /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     1502..1524
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     1521..1537
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     1521..1537
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     polyA_signal    1586..1707
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1597..1616)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     1651..1670
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     CDS             complement(1837..2859)
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="KKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRGY
                     VLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLPE
                     TELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQT
                     VMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMFG
                     DSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDGN
                     FDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAKE
                     "
     CDS             complement(1837..2859)
                     /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(2867..2914)
                     /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     3046..3065
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     polyA_signal    3237..3461
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     3450..3467
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     CDS             complement(3523..3552)
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     primer_bind     complement(5639..5663)
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     protein_bind    5673..5691
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    5694..5712
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     promoter        complement(5714..5917)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(5743..5763)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        5918..6297
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     primer_bind     6464..6483
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        6558..6662
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6663..6731,6732..7147,1..376)
                     /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(6881..6900)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
ORIGIN
        1 ttgggaaccg gagctgaatg aagccatacc aaacgacgag cgtgacacca cgatgcctgt
       61 agcaatggca acaacgttgc gcaaactatt aactggcgaa ctacttactc tagcttcccg
      121 gcaacaatta atagactgga tggaggcgga taaagttgca ggaccacttc tgcgctcggc
      181 ccttccggct ggctggttta ttgctgataa atctggagcc ggtgagcgtg ggtctcgcgg
      241 tatcattgca gcactggggc cagatggtaa gccctcccgt atcgtagtta tctacacgac
      301 ggggagtcag gcaactatgg atgaacgaaa tagacagatc gctgagatag gtgcctcact
      361 gattaagcat tggtaactgt cagaccaagt ttactcatat atactttaga ttgatttaaa
      421 acttcatttt taatttaaaa ggatctaggt gaagatcctt tttgataatc tcatgaccaa
      481 aatcccttaa cgtgagtttt cgttccactg agcgtcagac cccgtagaaa agatcaaagg
      541 atcttcttga gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc
      601 gctaccagcg gtggtttgtt tgccggatca agagctacca actctttttc cgaaggtaac
      661 tggcttcagc agagcgcaga taccaaatac tgttcttcta gtgtagccgt agttaggcca
      721 ccacttcaag aactctgtag caccgcctac atacctcgct ctgctaatcc tgttaccagt
      781 ggctgctgcc agtggcgata agtcgtgtct taccgggttg gactcaagac gatagttacc
      841 ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc acacagccca gcttggagcg
      901 aacgacctac accgaactga gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc
      961 cgaagggaga aaggcggaca ggtatccggt aagcggcagg gtcggaacag gagagcgcac
     1021 gagggagctt ccagggggaa acgcctggta tctttatagt cctgtcgggt ttcgccacct
     1081 ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc
     1141 cagcaacgcg gcctttttac ggttcctggc cttttgctgg ccttttgctc acatgttctt
     1201 tcctgcgtta tcccctgatt ctgtggataa ccgtattacc gcctttgagt gagctgatac
     1261 cgctcgccgc agccgaacga ccgagcgcag cgagtcagtg agcgaggaag cggaagagcg
     1321 cccaatacgc aaaccgcctc tccccgcgcg ttggccgatt cattaatgca gctggcacga
     1381 caggtttccc gactggaaag cgggcagtga gcgcaacgca attaatgtga gttagctcac
     1441 tcattaggca ccccaggctt tacactttat gcttccggct cgtatgttgt gtggaattgt
     1501 gagcggataa caatttcaca caggaaacag ctatgaccat gattacgcca agctctagct
     1561 agaggtcgac ggtatacaga catgataaga tacattgatg agtttggaca aaccacaact
     1621 agaatgcagt gaaaaaaatg ctttatttgt gaaatttgtg atgctattgc tttatttgta
     1681 accattataa gctgcaataa acaagttggg gtgggcgaag aactccagca tgagatcccc
     1741 gcgctggagg atcatccagc cggcgtcccg gaaaacgatt ccgaagccca acctttcata
     1801 gaaggcggcg gtggaatcga aatctcgtag tacgtgctat tcctttgccc tcggacgagt
     1861 gctggggcgt cggtttccac tatcggcgag tacttctaca cagccatcgg tccagacggc
     1921 cgcgcttctg cgggcgattt gtgtacgccc gacagtcccg gctccggatc ggacgattgc
     1981 gtcgcatcga ccctgcgccc aagctgcatc atcgaaattg ccgtcaacca agctctgata
     2041 gagttggtca agaccaatgc ggagcatata cgcccggagc cgcggcgatc ctgcaagctc
     2101 cggatgcctc cgctcgaagt agcgcgtctg ctgctccata caagccaacc acggcctcca
     2161 gaagaagatg ttggcgacct cgtattggga atccccgaac atcgcctcgc tccagtcaat
     2221 gaccgctgtt atgcggccat tgtccgtcag gacattgttg gagccgaaat ccgcgtgcac
     2281 gaggtgccgg acttcggggc agtcctcggc ccaaagcatc agctcatcga gagcctgcgc
     2341 gacggacgca ctgacggtgt cgtccatcac agtttgccag tgatacacat ggggatcagc
     2401 aatcgcgcat atgaaatcac gccatgtagt gtattgaccg attccttgcg gtccgaatgg
     2461 gccgaacccg ctcgtctggc taagatcggc cgcagcgatc gcatccatgg cctccgcgac
     2521 cggctgcaga acagcgggca gttcggtttc aggcaggtct tgcaacgtga caccctgtgc
     2581 acggcgggag atgcaatagg tcaggctctc gctgaattcc ccaatgtcaa gcacttccgg
     2641 aatcgggagc gcggccgatg caaagtgccg ataaacataa cgatctttgt agaaaccatc
     2701 ggcgcagcta tttacccgca ggacatatcc acgccctcct acatcgaagc tgaaagcacg
     2761 agattcttcg ccctccgaga gctgcatcag gtcggagacg ctgtcgaact tttcgatcag
     2821 aaacttctcg acagacgtcg cggtgagttc aggctttttg gccaaggaag ttcctatact
     2881 ttctagagaa taggaacttc ggaataggaa cttctaggta cgtgaaccat caccctaatc
     2941 aagttttttg gggtcgaggt gccgtaaagc actaaatcgg aaccctaaag ggagcccccg
     3001 atttagagct tgacggggaa agccggcgaa cgtggcgaga aaggaaggga agaaagcgaa
     3061 aggagcgggc gctagggcgc tggcaagtgt agcggtcacg ctgcgcgtaa ccaccacacc
     3121 cgccgcgctt aatgcgccgc tacagggcgc gtggggatac cccctagagc cccagctggt
     3181 tctttccgcc tcagaagcca tagagcccac cgcatcccca gaacatcgat agatctccat
     3241 agagcccacc gcatccccag catgcctgct attgtcttcc caatcctccc ccttgctgtc
     3301 ctgccccacc ccacccccca gaatagaatg acacctactc agacaatgcg atgcaatttc
     3361 ctcattttat taggaaagga cagtgggagt ggcaccttcc agggtcaagg aaggcacggg
     3421 ggaggggcaa acaacagatg gctggcaact agaaggcaca gtcgaggctg atcagcgggt
     3481 ttaaactcaa tgatgattat gatgatggtc gacggcgcta ttcagatcct cttctgagat
     3541 gagtttttgt tcgggccctc ctcgagcggc tcagtaaatt ccgctattcg cagcgattgc
     3601 ggaaggtatc gggttgccat ctttcagaag tccttgcatg gcttttacaa taagctcgca
     3661 gtcgctattg gtcgcaaata tcggaatcgt caattccata ttcaagtatg agcgccatgc
     3721 acctttgggg acctccactt taatggtata cttccggttc tgggcactag attggcgtac
     3781 actacaagtc accttatagg cttgtgatct tgaattggaa gagatccact cggctatccc
     3841 attcgcaaag ttacttggag ctactgtgac gtctccagtg ccgccgttgt ccacgagcac
     3901 aaactgtgta aaattggagg ccatggaccc gccaccggtt ccgccaccat atattcctga
     3961 gttggcggcg atcgcagagg gaatcgggtt accgtctttc aaaagacctt gcatggcctt
     4021 gacaatcagc tcgcagtctg aattagtcgc aaagattggg atcgtaagct ccatattaag
     4081 ataagatcgc cacgcccctt ttggcacttc tacctttatg gtatattttc ggttctgtgc
     4141 ggaactttgg cgaactgagc atgtgacttt atatgcctga ctccggctgt tactacttat
     4201 ccattctgca atcccgttcg caaaattgct cggcgcaaca gttacgtctc cggtgccacc
     4261 attgtcaacc aacacaaatt gcgtaaagtt gctcgccata gccccgccac cggttccgcc
     4321 accggtaccg ggcgtgagtg agaactggtc ctgctcggtg ggcttctcca cccaggcccc
     4381 cagccccgcc ttgatgaagg ctgtgaacag acgcgccttg gcggcctggt actcctttgc
     4441 cgccagcttg gactcatggt acacgttggg tttggtaatc ttggagcgta gtaagtggga
     4501 gggaaccttg ccgtgcacac gcatccagcg acagtacaac gcgtgcttac acaggcggga
     4561 cgcgcggccc agctcatcct tcccagtcgt ggcgttgatg acctcaatag cggagtcgcc
     4621 taccgtccag ttgacactga agttgggggc cttccctggc tgccgtgctt ctgcattgct
     4681 gatgccactg agcaaaggct tgttgagggt gtagagaggt ggcaggtcct ctatgttgga
     4741 gatccgctgg tacatggccc tggaaaggtg gtccccgtgg taaaggctgc ccaggatgat
     4801 gctcgagaag taaatgggct ccacgaaaat gctgagcagg gatccctgga tgcccaccac
     4861 gttccagcgt gcaatcttgt cactgcagga catggtgagc agccgctccc cttgcagcac
     4921 cccgtcccac gcttggatgc tcgcattgga gcgcactgga atcgtcccct gaccagactc
     4981 tattttggtc cgtagctgtc ctcttgcttt acgatttggg tgtctatctg ctggttcttc
     5041 caggattggc tcatgtggtg agaagattct ggcatctcca cagggagagg tgctgatgta
     5101 cagatgaaac tggacattct ccttcagcct aaacccccct cgctctgatt tctgaaagat
     5161 ggatcttttt tgatcatctt tgttatttaa gtaaagctca agttgtgtat aaagaaatct
     5221 gagcaaggat ctccgagata ttatttctgc atggcagtca tttaatgcaa ggccacgatc
     5281 actcatgtat tcaccattaa tacattttgt tcctgtagaa acacttatca ccttggcatc
     5341 tttaacatct gtgcctgttg tcatgacgac tccagccagc acttttctgc gagcgtgagg
     5401 ggaggagaag ttgtcggtca ggtcaccaaa cttacccagg accaggcgtg agacagcgtc
     5461 agctaaaacc tgcggtaaat gcagctgaag accctcactg ggaataggct ggcgagatgg
     5521 cgtttgatcc aagtgcaaca tggtggcgaa gcttaagttt aaacgctaga gtccggaggc
     5581 tggatcggtc ccggtgtctt ctatggaggt caaaacagcg tggatggcgt ctccaggcga
     5641 tctgacggtt cactaaacga gctcgtcgac gatctctatc actgataggg agatctctat
     5701 cactgatagg gagagctctg cttatataga cctcccaccg tacacgccta ccgcccattt
     5761 gcgtcaatgg ggcggagttg ttacgacatt ttggaaagtc ccgttgattt tggtgccaaa
     5821 acaaactccc attgacgtca atggggtgga gacttggaaa tccccgtgag tcaaaccgct
     5881 atccacgccc attgatgtac tgccaaaacc gcatcaccat ggtaatagcg atgactaata
     5941 cgtagatgta ctgccaagta ggaaagtccc ataaggtcat gtactgggca taatgccagg
     6001 cgggccattt accgtcattg acgtcaatag ggggcgtact tggcatatga tacacttgat
     6061 gtactgccaa gtgggcagtt taccgtaaat actccaccca ttgacgtcaa tggaaagtcc
     6121 ctattggcgt tactatggga acatacgtca ttattgacgt caatgggcgg gggtcgttgg
     6181 gcggtcagcc aggcgggcca tttaccgtaa gttatgtaac gcggaactcc atatatgggc
     6241 tatgaactaa tgaccccgta attgattact attaataact agtcaataat caatgtcaac
     6301 gcgtatatct ggcccgtacg aagcagcgca aaacgcctaa ccctaagcag attcttcatg
     6361 caattgtcgg tcaagccttg ccttgttgta gcttaaattt tgctcgcgca ctactcagcg
     6421 acctccaaca cacaagcagg gagcagatac tggcttaact atgcggcatc agagcagatt
     6481 gtactgagag tgcaccatag gggatcggga gatctcccga tccgtcgacg tcaggtggca
     6541 cttttcgggg aaatgtgcgc ggaaccccta tttgtttatt tttctaaata cattcaaata
     6601 tgtatccgct catgagacaa taaccctgat aaatgcttca ataatattga aaaaggaaga
     6661 gtatgagtat tcaacatttc cgtgtcgccc ttattccctt ttttgcggca ttttgccttc
     6721 ctgtttttgc tcacccagaa acgctggtga aagtaaaaga tgctgaagat cagttgggtg
     6781 cacgagtggg ttacatcgaa ctggatctca acagcggtaa gatccttgag agttttcgcc
     6841 ccgaagaacg ttttccaatg atgagcactt ttaaagttct gctatgtggc gcggtattat
     6901 cccgtattga cgccgggcaa gagcaactcg gtcgccgcat acactattct cagaatgact
     6961 tggttgagta ctcaccagtc acagaaaagc atcttacgga tggcatgaca gtaagagaat
     7021 tatgcagtgc tgccataacc atgagtgata acactgcggc caacttactt ctgacaacga
     7081 tcggaggacc gaaggagcta accgcttttt tgcacaacat gggggatcat gtaactcgcc
     7141 ttgatcg
//
