LOCUS       sequence-466406-        7690 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  CCR6-V2-ADAR2dd(E488Q, T501A) with a short 4 amino acid linker and 
            V2 tail.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7690)
  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 7690)
  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: XZ032 CMV-TO-CCR6-V2-ADAR2dd(E488Q, T501A)
FEATURES             Location/Qualifiers
     source          1..7690
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      779..1367
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1268..1287
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     1521..1538
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    1655..1676
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(1691..1696,1697..1714,1715..1721)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1729..1745
                     /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     1734..1756
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     1753..1769
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     1753..1769
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     polyA_signal    1818..1939
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1829..1848)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     1883..1902
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     CDS             complement(2069..3091)
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="KKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRGY
                     VLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLPE
                     TELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQT
                     VMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMFG
                     DSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDGN
                     FDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAKE
                     "
     CDS             complement(2069..3091)
                     /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(3099..3146)
                     /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     3278..3297
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     polyA_signal    3469..3693
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     3682..3699
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     CDS             complement(3755..3784)
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     primer_bind     complement(6414..6438)
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     protein_bind    6448..6466
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    6469..6487
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     promoter        complement(6489..6692)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(6518..6538)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        6693..7072
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     primer_bind     7239..7258
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        7333..7437
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(7438..7506,7507..7690,1..608)
                     /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(7656..7675)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
ORIGIN
        1 ggcaagagca actcggtcgc cgcatacact attctcagaa tgacttggtt gagtactcac
       61 cagtcacaga aaagcatctt acggatggca tgacagtaag agaattatgc agtgctgcca
      121 taaccatgag tgataacact gcggccaact tacttctgac aacgatcgga ggaccgaagg
      181 agctaaccgc ttttttgcac aacatggggg atcatgtaac tcgccttgat cgttgggaac
      241 cggagctgaa tgaagccata ccaaacgacg agcgtgacac cacgatgcct gtagcaatgg
      301 caacaacgtt gcgcaaacta ttaactggcg aactacttac tctagcttcc cggcaacaat
      361 taatagactg gatggaggcg gataaagttg caggaccact tctgcgctcg gcccttccgg
      421 ctggctggtt tattgctgat aaatctggag ccggtgagcg tgggtctcgc ggtatcattg
      481 cagcactggg gccagatggt aagccctccc gtatcgtagt tatctacacg acggggagtc
      541 aggcaactat ggatgaacga aatagacaga tcgctgagat aggtgcctca ctgattaagc
      601 attggtaact gtcagaccaa gtttactcat atatacttta gattgattta aaacttcatt
      661 tttaatttaa aaggatctag gtgaagatcc tttttgataa tctcatgacc aaaatccctt
      721 aacgtgagtt ttcgttccac tgagcgtcag accccgtaga aaagatcaaa ggatcttctt
      781 gagatccttt ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag
      841 cggtggtttg tttgccggat caagagctac caactctttt tccgaaggta actggcttca
      901 gcagagcgca gataccaaat actgttcttc tagtgtagcc gtagttaggc caccacttca
      961 agaactctgt agcaccgcct acatacctcg ctctgctaat cctgttacca gtggctgctg
     1021 ccagtggcga taagtcgtgt cttaccgggt tggactcaag acgatagtta ccggataagg
     1081 cgcagcggtc gggctgaacg gggggttcgt gcacacagcc cagcttggag cgaacgacct
     1141 acaccgaact gagataccta cagcgtgagc tatgagaaag cgccacgctt cccgaaggga
     1201 gaaaggcgga caggtatccg gtaagcggca gggtcggaac aggagagcgc acgagggagc
     1261 ttccaggggg aaacgcctgg tatctttata gtcctgtcgg gtttcgccac ctctgacttg
     1321 agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac gccagcaacg
     1381 cggccttttt acggttcctg gccttttgct ggccttttgc tcacatgttc tttcctgcgt
     1441 tatcccctga ttctgtggat aaccgtatta ccgcctttga gtgagctgat accgctcgcc
     1501 gcagccgaac gaccgagcgc agcgagtcag tgagcgagga agcggaagag cgcccaatac
     1561 gcaaaccgcc tctccccgcg cgttggccga ttcattaatg cagctggcac gacaggtttc
     1621 ccgactggaa agcgggcagt gagcgcaacg caattaatgt gagttagctc actcattagg
     1681 caccccaggc tttacacttt atgcttccgg ctcgtatgtt gtgtggaatt gtgagcggat
     1741 aacaatttca cacaggaaac agctatgacc atgattacgc caagctctag ctagaggtcg
     1801 acggtataca gacatgataa gatacattga tgagtttgga caaaccacaa ctagaatgca
     1861 gtgaaaaaaa tgctttattt gtgaaatttg tgatgctatt gctttatttg taaccattat
     1921 aagctgcaat aaacaagttg gggtgggcga agaactccag catgagatcc ccgcgctgga
     1981 ggatcatcca gccggcgtcc cggaaaacga ttccgaagcc caacctttca tagaaggcgg
     2041 cggtggaatc gaaatctcgt agtacgtgct attcctttgc cctcggacga gtgctggggc
     2101 gtcggtttcc actatcggcg agtacttcta cacagccatc ggtccagacg gccgcgcttc
     2161 tgcgggcgat ttgtgtacgc ccgacagtcc cggctccgga tcggacgatt gcgtcgcatc
     2221 gaccctgcgc ccaagctgca tcatcgaaat tgccgtcaac caagctctga tagagttggt
     2281 caagaccaat gcggagcata tacgcccgga gccgcggcga tcctgcaagc tccggatgcc
     2341 tccgctcgaa gtagcgcgtc tgctgctcca tacaagccaa ccacggcctc cagaagaaga
     2401 tgttggcgac ctcgtattgg gaatccccga acatcgcctc gctccagtca atgaccgctg
     2461 ttatgcggcc attgtccgtc aggacattgt tggagccgaa atccgcgtgc acgaggtgcc
     2521 ggacttcggg gcagtcctcg gcccaaagca tcagctcatc gagagcctgc gcgacggacg
     2581 cactgacggt gtcgtccatc acagtttgcc agtgatacac atggggatca gcaatcgcgc
     2641 atatgaaatc acgccatgta gtgtattgac cgattccttg cggtccgaat gggccgaacc
     2701 cgctcgtctg gctaagatcg gccgcagcga tcgcatccat ggcctccgcg accggctgca
     2761 gaacagcggg cagttcggtt tcaggcaggt cttgcaacgt gacaccctgt gcacggcggg
     2821 agatgcaata ggtcaggctc tcgctgaatt ccccaatgtc aagcacttcc ggaatcggga
     2881 gcgcggccga tgcaaagtgc cgataaacat aacgatcttt gtagaaacca tcggcgcagc
     2941 tatttacccg caggacatat ccacgccctc ctacatcgaa gctgaaagca cgagattctt
     3001 cgccctccga gagctgcatc aggtcggaga cgctgtcgaa cttttcgatc agaaacttct
     3061 cgacagacgt cgcggtgagt tcaggctttt tggccaagga agttcctata ctttctagag
     3121 aataggaact tcggaatagg aacttctagg tacgtgaacc atcaccctaa tcaagttttt
     3181 tggggtcgag gtgccgtaaa gcactaaatc ggaaccctaa agggagcccc cgatttagag
     3241 cttgacgggg aaagccggcg aacgtggcga gaaaggaagg gaagaaagcg aaaggagcgg
     3301 gcgctagggc gctggcaagt gtagcggtca cgctgcgcgt aaccaccaca cccgccgcgc
     3361 ttaatgcgcc gctacagggc gcgtggggat accccctaga gccccagctg gttctttccg
     3421 cctcagaagc catagagccc accgcatccc cagaacatcg atagatctcc atagagccca
     3481 ccgcatcccc agcatgcctg ctattgtctt cccaatcctc ccccttgctg tcctgcccca
     3541 ccccaccccc cagaatagaa tgacacctac tcagacaatg cgatgcaatt tcctcatttt
     3601 attaggaaag gacagtggga gtggcacctt ccagggtcaa ggaaggcacg ggggaggggc
     3661 aaacaacaga tggctggcaa ctagaaggca cagtcgaggc tgatcagcgg gtttaaactc
     3721 aatgatgatt atgatgatgg tcgacggcgc tattcagatc ctcttctgag atgagttttt
     3781 gttcgggccc tcctcgagcg gctcagggcg tgagtgagaa ctggtcctgc tcggtgggct
     3841 tctccaccca ggcccccagc cccgccttga tgaaggctgt gaacagacgc gccttggcgg
     3901 cctggtactc ctttgccgcc agcttggact catggtacac gttgggtttg gtaatcttgg
     3961 agcgtagtaa gtgggaggga accttgccgt gcacacgcat ccagcgacag tacaacgcgt
     4021 gcttacacag gcgggacgcg cggcccagct catccttccc agtcgtggcg ttgatgacct
     4081 caatagcgga gtcgcctacc gtccagttga cactgaagtt gggggccttc cctggctgcc
     4141 gtgcttctgc attgctgatg ccactgagca aaggcttgtt gagggtgtag agaggtggca
     4201 ggtcctctat gttggagatc cgctggtaca tggccctgga aaggtggtcc ccgtggtaaa
     4261 ggctgcccag gatgatgctc gagaagtaaa tgggctccac gaaaatgctg agcagggatc
     4321 cctggatgcc caccacgttc cagcgtgcaa tcttgtcact gcaggacatg gtgagcagcc
     4381 gctccccttg cagcaccccg tcccacgctt ggatgctcgc attggagcgc actggaatcg
     4441 tcccctgacc agactctatt ttggtccgta gctgtcctct tgctttacga tttgggtgtc
     4501 tatctgctgg ttcttccagg attggctcat gtggtgagaa gattctggca tctccacagg
     4561 gagaggtgct gatgtacaga tgaaactgga cattctcctt cagcctaaac ccccctcgct
     4621 ctgatttctg aaagatggat cttttttgat catctttgtt atttaagtaa agctcaagtt
     4681 gtgtataaag aaatctgagc aaggatctcc gagatattat ttctgcatgg cagtcattta
     4741 atgcaaggcc acgatcactc atgtattcac cattaataca ttttgttcct gtagaaacac
     4801 ttatcacctt ggcatcttta acatctgtgc ctgttgtcat gacgactcca gccagcactt
     4861 ttctgcgagc gtgaggggag gagaagttgt cggtcaggtc accaaactta cccaggacca
     4921 ggcgtgagac agcgtcagct aaaacctgcg gtaaatgcag ctgaagaccc tcactgggaa
     4981 taggctggcg agatggcgtt tgatccaagt gcaaagcacc ggtacccgat gaagtgtcct
     5041 tggccaggga ggagctggcg gtggtgcagg actcatcttg gggacccagg ctgggtgggg
     5101 tgcgtccacc ggtcatggta aaggaggagg cgttatcgtt atcggccgtc tcactggtct
     5161 gtctggaaat gttttcggag tagcgtcctg cgcatgaaaa tccactactt ttatactttc
     5221 ttcttacaca ccagaggtct ttgagaatct tcaggaagta gtttcgaaac ttctgaccta
     5281 taaatgcgta cagcacaggg ttaaggcaac aatgcaggaa cgccagaacc tccgtaaccg
     5341 ttttggtgta cccgataagc ttttctgact gacatgagcg attcattttg cccaggttgg
     5401 ctgccgttac cagaaggacc atattatgag ggatctggca agccagaaat accagcacaa
     5461 cagcgatgat tacccggatg gccttgtgtc gcttagagtt ttgtgcctgg accagggtct
     5521 tcacgatgaa ggtataacaa aaaatcataa acatgagtgg gataaaaaag ccgaacagca
     5581 attccaaacc cagcatcaac agcttccacc tgataggctc agataccgtc tggtacttgg
     5641 gctcacacac atcgctgccc tgcgtgttgt acttctggtt aaacacgaaa gtactgctgg
     5701 aaataatcac gctcaagccc catacaacaa gacaaatgat tttagaccta gggagcgttc
     5761 gagacctcag tctgaaggat tttgttgcct gtactatggc gatgtatctg tccatactta
     5821 tgcaagtcag cagcagcata ccgcaattaa agttgatggc ataaattccc ttcaacagct
     5881 tgcaggttgc attactgaac acccaggccc cggtagcgtg actcacagcc cagaatggga
     5941 gcgtcaggac gaacaggatg tctgcaatgg ccatattaag caggtagacg tctgtcatgg
     6001 atctggcctt tttataaaaa gcaaatgtga tgactaccaa gatattcccc aggaggccaa
     6061 agacgcagat gagtgaatac gcgatgggaa cgaagagtcg ggagaactgg cgcacctcct
     6121 gcagggagca aagcagcatc tcagaatcta cgctgtagta ggatgtattg acggacacga
     6181 agtagtcctc actggaatcg aacacatcac tgaagttcat gctctctccg ctcatatcga
     6241 tgctggcgtc atcatcgtcc ttgtagtcgg cgaataccag gcagaagatg tagctcaggg
     6301 cgatgatcgt cttcatggtg gcgaagctta agtttaaacg ctagagtccg gaggctggat
     6361 cggtcccggt gtcttctatg gaggtcaaaa cagcgtggat ggcgtctcca ggcgatctga
     6421 cggttcacta aacgagctcg tcgacgatct ctatcactga tagggagatc tctatcactg
     6481 atagggagag ctctgcttat atagacctcc caccgtacac gcctaccgcc catttgcgtc
     6541 aatggggcgg agttgttacg acattttgga aagtcccgtt gattttggtg ccaaaacaaa
     6601 ctcccattga cgtcaatggg gtggagactt ggaaatcccc gtgagtcaaa ccgctatcca
     6661 cgcccattga tgtactgcca aaaccgcatc accatggtaa tagcgatgac taatacgtag
     6721 atgtactgcc aagtaggaaa gtcccataag gtcatgtact gggcataatg ccaggcgggc
     6781 catttaccgt cattgacgtc aatagggggc gtacttggca tatgatacac ttgatgtact
     6841 gccaagtggg cagtttaccg taaatactcc acccattgac gtcaatggaa agtccctatt
     6901 ggcgttacta tgggaacata cgtcattatt gacgtcaatg ggcgggggtc gttgggcggt
     6961 cagccaggcg ggccatttac cgtaagttat gtaacgcgga actccatata tgggctatga
     7021 actaatgacc ccgtaattga ttactattaa taactagtca ataatcaatg tcaacgcgta
     7081 tatctggccc gtacgaagca gcgcaaaacg cctaacccta agcagattct tcatgcaatt
     7141 gtcggtcaag ccttgccttg ttgtagctta aattttgctc gcgcactact cagcgacctc
     7201 caacacacaa gcagggagca gatactggct taactatgcg gcatcagagc agattgtact
     7261 gagagtgcac cataggggat cgggagatct cccgatccgt cgacgtcagg tggcactttt
     7321 cggggaaatg tgcgcggaac ccctatttgt ttatttttct aaatacattc aaatatgtat
     7381 ccgctcatga gacaataacc ctgataaatg cttcaataat attgaaaaag gaagagtatg
     7441 agtattcaac atttccgtgt cgcccttatt cccttttttg cggcattttg ccttcctgtt
     7501 tttgctcacc cagaaacgct ggtgaaagta aaagatgctg aagatcagtt gggtgcacga
     7561 gtgggttaca tcgaactgga tctcaacagc ggtaagatcc ttgagagttt tcgccccgaa
     7621 gaacgttttc caatgatgag cacttttaaa gttctgctat gtggcgcggt attatcccgt
     7681 attgacgccg
//
