LOCUS       sequence-467421-        8162 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  IVT template for CCR6-V2-ADAR2dd(E488Q, T501A).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8162)
  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 8162)
  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: XZ088 (IVT) CMV-TO-T7-CCR6-V2-ADAR2dd-hybridUTR3
FEATURES             Location/Qualifiers
     source          1..8162
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(join(171..962,963..1031))
                     /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     794..813
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(1032..1136)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(1211..1230)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        1397..1776
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        1777..1980
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     1931..1951
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     protein_bind    1982..2000
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    2003..2021
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     primer_bind     2031..2055
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     primer_bind     2165..2184
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        2165..2183
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             5157..5186
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     primer_bind     complement(5242..5259)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    5248..5472
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(5644..5663)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     protein_bind    5795..5842
                     /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             5850..6872
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="KKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRGY
                     VLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLPE
                     TELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQT
                     VMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMFG
                     DSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDGN
                     FDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAKE
                     "
     CDS             5850..6872
                     /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
                     "
     polyA_signal    7002..7123
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(7039..7058)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     7093..7112
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(7172..7188)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(7172..7188)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(7185..7207)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    7196..7212
                     /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(7220..7226,7227..7244,7245..7250))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7265..7286
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(7403..7420)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(7574..8162)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(7654..7673)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 gaagatcctt tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa
       61 gggattttgg tcatgagatt atcaaaaagg atcttcacct agatcctttt aaattaaaaa
      121 tgaagtttta aatcaatcta aagtatatat gagtaaactt ggtctgacag ttaccaatgc
      181 ttaatcagtg aggcacctat ctcagcgatc tgtctatttc gttcatccat agttgcctga
      241 ctccccgtcg tgtagataac tacgatacgg gagggcttac catctggccc cagtgctgca
      301 atgataccgc gagacccacg ctcaccggct ccagatttat cagcaataaa ccagccagcc
      361 ggaagggccg agcgcagaag tggtcctgca actttatccg cctccatcca gtctattaat
      421 tgttgccggg aagctagagt aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc
      481 attgctacag gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt cagctccggt
      541 tcccaacgat caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc
      601 ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag tgttatcact catggttatg
      661 gcagcactgc ataattctct tactgtcatg ccatccgtaa gatgcttttc tgtgactggt
      721 gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg
      781 gcgtcaatac gggataatac cgcgccacat agcagaactt taaaagtgct catcattgga
      841 aaacgttctt cggggcgaaa actctcaagg atcttaccgc tgttgagatc cagttcgatg
      901 taacccactc gtgcacccaa ctgatcttca gcatctttta ctttcaccag cgtttctggg
      961 tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa taagggcgac acggaaatgt
     1021 tgaatactca tactcttcct ttttcaatat tattgaagca tttatcaggg ttattgtctc
     1081 atgagcggat acatatttga atgtatttag aaaaataaac aaataggggt tccgcgcaca
     1141 tttccccgaa aagtgccacc tgacgtcgac ggatcgggag atctcccgat cccctatggt
     1201 gcactctcag tacaatctgc tctgatgccg catagttaag ccagtatctg ctccctgctt
     1261 gtgtgttgga ggtcgctgag tagtgcgcga gcaaaattta agctacaaca aggcaaggct
     1321 tgaccgacaa ttgcatgaag aatctgctta gggttaggcg ttttgcgctg cttcgtacgg
     1381 gccagatata cgcgttgaca ttgattattg actagttatt aatagtaatc aattacgggg
     1441 tcattagttc atagcccata tatggagttc cgcgttacat aacttacggt aaatggcccg
     1501 cctggctgac cgcccaacga cccccgccca ttgacgtcaa taatgacgta tgttcccata
     1561 gtaacgccaa tagggacttt ccattgacgt caatgggtgg agtatttacg gtaaactgcc
     1621 cacttggcag tacatcaagt gtatcatatg ccaagtacgc cccctattga cgtcaatgac
     1681 ggtaaatggc ccgcctggca ttatgcccag tacatgacct tatgggactt tcctacttgg
     1741 cagtacatct acgtattagt catcgctatt accatggtga tgcggttttg gcagtacatc
     1801 aatgggcgtg gatagcggtt tgactcacgg ggatttccaa gtctccaccc cattgacgtc
     1861 aatgggagtt tgttttggca ccaaaatcaa cgggactttc caaaatgtcg taacaactcc
     1921 gccccattga cgcaaatggg cggtaggcgt gtacggtggg aggtctatat aagcagagct
     1981 ctccctatca gtgatagaga tctccctatc agtgatagag atcgtcgacg agctcgttta
     2041 gtgaaccgtc agatcgcctg gagacgccat ccacgctgtt ttgacctcca tagaagacac
     2101 cgggaccgat ccagcctccg gactctagcg tttaaactta agctcgctag agatccgcgg
     2161 ccgctaatac gactcactat agggagaaaa ataacaaatc tcaacacaac atatacaaaa
     2221 caaacgaatc tcaagcaatc aagcattcta cttctattgc agcaatttaa atcatttctt
     2281 ttaaagcaaa agcaattttc tgaaaatttt caccatttac gaacgataaa gcttcgccac
     2341 catgaagacg atcatcgccc tgagctacat cttctgcctg gtattcgccg actacaagga
     2401 cgatgatgac gccagcatcg atatgagcgg agagagcatg aacttcagtg atgtgttcga
     2461 ttccagtgag gactacttcg tgtccgtcaa tacatcctac tacagcgtag attctgagat
     2521 gctgctttgc tccctgcagg aggtgcgcca gttctcccga ctcttcgttc ccatcgcgta
     2581 ttcactcatc tgcgtctttg gcctcctggg gaatatcttg gtagtcatca catttgcttt
     2641 ttataaaaag gccagatcca tgacagacgt ctacctgctt aatatggcca ttgcagacat
     2701 cctgttcgtc ctgacgctcc cattctgggc tgtgagtcac gctaccgggg cctgggtgtt
     2761 cagtaatgca acctgcaagc tgttgaaggg aatttatgcc atcaacttta attgcggtat
     2821 gctgctgctg acttgcataa gtatggacag atacatcgcc atagtacagg caacaaaatc
     2881 cttcagactg aggtctcgaa cgctccctag gtctaaaatc atttgtcttg ttgtatgggg
     2941 cttgagcgtg attatttcca gcagtacttt cgtgtttaac cagaagtaca acacgcaggg
     3001 cagcgatgtg tgtgagccca agtaccagac ggtatctgag cctatcaggt ggaagctgtt
     3061 gatgctgggt ttggaattgc tgttcggctt ttttatccca ctcatgttta tgattttttg
     3121 ttataccttc atcgtgaaga ccctggtcca ggcacaaaac tctaagcgac acaaggccat
     3181 ccgggtaatc atcgctgttg tgctggtatt tctggcttgc cagatccctc ataatatggt
     3241 ccttctggta acggcagcca acctgggcaa aatgaatcgc tcatgtcagt cagaaaagct
     3301 tatcgggtac accaaaacgg ttacggaggt tctggcgttc ctgcattgtt gccttaaccc
     3361 tgtgctgtac gcatttatag gtcagaagtt tcgaaactac ttcctgaaga ttctcaaaga
     3421 cctctggtgt gtaagaagaa agtataaaag tagtggattt tcatgcgcag gacgctactc
     3481 cgaaaacatt tccagacaga ccagtgagac ggccgataac gataacgcct cctcctttac
     3541 catgaccggt ggacgcaccc cacccagcct gggtccccaa gatgagtcct gcaccaccgc
     3601 cagctcctcc ctggccaagg acacttcatc gggtaccggt gctttgcact tggatcaaac
     3661 gccatctcgc cagcctattc ccagtgaggg tcttcagctg catttaccgc aggttttagc
     3721 tgacgctgtc tcacgcctgg tcctgggtaa gtttggtgac ctgaccgaca acttctcctc
     3781 ccctcacgct cgcagaaaag tgctggctgg agtcgtcatg acaacaggca cagatgttaa
     3841 agatgccaag gtgataagtg tttctacagg aacaaaatgt attaatggtg aatacatgag
     3901 tgatcgtggc cttgcattaa atgactgcca tgcagaaata atatctcgga gatccttgct
     3961 cagatttctt tatacacaac ttgagcttta cttaaataac aaagatgatc aaaaaagatc
     4021 catctttcag aaatcagagc gaggggggtt taggctgaag gagaatgtcc agtttcatct
     4081 gtacatcagc acctctccct gtggagatgc cagaatcttc tcaccacatg agccaatcct
     4141 ggaagaacca gcagatagac acccaaatcg taaagcaaga ggacagctac ggaccaaaat
     4201 agagtctggt caggggacga ttccagtgcg ctccaatgcg agcatccaag cgtgggacgg
     4261 ggtgctgcaa ggggagcggc tgctcaccat gtcctgcagt gacaagattg cacgctggaa
     4321 cgtggtgggc atccagggat ccctgctcag cattttcgtg gagcccattt acttctcgag
     4381 catcatcctg ggcagccttt accacgggga ccacctttcc agggccatgt accagcggat
     4441 ctccaacata gaggacctgc cacctctcta caccctcaac aagcctttgc tcagtggcat
     4501 cagcaatgca gaagcacggc agccagggaa ggcccccaac ttcagtgtca actggacggt
     4561 aggcgactcc gctattgagg tcatcaacgc cacgactggg aaggatgagc tgggccgcgc
     4621 gtcccgcctg tgtaagcacg cgttgtactg tcgctggatg cgtgtgcacg gcaaggttcc
     4681 ctcccactta ctacgctcca agattaccaa acccaacgtg taccatgagt ccaagctggc
     4741 ggcaaaggag taccaggccg ccaaggcgcg tctgttcaca gccttcatca aggcggggct
     4801 gggggcctgg gtggagaagc ccaccgagca ggaccagttc tcactcacgc cctgactcga
     4861 gctggtactg catgcacgca atgctagctg cccctttccc gtcctgggta ccccgagtct
     4921 cccccgacct cgggtcccag gtatgctccc acctccacct gccccactca ccacctctgc
     4981 tagttccaga cacctcccaa gcacgcagca atgcagctca aaacgcttag cctagccaca
     5041 cccccacggg aaacagcagt gattaacctt tagcaataaa cgaaagttta actaagctat
     5101 actaacccca gggttggtca atttcgtgcc agccacaccc tggagctagc gggcccgaac
     5161 aaaaactcat ctcagaagag gatctgaata gcgccgtcga ccatcatcat aatcatcatt
     5221 gagtttaaac ccgctgatca gcctcgactg tgccttctag ttgccagcca tctgttgttt
     5281 gcccctcccc cgtgccttcc ttgaccctgg aaggtgccac tcccactgtc ctttcctaat
     5341 aaaatgagga aattgcatcg cattgtctga gtaggtgtca ttctattctg gggggtgggg
     5401 tggggcagga cagcaagggg gaggattggg aagacaatag caggcatgct ggggatgcgg
     5461 tgggctctat ggagatctat cgatgttctg gggatgcggt gggctctatg gcttctgagg
     5521 cggaaagaac cagctggggc tctagggggt atccccacgc gccctgtagc ggcgcattaa
     5581 gcgcggcggg tgtggtggtt acgcgcagcg tgaccgctac acttgccagc gccctagcgc
     5641 ccgctccttt cgctttcttc ccttcctttc tcgccacgtt cgccggcttt ccccgtcaag
     5701 ctctaaatcg ggggctccct ttagggttcc gatttagtgc tttacggcac ctcgacccca
     5761 aaaaacttga ttagggtgat ggttcacgta cctagaagtt cctattccga agttcctatt
     5821 ctctagaaag tataggaact tccttggcca aaaagcctga actcaccgcg acgtctgtcg
     5881 agaagtttct gatcgaaaag ttcgacagcg tctccgacct gatgcagctc tcggagggcg
     5941 aagaatctcg tgctttcagc ttcgatgtag gagggcgtgg atatgtcctg cgggtaaata
     6001 gctgcgccga tggtttctac aaagatcgtt atgtttatcg gcactttgca tcggccgcgc
     6061 tcccgattcc ggaagtgctt gacattgggg aattcagcga gagcctgacc tattgcatct
     6121 cccgccgtgc acagggtgtc acgttgcaag acctgcctga aaccgaactg cccgctgttc
     6181 tgcagccggt cgcggaggcc atggatgcga tcgctgcggc cgatcttagc cagacgagcg
     6241 ggttcggccc attcggaccg caaggaatcg gtcaatacac tacatggcgt gatttcatat
     6301 gcgcgattgc tgatccccat gtgtatcact ggcaaactgt gatggacgac accgtcagtg
     6361 cgtccgtcgc gcaggctctc gatgagctga tgctttgggc cgaggactgc cccgaagtcc
     6421 ggcacctcgt gcacgcggat ttcggctcca acaatgtcct gacggacaat ggccgcataa
     6481 cagcggtcat tgactggagc gaggcgatgt tcggggattc ccaatacgag gtcgccaaca
     6541 tcttcttctg gaggccgtgg ttggcttgta tggagcagca gacgcgctac ttcgagcgga
     6601 ggcatccgga gcttgcagga tcgccgcggc tccgggcgta tatgctccgc attggtcttg
     6661 accaactcta tcagagcttg gttgacggca atttcgatga tgcagcttgg gcgcagggtc
     6721 gatgcgacgc aatcgtccga tccggagccg ggactgtcgg gcgtacacaa atcgcccgca
     6781 gaagcgcggc cgtctggacc gatggctgtg tagaagtact cgccgatagt ggaaaccgac
     6841 gccccagcac tcgtccgagg gcaaaggaat agcacgtact acgagatttc gattccaccg
     6901 ccgccttcta tgaaaggttg ggcttcggaa tcgttttccg ggacgccggc tggatgatcc
     6961 tccagcgcgg ggatctcatg ctggagttct tcgcccaccc caacttgttt attgcagctt
     7021 ataatggtta caaataaagc aatagcatca caaatttcac aaataaagca tttttttcac
     7081 tgcattctag ttgtggtttg tccaaactca tcaatgtatc ttatcatgtc tgtataccgt
     7141 cgacctctag ctagagcttg gcgtaatcat ggtcatagct gtttcctgtg tgaaattgtt
     7201 atccgctcac aattccacac aacatacgag ccggaagcat aaagtgtaaa gcctggggtg
     7261 cctaatgagt gagctaactc acattaattg cgttgcgctc actgcccgct ttccagtcgg
     7321 gaaacctgtc gtgccagctg cattaatgaa tcggccaacg cgcggggaga ggcggtttgc
     7381 gtattgggcg ctcttccgct tcctcgctca ctgactcgct gcgctcggtc gttcggctgc
     7441 ggcgagcggt atcagctcac tcaaaggcgg taatacggtt atccacagaa tcaggggata
     7501 acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg
     7561 cgttgctggc gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct
     7621 caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa
     7681 gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc
     7741 tcccttcggg aagcgtggcg ctttctcata gctcacgctg taggtatctc agttcggtgt
     7801 aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg
     7861 ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg
     7921 cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct
     7981 tgaagtggtg gcctaactac ggctacacta gaagaacagt atttggtatc tgcgctctgc
     8041 tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg
     8101 ctggtagcgg tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc
     8161 aa
//
