LOCUS       sequence-466409-        7465 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  CX3CR1-ADAR2dd(E488Q, T501A) with a short 4 amino acid linker and no
            V2 tail.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7465)
  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 7465)
  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: LSM105 CMV-TO-CX3CR1-(noV2)-ADAR2dd(E488Q, T50...
FEATURES             Location/Qualifiers
     source          1..7465
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     regulatory      417..426
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     primer_bind     complement(518..542)
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     protein_bind    552..570
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    573..591
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     promoter        complement(593..796)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(622..642)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        797..1176
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     primer_bind     1343..1362
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        1437..1541
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1542..1610,1611..2402)
                     /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(1760..1779)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2573..3161
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3062..3081
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3315..3332
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    3449..3470
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(3485..3490,3491..3508,3509..3515)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3523..3539
                     /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     3528..3550
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     3547..3563
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     3547..3563
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     polyA_signal    3612..3733
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3623..3642)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     3677..3696
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     CDS             complement(3863..4885)
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="KKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRGY
                     VLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLPE
                     TELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQT
                     VMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMFG
                     DSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDGN
                     FDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAKE
                     "
     CDS             complement(3863..4885)
                     /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(4893..4940)
                     /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     5072..5091
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     polyA_signal    5263..5487
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     5476..5493
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     CDS             complement(5549..5578)
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
ORIGIN
        1 attattcatg ctgttggcag ccagaacgat ggccagatat ctatcaattg atatgactgt
       61 tataaagaag attgacccaa agaaccctat gaagaagaag gctgtagtaa atttacacat
      121 cgcgttgtga agtcccttct cattaattaa atagtgggtc caaaatggca gggtggcgac
      181 aaataacaag tctgacaatg cgaggttaag aagataaatg tctgtcacgc ttttaggctt
      241 cttactattg gtgagcgcga acaccaccag aagatttcca acgaggccaa tggcgaatat
      301 aacggaatag aaaatgctca ggaaaacagt gccgaagaca acaatatcgc cgatgtagca
      361 ggcctctgcc aggtcgtcat actcaaaatt ttcagtcaca ctttctggaa attgatccat
      421 ggtggcgaag cttaagttta aacgctagag tccggaggct ggatcggtcc cggtgtcttc
      481 tatggaggtc aaaacagcgt ggatggcgtc tccaggcgat ctgacggttc actaaacgag
      541 ctcgtcgacg atctctatca ctgataggga gatctctatc actgataggg agagctctgc
      601 ttatatagac ctcccaccgt acacgcctac cgcccatttg cgtcaatggg gcggagttgt
      661 tacgacattt tggaaagtcc cgttgatttt ggtgccaaaa caaactccca ttgacgtcaa
      721 tggggtggag acttggaaat ccccgtgagt caaaccgcta tccacgccca ttgatgtact
      781 gccaaaaccg catcaccatg gtaatagcga tgactaatac gtagatgtac tgccaagtag
      841 gaaagtccca taaggtcatg tactgggcat aatgccaggc gggccattta ccgtcattga
      901 cgtcaatagg gggcgtactt ggcatatgat acacttgatg tactgccaag tgggcagttt
      961 accgtaaata ctccacccat tgacgtcaat ggaaagtccc tattggcgtt actatgggaa
     1021 catacgtcat tattgacgtc aatgggcggg ggtcgttggg cggtcagcca ggcgggccat
     1081 ttaccgtaag ttatgtaacg cggaactcca tatatgggct atgaactaat gaccccgtaa
     1141 ttgattacta ttaataacta gtcaataatc aatgtcaacg cgtatatctg gcccgtacga
     1201 agcagcgcaa aacgcctaac cctaagcaga ttcttcatgc aattgtcggt caagccttgc
     1261 cttgttgtag cttaaatttt gctcgcgcac tactcagcga cctccaacac acaagcaggg
     1321 agcagatact ggcttaacta tgcggcatca gagcagattg tactgagagt gcaccatagg
     1381 ggatcgggag atctcccgat ccgtcgacgt caggtggcac ttttcgggga aatgtgcgcg
     1441 gaacccctat ttgtttattt ttctaaatac attcaaatat gtatccgctc atgagacaat
     1501 aaccctgata aatgcttcaa taatattgaa aaaggaagag tatgagtatt caacatttcc
     1561 gtgtcgccct tattcccttt tttgcggcat tttgccttcc tgtttttgct cacccagaaa
     1621 cgctggtgaa agtaaaagat gctgaagatc agttgggtgc acgagtgggt tacatcgaac
     1681 tggatctcaa cagcggtaag atccttgaga gttttcgccc cgaagaacgt tttccaatga
     1741 tgagcacttt taaagttctg ctatgtggcg cggtattatc ccgtattgac gccgggcaag
     1801 agcaactcgg tcgccgcata cactattctc agaatgactt ggttgagtac tcaccagtca
     1861 cagaaaagca tcttacggat ggcatgacag taagagaatt atgcagtgct gccataacca
     1921 tgagtgataa cactgcggcc aacttacttc tgacaacgat cggaggaccg aaggagctaa
     1981 ccgctttttt gcacaacatg ggggatcatg taactcgcct tgatcgttgg gaaccggagc
     2041 tgaatgaagc cataccaaac gacgagcgtg acaccacgat gcctgtagca atggcaacaa
     2101 cgttgcgcaa actattaact ggcgaactac ttactctagc ttcccggcaa caattaatag
     2161 actggatgga ggcggataaa gttgcaggac cacttctgcg ctcggccctt ccggctggct
     2221 ggtttattgc tgataaatct ggagccggtg agcgtgggtc tcgcggtatc attgcagcac
     2281 tggggccaga tggtaagccc tcccgtatcg tagttatcta cacgacgggg agtcaggcaa
     2341 ctatggatga acgaaataga cagatcgctg agataggtgc ctcactgatt aagcattggt
     2401 aactgtcaga ccaagtttac tcatatatac tttagattga tttaaaactt catttttaat
     2461 ttaaaaggat ctaggtgaag atcctttttg ataatctcat gaccaaaatc ccttaacgtg
     2521 agttttcgtt ccactgagcg tcagaccccg tagaaaagat caaaggatct tcttgagatc
     2581 ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg
     2641 tttgtttgcc ggatcaagag ctaccaactc tttttccgaa ggtaactggc ttcagcagag
     2701 cgcagatacc aaatactgtt cttctagtgt agccgtagtt aggccaccac ttcaagaact
     2761 ctgtagcacc gcctacatac ctcgctctgc taatcctgtt accagtggct gctgccagtg
     2821 gcgataagtc gtgtcttacc gggttggact caagacgata gttaccggat aaggcgcagc
     2881 ggtcgggctg aacggggggt tcgtgcacac agcccagctt ggagcgaacg acctacaccg
     2941 aactgagata cctacagcgt gagctatgag aaagcgccac gcttcccgaa gggagaaagg
     3001 cggacaggta tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg gagcttccag
     3061 ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg ccacctctga cttgagcgtc
     3121 gatttttgtg atgctcgtca ggggggcgga gcctatggaa aaacgccagc aacgcggcct
     3181 ttttacggtt cctggccttt tgctggcctt ttgctcacat gttctttcct gcgttatccc
     3241 ctgattctgt ggataaccgt attaccgcct ttgagtgagc tgataccgct cgccgcagcc
     3301 gaacgaccga gcgcagcgag tcagtgagcg aggaagcgga agagcgccca atacgcaaac
     3361 cgcctctccc cgcgcgttgg ccgattcatt aatgcagctg gcacgacagg tttcccgact
     3421 ggaaagcggg cagtgagcgc aacgcaatta atgtgagtta gctcactcat taggcacccc
     3481 aggctttaca ctttatgctt ccggctcgta tgttgtgtgg aattgtgagc ggataacaat
     3541 ttcacacagg aaacagctat gaccatgatt acgccaagct ctagctagag gtcgacggta
     3601 tacagacatg ataagataca ttgatgagtt tggacaaacc acaactagaa tgcagtgaaa
     3661 aaaatgcttt atttgtgaaa tttgtgatgc tattgcttta tttgtaacca ttataagctg
     3721 caataaacaa gttggggtgg gcgaagaact ccagcatgag atccccgcgc tggaggatca
     3781 tccagccggc gtcccggaaa acgattccga agcccaacct ttcatagaag gcggcggtgg
     3841 aatcgaaatc tcgtagtacg tgctattcct ttgccctcgg acgagtgctg gggcgtcggt
     3901 ttccactatc ggcgagtact tctacacagc catcggtcca gacggccgcg cttctgcggg
     3961 cgatttgtgt acgcccgaca gtcccggctc cggatcggac gattgcgtcg catcgaccct
     4021 gcgcccaagc tgcatcatcg aaattgccgt caaccaagct ctgatagagt tggtcaagac
     4081 caatgcggag catatacgcc cggagccgcg gcgatcctgc aagctccgga tgcctccgct
     4141 cgaagtagcg cgtctgctgc tccatacaag ccaaccacgg cctccagaag aagatgttgg
     4201 cgacctcgta ttgggaatcc ccgaacatcg cctcgctcca gtcaatgacc gctgttatgc
     4261 ggccattgtc cgtcaggaca ttgttggagc cgaaatccgc gtgcacgagg tgccggactt
     4321 cggggcagtc ctcggcccaa agcatcagct catcgagagc ctgcgcgacg gacgcactga
     4381 cggtgtcgtc catcacagtt tgccagtgat acacatgggg atcagcaatc gcgcatatga
     4441 aatcacgcca tgtagtgtat tgaccgattc cttgcggtcc gaatgggccg aacccgctcg
     4501 tctggctaag atcggccgca gcgatcgcat ccatggcctc cgcgaccggc tgcagaacag
     4561 cgggcagttc ggtttcaggc aggtcttgca acgtgacacc ctgtgcacgg cgggagatgc
     4621 aataggtcag gctctcgctg aattccccaa tgtcaagcac ttccggaatc gggagcgcgg
     4681 ccgatgcaaa gtgccgataa acataacgat ctttgtagaa accatcggcg cagctattta
     4741 cccgcaggac atatccacgc cctcctacat cgaagctgaa agcacgagat tcttcgccct
     4801 ccgagagctg catcaggtcg gagacgctgt cgaacttttc gatcagaaac ttctcgacag
     4861 acgtcgcggt gagttcaggc tttttggcca aggaagttcc tatactttct agagaatagg
     4921 aacttcggaa taggaacttc taggtacgtg aaccatcacc ctaatcaagt tttttggggt
     4981 cgaggtgccg taaagcacta aatcggaacc ctaaagggag cccccgattt agagcttgac
     5041 ggggaaagcc ggcgaacgtg gcgagaaagg aagggaagaa agcgaaagga gcgggcgcta
     5101 gggcgctggc aagtgtagcg gtcacgctgc gcgtaaccac cacacccgcc gcgcttaatg
     5161 cgccgctaca gggcgcgtgg ggataccccc tagagcccca gctggttctt tccgcctcag
     5221 aagccataga gcccaccgca tccccagaac atcgatagat ctccatagag cccaccgcat
     5281 ccccagcatg cctgctattg tcttcccaat cctccccctt gctgtcctgc cccaccccac
     5341 cccccagaat agaatgacac ctactcagac aatgcgatgc aatttcctca ttttattagg
     5401 aaaggacagt gggagtggca ccttccaggg tcaaggaagg cacgggggag gggcaaacaa
     5461 cagatggctg gcaactagaa ggcacagtcg aggctgatca gcgggtttaa actcaatgat
     5521 gattatgatg atggtcgacg gcgctattca gatcctcttc tgagatgagt ttttgttcgg
     5581 gccctcctcg agcggctcag ggcgtgagtg agaactggtc ctgctcggtg ggcttctcca
     5641 cccaggcccc cagccccgcc ttgatgaagg ctgtgaacag acgcgccttg gcggcctggt
     5701 actcctttgc cgccagcttg gactcatggt acacgttggg tttggtaatc ttggagcgta
     5761 gtaagtggga gggaaccttg ccgtgcacac gcatccagcg acagtacaac gcgtgcttac
     5821 acaggcggga cgcgcggccc agctcatcct tcccagtcgt ggcgttgatg acctcaatag
     5881 cggagtcgcc taccgtccag ttgacactga agttgggggc cttccctggc tgccgtgctt
     5941 ctgcattgct gatgccactg agcaaaggct tgttgagggt gtagagaggt ggcaggtcct
     6001 ctatgttgga gatccgctgg tacatggccc tggaaaggtg gtccccgtgg taaaggctgc
     6061 ccaggatgat gctcgagaag taaatgggct ccacgaaaat gctgagcagg gatccctgga
     6121 tgcccaccac gttccagcgt gcaatcttgt cactgcagga catggtgagc agccgctccc
     6181 cttgcagcac cccgtcccac gcttggatgc tcgcattgga gcgcactgga atcgtcccct
     6241 gaccagactc tattttggtc cgtagctgtc ctcttgcttt acgatttggg tgtctatctg
     6301 ctggttcttc caggattggc tcatgtggtg agaagattct ggcatctcca cagggagagg
     6361 tgctgatgta cagatgaaac tggacattct ccttcagcct aaacccccct cgctctgatt
     6421 tctgaaagat ggatcttttt tgatcatctt tgttatttaa gtaaagctca agttgtgtat
     6481 aaagaaatct gagcaaggat ctccgagata ttatttctgc atggcagtca tttaatgcaa
     6541 ggccacgatc actcatgtat tcaccattaa tacattttgt tcctgtagaa acacttatca
     6601 ccttggcatc tttaacatct gtgcctgttg tcatgacgac tccagccagc acttttctgc
     6661 gagcgtgagg ggaggagaag ttgtcggtca ggtcaccaaa cttacccagg accaggcgtg
     6721 agacagcgtc agctaaaacc tgcggtaaat gcagctgaag accctcactg ggaataggct
     6781 ggcgagatgg cgtttgatcc aagtgcaaag caccggtacc cagcaggagc agagcatcac
     6841 catcagaggt gtggtaagtg aagttggaac tcaaaacgct cccgtgtctg gatctttgac
     6901 tttcactgga gctgaaatcg acatggacgg accgaccgca gagcaccgca aggcacttgc
     6961 cataaagatg gtagagataa cgcctgaact tttctccggc aaatgcatag atcaacgggt
     7021 ttaagcagca gtgactgaaa gcaactgttt ctgttacaga cagagcaagc ctcaaatcct
     7081 tgcgcatatc gcaagatgga aagaaatcat agagcttgag tgtctccaag aaaatcatga
     7141 cattataagg cgtccagaag agaaagaaga caattactac cagtaatatg agcttgatag
     7201 ctttggcttt cttgtgattc ttacagctaa agagggtttg gattatccta aagtagcaat
     7261 aggacataat cagaagtggc aacagaaacc caagaaagtt tgtctccacg tttctgagta
     7321 cgggccaaat ttcctgcagt acttctggat aatctcccaa gcattcgttt tccttttgtt
     7381 tggtaaacat gaattgggga gcggccacta aaatagcggc ggcccatacg cccagactaa
     7441 tagttacacc atgttgcact gtccg
//
