LOCUS       sequence-466411-        7552 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  CX3CR1-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 7552)
  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 7552)
  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: LSM068 CMV-TO-CX3CR1-ADAR2dd(E488Q, T501A) (FL...
FEATURES             Location/Qualifiers
     source          1..7552
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(96..115)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     protein_bind    247..294
                     /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             302..1324
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="KKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRGY
                     VLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLPE
                     TELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQT
                     VMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMFG
                     DSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDGN
                     FDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAKE
                     "
     CDS             302..1324
                     /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    1454..1575
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1491..1510)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     1545..1564
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(1624..1640)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1624..1640)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1637..1659)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1648..1664
                     /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(1672..1678,1679..1696,1697..1702))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1717..1738
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(1855..1872)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2026..2614)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2106..2125)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2785..3576,3577..3645))
                     /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     3408..3427
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3646..3750)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(3825..3844)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        4011..4390
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        4391..4594
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     4545..4565
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     protein_bind    4596..4614
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    4617..4635
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     primer_bind     4645..4669
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     regulatory      4761..4770
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             7161..7190
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     primer_bind     complement(7246..7263)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    7252..7476
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
ORIGIN
        1 gtatccccac gcgccctgta gcggcgcatt aagcgcggcg ggtgtggtgg ttacgcgcag
       61 cgtgaccgct acacttgcca gcgccctagc gcccgctcct ttcgctttct tcccttcctt
      121 tctcgccacg ttcgccggct ttccccgtca agctctaaat cgggggctcc ctttagggtt
      181 ccgatttagt gctttacggc acctcgaccc caaaaaactt gattagggtg atggttcacg
      241 tacctagaag ttcctattcc gaagttccta ttctctagaa agtataggaa cttccttggc
      301 caaaaagcct gaactcaccg cgacgtctgt cgagaagttt ctgatcgaaa agttcgacag
      361 cgtctccgac ctgatgcagc tctcggaggg cgaagaatct cgtgctttca gcttcgatgt
      421 aggagggcgt ggatatgtcc tgcgggtaaa tagctgcgcc gatggtttct acaaagatcg
      481 ttatgtttat cggcactttg catcggccgc gctcccgatt ccggaagtgc ttgacattgg
      541 ggaattcagc gagagcctga cctattgcat ctcccgccgt gcacagggtg tcacgttgca
      601 agacctgcct gaaaccgaac tgcccgctgt tctgcagccg gtcgcggagg ccatggatgc
      661 gatcgctgcg gccgatctta gccagacgag cgggttcggc ccattcggac cgcaaggaat
      721 cggtcaatac actacatggc gtgatttcat atgcgcgatt gctgatcccc atgtgtatca
      781 ctggcaaact gtgatggacg acaccgtcag tgcgtccgtc gcgcaggctc tcgatgagct
      841 gatgctttgg gccgaggact gccccgaagt ccggcacctc gtgcacgcgg atttcggctc
      901 caacaatgtc ctgacggaca atggccgcat aacagcggtc attgactgga gcgaggcgat
      961 gttcggggat tcccaatacg aggtcgccaa catcttcttc tggaggccgt ggttggcttg
     1021 tatggagcag cagacgcgct acttcgagcg gaggcatccg gagcttgcag gatcgccgcg
     1081 gctccgggcg tatatgctcc gcattggtct tgaccaactc tatcagagct tggttgacgg
     1141 caatttcgat gatgcagctt gggcgcaggg tcgatgcgac gcaatcgtcc gatccggagc
     1201 cgggactgtc gggcgtacac aaatcgcccg cagaagcgcg gccgtctgga ccgatggctg
     1261 tgtagaagta ctcgccgata gtggaaaccg acgccccagc actcgtccga gggcaaagga
     1321 atagcacgta ctacgagatt tcgattccac cgccgccttc tatgaaaggt tgggcttcgg
     1381 aatcgttttc cgggacgccg gctggatgat cctccagcgc ggggatctca tgctggagtt
     1441 cttcgcccac cccaacttgt ttattgcagc ttataatggt tacaaataaa gcaatagcat
     1501 cacaaatttc acaaataaag catttttttc actgcattct agttgtggtt tgtccaaact
     1561 catcaatgta tcttatcatg tctgtatacc gtcgacctct agctagagct tggcgtaatc
     1621 atggtcatag ctgtttcctg tgtgaaattg ttatccgctc acaattccac acaacatacg
     1681 agccggaagc ataaagtgta aagcctgggg tgcctaatga gtgagctaac tcacattaat
     1741 tgcgttgcgc tcactgcccg ctttccagtc gggaaacctg tcgtgccagc tgcattaatg
     1801 aatcggccaa cgcgcgggga gaggcggttt gcgtattggg cgctcttccg cttcctcgct
     1861 cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc actcaaaggc
     1921 ggtaatacgg ttatccacag aatcagggga taacgcagga aagaacatgt gagcaaaagg
     1981 ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg
     2041 cccccctgac gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg
     2101 actataaaga taccaggcgt ttccccctgg aagctccctc gtgcgctctc ctgttccgac
     2161 cctgccgctt accggatacc tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca
     2221 tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt
     2281 gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc
     2341 caacccggta agacacgact tatcgccact ggcagcagcc actggtaaca ggattagcag
     2401 agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg tggcctaact acggctacac
     2461 tagaagaaca gtatttggta tctgcgctct gctgaagcca gttaccttcg gaaaaagagt
     2521 tggtagctct tgatccggca aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa
     2581 gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat cctttgatct tttctacggg
     2641 gtctgacgct cagtggaacg aaaactcacg ttaagggatt ttggtcatga gattatcaaa
     2701 aaggatcttc acctagatcc ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat
     2761 atatgagtaa acttggtctg acagttacca atgcttaatc agtgaggcac ctatctcagc
     2821 gatctgtcta tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga taactacgat
     2881 acgggagggc ttaccatctg gccccagtgc tgcaatgata ccgcgagacc cacgctcacc
     2941 ggctccagat ttatcagcaa taaaccagcc agccggaagg gccgagcgca gaagtggtcc
     3001 tgcaacttta tccgcctcca tccagtctat taattgttgc cgggaagcta gagtaagtag
     3061 ttcgccagtt aatagtttgc gcaacgttgt tgccattgct acaggcatcg tggtgtcacg
     3121 ctcgtcgttt ggtatggctt cattcagctc cggttcccaa cgatcaaggc gagttacatg
     3181 atcccccatg ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg ttgtcagaag
     3241 taagttggcc gcagtgttat cactcatggt tatggcagca ctgcataatt ctcttactgt
     3301 catgccatcc gtaagatgct tttctgtgac tggtgagtac tcaaccaagt cattctgaga
     3361 atagtgtatg cggcgaccga gttgctcttg cccggcgtca atacgggata ataccgcgcc
     3421 acatagcaga actttaaaag tgctcatcat tggaaaacgt tcttcggggc gaaaactctc
     3481 aaggatctta ccgctgttga gatccagttc gatgtaaccc actcgtgcac ccaactgatc
     3541 ttcagcatct tttactttca ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc
     3601 cgcaaaaaag ggaataaggg cgacacggaa atgttgaata ctcatactct tcctttttca
     3661 atattattga agcatttatc agggttattg tctcatgagc ggatacatat ttgaatgtat
     3721 ttagaaaaat aaacaaatag gggttccgcg cacatttccc cgaaaagtgc cacctgacgt
     3781 cgacggatcg ggagatctcc cgatccccta tggtgcactc tcagtacaat ctgctctgat
     3841 gccgcatagt taagccagta tctgctccct gcttgtgtgt tggaggtcgc tgagtagtgc
     3901 gcgagcaaaa tttaagctac aacaaggcaa ggcttgaccg acaattgcat gaagaatctg
     3961 cttagggtta ggcgttttgc gctgcttcgt acgggccaga tatacgcgtt gacattgatt
     4021 attgactagt tattaatagt aatcaattac ggggtcatta gttcatagcc catatatgga
     4081 gttccgcgtt acataactta cggtaaatgg cccgcctggc tgaccgccca acgacccccg
     4141 cccattgacg tcaataatga cgtatgttcc catagtaacg ccaataggga ctttccattg
     4201 acgtcaatgg gtggagtatt tacggtaaac tgcccacttg gcagtacatc aagtgtatca
     4261 tatgccaagt acgcccccta ttgacgtcaa tgacggtaaa tggcccgcct ggcattatgc
     4321 ccagtacatg accttatggg actttcctac ttggcagtac atctacgtat tagtcatcgc
     4381 tattaccatg gtgatgcggt tttggcagta catcaatggg cgtggatagc ggtttgactc
     4441 acggggattt ccaagtctcc accccattga cgtcaatggg agtttgtttt ggcaccaaaa
     4501 tcaacgggac tttccaaaat gtcgtaacaa ctccgcccca ttgacgcaaa tgggcggtag
     4561 gcgtgtacgg tgggaggtct atataagcag agctctccct atcagtgata gagatctccc
     4621 tatcagtgat agagatcgtc gacgagctcg tttagtgaac cgtcagatcg cctggagacg
     4681 ccatccacgc tgttttgacc tccatagaag acaccgggac cgatccagcc tccggactct
     4741 agcgtttaaa cttaagcttc gccaccatgg atcaatttcc agaaagtgtg actgaaaatt
     4801 ttgagtatga cgacctggca gaggcctgct acatcggcga tattgttgtc ttcggcactg
     4861 ttttcctgag cattttctat tccgttatat tcgccattgg cctcgttgga aatcttctgg
     4921 tggtgttcgc gctcaccaat agtaagaagc ctaaaagcgt gacagacatt tatcttctta
     4981 acctcgcatt gtcagacttg ttatttgtcg ccaccctgcc attttggacc cactatttaa
     5041 ttaatgagaa gggacttcac aacgcgatgt gtaaatttac tacagccttc ttcttcatag
     5101 ggttctttgg gtcaatcttc tttataacag tcatatcaat tgatagatat ctggccatcg
     5161 ttctggctgc caacagcatg aataatcgga cagtgcaaca tggtgtaact attagtctgg
     5221 gcgtatgggc cgccgctatt ttagtggccg ctccccaatt catgtttacc aaacaaaagg
     5281 aaaacgaatg cttgggagat tatccagaag tactgcagga aatttggccc gtactcagaa
     5341 acgtggagac aaactttctt gggtttctgt tgccacttct gattatgtcc tattgctact
     5401 ttaggataat ccaaaccctc tttagctgta agaatcacaa gaaagccaaa gctatcaagc
     5461 tcatattact ggtagtaatt gtcttctttc tcttctggac gccttataat gtcatgattt
     5521 tcttggagac actcaagctc tatgatttct ttccatcttg cgatatgcgc aaggatttga
     5581 ggcttgctct gtctgtaaca gaaacagttg ctttcagtca ctgctgctta aacccgttga
     5641 tctatgcatt tgccggagaa aagttcaggc gttatctcta ccatctttat ggcaagtgcc
     5701 ttgcggtgct ctgcggtcgg tccgtccatg tcgatttcag ctccagtgaa agtcaaagat
     5761 ccagacacgg gagcgttttg agttccaact tcacttacca cacctctgat ggtgatgctc
     5821 tgctcctgct gacaggcggt cgcactcccc ctagcctcgg tccacaggat gagtcatgca
     5881 cgaccgcgag tagctccctc gccaaagaca cctcttccgg taccggtgct ttgcacttgg
     5941 atcaaacgcc atctcgccag cctattccca gtgagggtct tcagctgcat ttaccgcagg
     6001 ttttagctga cgctgtctca cgcctggtcc tgggtaagtt tggtgacctg accgacaact
     6061 tctcctcccc tcacgctcgc agaaaagtgc tggctggagt cgtcatgaca acaggcacag
     6121 atgttaaaga tgccaaggtg ataagtgttt ctacaggaac aaaatgtatt aatggtgaat
     6181 acatgagtga tcgtggcctt gcattaaatg actgccatgc agaaataata tctcggagat
     6241 ccttgctcag atttctttat acacaacttg agctttactt aaataacaaa gatgatcaaa
     6301 aaagatccat ctttcagaaa tcagagcgag gggggtttag gctgaaggag aatgtccagt
     6361 ttcatctgta catcagcacc tctccctgtg gagatgccag aatcttctca ccacatgagc
     6421 caatcctgga agaaccagca gatagacacc caaatcgtaa agcaagagga cagctacgga
     6481 ccaaaataga gtctggtcag gggacgattc cagtgcgctc caatgcgagc atccaagcgt
     6541 gggacggggt gctgcaaggg gagcggctgc tcaccatgtc ctgcagtgac aagattgcac
     6601 gctggaacgt ggtgggcatc cagggatccc tgctcagcat tttcgtggag cccatttact
     6661 tctcgagcat catcctgggc agcctttacc acggggacca cctttccagg gccatgtacc
     6721 agcggatctc caacatagag gacctgccac ctctctacac cctcaacaag cctttgctca
     6781 gtggcatcag caatgcagaa gcacggcagc cagggaaggc ccccaacttc agtgtcaact
     6841 ggacggtagg cgactccgct attgaggtca tcaacgccac gactgggaag gatgagctgg
     6901 gccgcgcgtc ccgcctgtgt aagcacgcgt tgtactgtcg ctggatgcgt gtgcacggca
     6961 aggttccctc ccacttacta cgctccaaga ttaccaaacc caacgtgtac catgagtcca
     7021 agctggcggc aaaggagtac caggccgcca aggcgcgtct gttcacagcc ttcatcaagg
     7081 cggggctggg ggcctgggtg gagaagccca ccgagcagga ccagttctca ctcacgccct
     7141 gagccgctcg aggagggccc gaacaaaaac tcatctcaga agaggatctg aatagcgccg
     7201 tcgaccatca tcataatcat cattgagttt aaacccgctg atcagcctcg actgtgcctt
     7261 ctagttgcca gccatctgtt gtttgcccct cccccgtgcc ttccttgacc ctggaaggtg
     7321 ccactcccac tgtcctttcc taataaaatg aggaaattgc atcgcattgt ctgagtaggt
     7381 gtcattctat tctggggggt ggggtggggc aggacagcaa gggggaggat tgggaagaca
     7441 atagcaggca tgctggggat gcggtgggct ctatggagat ctatcgatgt tctggggatg
     7501 cggtgggctc tatggcttct gaggcggaaa gaaccagctg gggctctagg gg
//
