LOCUS       sequence-476096-        7123 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  CX3CR1-V2-PCP with a 10 amino acid linker and V2 tail.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7123)
  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 7123)
  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: LSM102 CMV-TO-CX3CR1-10aa-PCP (FLP-IN)
FEATURES             Location/Qualifiers
     source          1..7123
                     /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"
     regulatory      2147..2156
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             4118..4147
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     primer_bind     complement(4203..4220)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    4209..4433
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(4605..4624)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     protein_bind    4756..4803
                     /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             4811..5833
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="KKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRGY
                     VLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLPE
                     TELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQT
                     VMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMFG
                     DSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDGN
                     FDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAKE
                     "
     CDS             4811..5833
                     /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    5963..6084
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6000..6019)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     6054..6073
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(6133..6149)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6133..6149)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(6146..6168)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    6157..6173
                     /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(6181..6187,6188..6205,6206..6211))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6226..6247
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(6364..6381)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6535..7123)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6615..6634)
                     /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 agcttcgcca ccatggatca
     2161 atttccagaa agtgtgactg aaaattttga gtatgacgac ctggcagagg cctgctacat
     2221 cggcgatatt gttgtcttcg gcactgtttt cctgagcatt ttctattccg ttatattcgc
     2281 cattggcctc gttggaaatc ttctggtggt gttcgcgctc accaatagta agaagcctaa
     2341 aagcgtgaca gacatttatc ttcttaacct cgcattgtca gacttgttat ttgtcgccac
     2401 cctgccattt tggacccact atttaattaa tgagaaggga cttcacaacg cgatgtgtaa
     2461 atttactaca gccttcttct tcatagggtt ctttgggtca atcttcttta taacagtcat
     2521 atcaattgat agatatctgg ccatcgttct ggctgccaac agcatgaata atcggacagt
     2581 gcaacatggt gtaactatta gtctgggcgt atgggccgcc gctattttag tggccgctcc
     2641 ccaattcatg tttaccaaac aaaaggaaaa cgaatgcttg ggagattatc cagaagtact
     2701 gcaggaaatt tggcccgtac tcagaaacgt ggagacaaac tttcttgggt ttctgttgcc
     2761 acttctgatt atgtcctatt gctactttag gataatccaa accctcttta gctgtaagaa
     2821 tcacaagaaa gccaaagcta tcaagctcat attactggta gtaattgtct tctttctctt
     2881 ctggacgcct tataatgtca tgattttctt ggagacactc aagctctatg atttctttcc
     2941 atcttgcgat atgcgcaagg atttgaggct tgctctgtct gtaacagaaa cagttgcttt
     3001 cagtcactgc tgcttaaacc cgttgatcta tgcatttgcc ggagaaaagt tcaggcgtta
     3061 tctctaccat ctttatggca agtgccttgc ggtgctctgc ggtcggtccg tccatgtcga
     3121 tttcagctcc agtgaaagtc aaagatccag acacgggagc gttttgagtt ccaacttcac
     3181 ttaccacacc tctgatggtg atgctctgct cctgctgaca ggcggtcgca ctccccctag
     3241 cctcggtcca caggatgagt catgcacgac cgcgagtagc tccctcgcca aagacacctc
     3301 ttccggtacc ggtggcggaa ccggtggcgg ggctctagct tctaaaacaa tagtgttgtc
     3361 tgtaggagaa gccaccagaa ccctcacgga gatccaaagc acagcggata gacaaatttt
     3421 cgaagagaaa gtgggaccgc ttgtcggaag actcaggttg acagcttccc tgcggcagaa
     3481 tggcgctaag accgcttata gggtgaatct caaacttgat caagcagatg ttgttgatag
     3541 cggcttgccc aaagtacgat atactcaagt ttggtcacat gatgtgacaa tcgttgcaaa
     3601 ctcaacggaa gcgagtagga aaagccttta tgatctcact aagtcactcg tcgccacctc
     3661 tcaagttgag gaccttgtcg tcaatttggt gcctcttggt cggggaggcg gttccggcgg
     3721 gggtaccctt gccagtaaga ccatcgtgtt gagcgtggga gaggcaaccc gcacactgac
     3781 agaaatccag tcaacagcag acagacagat ttttgaggag aaagtgggtc cgcttgtcgg
     3841 gcggctgcgg ctgacggctt cattgcgaca gaacggagcc aaaactgcct atcgagtcaa
     3901 ccttaaactc gatcaagcgg atgttgtgga ttccggtctg ccaaaagtac ggtacacgca
     3961 ggtgtggtcc catgacgtga ccattgtggc gaacagtacc gaggcgtctc gcaaaagcct
     4021 ttacgatttg accaaatcac tcgttgcaac ttcacaggtg gaagacctgg ttgtcaatct
     4081 tgtacctttg ggcaggtgag ccgctcgagg agggcccgaa caaaaactca tctcagaaga
     4141 ggatctgaat agcgccgtcg accatcatca taatcatcat tgagtttaaa cccgctgatc
     4201 agcctcgact gtgccttcta gttgccagcc atctgttgtt tgcccctccc ccgtgccttc
     4261 cttgaccctg gaaggtgcca ctcccactgt cctttcctaa taaaatgagg aaattgcatc
     4321 gcattgtctg agtaggtgtc attctattct ggggggtggg gtggggcagg acagcaaggg
     4381 ggaggattgg gaagacaata gcaggcatgc tggggatgcg gtgggctcta tggagatcta
     4441 tcgatgttct ggggatgcgg tgggctctat ggcttctgag gcggaaagaa ccagctgggg
     4501 ctctaggggg tatccccacg cgccctgtag cggcgcatta agcgcggcgg gtgtggtggt
     4561 tacgcgcagc gtgaccgcta cacttgccag cgccctagcg cccgctcctt tcgctttctt
     4621 cccttccttt ctcgccacgt tcgccggctt tccccgtcaa gctctaaatc gggggctccc
     4681 tttagggttc cgatttagtg ctttacggca cctcgacccc aaaaaacttg attagggtga
     4741 tggttcacgt acctagaagt tcctattccg aagttcctat tctctagaaa gtataggaac
     4801 ttccttggcc aaaaagcctg aactcaccgc gacgtctgtc gagaagtttc tgatcgaaaa
     4861 gttcgacagc gtctccgacc tgatgcagct ctcggagggc gaagaatctc gtgctttcag
     4921 cttcgatgta ggagggcgtg gatatgtcct gcgggtaaat agctgcgccg atggtttcta
     4981 caaagatcgt tatgtttatc ggcactttgc atcggccgcg ctcccgattc cggaagtgct
     5041 tgacattggg gaattcagcg agagcctgac ctattgcatc tcccgccgtg cacagggtgt
     5101 cacgttgcaa gacctgcctg aaaccgaact gcccgctgtt ctgcagccgg tcgcggaggc
     5161 catggatgcg atcgctgcgg ccgatcttag ccagacgagc gggttcggcc cattcggacc
     5221 gcaaggaatc ggtcaataca ctacatggcg tgatttcata tgcgcgattg ctgatcccca
     5281 tgtgtatcac tggcaaactg tgatggacga caccgtcagt gcgtccgtcg cgcaggctct
     5341 cgatgagctg atgctttggg ccgaggactg ccccgaagtc cggcacctcg tgcacgcgga
     5401 tttcggctcc aacaatgtcc tgacggacaa tggccgcata acagcggtca ttgactggag
     5461 cgaggcgatg ttcggggatt cccaatacga ggtcgccaac atcttcttct ggaggccgtg
     5521 gttggcttgt atggagcagc agacgcgcta cttcgagcgg aggcatccgg agcttgcagg
     5581 atcgccgcgg ctccgggcgt atatgctccg cattggtctt gaccaactct atcagagctt
     5641 ggttgacggc aatttcgatg atgcagcttg ggcgcagggt cgatgcgacg caatcgtccg
     5701 atccggagcc gggactgtcg ggcgtacaca aatcgcccgc agaagcgcgg ccgtctggac
     5761 cgatggctgt gtagaagtac tcgccgatag tggaaaccga cgccccagca ctcgtccgag
     5821 ggcaaaggaa tagcacgtac tacgagattt cgattccacc gccgccttct atgaaaggtt
     5881 gggcttcgga atcgttttcc gggacgccgg ctggatgatc ctccagcgcg gggatctcat
     5941 gctggagttc ttcgcccacc ccaacttgtt tattgcagct tataatggtt acaaataaag
     6001 caatagcatc acaaatttca caaataaagc atttttttca ctgcattcta gttgtggttt
     6061 gtccaaactc atcaatgtat cttatcatgt ctgtataccg tcgacctcta gctagagctt
     6121 ggcgtaatca tggtcatagc tgtttcctgt gtgaaattgt tatccgctca caattccaca
     6181 caacatacga gccggaagca taaagtgtaa agcctggggt gcctaatgag tgagctaact
     6241 cacattaatt gcgttgcgct cactgcccgc tttccagtcg ggaaacctgt cgtgccagct
     6301 gcattaatga atcggccaac gcgcggggag aggcggtttg cgtattgggc gctcttccgc
     6361 ttcctcgctc actgactcgc tgcgctcggt cgttcggctg cggcgagcgg tatctgctca
     6421 ctcaaaggcg gtaatacggt tatccacaga atcaggggat aacgcaggaa agaacatgtg
     6481 agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca
     6541 taggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa
     6601 cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg tgcgctctcc
     6661 tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg gaagcgtggc
     6721 gctttctcat agctcacgct gtaggtatct cagttcggtg taggtcgttc gctccaagct
     6781 gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg gtaactatcg
     6841 tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca ctggtaacag
     6901 gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt ggcctaacta
     6961 cggctacact agaagaacag tatttggtat ctgcgctctg ctgaagccag ttaccttcgg
     7021 aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg gtggtttttt
     7081 tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct caa
//
