LOCUS       sequence-466399-        7534 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  CCR7-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 7534)
  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 7534)
  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: LSM044-3 CMV-TO-CCR7-ADAR2dd(E488Q, T501A) (FL...
FEATURES             Location/Qualifiers
     source          1..7534
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(87..106)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     protein_bind    238..285
                     /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             293..1315
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="KKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRGY
                     VLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLPE
                     TELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQT
                     VMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMFG
                     DSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDGN
                     FDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAKE
                     "
     CDS             293..1315
                     /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    1445..1566
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1482..1501)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     1536..1555
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(1615..1631)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1615..1631)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1628..1650)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1639..1655
                     /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(1663..1669,1670..1687,1688..1693))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1708..1729
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(1846..1863)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2017..2605)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2097..2116)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2776..3567,3568..3636))
                     /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     3399..3418
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3637..3741)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(3816..3835)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        4002..4381
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        4382..4585
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     4536..4556
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     protein_bind    4587..4605
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    4608..4626
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     primer_bind     4636..4660
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     regulatory      4752..4761
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             7134..7163
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     primer_bind     complement(7219..7236)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    7225..7449
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
ORIGIN
        1 cgcgccctgt agcggcgcat taagcgcggc gggtgtggtg gttacgcgca gcgtgaccgc
       61 tacacttgcc agcgccctag cgcccgctcc tttcgctttc ttcccttcct ttctcgccac
      121 gttcgccggc tttccccgtc aagctctaaa tcgggggctc cctttagggt tccgatttag
      181 tgctttacgg cacctcgacc ccaaaaaact tgattagggt gatggttcac gtacctagaa
      241 gttcctattc cgaagttcct attctctaga aagtatagga acttccttgg ccaaaaagcc
      301 tgaactcacc gcgacgtctg tcgagaagtt tctgatcgaa aagttcgaca gcgtctccga
      361 cctgatgcag ctctcggagg gcgaagaatc tcgtgctttc agcttcgatg taggagggcg
      421 tggatatgtc ctgcgggtaa atagctgcgc cgatggtttc tacaaagatc gttatgttta
      481 tcggcacttt gcatcggccg cgctcccgat tccggaagtg cttgacattg gggaattcag
      541 cgagagcctg acctattgca tctcccgccg tgcacagggt gtcacgttgc aagacctgcc
      601 tgaaaccgaa ctgcccgctg ttctgcagcc ggtcgcggag gccatggatg cgatcgctgc
      661 ggccgatctt agccagacga gcgggttcgg cccattcgga ccgcaaggaa tcggtcaata
      721 cactacatgg cgtgatttca tatgcgcgat tgctgatccc catgtgtatc actggcaaac
      781 tgtgatggac gacaccgtca gtgcgtccgt cgcgcaggct ctcgatgagc tgatgctttg
      841 ggccgaggac tgccccgaag tccggcacct cgtgcacgcg gatttcggct ccaacaatgt
      901 cctgacggac aatggccgca taacagcggt cattgactgg agcgaggcga tgttcgggga
      961 ttcccaatac gaggtcgcca acatcttctt ctggaggccg tggttggctt gtatggagca
     1021 gcagacgcgc tacttcgagc ggaggcatcc ggagcttgca ggatcgccgc ggctccgggc
     1081 gtatatgctc cgcattggtc ttgaccaact ctatcagagc ttggttgacg gcaatttcga
     1141 tgatgcagct tgggcgcagg gtcgatgcga cgcaatcgtc cgatccggag ccgggactgt
     1201 cgggcgtaca caaatcgccc gcagaagcgc ggccgtctgg accgatggct gtgtagaagt
     1261 actcgccgat agtggaaacc gacgccccag cactcgtccg agggcaaagg aatagcacgt
     1321 actacgagat ttcgattcca ccgccgcctt ctatgaaagg ttgggcttcg gaatcgtttt
     1381 ccgggacgcc ggctggatga tcctccagcg cggggatctc atgctggagt tcttcgccca
     1441 ccccaacttg tttattgcag cttataatgg ttacaaataa agcaatagca tcacaaattt
     1501 cacaaataaa gcattttttt cactgcattc tagttgtggt ttgtccaaac tcatcaatgt
     1561 atcttatcat gtctgtatac cgtcgacctc tagctagagc ttggcgtaat catggtcata
     1621 gctgtttcct gtgtgaaatt gttatccgct cacaattcca cacaacatac gagccggaag
     1681 cataaagtgt aaagcctggg gtgcctaatg agtgagctaa ctcacattaa ttgcgttgcg
     1741 ctcactgccc gctttccagt cgggaaacct gtcgtgccag ctgcattaat gaatcggcca
     1801 acgcgcgggg agaggcggtt tgcgtattgg gcgctcttcc gcttcctcgc tcactgactc
     1861 gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg cggtaatacg
     1921 gttatccaca gaatcagggg ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa
     1981 ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc gcccccctga
     2041 cgagcatcac aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag
     2101 ataccaggcg tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct
     2161 taccggatac ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc atagctcacg
     2221 ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc
     2281 ccccgttcag cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt
     2341 aagacacgac ttatcgccac tggcagcagc cactggtaac aggattagca gagcgaggta
     2401 tgtaggcggt gctacagagt tcttgaagtg gtggcctaac tacggctaca ctagaagaac
     2461 agtatttggt atctgcgctc tgctgaagcc agttaccttc ggaaaaagag ttggtagctc
     2521 ttgatccggc aaacaaacca ccgctggtag cggtggtttt tttgtttgca agcagcagat
     2581 tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc
     2641 tcagtggaac gaaaactcac gttaagggat tttggtcatg agattatcaa aaaggatctt
     2701 cacctagatc cttttaaatt aaaaatgaag ttttaaatca atctaaagta tatatgagta
     2761 aacttggtct gacagttacc aatgcttaat cagtgaggca cctatctcag cgatctgtct
     2821 atttcgttca tccatagttg cctgactccc cgtcgtgtag ataactacga tacgggaggg
     2881 cttaccatct ggccccagtg ctgcaatgat accgcgagac ccacgctcac cggctccaga
     2941 tttatcagca ataaaccagc cagccggaag ggccgagcgc agaagtggtc ctgcaacttt
     3001 atccgcctcc atccagtcta ttaattgttg ccgggaagct agagtaagta gttcgccagt
     3061 taatagtttg cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt
     3121 tggtatggct tcattcagct ccggttccca acgatcaagg cgagttacat gatcccccat
     3181 gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc gttgtcagaa gtaagttggc
     3241 cgcagtgtta tcactcatgg ttatggcagc actgcataat tctcttactg tcatgccatc
     3301 cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag tcattctgag aatagtgtat
     3361 gcggcgaccg agttgctctt gcccggcgtc aatacgggat aataccgcgc cacatagcag
     3421 aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct caaggatctt
     3481 accgctgttg agatccagtt cgatgtaacc cactcgtgca cccaactgat cttcagcatc
     3541 ttttactttc accagcgttt ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa
     3601 gggaataagg gcgacacgga aatgttgaat actcatactc ttcctttttc aatattattg
     3661 aagcatttat cagggttatt gtctcatgag cggatacata tttgaatgta tttagaaaaa
     3721 taaacaaata ggggttccgc gcacatttcc ccgaaaagtg ccacctgacg tcgacggatc
     3781 gggagatctc ccgatcccct atggtgcact ctcagtacaa tctgctctga tgccgcatag
     3841 ttaagccagt atctgctccc tgcttgtgtg ttggaggtcg ctgagtagtg cgcgagcaaa
     3901 atttaagcta caacaaggca aggcttgacc gacaattgca tgaagaatct gcttagggtt
     3961 aggcgttttg cgctgcttcg tacgggccag atatacgcgt tgacattgat tattgactag
     4021 ttattaatag taatcaatta cggggtcatt agttcatagc ccatatatgg agttccgcgt
     4081 tacataactt acggtaaatg gcccgcctgg ctgaccgccc aacgaccccc gcccattgac
     4141 gtcaataatg acgtatgttc ccatagtaac gccaataggg actttccatt gacgtcaatg
     4201 ggtggagtat ttacggtaaa ctgcccactt ggcagtacat caagtgtatc atatgccaag
     4261 tacgccccct attgacgtca atgacggtaa atggcccgcc tggcattatg cccagtacat
     4321 gaccttatgg gactttccta cttggcagta catctacgta ttagtcatcg ctattaccat
     4381 ggtgatgcgg ttttggcagt acatcaatgg gcgtggatag cggtttgact cacggggatt
     4441 tccaagtctc caccccattg acgtcaatgg gagtttgttt tggcaccaaa atcaacggga
     4501 ctttccaaaa tgtcgtaaca actccgcccc attgacgcaa atgggcggta ggcgtgtacg
     4561 gtgggaggtc tatataagca gagctctccc tatcagtgat agagatctcc ctatcagtga
     4621 tagagatcgt cgacgagctc gtttagtgaa ccgtcagatc gcctggagac gccatccacg
     4681 ctgttttgac ctccatagaa gacaccggga ccgatccagc ctccggactc tagcgtttaa
     4741 acttaagctt cgccaccatg gatctcggga agcctatgaa aagcgtactg gttgtggctt
     4801 tgctggtgat attccaggta tgtctgtgtc aggatgaagt tacagacgac tacataggcg
     4861 acaataccac agtagattac acccttttcg agtccctctg cagcaagaag gacgtgagga
     4921 atttcaaggc atggtttctg cctattatgt attccataat ctgctttgtc gggctgctgg
     4981 ggaacggttt ggtcgtcttg acgtatattt acttcaaacg cctgaagaca atgacagata
     5041 cctacctttt gaacctcgcc gtggccgata ttctgtttct tctcactctc cctttctggg
     5101 catattccgc agcaaagtca tgggtcttcg gagtccactt ttgcaagctg atctttgcta
     5161 tatacaagat gagctttttc tctgggatgc tgctgctgct gtgcatcagc atagatagat
     5221 atgtggccat tgtgcaggcg gtctctgcac atagacaccg ggctagggtg cttctgataa
     5281 gtaaattgag ctgcgttggg atctggattt tggccacagt cttgtccatt cctgaactgt
     5341 tgtactcaga tttgcagagg agttcctctg agcaagctat gcgatgttca ctgatcactg
     5401 agcatgtgga agcgttcatt acgatccaag tcgcccaaat ggtgattggc ttccttgttc
     5461 ctttgcttgc tatgagcttt tgctacctcg taattatacg cacactcctg caggcccgga
     5521 attttgagcg gaacaaggct attaaggtga taattgccgt agtggttgta ttcattgtct
     5581 ttcagctgcc ttataatggg gtagttttgg cccaaacagt tgctaacttt aacatcactt
     5641 cttcaacatg tgagctgtca aagcagctga atatcgcata cgatgtgaca tactccttgg
     5701 cgtgcgtcag gtgttgcgtg aatcctttcc tgtacgcttt tattggtgtg aagtttcgaa
     5761 atgacctgtt taaacttttc aaagacttgg gctgtctgtc tcaggagcag cttcgccagt
     5821 ggtcatcatg cagacacatt aggcgctcca gtatgagtgt ggaggcagag accactacta
     5881 cgttctcccc tggtaccggt gctttgcact tggatcaaac gccatctcgc cagcctattc
     5941 ccagtgaggg tcttcagctg catttaccgc aggttttagc tgacgctgtc tcacgcctgg
     6001 tcctgggtaa gtttggtgac ctgaccgaca acttctcctc ccctcacgct cgcagaaaag
     6061 tgctggctgg agtcgtcatg acaacaggca cagatgttaa agatgccaag gtgataagtg
     6121 tttctacagg aacaaaatgt attaatggtg aatacatgag tgatcgtggc cttgcattaa
     6181 atgactgcca tgcagaaata atatctcgga gatccttgct cagatttctt tatacacaac
     6241 ttgagcttta cttaaataac aaagatgatc aaaaaagatc catctttcag aaatcagagc
     6301 gaggggggtt taggctgaag gagaatgtcc agtttcatct gtacatcagc acctctccct
     6361 gtggagatgc cagaatcttc tcaccacatg agccaatcct ggaagaacca gcagatagac
     6421 acccaaatcg taaagcaaga ggacagctac ggaccaaaat agagtctggt caggggacga
     6481 ttccagtgcg ctccaatgcg agcatccaag cgtgggacgg ggtgctgcaa ggggagcggc
     6541 tgctcaccat gtcctgcagt gacaagattg cacgctggaa cgtggtgggc atccagggat
     6601 ccctgctcag cattttcgtg gagcccattt acttctcgag catcatcctg ggcagccttt
     6661 accacgggga ccacctttcc agggccatgt accagcggat ctccaacata gaggacctgc
     6721 cacctctcta caccctcaac aagcctttgc tcagtggcat cagcaatgca gaagcacggc
     6781 agccagggaa ggcccccaac ttcagtgtca actggacggt aggcgactcc gctattgagg
     6841 tcatcaacgc cacgactggg aaggatgagc tgggccgcgc gtcccgcctg tgtaagcacg
     6901 cgttgtactg tcgctggatg cgtgtgcacg gcaaggttcc ctcccactta ctacgctcca
     6961 agattaccaa acccaacgtg taccatgagt ccaagctggc ggcaaaggag taccaggccg
     7021 ccaaggcgcg tctgttcaca gccttcatca aggcggggct gggggcctgg gtggagaagc
     7081 ccaccgagca ggaccagttc tcactcacgc cctgagccgc tcgaggaggg cccgaacaaa
     7141 aactcatctc agaagaggat ctgaatagcg ccgtcgacca tcatcataat catcattgag
     7201 tttaaacccg ctgatcagcc tcgactgtgc cttctagttg ccagccatct gttgtttgcc
     7261 cctcccccgt gccttccttg accctggaag gtgccactcc cactgtcctt tcctaataaa
     7321 atgaggaaat tgcatcgcat tgtctgagta ggtgtcattc tattctgggg ggtggggtgg
     7381 ggcaggacag caagggggag gattgggaag acaatagcag gcatgctggg gatgcggtgg
     7441 gctctatgga gatctatcga tgttctgggg atgcggtggg ctctatggct tctgaggcgg
     7501 aaagaaccag ctggggctct agggggtatc ccca
//
