LOCUS       sequence-476095-        7621 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  CCR7-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 7621)
  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 7621)
  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: LSM044-2 CMV-TO-CCR7-V2-ADAR2dd(E488Q, T501A)
            ...
FEATURES             Location/Qualifiers
     source          1..7621
                     /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             4616..4645
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     primer_bind     complement(4701..4718)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    4707..4931
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(5103..5122)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     protein_bind    5254..5301
                     /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             5309..6331
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="KKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRGY
                     VLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLPE
                     TELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQT
                     VMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMFG
                     DSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDGN
                     FDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAKE
                     "
     CDS             5309..6331
                     /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    6461..6582
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6498..6517)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     6552..6571
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(6631..6647)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6631..6647)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(6644..6666)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    6655..6671
                     /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(6679..6685,6686..6703,6704..6709))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6724..6745
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(6862..6879)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(7033..7621)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(7113..7132)
                     /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 ccatggatct
     2161 cgggaagcct atgaaaagcg tactggttgt ggctttgctg gtgatattcc aggtatgtct
     2221 gtgtcaggat gaagttacag acgactacat aggcgacaat accacagtag attacaccct
     2281 tttcgagtcc ctctgcagca agaaggacgt gaggaatttc aaggcatggt ttctgcctat
     2341 tatgtattcc ataatctgct ttgtcgggct gctggggaac ggtttggtcg tcttgacgta
     2401 tatttacttc aaacgcctga agacaatgac agatacctac cttttgaacc tcgccgtggc
     2461 cgatattctg tttcttctca ctctcccttt ctgggcatat tccgcagcaa agtcatgggt
     2521 cttcggagtc cacttttgca agctgatctt tgctatatac aagatgagct ttttctctgg
     2581 gatgctgctg ctgctgtgca tcagcataga tagatatgtg gccattgtgc aggcggtctc
     2641 tgcacataga caccgggcta gggtgcttct gataagtaaa ttgagctgcg ttgggatctg
     2701 gattttggcc acagtcttgt ccattcctga actgttgtac tcagatttgc agaggagttc
     2761 ctctgagcaa gctatgcgat gttcactgat cactgagcat gtggaagcgt tcattacgat
     2821 ccaagtcgcc caaatggtga ttggcttcct tgttcctttg cttgctatga gcttttgcta
     2881 cctcgtaatt atacgcacac tcctgcaggc ccggaatttt gagcggaaca aggctattaa
     2941 ggtgataatt gccgtagtgg ttgtattcat tgtctttcag ctgccttata atggggtagt
     3001 tttggcccaa acagttgcta actttaacat cacttcttca acatgtgagc tgtcaaagca
     3061 gctgaatatc gcatacgatg tgacatactc cttggcgtgc gtcaggtgtt gcgtgaatcc
     3121 tttcctgtac gcttttattg gtgtgaagtt tcgaaatgac ctgtttaaac ttttcaaaga
     3181 cttgggctgt ctgtctcagg agcagcttcg ccagtggtca tcatgcagac acattaggcg
     3241 ctccagtatg agtgtggagg cagagaccac tactacgttc tcccctaccg gtggacgcac
     3301 cccacccagc ctgggtcccc aagatgagtc ctgcaccacc gccagctcct ccctggccaa
     3361 ggacacttca tcgggtaccg gtgctttgca cttggatcaa acgccatctc gccagcctat
     3421 tcccagtgag ggtcttcagc tgcatttacc gcaggtttta gctgacgctg tctcacgcct
     3481 ggtcctgggt aagtttggtg acctgaccga caacttctcc tcccctcacg ctcgcagaaa
     3541 agtgctggct ggagtcgtca tgacaacagg cacagatgtt aaagatgcca aggtgataag
     3601 tgtttctaca ggaacaaaat gtattaatgg tgaatacatg agtgatcgtg gccttgcatt
     3661 aaatgactgc catgcagaaa taatatctcg gagatccttg ctcagatttc tttatacaca
     3721 acttgagctt tacttaaata acaaagatga tcaaaaaaga tccatctttc agaaatcaga
     3781 gcgagggggg tttaggctga aggagaatgt ccagtttcat ctgtacatca gcacctctcc
     3841 ctgtggagat gccagaatct tctcaccaca tgagccaatc ctggaagaac cagcagatag
     3901 acacccaaat cgtaaagcaa gaggacagct acggaccaaa atagagtctg gtcaggggac
     3961 gattccagtg cgctccaatg cgagcatcca agcgtgggac ggggtgctgc aaggggagcg
     4021 gctgctcacc atgtcctgca gtgacaagat tgcacgctgg aacgtggtgg gcatccaggg
     4081 atccctgctc agcattttcg tggagcccat ttacttctcg agcatcatcc tgggcagcct
     4141 ttaccacggg gaccaccttt ccagggccat gtaccagcgg atctccaaca tagaggacct
     4201 gccacctctc tacaccctca acaagccttt gctcagtggc atcagcaatg cagaagcacg
     4261 gcagccaggg aaggccccca acttcagtgt caactggacg gtaggcgact ccgctattga
     4321 ggtcatcaac gccacgactg ggaaggatga gctgggccgc gcgtcccgcc tgtgtaagca
     4381 cgcgttgtac tgtcgctgga tgcgtgtgca cggcaaggtt ccctcccact tactacgctc
     4441 caagattacc aaacccaacg tgtaccatga gtccaagctg gcggcaaagg agtaccaggc
     4501 cgccaaggcg cgtctgttca cagccttcat caaggcgggg ctgggggcct gggtggagaa
     4561 gcccaccgag caggaccagt tctcactcac gccctgagcc gctcgaggag ggcccgaaca
     4621 aaaactcatc tcagaagagg atctgaatag cgccgtcgac catcatcata atcatcattg
     4681 agtttaaacc cgctgatcag cctcgactgt gccttctagt tgccagccat ctgttgtttg
     4741 cccctccccc gtgccttcct tgaccctgga aggtgccact cccactgtcc tttcctaata
     4801 aaatgaggaa attgcatcgc attgtctgag taggtgtcat tctattctgg ggggtggggt
     4861 ggggcaggac agcaaggggg aggattggga agacaatagc aggcatgctg gggatgcggt
     4921 gggctctatg gagatctatc gatgttctgg ggatgcggtg ggctctatgg cttctgaggc
     4981 ggaaagaacc agctggggct ctagggggta tccccacgcg ccctgtagcg gcgcattaag
     5041 cgcggcgggt gtggtggtta cgcgcagcgt gaccgctaca cttgccagcg ccctagcgcc
     5101 cgctcctttc gctttcttcc cttcctttct cgccacgttc gccggctttc cccgtcaagc
     5161 tctaaatcgg gggctccctt tagggttccg atttagtgct ttacggcacc tcgaccccaa
     5221 aaaacttgat tagggtgatg gttcacgtac ctagaagttc ctattccgaa gttcctattc
     5281 tctagaaagt ataggaactt ccttggccaa aaagcctgaa ctcaccgcga cgtctgtcga
     5341 gaagtttctg atcgaaaagt tcgacagcgt ctccgacctg atgcagctct cggagggcga
     5401 agaatctcgt gctttcagct tcgatgtagg agggcgtgga tatgtcctgc gggtaaatag
     5461 ctgcgccgat ggtttctaca aagatcgtta tgtttatcgg cactttgcat cggccgcgct
     5521 cccgattccg gaagtgcttg acattgggga attcagcgag agcctgacct attgcatctc
     5581 ccgccgtgca cagggtgtca cgttgcaaga cctgcctgaa accgaactgc ccgctgttct
     5641 gcagccggtc gcggaggcca tggatgcgat cgctgcggcc gatcttagcc agacgagcgg
     5701 gttcggccca ttcggaccgc aaggaatcgg tcaatacact acatggcgtg atttcatatg
     5761 cgcgattgct gatccccatg tgtatcactg gcaaactgtg atggacgaca ccgtcagtgc
     5821 gtccgtcgcg caggctctcg atgagctgat gctttgggcc gaggactgcc ccgaagtccg
     5881 gcacctcgtg cacgcggatt tcggctccaa caatgtcctg acggacaatg gccgcataac
     5941 agcggtcatt gactggagcg aggcgatgtt cggggattcc caatacgagg tcgccaacat
     6001 cttcttctgg aggccgtggt tggcttgtat ggagcagcag acgcgctact tcgagcggag
     6061 gcatccggag cttgcaggat cgccgcggct ccgggcgtat atgctccgca ttggtcttga
     6121 ccaactctat cagagcttgg ttgacggcaa tttcgatgat gcagcttggg cgcagggtcg
     6181 atgcgacgca atcgtccgat ccggagccgg gactgtcggg cgtacacaaa tcgcccgcag
     6241 aagcgcggcc gtctggaccg atggctgtgt agaagtactc gccgatagtg gaaaccgacg
     6301 ccccagcact cgtccgaggg caaaggaata gcacgtacta cgagatttcg attccaccgc
     6361 cgccttctat gaaaggttgg gcttcggaat cgttttccgg gacgccggct ggatgatcct
     6421 ccagcgcggg gatctcatgc tggagttctt cgcccacccc aacttgttta ttgcagctta
     6481 taatggttac aaataaagca atagcatcac aaatttcaca aataaagcat ttttttcact
     6541 gcattctagt tgtggtttgt ccaaactcat caatgtatct tatcatgtct gtataccgtc
     6601 gacctctagc tagagcttgg cgtaatcatg gtcatagctg tttcctgtgt gaaattgtta
     6661 tccgctcaca attccacaca acatacgagc cggaagcata aagtgtaaag cctggggtgc
     6721 ctaatgagtg agctaactca cattaattgc gttgcgctca ctgcccgctt tccagtcggg
     6781 aaacctgtcg tgccagctgc attaatgaat cggccaacgc gcggggagag gcggtttgcg
     6841 tattgggcgc tcttccgctt cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg
     6901 gcgagcggta tcagctcact caaaggcggt aatacggtta tccacagaat caggggataa
     6961 cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc
     7021 gttgctggcg tttttccata ggctccgccc ccctgacgag catcacaaaa atcgacgctc
     7081 aagtcagagg tggcgaaacc cgacaggact ataaagatac caggcgtttc cccctggaag
     7141 ctccctcgtg cgctctcctg ttccgaccct gccgcttacc ggatacctgt ccgcctttct
     7201 cccttcggga agcgtggcgc tttctcatag ctcacgctgt aggtatctca gttcggtgta
     7261 ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc gttcagcccg accgctgcgc
     7321 cttatccggt aactatcgtc ttgagtccaa cccggtaaga cacgacttat cgccactggc
     7381 agcagccact ggtaacagga ttagcagagc gaggtatgta ggcggtgcta cagagttctt
     7441 gaagtggtgg cctaactacg gctacactag aagaacagta tttggtatct gcgctctgct
     7501 gaagccagtt accttcggaa aaagagttgg tagctcttga tccggcaaac aaaccaccgc
     7561 tggtagcggt ggtttttttg tttgcaagca gcagattacg cgcagaaaaa aaggatctca
     7621 a
//
