LOCUS       sequence-476097-        7708 bp    DNA     circular SYN 13-JUN-2025
DEFINITION  ARRB2-ADAR2dd(E488Q, T501A) with a long 31 amino acid linker.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7708)
  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 7708)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Jun 13, 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: LSM059 CMV-TO-bArrestin2-(31-linker)-ADAR2dd(E...
FEATURES             Location/Qualifiers
     source          1..7708
                     /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             4703..4732
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     primer_bind     complement(4788..4805)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    4794..5018
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(5190..5209)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     protein_bind    5341..5388
                     /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             5396..6418
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="KKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRGY
                     VLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLPE
                     TELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQT
                     VMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMFG
                     DSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDGN
                     FDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAKE
                     "
     CDS             5396..6418
                     /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    6548..6669
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6585..6604)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     6639..6658
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(6718..6734)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6718..6734)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(6731..6753)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    6742..6758
                     /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(6766..6772,6773..6790,6791..6796))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6811..6832
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(6949..6966)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(7120..7708)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(7200..7219)
                     /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 ccatggggaa
     2161 gaaacccggg accagggtct tcaagaagtc gagccctaac tgcaagctca ccgtgtactt
     2221 gggcaagcgg gacttcgtag atcacctgga caaagtggac cctgtagatg gcgtggtgct
     2281 tgtggaccct gactacctga aggaccgcaa agtgtttgtg accctcacct gcgccttccg
     2341 ctatggccgt gaagacctgg atgtgctggg cttgtccttc cgcaaagacc tgttcatcgc
     2401 cacctaccag gccttccccc cggtgcccaa cccaccccgg ccccccaccc gcctgcagga
     2461 ccggctgctg aggaagctgg gccagcatgc ccaccccttc ttcttcacca taccccagaa
     2521 tcttccatgc tccgtcacac tgcagccagg cccagaggat acaggaaagg cctgcggcgt
     2581 agactttgag attcgagcct tctgtgctaa atcactagaa gagaaaagcc acaaaaggaa
     2641 ctctgtgcgg ctggtgatcc gaaaggtgca gttcgccccg gagaaacccg gcccccagcc
     2701 ttcagccgaa accacacgcc acttcctcat gtctgaccgg tccctgcacc tcgaggcttc
     2761 cctggacaag gagctgtact accatgggga gcccctcaat gtaaatgtcc acgtcaccaa
     2821 caactccacc aagaccgtca agaagatcaa agtctctgtg agacagtacg ccgacatctg
     2881 cctcttcagc accgcccagt acaagtgtcc tgtggctcaa ctcgaacaag atgaccaggt
     2941 atctcccagc tccacattct gtaaggtgta caccataacc ccactgctca gcgacaaccg
     3001 ggagaagcgg ggtctcgccc tggatgggaa actcaagcac gaggacacca acctggcttc
     3061 cagcaccatc gtgaaggagg gtgccaacaa ggaggtgctg ggaatcctgg tgtcctacag
     3121 ggtcaaggtg aagctggtgg tgtctcgagg cggggatgtc tctgtggagc tgccttttgt
     3181 tcttatgcac cccaagcccc acgaccacat ccccctcccc agaccccagt cagccgctcc
     3241 ggagacagat gtccctgtgg acaccaacct cattgaattt gataccaact atgccacaga
     3301 tgatgacatt gtgtttgagg actttgcccg gcttcggctg aaggggatga aggatgacga
     3361 ctatgatgat caactctgcg gtacctcagg cggttccggc agcggggcgg gtagcgggtc
     3421 cagtgggtca gaaacacccg gcacgagtga gtcagcaacg ccagagagcg ctttgcactt
     3481 ggatcaaacg ccatctcgcc agcctattcc cagtgagggt cttcagctgc atttaccgca
     3541 ggttttagct gacgctgtct cacgcctggt cctgggtaag tttggtgacc tgaccgacaa
     3601 cttctcctcc cctcacgctc gcagaaaagt gctggctgga gtcgtcatga caacaggcac
     3661 agatgttaaa gatgccaagg tgataagtgt ttctacagga acaaaatgta ttaatggtga
     3721 atacatgagt gatcgtggcc ttgcattaaa tgactgccat gcagaaataa tatctcggag
     3781 atccttgctc agatttcttt atacacaact tgagctttac ttaaataaca aagatgatca
     3841 aaaaagatcc atctttcaga aatcagagcg aggggggttt aggctgaagg agaatgtcca
     3901 gtttcatctg tacatcagca cctctccctg tggagatgcc agaatcttct caccacatga
     3961 gccaatcctg gaagaaccag cagatagaca cccaaatcgt aaagcaagag gacagctacg
     4021 gaccaaaata gagtctggtc aggggacgat tccagtgcgc tccaatgcga gcatccaagc
     4081 gtgggacggg gtgctgcaag gggagcggct gctcaccatg tcctgcagtg acaagattgc
     4141 acgctggaac gtggtgggca tccagggatc cctgctcagc attttcgtgg agcccattta
     4201 cttctcgagc atcatcctgg gcagccttta ccacggggac cacctttcca gggccatgta
     4261 ccagcggatc tccaacatag aggacctgcc acctctctac accctcaaca agcctttgct
     4321 cagtggcatc agcaatgcag aagcacggca gccagggaag gcccccaact tcagtgtcaa
     4381 ctggacggta ggcgactccg ctattgaggt catcaacgcc acgactggga aggatgagct
     4441 gggccgcgcg tcccgcctgt gtaagcacgc gttgtactgt cgctggatgc gtgtgcacgg
     4501 caaggttccc tcccacttac tacgctccaa gattaccaaa cccaacgtgt accatgagtc
     4561 caagctggcg gcaaaggagt accaggccgc caaggcgcgt ctgttcacag ccttcatcaa
     4621 ggcggggctg ggggcctggg tggagaagcc caccgagcag gaccagttct cactcacgcc
     4681 ctgagccgct cgaggagggc ccgaacaaaa actcatctca gaagaggatc tgaatagcgc
     4741 cgtcgaccat catcataatc atcattgagt ttaaacccgc tgatcagcct cgactgtgcc
     4801 ttctagttgc cagccatctg ttgtttgccc ctcccccgtg ccttccttga ccctggaagg
     4861 tgccactccc actgtccttt cctaataaaa tgaggaaatt gcatcgcatt gtctgagtag
     4921 gtgtcattct attctggggg gtggggtggg gcaggacagc aagggggagg attgggaaga
     4981 caatagcagg catgctgggg atgcggtggg ctctatggag atctatcgat gttctgggga
     5041 tgcggtgggc tctatggctt ctgaggcgga aagaaccagc tggggctcta gggggtatcc
     5101 ccacgcgccc tgtagcggcg cattaagcgc ggcgggtgtg gtggttacgc gcagcgtgac
     5161 cgctacactt gccagcgccc tagcgcccgc tcctttcgct ttcttccctt cctttctcgc
     5221 cacgttcgcc ggctttcccc gtcaagctct aaatcggggg ctccctttag ggttccgatt
     5281 tagtgcttta cggcacctcg accccaaaaa acttgattag ggtgatggtt cacgtaccta
     5341 gaagttccta ttccgaagtt cctattctct agaaagtata ggaacttcct tggccaaaaa
     5401 gcctgaactc accgcgacgt ctgtcgagaa gtttctgatc gaaaagttcg acagcgtctc
     5461 cgacctgatg cagctctcgg agggcgaaga atctcgtgct ttcagcttcg atgtaggagg
     5521 gcgtggatat gtcctgcggg taaatagctg cgccgatggt ttctacaaag atcgttatgt
     5581 ttatcggcac tttgcatcgg ccgcgctccc gattccggaa gtgcttgaca ttggggaatt
     5641 cagcgagagc ctgacctatt gcatctcccg ccgtgcacag ggtgtcacgt tgcaagacct
     5701 gcctgaaacc gaactgcccg ctgttctgca gccggtcgcg gaggccatgg atgcgatcgc
     5761 tgcggccgat cttagccaga cgagcgggtt cggcccattc ggaccgcaag gaatcggtca
     5821 atacactaca tggcgtgatt tcatatgcgc gattgctgat ccccatgtgt atcactggca
     5881 aactgtgatg gacgacaccg tcagtgcgtc cgtcgcgcag gctctcgatg agctgatgct
     5941 ttgggccgag gactgccccg aagtccggca cctcgtgcac gcggatttcg gctccaacaa
     6001 tgtcctgacg gacaatggcc gcataacagc ggtcattgac tggagcgagg cgatgttcgg
     6061 ggattcccaa tacgaggtcg ccaacatctt cttctggagg ccgtggttgg cttgtatgga
     6121 gcagcagacg cgctacttcg agcggaggca tccggagctt gcaggatcgc cgcggctccg
     6181 ggcgtatatg ctccgcattg gtcttgacca actctatcag agcttggttg acggcaattt
     6241 cgatgatgca gcttgggcgc agggtcgatg cgacgcaatc gtccgatccg gagccgggac
     6301 tgtcgggcgt acacaaatcg cccgcagaag cgcggccgtc tggaccgatg gctgtgtaga
     6361 agtactcgcc gatagtggaa accgacgccc cagcactcgt ccgagggcaa aggaatagca
     6421 cgtactacga gatttcgatt ccaccgccgc cttctatgaa aggttgggct tcggaatcgt
     6481 tttccgggac gccggctgga tgatcctcca gcgcggggat ctcatgctgg agttcttcgc
     6541 ccaccccaac ttgtttattg cagcttataa tggttacaaa taaagcaata gcatcacaaa
     6601 tttcacaaat aaagcatttt tttcactgca ttctagttgt ggtttgtcca aactcatcaa
     6661 tgtatcttat catgtctgta taccgtcgac ctctagctag agcttggcgt aatcatggtc
     6721 atagctgttt cctgtgtgaa attgttatcc gctcacaatt ccacacaaca tacgagccgg
     6781 aagcataaag tgtaaagcct ggggtgccta atgagtgagc taactcacat taattgcgtt
     6841 gcgctcactg cccgctttcc agtcgggaaa cctgtcgtgc cagctgcatt aatgaatcgg
     6901 ccaacgcgcg gggagaggcg gtttgcgtat tgggcgctct tccgcttcct cgctcactga
     6961 ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca gctcactcaa aggcggtaat
     7021 acggttatcc acagaatcag gggataacgc aggaaagaac atgtgagcaa aaggccagca
     7081 aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc tccgcccccc
     7141 tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga caggactata
     7201 aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc
     7261 gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt ctcatagctc
     7321 acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga
     7381 accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc
     7441 ggtaagacac gacttatcgc cactggcagc agccactggt aacaggatta gcagagcgag
     7501 gtatgtaggc ggtgctacag agttcttgaa gtggtggcct aactacggct acactagaag
     7561 aacagtattt ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa gagttggtag
     7621 ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt gcaagcagca
     7681 gattacgcgc agaaaaaaag gatctcaa
//
