LOCUS       sequence-466390-        7147 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  ARRB2-tdMCP with a short linker.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7147)
  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 7147)
  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: XZ024 CMV-TO-bArrestin2-MCP (FLP-IN)
FEATURES             Location/Qualifiers
     source          1..7147
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(19..1041)
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="KKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRGY
                     VLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLPE
                     TELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQT
                     VMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMFG
                     DSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDGN
                     FDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAKE
                     "
     CDS             complement(19..1041)
                     /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
                     "
     protein_bind    complement(1049..1096)
                     /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)."
     primer_bind     1228..1247
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     polyA_signal    1419..1643
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     1632..1649
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     CDS             complement(1705..1734)
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     regulatory      3720..3729
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     primer_bind     complement(3821..3845)
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     protein_bind    3855..3873
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    3876..3894
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     promoter        complement(3896..4099)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(3925..3945)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        4100..4479
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     primer_bind     4646..4665
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        4740..4844
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4845..4913,4914..5705)
                     /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     complement(5063..5082)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5876..6464
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6365..6384
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6618..6635
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    6752..6773
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(6788..6793,6794..6811,6812..6818)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6826..6842
                     /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)."
     primer_bind     6831..6853
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     6850..6866
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     6850..6866
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     polyA_signal    6915..7036
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6926..6945)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     6980..6999
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
ORIGIN
        1 gaaatctcgt agtacgtgct attcctttgc cctcggacga gtgctggggc gtcggtttcc
       61 actatcggcg agtacttcta cacagccatc ggtccagacg gccgcgcttc tgcgggcgat
      121 ttgtgtacgc ccgacagtcc cggctccgga tcggacgatt gcgtcgcatc gaccctgcgc
      181 ccaagctgca tcatcgaaat tgccgtcaac caagctctga tagagttggt caagaccaat
      241 gcggagcata tacgcccgga gccgcggcga tcctgcaagc tccggatgcc tccgctcgaa
      301 gtagcgcgtc tgctgctcca tacaagccaa ccacggcctc cagaagaaga tgttggcgac
      361 ctcgtattgg gaatccccga acatcgcctc gctccagtca atgaccgctg ttatgcggcc
      421 attgtccgtc aggacattgt tggagccgaa atccgcgtgc acgaggtgcc ggacttcggg
      481 gcagtcctcg gcccaaagca tcagctcatc gagagcctgc gcgacggacg cactgacggt
      541 gtcgtccatc acagtttgcc agtgatacac atggggatca gcaatcgcgc atatgaaatc
      601 acgccatgta gtgtattgac cgattccttg cggtccgaat gggccgaacc cgctcgtctg
      661 gctaagatcg gccgcagcga tcgcatccat ggcctccgcg accggctgca gaacagcggg
      721 cagttcggtt tcaggcaggt cttgcaacgt gacaccctgt gcacggcggg agatgcaata
      781 ggtcaggctc tcgctgaatt ccccaatgtc aagcacttcc ggaatcggga gcgcggccga
      841 tgcaaagtgc cgataaacat aacgatcttt gtagaaacca tcggcgcagc tatttacccg
      901 caggacatat ccacgccctc ctacatcgaa gctgaaagca cgagattctt cgccctccga
      961 gagctgcatc aggtcggaga cgctgtcgaa cttttcgatc agaaacttct cgacagacgt
     1021 cgcggtgagt tcaggctttt tggccaagga agttcctata ctttctagag aataggaact
     1081 tcggaatagg aacttctagg tacgtgaacc atcaccctaa tcaagttttt tggggtcgag
     1141 gtgccgtaaa gcactaaatc ggaaccctaa agggagcccc cgatttagag cttgacgggg
     1201 aaagccggcg aacgtggcga gaaaggaagg gaagaaagcg aaaggagcgg gcgctagggc
     1261 gctggcaagt gtagcggtca cgctgcgcgt aaccaccaca cccgccgcgc ttaatgcgcc
     1321 gctacagggc gcgtggggat accccctaga gccccagctg gttctttccg cctcagaagc
     1381 catagagccc accgcatccc cagaacatcg atagatctcc atagagccca ccgcatcccc
     1441 agcatgcctg ctattgtctt cccaatcctc ccccttgctg tcctgcccca ccccaccccc
     1501 cagaatagaa tgacacctac tcagacaatg cgatgcaatt tcctcatttt attaggaaag
     1561 gacagtggga gtggcacctt ccagggtcaa ggaaggcacg ggggaggggc aaacaacaga
     1621 tggctggcaa ctagaaggca cagtcgaggc tgatcagcgg gtttaaactc aatgatgatt
     1681 atgatgatgg tcgacggcgc tattcagatc ctcttctgag atgagttttt gttcgggccc
     1741 tcctcgagcg gctcagtaaa ttccgctatt cgcagcgatt gcggaaggta tcgggttgcc
     1801 atctttcaga agtccttgca tggcttttac aataagctcg cagtcgctat tggtcgcaaa
     1861 tatcggaatc gtcaattcca tattcaagta tgagcgccat gcacctttgg ggacctccac
     1921 tttaatggta tacttccggt tctgggcact agattggcgt acactacaag tcaccttata
     1981 ggcttgtgat cttgaattgg aagagatcca ctcggctatc ccattcgcaa agttacttgg
     2041 agctactgtg acgtctccag tgccgccgtt gtccacgagc acaaactgtg taaaattgga
     2101 ggccatggac ccgccaccgg ttccgccacc atatattcct gagttggcgg cgatcgcaga
     2161 gggaatcggg ttaccgtctt tcaaaagacc ttgcatggcc ttgacaatca gctcgcagtc
     2221 tgaattagtc gcaaagattg ggatcgtaag ctccatatta agataagatc gccacgcccc
     2281 ttttggcact tctaccttta tggtatattt tcggttctgt gcggaacttt ggcgaactga
     2341 gcatgtgact ttatatgcct gactccggct gttactactt atccattctg caatcccgtt
     2401 cgcaaaattg ctcggcgcaa cagttacgtc tccggtgcca ccattgtcaa ccaacacaaa
     2461 ttgcgtaaag ttgctcgcca tagcgctacc accgccgcag agttgatcat catagtcgtc
     2521 atccttcatc cccttcagcc gaagccgggc aaagtcctca aacacaatgt catcatctgt
     2581 ggcatagttg gtatcaaatt caatgaggtt ggtgtccaca gggacatctg tctccggagc
     2641 ggctgactgg ggtctgggga gggggatgtg gtcgtggggc ttggggtgca taagaacaaa
     2701 aggcagctcc acagagacat ccccgcctcg agacaccacc agcttcacct tgaccctgta
     2761 ggacaccagg attcccagca cctccttgtt ggcaccctcc ttcacgatgg tgctggaagc
     2821 caggttggtg tcctcgtgct tgagtttccc atccagggcg agaccccgct tctcccggtt
     2881 gtcgctgagc agtggggtta tggtgtacac cttacagaat gtggagctgg gagatacctg
     2941 gtcatcttgt tcgagttgag ccacaggaca cttgtactgg gcggtgctga agaggcagat
     3001 gtcggcgtac tgtctcacag agactttgat cttcttgacg gtcttggtgg agttgttggt
     3061 gacgtggaca tttacattga ggggctcccc atggtagtac agctccttgt ccagggaagc
     3121 ctcgaggtgc agggaccggt cagacatgag gaagtggcgt gtggtttcgg ctgaaggctg
     3181 ggggccgggt ttctccgggg cgaactgcac ctttcggatc accagccgca cagagttcct
     3241 tttgtggctt ttctcttcta gtgatttagc acagaaggct cgaatctcaa agtctacgcc
     3301 gcaggccttt cctgtatcct ctgggcctgg ctgcagtgtg acggagcatg gaagattctg
     3361 gggtatggtg aagaagaagg ggtgggcatg ctggcccagc ttcctcagca gccggtcctg
     3421 caggcgggtg gggggccggg gtgggttggg caccgggggg aaggcctggt aggtggcgat
     3481 gaacaggtct ttgcggaagg acaagcccag cacatccagg tcttcacggc catagcggaa
     3541 ggcgcaggtg agggtcacaa acactttgcg gtccttcagg tagtcagggt ccacaagcac
     3601 cacgccatct acagggtcca ctttgtccag gtgatctacg aagtcccgct tgcccaagta
     3661 cacggtgagc ttgcagttag ggctcgactt cttgaagacc ctggtcccgg gtttctcccc
     3721 catggtggcg aagcttaagt ttaaacgcta gagtccggag gctggatcgg tcccggtgtc
     3781 ttctatggag gtcaaaacag cgtggatggc gtctccaggc gatctgacgg ttcactaaac
     3841 gagctcgtcg acgatctcta tcactgatag ggagatctct atcactgata gggagagctc
     3901 tgcttatata gacctcccac cgtacacgcc taccgcccat ttgcgtcaat ggggcggagt
     3961 tgttacgaca ttttggaaag tcccgttgat tttggtgcca aaacaaactc ccattgacgt
     4021 caatggggtg gagacttgga aatccccgtg agtcaaaccg ctatccacgc ccattgatgt
     4081 actgccaaaa ccgcatcacc atggtaatag cgatgactaa tacgtagatg tactgccaag
     4141 taggaaagtc ccataaggtc atgtactggg cataatgcca ggcgggccat ttaccgtcat
     4201 tgacgtcaat agggggcgta cttggcatat gatacacttg atgtactgcc aagtgggcag
     4261 tttaccgtaa atactccacc cattgacgtc aatggaaagt ccctattggc gttactatgg
     4321 gaacatacgt cattattgac gtcaatgggc gggggtcgtt gggcggtcag ccaggcgggc
     4381 catttaccgt aagttatgta acgcggaact ccatatatgg gctatgaact aatgaccccg
     4441 taattgatta ctattaataa ctagtcaata atcaatgtca acgcgtatat ctggcccgta
     4501 cgaagcagcg caaaacgcct aaccctaagc agattcttca tgcaattgtc ggtcaagcct
     4561 tgccttgttg tagcttaaat tttgctcgcg cactactcag cgacctccaa cacacaagca
     4621 gggagcagat actggcttaa ctatgcggca tcagagcaga ttgtactgag agtgcaccat
     4681 aggggatcgg gagatctccc gatccgtcga cgtcaggtgg cacttttcgg ggaaatgtgc
     4741 gcggaacccc tatttgttta tttttctaaa tacattcaaa tatgtatccg ctcatgagac
     4801 aataaccctg ataaatgctt caataatatt gaaaaaggaa gagtatgagt attcaacatt
     4861 tccgtgtcgc ccttattccc ttttttgcgg cattttgcct tcctgttttt gctcacccag
     4921 aaacgctggt gaaagtaaaa gatgctgaag atcagttggg tgcacgagtg ggttacatcg
     4981 aactggatct caacagcggt aagatccttg agagttttcg ccccgaagaa cgttttccaa
     5041 tgatgagcac ttttaaagtt ctgctatgtg gcgcggtatt atcccgtatt gacgccgggc
     5101 aagagcaact cggtcgccgc atacactatt ctcagaatga cttggttgag tactcaccag
     5161 tcacagaaaa gcatcttacg gatggcatga cagtaagaga attatgcagt gctgccataa
     5221 ccatgagtga taacactgcg gccaacttac ttctgacaac gatcggagga ccgaaggagc
     5281 taaccgcttt tttgcacaac atgggggatc atgtaactcg ccttgatcgt tgggaaccgg
     5341 agctgaatga agccatacca aacgacgagc gtgacaccac gatgcctgta gcaatggcaa
     5401 caacgttgcg caaactatta actggcgaac tacttactct agcttcccgg caacaattaa
     5461 tagactggat ggaggcggat aaagttgcag gaccacttct gcgctcggcc cttccggctg
     5521 gctggtttat tgctgataaa tctggagccg gtgagcgtgg gtctcgcggt atcattgcag
     5581 cactggggcc agatggtaag ccctcccgta tcgtagttat ctacacgacg gggagtcagg
     5641 caactatgga tgaacgaaat agacagatcg ctgagatagg tgcctcactg attaagcatt
     5701 ggtaactgtc agaccaagtt tactcatata tactttagat tgatttaaaa cttcattttt
     5761 aatttaaaag gatctaggtg aagatccttt ttgataatct catgaccaaa atcccttaac
     5821 gtgagttttc gttccactga gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag
     5881 atcctttttt tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg
     5941 tggtttgttt gccggatcaa gagctaccaa ctctttttcc gaaggtaact ggcttcagca
     6001 gagcgcagat accaaatact gttcttctag tgtagccgta gttaggccac cacttcaaga
     6061 actctgtagc accgcctaca tacctcgctc tgctaatcct gttaccagtg gctgctgcca
     6121 gtggcgataa gtcgtgtctt accgggttgg actcaagacg atagttaccg gataaggcgc
     6181 agcggtcggg ctgaacgggg ggttcgtgca cacagcccag cttggagcga acgacctaca
     6241 ccgaactgag atacctacag cgtgagctat gagaaagcgc cacgcttccc gaagggagaa
     6301 aggcggacag gtatccggta agcggcaggg tcggaacagg agagcgcacg agggagcttc
     6361 cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc
     6421 gtcgattttt gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg
     6481 cctttttacg gttcctggcc ttttgctggc cttttgctca catgttcttt cctgcgttat
     6541 cccctgattc tgtggataac cgtattaccg cctttgagtg agctgatacc gctcgccgca
     6601 gccgaacgac cgagcgcagc gagtcagtga gcgaggaagc ggaagagcgc ccaatacgca
     6661 aaccgcctct ccccgcgcgt tggccgattc attaatgcag ctggcacgac aggtttcccg
     6721 actggaaagc gggcagtgag cgcaacgcaa ttaatgtgag ttagctcact cattaggcac
     6781 cccaggcttt acactttatg cttccggctc gtatgttgtg tggaattgtg agcggataac
     6841 aatttcacac aggaaacagc tatgaccatg attacgccaa gctctagcta gaggtcgacg
     6901 gtatacagac atgataagat acattgatga gtttggacaa accacaacta gaatgcagtg
     6961 aaaaaaatgc tttatttgtg aaatttgtga tgctattgct ttatttgtaa ccattataag
     7021 ctgcaataaa caagttgggg tgggcgaaga actccagcat gagatccccg cgctggagga
     7081 tcatccagcc ggcgtcccgg aaaacgattc cgaagcccaa cctttcatag aaggcggcgg
     7141 tggaatc
//
