LOCUS       sequence-466403-        7619 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  IVT template for ARRB2-tdMCP.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7619)
  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 7619)
  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: XZ089 (IVT) CMV-TO-T7-ARRB2-MCP-hybridUTR3
FEATURES             Location/Qualifiers
     source          1..7619
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    39..60
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(177..194)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(348..936)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(428..447)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(1107..1898,1899..1967))
                     /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     1730..1749
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(1968..2072)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(2147..2166)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        2333..2712
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        2713..2916
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     2867..2887
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     protein_bind    2918..2936
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    2939..2957
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     primer_bind     2967..2991
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     primer_bind     3101..3120
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        3101..3119
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      3272..3281
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             5550..5579
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     primer_bind     complement(5635..5652)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    5641..5865
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(6037..6056)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     protein_bind    6188..6235
                     /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             6243..7265
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="KKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRGY
                     VLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLPE
                     TELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQT
                     VMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMFG
                     DSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDGN
                     FDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAKE
                     "
     CDS             6243..7265
                     /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    7395..7516
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(7432..7451)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     7486..7505
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(7565..7581)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(7565..7581)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(7578..7600)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    7589..7605
                     /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(7613..7619,1..18,19..24))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
ORIGIN
        1 cgagccggaa gcataaagtg taaagcctgg ggtgcctaat gagtgagcta actcacatta
       61 attgcgttgc gctcactgcc cgctttccag tcgggaaacc tgtcgtgcca gctgcattaa
      121 tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg ggcgctcttc cgcttcctcg
      181 ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag
      241 gcggtaatac ggttatccac agaatcaggg gataacgcag gaaagaacat gtgagcaaaa
      301 ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc
      361 cgcccccctg acgagcatca caaaaatcga cgctcaagtc agaggtggcg aaacccgaca
      421 ggactataaa gataccaggc gtttccccct ggaagctccc tcgtgcgctc tcctgttccg
      481 accctgccgc ttaccggata cctgtccgcc tttctccctt cgggaagcgt ggcgctttct
      541 catagctcac gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt
      601 gtgcacgaac cccccgttca gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag
      661 tccaacccgg taagacacga cttatcgcca ctggcagcag ccactggtaa caggattagc
      721 agagcgaggt atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa ctacggctac
      781 actagaagaa cagtatttgg tatctgcgct ctgctgaagc cagttacctt cggaaaaaga
      841 gttggtagct cttgatccgg caaacaaacc accgctggta gcggtggttt ttttgtttgc
      901 aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag atcctttgat cttttctacg
      961 gggtctgacg ctcagtggaa cgaaaactca cgttaaggga ttttggtcat gagattatca
     1021 aaaaggatct tcacctagat ccttttaaat taaaaatgaa gttttaaatc aatctaaagt
     1081 atatatgagt aaacttggtc tgacagttac caatgcttaa tcagtgaggc acctatctca
     1141 gcgatctgtc tatttcgttc atccatagtt gcctgactcc ccgtcgtgta gataactacg
     1201 atacgggagg gcttaccatc tggccccagt gctgcaatga taccgcgaga cccacgctca
     1261 ccggctccag atttatcagc aataaaccag ccagccggaa gggccgagcg cagaagtggt
     1321 cctgcaactt tatccgcctc catccagtct attaattgtt gccgggaagc tagagtaagt
     1381 agttcgccag ttaatagttt gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca
     1441 cgctcgtcgt ttggtatggc ttcattcagc tccggttccc aacgatcaag gcgagttaca
     1501 tgatccccca tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat cgttgtcaga
     1561 agtaagttgg ccgcagtgtt atcactcatg gttatggcag cactgcataa ttctcttact
     1621 gtcatgccat ccgtaagatg cttttctgtg actggtgagt actcaaccaa gtcattctga
     1681 gaatagtgta tgcggcgacc gagttgctct tgcccggcgt caatacggga taataccgcg
     1741 ccacatagca gaactttaaa agtgctcatc attggaaaac gttcttcggg gcgaaaactc
     1801 tcaaggatct taccgctgtt gagatccagt tcgatgtaac ccactcgtgc acccaactga
     1861 tcttcagcat cttttacttt caccagcgtt tctgggtgag caaaaacagg aaggcaaaat
     1921 gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa tactcatact cttccttttt
     1981 caatattatt gaagcattta tcagggttat tgtctcatga gcggatacat atttgaatgt
     2041 atttagaaaa ataaacaaat aggggttccg cgcacatttc cccgaaaagt gccacctgac
     2101 gtcgacggat cgggagatct cccgatcccc tatggtgcac tctcagtaca atctgctctg
     2161 atgccgcata gttaagccag tatctgctcc ctgcttgtgt gttggaggtc gctgagtagt
     2221 gcgcgagcaa aatttaagct acaacaaggc aaggcttgac cgacaattgc atgaagaatc
     2281 tgcttagggt taggcgtttt gcgctgcttc gtacgggcca gatatacgcg ttgacattga
     2341 ttattgacta gttattaata gtaatcaatt acggggtcat tagttcatag cccatatatg
     2401 gagttccgcg ttacataact tacggtaaat ggcccgcctg gctgaccgcc caacgacccc
     2461 cgcccattga cgtcaataat gacgtatgtt cccatagtaa cgccaatagg gactttccat
     2521 tgacgtcaat gggtggagta tttacggtaa actgcccact tggcagtaca tcaagtgtat
     2581 catatgccaa gtacgccccc tattgacgtc aatgacggta aatggcccgc ctggcattat
     2641 gcccagtaca tgaccttatg ggactttcct acttggcagt acatctacgt attagtcatc
     2701 gctattacca tggtgatgcg gttttggcag tacatcaatg ggcgtggata gcggtttgac
     2761 tcacggggat ttccaagtct ccaccccatt gacgtcaatg ggagtttgtt ttggcaccaa
     2821 aatcaacggg actttccaaa atgtcgtaac aactccgccc cattgacgca aatgggcggt
     2881 aggcgtgtac ggtgggaggt ctatataagc agagctctcc ctatcagtga tagagatctc
     2941 cctatcagtg atagagatcg tcgacgagct cgtttagtga accgtcagat cgcctggaga
     3001 cgccatccac gctgttttga cctccataga agacaccggg accgatccag cctccggact
     3061 ctagcgttta aacttaagct cgctagagat ccgcggccgc taatacgact cactataggg
     3121 agaaaaataa caaatctcaa cacaacatat acaaaacaaa cgaatctcaa gcaatcaagc
     3181 attctacttc tattgcagca atttaaatca tttcttttaa agcaaaagca attttctgaa
     3241 aattttcacc atttacgaac gataaagctt cgccaccatg ggggagaaac ccgggaccag
     3301 ggtcttcaag aagtcgagcc ctaactgcaa gctcaccgtg tacttgggca agcgggactt
     3361 cgtagatcac ctggacaaag tggaccctgt agatggcgtg gtgcttgtgg accctgacta
     3421 cctgaaggac cgcaaagtgt ttgtgaccct cacctgcgcc ttccgctatg gccgtgaaga
     3481 cctggatgtg ctgggcttgt ccttccgcaa agacctgttc atcgccacct accaggcctt
     3541 ccccccggtg cccaacccac cccggccccc cacccgcctg caggaccggc tgctgaggaa
     3601 gctgggccag catgcccacc ccttcttctt caccataccc cagaatcttc catgctccgt
     3661 cacactgcag ccaggcccag aggatacagg aaaggcctgc ggcgtagact ttgagattcg
     3721 agccttctgt gctaaatcac tagaagagaa aagccacaaa aggaactctg tgcggctggt
     3781 gatccgaaag gtgcagttcg ccccggagaa acccggcccc cagccttcag ccgaaaccac
     3841 acgccacttc ctcatgtctg accggtccct gcacctcgag gcttccctgg acaaggagct
     3901 gtactaccat ggggagcccc tcaatgtaaa tgtccacgtc accaacaact ccaccaagac
     3961 cgtcaagaag atcaaagtct ctgtgagaca gtacgccgac atctgcctct tcagcaccgc
     4021 ccagtacaag tgtcctgtgg ctcaactcga acaagatgac caggtatctc ccagctccac
     4081 attctgtaag gtgtacacca taaccccact gctcagcgac aaccgggaga agcggggtct
     4141 cgccctggat gggaaactca agcacgagga caccaacctg gcttccagca ccatcgtgaa
     4201 ggagggtgcc aacaaggagg tgctgggaat cctggtgtcc tacagggtca aggtgaagct
     4261 ggtggtgtct cgaggcgggg atgtctctgt ggagctgcct tttgttctta tgcaccccaa
     4321 gccccacgac cacatccccc tccccagacc ccagtcagcc gctccggaga cagatgtccc
     4381 tgtggacacc aacctcattg aatttgatac caactatgcc acagatgatg acattgtgtt
     4441 tgaggacttt gcccggcttc ggctgaaggg gatgaaggat gacgactatg atgatcaact
     4501 ctgcggcggt ggtagcgcta tggcgagcaa ctttacgcaa tttgtgttgg ttgacaatgg
     4561 tggcaccgga gacgtaactg ttgcgccgag caattttgcg aacgggattg cagaatggat
     4621 aagtagtaac agccggagtc aggcatataa agtcacatgc tcagttcgcc aaagttccgc
     4681 acagaaccga aaatatacca taaaggtaga agtgccaaaa ggggcgtggc gatcttatct
     4741 taatatggag cttacgatcc caatctttgc gactaattca gactgcgagc tgattgtcaa
     4801 ggccatgcaa ggtcttttga aagacggtaa cccgattccc tctgcgatcg ccgccaactc
     4861 aggaatatat ggtggcggaa ccggtggcgg gtccatggcc tccaatttta cacagtttgt
     4921 gctcgtggac aacggcggca ctggagacgt cacagtagct ccaagtaact ttgcgaatgg
     4981 gatagccgag tggatctctt ccaattcaag atcacaagcc tataaggtga cttgtagtgt
     5041 acgccaatct agtgcccaga accggaagta taccattaaa gtggaggtcc ccaaaggtgc
     5101 atggcgctca tacttgaata tggaattgac gattccgata tttgcgacca atagcgactg
     5161 cgagcttatt gtaaaagcca tgcaaggact tctgaaagat ggcaacccga taccttccgc
     5221 aatcgctgcg aatagcggaa tttactgact cgagctggta ctgcatgcac gcaatgctag
     5281 ctgccccttt cccgtcctgg gtaccccgag tctcccccga cctcgggtcc caggtatgct
     5341 cccacctcca cctgccccac tcaccacctc tgctagttcc agacacctcc caagcacgca
     5401 gcaatgcagc tcaaaacgct tagcctagcc acacccccac gggaaacagc agtgattaac
     5461 ctttagcaat aaacgaaagt ttaactaagc tatactaacc ccagggttgg tcaatttcgt
     5521 gccagccaca ccctggagct agcgggcccg aacaaaaact catctcagaa gaggatctga
     5581 atagcgccgt cgaccatcat cataatcatc attgagttta aacccgctga tcagcctcga
     5641 ctgtgccttc tagttgccag ccatctgttg tttgcccctc ccccgtgcct tccttgaccc
     5701 tggaaggtgc cactcccact gtcctttcct aataaaatga ggaaattgca tcgcattgtc
     5761 tgagtaggtg tcattctatt ctggggggtg gggtggggca ggacagcaag ggggaggatt
     5821 gggaagacaa tagcaggcat gctggggatg cggtgggctc tatggagatc tatcgatgtt
     5881 ctggggatgc ggtgggctct atggcttctg aggcggaaag aaccagctgg ggctctaggg
     5941 ggtatcccca cgcgccctgt agcggcgcat taagcgcggc gggtgtggtg gttacgcgca
     6001 gcgtgaccgc tacacttgcc agcgccctag cgcccgctcc tttcgctttc ttcccttcct
     6061 ttctcgccac gttcgccggc tttccccgtc aagctctaaa tcgggggctc cctttagggt
     6121 tccgatttag tgctttacgg cacctcgacc ccaaaaaact tgattagggt gatggttcac
     6181 gtacctagaa gttcctattc cgaagttcct attctctaga aagtatagga acttccttgg
     6241 ccaaaaagcc tgaactcacc gcgacgtctg tcgagaagtt tctgatcgaa aagttcgaca
     6301 gcgtctccga cctgatgcag ctctcggagg gcgaagaatc tcgtgctttc agcttcgatg
     6361 taggagggcg tggatatgtc ctgcgggtaa atagctgcgc cgatggtttc tacaaagatc
     6421 gttatgttta tcggcacttt gcatcggccg cgctcccgat tccggaagtg cttgacattg
     6481 gggaattcag cgagagcctg acctattgca tctcccgccg tgcacagggt gtcacgttgc
     6541 aagacctgcc tgaaaccgaa ctgcccgctg ttctgcagcc ggtcgcggag gccatggatg
     6601 cgatcgctgc ggccgatctt agccagacga gcgggttcgg cccattcgga ccgcaaggaa
     6661 tcggtcaata cactacatgg cgtgatttca tatgcgcgat tgctgatccc catgtgtatc
     6721 actggcaaac tgtgatggac gacaccgtca gtgcgtccgt cgcgcaggct ctcgatgagc
     6781 tgatgctttg ggccgaggac tgccccgaag tccggcacct cgtgcacgcg gatttcggct
     6841 ccaacaatgt cctgacggac aatggccgca taacagcggt cattgactgg agcgaggcga
     6901 tgttcgggga ttcccaatac gaggtcgcca acatcttctt ctggaggccg tggttggctt
     6961 gtatggagca gcagacgcgc tacttcgagc ggaggcatcc ggagcttgca ggatcgccgc
     7021 ggctccgggc gtatatgctc cgcattggtc ttgaccaact ctatcagagc ttggttgacg
     7081 gcaatttcga tgatgcagct tgggcgcagg gtcgatgcga cgcaatcgtc cgatccggag
     7141 ccgggactgt cgggcgtaca caaatcgccc gcagaagcgc ggccgtctgg accgatggct
     7201 gtgtagaagt actcgccgat agtggaaacc gacgccccag cactcgtccg agggcaaagg
     7261 aatagcacgt actacgagat ttcgattcca ccgccgcctt ctatgaaagg ttgggcttcg
     7321 gaatcgtttt ccgggacgcc ggctggatga tcctccagcg cggggatctc atgctggagt
     7381 tcttcgccca ccccaacttg tttattgcag cttataatgg ttacaaataa agcaatagca
     7441 tcacaaattt cacaaataaa gcattttttt cactgcattc tagttgtggt ttgtccaaac
     7501 tcatcaatgt atcttatcat gtctgtatac cgtcgacctc tagctagagc ttggcgtaat
     7561 catggtcata gctgtttcct gtgtgaaatt gttatccgct cacaattcca cacaacata
//
