LOCUS       sequence-467285-        6833 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  SFFV driven ARRB2-tdMCP with a short linker.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6833)
  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 6833)
  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: APL0030 SFFV-bArrestin2-MCP (FLP-IN)
FEATURES             Location/Qualifiers
     source          1..6833
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(join(145..936,937..1005))
                     /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     768..787
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(1006..1110)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(1185..1204)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        1392..1799
                     /label=SFFV promoter
                     /note="spleen focus-forming virus long terminal repeat 
                     (LTR) promoter"
     regulatory      1807..1816
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             3802..3831
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     primer_bind     complement(3887..3904)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3893..4117
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(4289..4308)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     protein_bind    4440..4487
                     /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             4495..5517
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="KKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRGY
                     VLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLPE
                     TELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQT
                     VMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMFG
                     DSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDGN
                     FDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAKE
                     "
     CDS             4495..5517
                     /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    5647..5768
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5684..5703)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5738..5757
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5817..5833)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5817..5833)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5830..5852)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5841..5857
                     /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(5865..5871,5872..5889,5890..5895))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5910..5931
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(6048..6065)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6219..6807)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6299..6318)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 gtctgacgct cagtggaacg aaaactcacg ttaagggatt ttggtcatga gattatcaaa
       61 aaggatcttc acctagatcc ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat
      121 atatgagtaa acttggtctg acagttacca atgcttaatc agtgaggcac ctatctcagc
      181 gatctgtcta tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga taactacgat
      241 acgggagggc ttaccatctg gccccagtgc tgcaatgata ccgcgagacc cacgctcacc
      301 ggctccagat ttatcagcaa taaaccagcc agccggaagg gccgagcgca gaagtggtcc
      361 tgcaacttta tccgcctcca tccagtctat taattgttgc cgggaagcta gagtaagtag
      421 ttcgccagtt aatagtttgc gcaacgttgt tgccattgct acaggcatcg tggtgtcacg
      481 ctcgtcgttt ggtatggctt cattcagctc cggttcccaa cgatcaaggc gagttacatg
      541 atcccccatg ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg ttgtcagaag
      601 taagttggcc gcagtgttat cactcatggt tatggcagca ctgcataatt ctcttactgt
      661 catgccatcc gtaagatgct tttctgtgac tggtgagtac tcaaccaagt cattctgaga
      721 atagtgtatg cggcgaccga gttgctcttg cccggcgtca atacgggata ataccgcgcc
      781 acatagcaga actttaaaag tgctcatcat tggaaaacgt tcttcggggc gaaaactctc
      841 aaggatctta ccgctgttga gatccagttc gatgtaaccc actcgtgcac ccaactgatc
      901 ttcagcatct tttactttca ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc
      961 cgcaaaaaag ggaataaggg cgacacggaa atgttgaata ctcatactct tcctttttca
     1021 atattattga agcatttatc agggttattg tctcatgagc ggatacatat ttgaatgtat
     1081 ttagaaaaat aaacaaatag gggttccgcg cacatttccc cgaaaagtgc cacctgacgt
     1141 cgacggatcg ggagatctcc cgatccccta tggtgcactc tcagtacaat ctgctctgat
     1201 gccgcatagt taagccagta tctgctccct gcttgtgtgt tggaggtcgc tgagtagtgc
     1261 gcgagcaaaa tttaagctac aacaaggcaa ggcttgaccg acaattgccg ataaaataaa
     1321 agattttatt tagtctccag aaaaaggggg gaatgaaaga ccccacctgt aggtttggca
     1381 agctagctgc agtaacgcca ttttgcaagg catggaaaaa taccaaacca agaatagaga
     1441 agttcagatc aagggcgggt acatgaaaat agctaacgtt gggccaaaca ggatatctgc
     1501 ggtgagcagt ttcggccccg gcccggggcc aagaacagat ggtcaccgca gtttcggccc
     1561 cggcccgagg ccaagaacag atggtcccca gatatggccc aaccctcagc agtttcttaa
     1621 gacccatcag atgtttccag gctcccccaa ggacctgaaa tgaccctgcg ccttatttga
     1681 attaaccaat cagcctgctt ctcgcttctg ttcgcgcgct tctgcttccc gagctctata
     1741 aaagagctca caacccctca ctcggcgcgc cagtcctccg acagactgag tcgcccggga
     1801 agcttcgcca ccatggggga gaaacccggg accagggtct tcaagaagtc gagccctaac
     1861 tgcaagctca ccgtgtactt gggcaagcgg gacttcgtag atcacctgga caaagtggac
     1921 cctgtagatg gcgtggtgct tgtggaccct gactacctga aggaccgcaa agtgtttgtg
     1981 accctcacct gcgccttccg ctatggccgt gaagacctgg atgtgctggg cttgtccttc
     2041 cgcaaagacc tgttcatcgc cacctaccag gccttccccc cggtgcccaa cccaccccgg
     2101 ccccccaccc gcctgcagga ccggctgctg aggaagctgg gccagcatgc ccaccccttc
     2161 ttcttcacca taccccagaa tcttccatgc tccgtcacac tgcagccagg cccagaggat
     2221 acaggaaagg cctgcggcgt agactttgag attcgagcct tctgtgctaa atcactagaa
     2281 gagaaaagcc acaaaaggaa ctctgtgcgg ctggtgatcc gaaaggtgca gttcgccccg
     2341 gagaaacccg gcccccagcc ttcagccgaa accacacgcc acttcctcat gtctgaccgg
     2401 tccctgcacc tcgaggcttc cctggacaag gagctgtact accatgggga gcccctcaat
     2461 gtaaatgtcc acgtcaccaa caactccacc aagaccgtca agaagatcaa agtctctgtg
     2521 agacagtacg ccgacatctg cctcttcagc accgcccagt acaagtgtcc tgtggctcaa
     2581 ctcgaacaag atgaccaggt atctcccagc tccacattct gtaaggtgta caccataacc
     2641 ccactgctca gcgacaaccg ggagaagcgg ggtctcgccc tggatgggaa actcaagcac
     2701 gaggacacca acctggcttc cagcaccatc gtgaaggagg gtgccaacaa ggaggtgctg
     2761 ggaatcctgg tgtcctacag ggtcaaggtg aagctggtgg tgtctcgagg cggggatgtc
     2821 tctgtggagc tgccttttgt tcttatgcac cccaagcccc acgaccacat ccccctcccc
     2881 agaccccagt cagccgctcc ggagacagat gtccctgtgg acaccaacct cattgaattt
     2941 gataccaact atgccacaga tgatgacatt gtgtttgagg actttgcccg gcttcggctg
     3001 aaggggatga aggatgacga ctatgatgat caactctgcg gcggtggtag cgctatggcg
     3061 agcaacttta cgcaatttgt gttggttgac aatggtggca ccggagacgt aactgttgcg
     3121 ccgagcaatt ttgcgaacgg gattgcagaa tggataagta gtaacagccg gagtcaggca
     3181 tataaagtca catgctcagt tcgccaaagt tccgcacaga accgaaaata taccataaag
     3241 gtagaagtgc caaaaggggc gtggcgatct tatcttaata tggagcttac gatcccaatc
     3301 tttgcgacta attcagactg cgagctgatt gtcaaggcca tgcaaggtct tttgaaagac
     3361 ggtaacccga ttccctctgc gatcgccgcc aactcaggaa tatatggtgg cggaaccggt
     3421 ggcgggtcca tggcctccaa ttttacacag tttgtgctcg tggacaacgg cggcactgga
     3481 gacgtcacag tagctccaag taactttgcg aatgggatag ccgagtggat ctcttccaat
     3541 tcaagatcac aagcctataa ggtgacttgt agtgtacgcc aatctagtgc ccagaaccgg
     3601 aagtatacca ttaaagtgga ggtccccaaa ggtgcatggc gctcatactt gaatatggaa
     3661 ttgacgattc cgatatttgc gaccaatagc gactgcgagc ttattgtaaa agccatgcaa
     3721 ggacttctga aagatggcaa cccgatacct tccgcaatcg ctgcgaatag cggaatttac
     3781 tgagccgctc gaggagggcc cgaacaaaaa ctcatctcag aagaggatct gaatagcgcc
     3841 gtcgaccatc atcataatca tcattgagtt taaacccgct gatcagcctc gactgtgcct
     3901 tctagttgcc agccatctgt tgtttgcccc tcccccgtgc cttccttgac cctggaaggt
     3961 gccactccca ctgtcctttc ctaataaaat gaggaaattg catcgcattg tctgagtagg
     4021 tgtcattcta ttctgggggg tggggtgggg caggacagca agggggagga ttgggaagac
     4081 aatagcaggc atgctgggga tgcggtgggc tctatggaga tctatcgatg ttctggggat
     4141 gcggtgggct ctatggcttc tgaggcggaa agaaccagct ggggctctag ggggtatccc
     4201 cacgcgccct gtagcggcgc attaagcgcg gcgggtgtgg tggttacgcg cagcgtgacc
     4261 gctacacttg ccagcgccct agcgcccgct cctttcgctt tcttcccttc ctttctcgcc
     4321 acgttcgccg gctttccccg tcaagctcta aatcgggggc tccctttagg gttccgattt
     4381 agtgctttac ggcacctcga ccccaaaaaa cttgattagg gtgatggttc acgtacctag
     4441 aagttcctat tccgaagttc ctattctcta gaaagtatag gaacttcctt ggccaaaaag
     4501 cctgaactca ccgcgacgtc tgtcgagaag tttctgatcg aaaagttcga cagcgtctcc
     4561 gacctgatgc agctctcgga gggcgaagaa tctcgtgctt tcagcttcga tgtaggaggg
     4621 cgtggatatg tcctgcgggt aaatagctgc gccgatggtt tctacaaaga tcgttatgtt
     4681 tatcggcact ttgcatcggc cgcgctcccg attccggaag tgcttgacat tggggaattc
     4741 agcgagagcc tgacctattg catctcccgc cgtgcacagg gtgtcacgtt gcaagacctg
     4801 cctgaaaccg aactgcccgc tgttctgcag ccggtcgcgg aggccatgga tgcgatcgct
     4861 gcggccgatc ttagccagac gagcgggttc ggcccattcg gaccgcaagg aatcggtcaa
     4921 tacactacat ggcgtgattt catatgcgcg attgctgatc cccatgtgta tcactggcaa
     4981 actgtgatgg acgacaccgt cagtgcgtcc gtcgcgcagg ctctcgatga gctgatgctt
     5041 tgggccgagg actgccccga agtccggcac ctcgtgcacg cggatttcgg ctccaacaat
     5101 gtcctgacgg acaatggccg cataacagcg gtcattgact ggagcgaggc gatgttcggg
     5161 gattcccaat acgaggtcgc caacatcttc ttctggaggc cgtggttggc ttgtatggag
     5221 cagcagacgc gctacttcga gcggaggcat ccggagcttg caggatcgcc gcggctccgg
     5281 gcgtatatgc tccgcattgg tcttgaccaa ctctatcaga gcttggttga cggcaatttc
     5341 gatgatgcag cttgggcgca gggtcgatgc gacgcaatcg tccgatccgg agccgggact
     5401 gtcgggcgta cacaaatcgc ccgcagaagc gcggccgtct ggaccgatgg ctgtgtagaa
     5461 gtactcgccg atagtggaaa ccgacgcccc agcactcgtc cgagggcaaa ggaatagcac
     5521 gtactacgag atttcgattc caccgccgcc ttctatgaaa ggttgggctt cggaatcgtt
     5581 ttccgggacg ccggctggat gatcctccag cgcggggatc tcatgctgga gttcttcgcc
     5641 caccccaact tgtttattgc agcttataat ggttacaaat aaagcaatag catcacaaat
     5701 ttcacaaata aagcattttt ttcactgcat tctagttgtg gtttgtccaa actcatcaat
     5761 gtatcttatc atgtctgtat accgtcgacc tctagctaga gcttggcgta atcatggtca
     5821 tagctgtttc ctgtgtgaaa ttgttatccg ctcacaattc cacacaacat acgagccgga
     5881 agcataaagt gtaaagcctg gggtgcctaa tgagtgagct aactcacatt aattgcgttg
     5941 cgctcactgc ccgctttcca gtcgggaaac ctgtcgtgcc agctgcatta atgaatcggc
     6001 caacgcgcgg ggagaggcgg tttgcgtatt gggcgctctt ccgcttcctc gctcactgac
     6061 tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa ggcggtaata
     6121 cggttatcca cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa
     6181 aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt tccataggct ccgcccccct
     6241 gacgagcatc acaaaaatcg acgctcaagt cagaggtggc gaaacccgac aggactataa
     6301 agataccagg cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg
     6361 cttaccggat acctgtccgc ctttctccct tcgggaagcg tggcgctttc tcatagctca
     6421 cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa
     6481 ccccccgttc agcccgaccg ctgcgcctta tccggtaact atcgtcttga gtccaacccg
     6541 gtaagacacg acttatcgcc actggcagca gccactggta acaggattag cagagcgagg
     6601 tatgtaggcg gtgctacaga gttcttgaag tggtggccta actacggcta cactagaaga
     6661 acagtatttg gtatctgcgc tctgctgaag ccagttacct tcggaaaaag agttggtagc
     6721 tcttgatccg gcaaacaaac caccgctggt agcggtggtt tttttgtttg caagcagcag
     6781 attacgcgca gaaaaaaagg atctcaagaa gatcctttga tcttttctac ggg
//
