LOCUS       sequence-467282-        6644 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  SFFV driven LIDAR reporter with an additional stop codon on the stem
            loop.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6644)
  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 6644)
  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: LSM081
            SFFV-mCherry-SGc(stem-1(3xMS2)2-stops)c...
FEATURES             Location/Qualifiers
     source          1..6644
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(join(172..963,964..1032))
                     /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     795..814
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(1033..1137)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(1212..1231)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        1419..1826
                     /label=SFFV promoter
                     /note="spleen focus-forming virus long terminal repeat 
                     (LTR) promoter"
     regulatory      1834..1843
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1840..2547
                     /codon_start=1
                     /product="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
                     /label=mCherry
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEG
                     TQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNF
                     EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK
                     GEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERA
                     EGRHSTGGMDELYK"
     primer_bind     complement(1985..2003)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     2253..2272
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     CDS             2593..2658
                     /codon_start=1
                     /product="2A peptide from foot-and-mouth disease virus 
                     polyprotein"
                     /label=F2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="VKQTLNFDLLKLAGDVESNPGP"
     CDS             2839..2904
                     /codon_start=1
                     /product="2A peptide from foot-and-mouth disease virus 
                     polyprotein"
                     /label=F2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="VKQTLNFDLLKLAGDVESNPGP"
     CDS             join(2911..2913,2914..3624)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELYK"
     CDS             3640..3669
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     primer_bind     complement(3725..3742)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3731..3955
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(4127..4146)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     protein_bind    4278..4325
                     /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             4333..5355
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="KKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRGY
                     VLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLPE
                     TELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQT
                     VMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMFG
                     DSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDGN
                     FDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAKE
                     "
     CDS             4333..5355
                     /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    5485..5606
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5522..5541)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5576..5595
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5655..5671)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5655..5671)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5668..5690)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5679..5695
                     /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(5703..5709,5710..5727,5728..5733))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5748..5769
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5886..5903)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(join(6057..6644,1))
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6137..6156)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 agaagatcct ttgatctttt ctacggggtc tgacgctcag tggaacgaaa actcacgtta
       61 agggattttg gtcatgagat tatcaaaaag gatcttcacc tagatccttt taaattaaaa
      121 atgaagtttt aaatcaatct aaagtatata tgagtaaact tggtctgaca gttaccaatg
      181 cttaatcagt gaggcaccta tctcagcgat ctgtctattt cgttcatcca tagttgcctg
      241 actccccgtc gtgtagataa ctacgatacg ggagggctta ccatctggcc ccagtgctgc
      301 aatgataccg cgagacccac gctcaccggc tccagattta tcagcaataa accagccagc
      361 cggaagggcc gagcgcagaa gtggtcctgc aactttatcc gcctccatcc agtctattaa
      421 ttgttgccgg gaagctagag taagtagttc gccagttaat agtttgcgca acgttgttgc
      481 cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat tcagctccgg
      541 ttcccaacga tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag cggttagctc
      601 cttcggtcct ccgatcgttg tcagaagtaa gttggccgca gtgttatcac tcatggttat
      661 ggcagcactg cataattctc ttactgtcat gccatccgta agatgctttt ctgtgactgg
      721 tgagtactca accaagtcat tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc
      781 ggcgtcaata cgggataata ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg
      841 aaaacgttct tcggggcgaa aactctcaag gatcttaccg ctgttgagat ccagttcgat
      901 gtaacccact cgtgcaccca actgatcttc agcatctttt actttcacca gcgtttctgg
      961 gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga ataagggcga cacggaaatg
     1021 ttgaatactc atactcttcc tttttcaata ttattgaagc atttatcagg gttattgtct
     1081 catgagcgga tacatatttg aatgtattta gaaaaataaa caaatagggg ttccgcgcac
     1141 atttccccga aaagtgccac ctgacgtcga cggatcggga gatctcccga tcccctatgg
     1201 tgcactctca gtacaatctg ctctgatgcc gcatagttaa gccagtatct gctccctgct
     1261 tgtgtgttgg aggtcgctga gtagtgcgcg agcaaaattt aagctacaac aaggcaaggc
     1321 ttgaccgaca attgccgata aaataaaaga ttttatttag tctccagaaa aaggggggaa
     1381 tgaaagaccc cacctgtagg tttggcaagc tagctgcagt aacgccattt tgcaaggcat
     1441 ggaaaaatac caaaccaaga atagagaagt tcagatcaag ggcgggtaca tgaaaatagc
     1501 taacgttggg ccaaacagga tatctgcggt gagcagtttc ggccccggcc cggggccaag
     1561 aacagatggt caccgcagtt tcggccccgg cccgaggcca agaacagatg gtccccagat
     1621 atggcccaac cctcagcagt ttcttaagac ccatcagatg tttccaggct cccccaagga
     1681 cctgaaatga ccctgcgcct tatttgaatt aaccaatcag cctgcttctc gcttctgttc
     1741 gcgcgcttct gcttcccgag ctctataaaa gagctcacaa cccctcactc ggcgcgccag
     1801 tcctccgaca gactgagtcg cccgggaagc ttcgccacca tggtgagcaa gggcgaggag
     1861 gataacatgg ccatcatcaa ggagttcatg cgcttcaagg tgcacatgga gggctccgtg
     1921 aacggccacg agttcgagat cgagggcgag ggcgagggcc gcccctacga gggcacccag
     1981 accgccaagc tgaaggtgac caagggtggc cccctgccct tcgcctggga catcctgtcc
     2041 cctcagttca tgtacggctc caaggcctac gtgaagcacc ccgccgacat tcccgactac
     2101 ttgaagctgt ccttccccga gggcttcaag tgggagcgcg tgatgaactt cgaggacggc
     2161 ggcgtggtga ccgtgaccca ggactcctcc ctgcaggacg gcgagttcat ctacaaggtg
     2221 aagctgcgcg gcaccaactt cccctccgac ggccccgtaa tgcagaagaa gaccatgggc
     2281 tgggaggcct cctccgagcg gatgtacccc gaggacggcg ccctgaaggg cgagatcaag
     2341 cagaggctga agctgaagga cggcggccac tacgacgctg aggtcaagac cacctacaag
     2401 gccaagaagc ccgtgcagct gcccggcgcc tacaacgtca acatcaagtt ggacatcacc
     2461 tcccacaacg aggactacac catcgtggaa cagtacgaac gcgccgaggg ccgccactcc
     2521 accggcggca tggacgagct gtacaagagc gacgcgttgg aagccagaca caaacagaaa
     2581 attgtggctc cggtgaaaca gactttgaat tttgaccttc tgaagctggc gggagacgtc
     2641 gagtccaacc ccggtcccgg taccggatcc tccgtttagg tttaggtggg tggaatagta
     2701 acgtacacca tcagggtacg tctcagacac catcagggtc tgtctggtac agcatcagcg
     2761 taccatagta tcccacctac ccaaacccaa acgaccggta ctagtgaggc acggcataag
     2821 caaaagatcg tagccccagt aaagcaaaca ctcaacttcg atctactcaa actcgcaggt
     2881 gatgtggaat ctaatccagg accctcgagt gtctcaaagg gcgaggaact ttttacgggc
     2941 gttgttccga tcctcgtcga attggacggg gacgttaatg ggcataaatt ctcagttagt
     3001 ggtgagggtg agggggacgc gacatatggg aaacttacgc tgaagttcat atgtaccacc
     3061 ggaaaattgc cagtgccgtg gcctactctc gtcaccaccc tgacatatgg tgttcagtgt
     3121 tttagccggt atccagacca tatgaaacag cacgattttt ttaaatccgc tatgcctgaa
     3181 ggttacgtac aggagcgaac gatatttttt aaagacgacg gaaactacaa aactagagcc
     3241 gaggtaaaat tcgagggtga cactctcgtg aaccggatcg aactgaaggg catagatttt
     3301 aaggaggatg gcaatatact gggccacaag ctcgaataca actataattc tcataatgtc
     3361 tatatcatgg cggacaaaca gaaaaacggc atcaaggtaa actttaaaat ccgacacaat
     3421 attgaggacg gctctgtcca gcttgcggac cattaccaac agaacacgcc gataggggat
     3481 ggtccagtac ttctccctga taatcactat ttgtctactc agtcagctct tagcaaggac
     3541 ccaaacgaaa aacgggatca catggtcctt cttgagttcg tcactgcggc tgggataaca
     3601 ctcgggatgg acgaactgta taagtaggcc ggagggcccg aacaaaaact catctcagaa
     3661 gaggatctga atagcgccgt cgaccatcat cataatcatc attgagttta aacccgctga
     3721 tcagcctcga ctgtgccttc tagttgccag ccatctgttg tttgcccctc ccccgtgcct
     3781 tccttgaccc tggaaggtgc cactcccact gtcctttcct aataaaatga ggaaattgca
     3841 tcgcattgtc tgagtaggtg tcattctatt ctggggggtg gggtggggca ggacagcaag
     3901 ggggaggatt gggaagacaa tagcaggcat gctggggatg cggtgggctc tatggagatc
     3961 tatcgatgtt ctggggatgc ggtgggctct atggcttctg aggcggaaag aaccagctgg
     4021 ggctctaggg ggtatcccca cgcgccctgt agcggcgcat taagcgcggc gggtgtggtg
     4081 gttacgcgca gcgtgaccgc tacacttgcc agcgccctag cgcccgctcc tttcgctttc
     4141 ttcccttcct ttctcgccac gttcgccggc tttccccgtc aagctctaaa tcgggggctc
     4201 cctttagggt tccgatttag tgctttacgg cacctcgacc ccaaaaaact tgattagggt
     4261 gatggttcac gtacctagaa gttcctattc cgaagttcct attctctaga aagtatagga
     4321 acttccttgg ccaaaaagcc tgaactcacc gcgacgtctg tcgagaagtt tctgatcgaa
     4381 aagttcgaca gcgtctccga cctgatgcag ctctcggagg gcgaagaatc tcgtgctttc
     4441 agcttcgatg taggagggcg tggatatgtc ctgcgggtaa atagctgcgc cgatggtttc
     4501 tacaaagatc gttatgttta tcggcacttt gcatcggccg cgctcccgat tccggaagtg
     4561 cttgacattg gggaattcag cgagagcctg acctattgca tctcccgccg tgcacagggt
     4621 gtcacgttgc aagacctgcc tgaaaccgaa ctgcccgctg ttctgcagcc ggtcgcggag
     4681 gccatggatg cgatcgctgc ggccgatctt agccagacga gcgggttcgg cccattcgga
     4741 ccgcaaggaa tcggtcaata cactacatgg cgtgatttca tatgcgcgat tgctgatccc
     4801 catgtgtatc actggcaaac tgtgatggac gacaccgtca gtgcgtccgt cgcgcaggct
     4861 ctcgatgagc tgatgctttg ggccgaggac tgccccgaag tccggcacct cgtgcacgcg
     4921 gatttcggct ccaacaatgt cctgacggac aatggccgca taacagcggt cattgactgg
     4981 agcgaggcga tgttcgggga ttcccaatac gaggtcgcca acatcttctt ctggaggccg
     5041 tggttggctt gtatggagca gcagacgcgc tacttcgagc ggaggcatcc ggagcttgca
     5101 ggatcgccgc ggctccgggc gtatatgctc cgcattggtc ttgaccaact ctatcagagc
     5161 ttggttgacg gcaatttcga tgatgcagct tgggcgcagg gtcgatgcga cgcaatcgtc
     5221 cgatccggag ccgggactgt cgggcgtaca caaatcgccc gcagaagcgc ggccgtctgg
     5281 accgatggct gtgtagaagt actcgccgat agtggaaacc gacgccccag cactcgtccg
     5341 agggcaaagg aatagcacgt actacgagat ttcgattcca ccgccgcctt ctatgaaagg
     5401 ttgggcttcg gaatcgtttt ccgggacgcc ggctggatga tcctccagcg cggggatctc
     5461 atgctggagt tcttcgccca ccccaacttg tttattgcag cttataatgg ttacaaataa
     5521 agcaatagca tcacaaattt cacaaataaa gcattttttt cactgcattc tagttgtggt
     5581 ttgtccaaac tcatcaatgt atcttatcat gtctgtatac cgtcgacctc tagctagagc
     5641 ttggcgtaat catggtcata gctgtttcct gtgtgaaatt gttatccgct cacaattcca
     5701 cacaacatac gagccggaag cataaagtgt aaagcctggg gtgcctaatg agtgagctaa
     5761 ctcacattaa ttgcgttgcg ctcactgccc gctttccagt cgggaaacct gtcgtgccag
     5821 ctgcattaat gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg gcgctcttcc
     5881 gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct
     5941 cactcaaagg cggtaatacg gttatccaca gaatcagggg ataacgcagg aaagaacatg
     6001 tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc
     6061 cataggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagtca gaggtggcga
     6121 aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct cgtgcgctct
     6181 cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc gggaagcgtg
     6241 gcgctttctc atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag
     6301 ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc cggtaactat
     6361 cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc cactggtaac
     6421 aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg gtggcctaac
     6481 tacggctaca ctagaagaac agtatttggt atctgcgctc tgctgaagcc agttaccttc
     6541 ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag cggtggtttt
     6601 tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ctca
//
