LOCUS       sequence-467288-        7316 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  LIDAR reporter RNA; secreted NanoLuc Luciferase as marker, SEAP as 
            output.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7316)
  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 7316)
  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: XZ392
            SFFV-secNLuc-SGc(stem-1(3xMS2))cSG(tight...
FEATURES             Location/Qualifiers
     source          1..7316
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        176..583
                     /label=SFFV promoter
                     /note="spleen focus-forming virus long terminal repeat 
                     (LTR) promoter"
     CDS             684..1193
                     /codon_start=1
                     /product="NanoLuc(R) luciferase"
                     /label=Nluc
                     /note="human codon-optimized"
                     /translation="VFTLEDFVGDWRQTAGYNLDQVLEQGGVSSLFQNLGVSVTPIQRI
                     VLSGENGLKIDIHVIIPYEGLSGDQMGQIEKIFKVVYPVDDHHFKVILHYGTLVIDGVT
                     PNMIDYFGRPYEGIAVFDGKKITVTGTLWNGNKIIDERLINPDGSLLFRVTINGVTGWR
                     LCERILA"
     CDS             1236..1301
                     /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             1470..1535
                     /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             1536..3038
                     /codon_start=1
                     /product="secreted alkaline phosphatase from human 
                     placenta"
                     /label=SEAP
                     /note="The natural enzyme has a C-terminal transmembrane 
                     sequence that is replaced by a GPI anchor."
                     /translation="MLLLLLLLGLRLQLSLGIIPVEEENPDFWNREAAEALGAAKKLQP
                     AQTAAKNLIIFLGDGMGVSTVTAARILKGQKKDKLGPEIPLAMDRFPYVALSKTYNVDK
                     HVPDSGATATAYLCGVKGNFQTIGLSAAARFNQCNTTRGNEVISVMNRAKKAGKSVGVV
                     TTTRVQHASPAGTYAHTVNRNWYSDADVPASARQEGCQDIATQLISNMDIDVILGGGRK
                     YMFPMGTPDPEYPDDYSQGGTRLDGKNLVQEWLAKRQGARYVWNRTELMQASLDPSVTH
                     LMGLFEPGDMKYEIHRDSTLDPSLMEMTEAALRLLSRNPRGFFLFVEGGRIDHGHHESR
                     AYRALTETIMFDDAIERAGQLTSEEDTLSLVTADHSHVFSFGGYPLRGSSIFGLAPGKA
                     RDRKAYTVLLYGNGPGYVLKDGARPDVTESESGSPEYRQQSAVPLDEETHAGEDVAVFA
                     RGPQAHLVHGVQEQTFIAHVMAFAACLEPYTACDLAPPAGTTD"
     CDS             3069..3098
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     primer_bind     complement(3154..3171)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3160..3384
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(3556..3575)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     protein_bind    3707..3754
                     /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             3762..4784
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="KKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRGY
                     VLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLPE
                     TELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQT
                     VMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMFG
                     DSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDGN
                     FDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAKE
                     "
     CDS             3762..4784
                     /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    4914..5035
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4951..4970)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5005..5024
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5084..5100)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5084..5100)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5097..5119)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5108..5124
                     /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(5132..5138,5139..5156,5157..5162))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5177..5198
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5315..5332)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5486..6074)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5566..5585)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6245..7036,7037..7105))
                     /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     6868..6887
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7106..7210)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(7285..7304)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 agtatctgct ccctgcttgt gtgttggagg tcgctgagta gtgcgcgagc aaaatttaag
       61 ctacaacaag gcaaggcttg accgacaatt gccgataaaa taaaagattt tatttagtct
      121 ccagaaaaag gggggaatga aagaccccac ctgtaggttt ggcaagctag ctgcagtaac
      181 gccattttgc aaggcatgga aaaataccaa accaagaata gagaagttca gatcaagggc
      241 gggtacatga aaatagctaa cgttgggcca aacaggatat ctgcggtgag cagtttcggc
      301 cccggcccgg ggccaagaac agatggtcac cgcagtttcg gccccggccc gaggccaaga
      361 acagatggtc cccagatatg gcccaaccct cagcagtttc ttaagaccca tcagatgttt
      421 ccaggctccc ccaaggacct gaaatgaccc tgcgccttat ttgaattaac caatcagcct
      481 gcttctcgct tctgttcgcg cgcttctgct tcccgagctc tataaaagag ctcacaaccc
      541 ctcactcggc gcgccagtcc tccgacagac tgagtcgccc gggaagcttc gccaccatga
      601 actccttctc cacaagcgcc ttcggtccag ttgccttctc cctgggcctg ctcctggtgt
      661 tgcctgctgc cttccctgcc ccagtcttca cactcgaaga tttcgttggg gactggcgac
      721 agacagccgg ctacaacctg gaccaagtcc ttgaacaggg aggtgtgtcc agtttgtttc
      781 agaatctcgg ggtgtccgta actccgatcc aaaggattgt cctgagcggt gaaaatgggc
      841 tgaagatcga catccatgtc atcatcccgt atgaaggtct gagcggcgac caaatgggcc
      901 agatcgaaaa aatttttaag gtggtgtacc ctgtggatga tcatcacttt aaggtgatcc
      961 tgcactatgg cacactggta atcgacgggg ttacgccgaa catgatcgac tatttcggac
     1021 ggccgtatga aggcatcgcc gtgttcgacg gcaaaaagat cactgtaaca gggaccctgt
     1081 ggaacggcaa caaaattatc gacgagcgcc tgatcaaccc cgacggctcc ctgctgttcc
     1141 gagtaaccat caacggagtg accggctggc ggctgtgcga acgcattctg gcggacgcgt
     1201 tggaagccag acacaaacag aaaattgtgg ctccggtgaa acagactttg aattttgacc
     1261 ttctgaagct ggcgggagac gtcgagtcca accccggtcc cggtaccgga tcctccgttt
     1321 aggtgggtgg aatagtaacg tacaccatca gggtacgtct cagacaccat cagggtctgt
     1381 ctggtacagc atcagcgtac catagtatcc cacctaccca gacgaccggt actagtgagg
     1441 cacggcataa gcaaaagatc gtagccccag taaagcaaac actcaacttc gatctactca
     1501 aactcgcagg tgatgtggaa tctaatccag gacccatgct gctcctgttg ctgctgctgg
     1561 gcctgaggct acagctctcc ctgggcatca tcccagttga ggaggagaac ccggacttct
     1621 ggaaccgcga ggcagccgag gccctgggtg ccgccaagaa gctgcagcct gcacagacag
     1681 ccgccaagaa cctcatcatc ttcctgggcg atgggatggg ggtgtctacg gtgacagctg
     1741 ccaggattct aaaagggcag aagaaggaca aactggggcc tgagataccc ctggccatgg
     1801 accgcttccc atatgtggct ctgtccaaga catacaatgt agacaaacat gtgccagaca
     1861 gtggagccac agccacggcc tacctgtgcg gggtcaaggg caacttccag accattggct
     1921 tgagtgcagc cgcccgcttt aaccagtgca acacgacacg cggcaacgag gtcatctccg
     1981 tgatgaatcg ggccaagaaa gcagggaagt cagtgggagt ggtaaccacc acacgagtgc
     2041 agcacgcctc gccagccggc acctacgccc acacggtgaa ccgcaactgg tactcggacg
     2101 ccgacgtgcc tgcctcggcc cgccaggagg ggtgccagga catcgctacg cagctcatct
     2161 ccaacatgga cattgacgtg atcctaggtg gaggccgaaa atacatgttt cccatgggaa
     2221 ccccagaccc tgagtaccca gatgactaca gccaaggtgg gaccaggctg gacgggaaga
     2281 atctggtgca ggaatggctg gcgaagcgcc agggtgcccg gtatgtgtgg aaccgcactg
     2341 agctcatgca ggcttccctg gacccgtctg tgacccatct catgggtctc tttgagcctg
     2401 gagacatgaa atacgagatc caccgagact ccacactgga cccctccctg atggagatga
     2461 cagaggctgc cctgcgcctg ctgagcagga acccccgcgg cttcttcctc ttcgtggagg
     2521 gtggtcgcat cgaccatggt catcatgaaa gcagggctta ccgggcactg actgagacga
     2581 tcatgttcga cgacgccatt gagagggcgg gccagctcac cagcgaggag gacacgctga
     2641 gcctcgtcac tgccgaccac tcccacgtct tctccttcgg aggctacccc ctgcgaggga
     2701 gctccatctt cgggctggcc cctggcaagg cccgggacag gaaggcctac acggtcctcc
     2761 tatacggaaa cggtccaggc tatgtgctca aggacggcgc tcggccggat gttaccgaga
     2821 gcgagagcgg gagccccgag tatcggcagc agtcagcagt gcccctggac gaagagaccc
     2881 acgcaggcga ggacgtggcg gtgttcgcgc gcggcccgca ggcgcacctg gttcacggcg
     2941 tgcaggagca gaccttcata gcgcacgtca tggccttcgc cgcctgcctg gagccctaca
     3001 ccgcctgcga cctggcaccc cccgccggca ccaccgacgc cgcgcacccg ggttaggccg
     3061 gagggcccga acaaaaactc atctcagaag aggatctgaa tagcgccgtc gaccatcatc
     3121 ataatcatca ttgagtttaa acccgctgat cagcctcgac tgtgccttct agttgccagc
     3181 catctgttgt ttgcccctcc cccgtgcctt ccttgaccct ggaaggtgcc actcccactg
     3241 tcctttccta ataaaatgag gaaattgcat cgcattgtct gagtaggtgt cattctattc
     3301 tggggggtgg ggtggggcag gacagcaagg gggaggattg ggaagacaat agcaggcatg
     3361 ctggggatgc ggtgggctct atggagatct atcgatgttc tggggatgcg gtgggctcta
     3421 tggcttctga ggcggaaaga accagctggg gctctagggg gtatccccac gcgccctgta
     3481 gcggcgcatt aagcgcggcg ggtgtggtgg ttacgcgcag cgtgaccgct acacttgcca
     3541 gcgccctagc gcccgctcct ttcgctttct tcccttcctt tctcgccacg ttcgccggct
     3601 ttccccgtca agctctaaat cgggggctcc ctttagggtt ccgatttagt gctttacggc
     3661 acctcgaccc caaaaaactt gattagggtg atggttcacg tacctagaag ttcctattcc
     3721 gaagttccta ttctctagaa agtataggaa cttccttggc caaaaagcct gaactcaccg
     3781 cgacgtctgt cgagaagttt ctgatcgaaa agttcgacag cgtctccgac ctgatgcagc
     3841 tctcggaggg cgaagaatct cgtgctttca gcttcgatgt aggagggcgt ggatatgtcc
     3901 tgcgggtaaa tagctgcgcc gatggtttct acaaagatcg ttatgtttat cggcactttg
     3961 catcggccgc gctcccgatt ccggaagtgc ttgacattgg ggaattcagc gagagcctga
     4021 cctattgcat ctcccgccgt gcacagggtg tcacgttgca agacctgcct gaaaccgaac
     4081 tgcccgctgt tctgcagccg gtcgcggagg ccatggatgc gatcgctgcg gccgatctta
     4141 gccagacgag cgggttcggc ccattcggac cgcaaggaat cggtcaatac actacatggc
     4201 gtgatttcat atgcgcgatt gctgatcccc atgtgtatca ctggcaaact gtgatggacg
     4261 acaccgtcag tgcgtccgtc gcgcaggctc tcgatgagct gatgctttgg gccgaggact
     4321 gccccgaagt ccggcacctc gtgcacgcgg atttcggctc caacaatgtc ctgacggaca
     4381 atggccgcat aacagcggtc attgactgga gcgaggcgat gttcggggat tcccaatacg
     4441 aggtcgccaa catcttcttc tggaggccgt ggttggcttg tatggagcag cagacgcgct
     4501 acttcgagcg gaggcatccg gagcttgcag gatcgccgcg gctccgggcg tatatgctcc
     4561 gcattggtct tgaccaactc tatcagagct tggttgacgg caatttcgat gatgcagctt
     4621 gggcgcaggg tcgatgcgac gcaatcgtcc gatccggagc cgggactgtc gggcgtacac
     4681 aaatcgcccg cagaagcgcg gccgtctgga ccgatggctg tgtagaagta ctcgccgata
     4741 gtggaaaccg acgccccagc actcgtccga gggcaaagga atagcacgta ctacgagatt
     4801 tcgattccac cgccgccttc tatgaaaggt tgggcttcgg aatcgttttc cgggacgccg
     4861 gctggatgat cctccagcgc ggggatctca tgctggagtt cttcgcccac cccaacttgt
     4921 ttattgcagc ttataatggt tacaaataaa gcaatagcat cacaaatttc acaaataaag
     4981 catttttttc actgcattct agttgtggtt tgtccaaact catcaatgta tcttatcatg
     5041 tctgtatacc gtcgacctct agctagagct tggcgtaatc atggtcatag ctgtttcctg
     5101 tgtgaaattg ttatccgctc acaattccac acaacatacg agccggaagc ataaagtgta
     5161 aagcctgggg tgcctaatga gtgagctaac tcacattaat tgcgttgcgc tcactgcccg
     5221 ctttccagtc gggaaacctg tcgtgccagc tgcattaatg aatcggccaa cgcgcgggga
     5281 gaggcggttt gcgtattggg cgctcttccg cttcctcgct cactgactcg ctgcgctcgg
     5341 tcgttcggct gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg ttatccacag
     5401 aatcagggga taacgcagga aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc
     5461 gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg cccccctgac gagcatcaca
     5521 aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg actataaaga taccaggcgt
     5581 ttccccctgg aagctccctc gtgcgctctc ctgttccgac cctgccgctt accggatacc
     5641 tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc
     5701 tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc
     5761 ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc caacccggta agacacgact
     5821 tatcgccact ggcagcagcc actggtaaca ggattagcag agcgaggtat gtaggcggtg
     5881 ctacagagtt cttgaagtgg tggcctaact acggctacac tagaagaaca gtatttggta
     5941 tctgcgctct gctgaagcca gttaccttcg gaaaaagagt tggtagctct tgatccggca
     6001 aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa
     6061 aaaaaggatc tcaagaagat cctttgatct tttctacggg gtctgacgct cagtggaacg
     6121 aaaactcacg ttaagggatt ttggtcatga gattatcaaa aaggatcttc acctagatcc
     6181 ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa acttggtctg
     6241 acagttacca atgcttaatc agtgaggcac ctatctcagc gatctgtcta tttcgttcat
     6301 ccatagttgc ctgactcccc gtcgtgtaga taactacgat acgggagggc ttaccatctg
     6361 gccccagtgc tgcaatgata ccgcgagacc cacgctcacc ggctccagat ttatcagcaa
     6421 taaaccagcc agccggaagg gccgagcgca gaagtggtcc tgcaacttta tccgcctcca
     6481 tccagtctat taattgttgc cgggaagcta gagtaagtag ttcgccagtt aatagtttgc
     6541 gcaacgttgt tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt
     6601 cattcagctc cggttcccaa cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa
     6661 aagcggttag ctccttcggt cctccgatcg ttgtcagaag taagttggcc gcagtgttat
     6721 cactcatggt tatggcagca ctgcataatt ctcttactgt catgccatcc gtaagatgct
     6781 tttctgtgac tggtgagtac tcaaccaagt cattctgaga atagtgtatg cggcgaccga
     6841 gttgctcttg cccggcgtca atacgggata ataccgcgcc acatagcaga actttaaaag
     6901 tgctcatcat tggaaaacgt tcttcggggc gaaaactctc aaggatctta ccgctgttga
     6961 gatccagttc gatgtaaccc actcgtgcac ccaactgatc ttcagcatct tttactttca
     7021 ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg
     7081 cgacacggaa atgttgaata ctcatactct tcctttttca atattattga agcatttatc
     7141 agggttattg tctcatgagc ggatacatat ttgaatgtat ttagaaaaat aaacaaatag
     7201 gggttccgcg cacatttccc cgaaaagtgc cacctgacgt cgacggatcg ggagatctcc
     7261 cgatccccta tggtgcactc tcagtacaat ctgctctgat gccgcatagt taagcc
//
