LOCUS       sequence-356646-        7729 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Express RADARS iCaspase that conditionally turn on iCaspase in the 
            presence of target RNA.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7729)
  TITLE     Unpublished, Abudayyeh Lab
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 7729)
  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: 
            "Unpublished"
COMMENT     Sequence Label: pDY0973-RADARS iCaspase
FEATURES             Location/Qualifiers
     source          1..7729
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     134..153
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     387..404
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    521..542
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(557..562,563..580,581..587)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    595..611
                     /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     600..622
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     619..635
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     619..635
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     polyA_signal    649..697
                     /note="synthetic polyadenylation signal"
     CDS             complement(698..2359)
                     /codon_start=1
                     /product="secreted Cypridina luciferase"
                     /label=CLuc
                     /note="contains 30 codon substitutions for efficient 
                     translation in mammalian cells"
                     /translation="MKTLILAVALVYCATVHCQDCPYEPDPPNTVPTSCEAKEGECIDS
                     SCGTCTRDILSDGLCENKPGKTCCRMCQYVIECRVEAAGWFRTFYGKRFQFQEPGTYVL
                     GQGTKGGDWKVSITLENLDGTKGAVLTKTRLEVAGDIIDIAQATENPITVNGGADPIIA
                     NPYTIGEVTIAVVEMPGFNITVIEFFKLIVIDILGGRSVRIAPDTANKGMISGLCGDLK
                     MMEDTDFTSDPEQLAIQPKINQEFDGCPLYGNPDDVAYCKGLLEPYKDSCRNPINFYYY
                     TISCAFARCMGGDERASHVLLDYRETCAAPETRGTCVLSGHTFYDTFDKARYQFQGPCK
                     EILMAADCFWNTWDVKVSHRNVDSYTEVEKVRIRKQSTVVELIVDGKQILVGGEAVSVP
                     YSSQNTSIYWQDGDILTTAILPEALVVKFNFKQLLVVHIRDPFDGKTCGICGNYNQDFS
                     DDSFDAEGACDLTPNPPGCTEEQKPEAERLCNSLFAGQSDLDQKCNVCHKPDRVERCMY
                     EYCLRGQQGFCDHAWEFKKECYIKHGDTLEVPDECK"
     primer_bind     complement(2384..2403)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2385..2403)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     promoter        complement(2448..2651)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(2477..2497)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        2652..3031
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        3145..4323
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation 
                     factor EF-1-alpha"
     intron          3376..4314
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
     primer_bind     4271..4291
                     /label=EF1a-F
                     /note="Human elongation factor-1a promoter, forward primer"
     CDS             4381..4437
                     /codon_start=1
                     /product="2A peptide from porcine teschovirus-1 
                     polyprotein"
                     /label=P2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="ATNFSLLKQAGDVEENPGP"
     CDS             4444..4764
                     /codon_start=1
                     /product="F36V mutant of FK506-binding protein FKBP12"
                     /label=DmrB
                     /note="binds synthetic ligands that do not bind wild-type 
                     FKBP"
                     /translation="GVQVETISPGDGRTFPKRGQTCVVHYTGMLEDGKKVDSSRDRNKP
                     FKFMLGKQEVIRGWEEGVAQMSVGQRAKLTISPDYAYGATGHPGIIPPHATLVFDVELL
                     KLE"
     CDS             complement(5116..5127)
                     /codon_start=1
                     /product="WELQut protease recognition and cleavage site"
                     /label=WELQut site
                     /translation="WELQ"
     CDS             5641..5667
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     polyA_signal    5677..5725
                     /note="synthetic polyadenylation signal"
     primer_bind     complement(5748..5765)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(5748..5764)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5757..5779)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     complement(5973..5992)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     6092..6114
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(6152..6170)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        6238..6342
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6343..6411,6412..7203)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSGGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(6561..6580)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      join(7374..7729,1..233)
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
ORIGIN
        1 cgacctacac cgaactgaga tacctacagc gtgagctatg agaaagcgcc acgcttcccg
       61 aagggagaaa ggcggacagg tatccggtaa gcggcagggt cggaacagga gagcgcacga
      121 gggagcttcc agggggaaac gcctggtatc tttatagtcc tgtcgggttt cgccacctct
      181 gacttgagcg tcgatttttg tgatgctcgt caggggggcg gagcctatgg aaaaacgcca
      241 gcaacgcggc ctttttacgg ttcctggcct tttgctggcc ttttgctcac atgttctttc
      301 ctgcgttatc ccctgattct gtggataacc gtattaccgc ctttgagtga gctgataccg
      361 ctcgccgcag ccgaacgacc gagcgcagcg agtcagtgag cgaggaagcg gaagagcgcc
      421 caatacgcaa accgcctctc cccgcgcgtt ggccgattca ttaatgcagc tggcacgaca
      481 ggtttcccga ctggaaagcg ggcagtgagc gcaacgcaat taatgtgagt tagctcactc
      541 attaggcacc ccaggcttta cactttatgc ttccggctcg tatgttgtgt ggaattgtga
      601 gcggataaca atttcacaca ggaaacagct atgaccatga ttacgccaca cacaaaaaac
      661 caacacacag atgtaatgaa aataaagata ttttatttta tttgcattca tctggtactt
      721 ctagggtgtc tccatgcttt atgtagcatt ctttcttgaa ctcccatgcg tggtcacaga
      781 aaccctgttg tcccctcagg caatactcgt acatgcatcg ttcgacacgg tcaggcttgt
      841 ggcacacgtt acatttctga tcaagatcac tttgaccggc gaagagacta ttgcagagtc
      901 gttcagcttc aggtttctgt tcttcggtgc atcccggtgg gttgggggtc agatcacagg
      961 ctccttcagc atcaaaagaa tcatcactga aatcctggtt gtagttaccg caaataccgc
     1021 aagtcttacc atcgaatgga tctctaatat gtacgacgag cagttgcttg aagttgaact
     1081 tgaccaccag agcttcaggt aggatggctg tagtcagtat gtcaccatct tgccagtaga
     1141 tggaagtgtt ctgagagctg tacgggacgg acacggcttc tcctccaacc agaatctgtt
     1201 ttccatcaac aatgagttct actacagtcg attgtttcct gattcgtact ttctctactt
     1261 cagtgtaaga gtcaacattc ctgtgtgaaa ccttcacatc ccaagtgttc cagaaacagt
     1321 cggcggccat aagaatctcc ttgcagggac cctggaattg gtatcttgct ttgtcaaatg
     1381 tatcgtagaa agtatgtcca gacaaaacgc aggttcctct agtttcggga gcagcgcacg
     1441 tctccctgta gtcaagcagc acgtgtgagg ctcgctcgtc tccacccata cagcgggcga
     1501 aggcgcagga gatggtgtag tagtagaagt tgatggggtt gcggcagctg tccttgtacg
     1561 gctccagaag acctttgcag tatgcaacgt catcaggatt tccatagagt ggacaaccgt
     1621 caaactcctg gttgatctta ggctgaatag cgagttgttc tggatctgaa gtgaagtctg
     1681 tatcttccat cattttaaga tctccacaga ggccagagat cattcctttg tttgctgtgt
     1741 ctggggcgat tcttacagat cttcctccga ggatgtcgat cacgatcagt ttgaagaact
     1801 caatgacggt gatgttgaag cctggcatct caacaacagc gatggtgacc tcgccgatgg
     1861 tgtacgggtt ggcgatgata gggtcagctc caccgtttac agtgatggga ttctcagtag
     1921 cttgagcgat gtcaatgatg tctccagcca cttccagtct tgtcttggtc agcacagccc
     1981 ccttggttcc atccaggttc tccagggtga tggacacctt ccagtcgccg cccttggttc
     2041 cttgacccaa cacgtatgta ccaggttcct ggaactggaa tctctttcca tagaatgttc
     2101 taaaccatcc tgcggcctct actctgcatt caattacata ctgacacatt cgacaacatg
     2161 tttttcctgg tttattttca cacagtccat ctgatagtat gtctctcgtg caggtgccac
     2221 agctgctatc aatacattct ccttctttag cttcacagga agttggaact gtgtttggtg
     2281 gatcaggttc gtaaggacag tcctggcaat gaacagtggc gcagtagact aatgcaacgg
     2341 caagaattaa ggtcttcatg gtggcggtac caagcttgcg tctccctata gtgagtcgta
     2401 ttaatttcga taagccagta agcagtgggt tctctagtta gccagagagc tctgcttata
     2461 tagacctccc accgtacacg cctaccgccc atttgcgtca atggggcgga gttgttacga
     2521 cattttggaa agtcccgttg attttggtgc caaaacaaac tcccattgac gtcaatgggg
     2581 tggagacttg gaaatccccg tgagtcaaac cgctatccac gcccattgat gtactgccaa
     2641 aaccgcatca ccatggtaat agcgatgact aatacgtaga tgtactgcca agtaggaaag
     2701 tcccataagg tcatgtactg ggcataatgc caggcgggcc atttaccgtc attgacgtca
     2761 atagggggcg tacttggcat atgatacact tgatgtactg ccaagtgggc agtttaccgt
     2821 aaatactcca cccattgacg tcaatggaaa gtccctattg gcgttactat gggaacatac
     2881 gtcattattg acgtcaatgg gcgggggtcg ttgggcggtc agccaggcgg gccatttacc
     2941 gtaagttatg taacgcggaa ctccatatat gggctatgaa ctaatgaccc cgtaattgat
     3001 tactattaat aactagtcaa taatcaatgt caacgcgtat atctggcccg tacatcgaag
     3061 ctgatgcaaa gatggataaa gttttaaaca gagaggaatc tttgcagcta atggaccttc
     3121 taggtcttga aaggagtggg aattggctcc ggtgcccgtc agtgggcaga gcgcacatcg
     3181 cccacagtcc ccgagaagtt ggggggaggg gtcggcaatt gaaccggtgc ctagagaagg
     3241 tggcgcgggg taaactggga aagtgatgtc gtgtactggc tccgcctttt tcccgagggt
     3301 gggggagaac cgtatataag tgcagtagtc gccgtgaacg ttctttttcg caacgggttt
     3361 gccgccagaa cacaggtaag tgccgtgtgt ggttcccgcg ggcctggcct ctttacgggt
     3421 tatggccctt gcgtgccttg aattacttcc acctggctgc agtacgtgat tcttgatccc
     3481 gagcttcggg ttggaagtgg gtgggagagt tcgaggcctt gcgcttaagg agccccttcg
     3541 cctcgtgctt gagttgaggc ctggcctggg cgctggggcc gccgcgtgcg aatctggtgg
     3601 caccttcgcg cctgtctcgc tgctttcgat aagtctctag ccatttaaaa tttttgatga
     3661 cctgctgcga cgcttttttt ctggcaagat agtcttgtaa atgcgggcca agatctgcac
     3721 actggtattt cggtttttgg ggccgcgggc ggcgacgggg cccgtgcgtc ccagcgcaca
     3781 tgttcggcga ggcggggcct gcgagcgcgg ccaccgagaa tcggacgggg gtagtctcaa
     3841 gctggccggc ctgctctggt gcctggcctc gcgccgccgt gtatcgcccc gccctgggcg
     3901 gcaaggctgg cccggtcggc accagttgcg tgagcggaaa gatggccgct tcccggccct
     3961 gctgcaggga gctcaaaatg gaggacgcgg cgctcgggag agcgggcggg tgagtcaccc
     4021 acacaaagga aaagggcctt tccgtcctca gccgtcgctt catgtgactc cacggagtac
     4081 cgggcgccgt ccaggcacct cgattagttc tcgagctttt ggagtacgtc gtctttaggt
     4141 tggggggagg ggttttatgc gatggagttt ccccacactg agtgggtgga gactgaagtt
     4201 aggccagctt ggcacttgat gtaattctcc ttggaatttg ccctttttga gtttggatct
     4261 tggttcattc tcaagcctca gacagtggtt caaagttttt ttcttccatt tcaggtgtcg
     4321 tgacgtacgg gatcctctag agccaccatg cgagaccatg tcaaggtctc aggaagcgga
     4381 gctactaact tcagcctgct gaagcaggct ggcgacgtgg aggagaaccc tggacctgtc
     4441 gagggagtgc aggtggagac tatctcccca ggagacgggc gcaccttccc caagcgcggc
     4501 cagacctgcg tggtgcacta caccgggatg cttgaagatg gaaagaaagt tgattcctcc
     4561 cgggacagaa acaagccctt taagtttatg ctaggcaagc aggaggtgat ccgaggctgg
     4621 gaagaagggg ttgcccagat gagtgtgggt cagagagcca aactgactat atctccagat
     4681 tatgcctatg gtgccactgg gcacccaggc atcatcccac cacatgccac tctcgtcttc
     4741 gatgtggagc ttctaaaact ggaatctggc ggtggatccg gagtcgacgg atttggtgat
     4801 gtcggtgctc ttgagagttt gaggggaaat gcagatttgg cttacatcct gagcatggag
     4861 ccctgtggcc actgcctcat tatcaacaat gtgaacttct gccgtgagtc cgggctccgc
     4921 acccgcactg gctccaacat cgactgtgag aagttgcggc gtcgcttctc ctcgctgcat
     4981 ttcatggtgg aggtgaaggg cgacctgact gccaagaaaa tggtgctggc tttgctggag
     5041 ctggcgcggc aggaccacgg tgctctggac tgctgcgtgg tggtcattct ctctcacggc
     5101 tgtcaggcca gccacctgca gttcccaggg gctgtctacg gcacagatgg atgccctgtg
     5161 tcggtcgaga agattgtgaa catcttcaat gggaccagct gccccagcct gggagggaag
     5221 cccaagctct ttttcatcca ggcctgtggt ggggagcaga aagaccatgg gtttgaggtg
     5281 gcctccactt cccctgaaga cgagtcccct ggcagtaacc ccgagccaga tgccaccccg
     5341 ttccaggaag gtttgaggac cttcgaccag ctggacgcca tatctagttt gcccacaccc
     5401 agtgacatct ttgtgtccta ctctactttc ccaggttttg tttcctggag ggaccccaag
     5461 agtggctcct ggtacgttga aaccctggac gacatctttg agcagtgggc tcactctgaa
     5521 gacctgcagt ccctcctgct tagggtcgct aatgctgttt cggtgaaagg gatttataaa
     5581 cagatgcctg gttgctttaa tttcctccgg aaaaaacttt tctttaaaac atcagtcgac
     5641 tatccgtacg acgtaccaga ctacgcactc gactaaaata aaatatcttt attttcatta
     5701 catctgtgtg ttggtttttt gtgtgcccgg gtaccgagct cgaattcact ggccgtcgtt
     5761 ttacaacgtc gtgactggga aaaccctggc gttacccaac ttaatcgcct tgcagcacat
     5821 ccccctttcg ccagctggcg taatagcgaa gaggcccgca ccgatcgccc ttcccaacag
     5881 ttgcgcagcc tgaatggcga atggcgcctg atgcggtatt ttctccttac gcatctgtgc
     5941 ggtatttcac accgcatatg gtgcactctc agtacaatct gctctgatgc cgcatagtta
     6001 agccagcccc gacacccgcc aacacccgct gacgcgccct gacgggcttg tctgctcccg
     6061 gcatccgctt acagacaagc tgtgaccgtc tccgggagct gcatgtgtca gaggttttca
     6121 ccgtcatcac cgaaacgcgc gagacgaaag ggcctcgtga tacgcctatt tttataggtt
     6181 aatgtcatga taataatggt ttcttagacg tcaggtggca cttttcgggg aaatgtgcgc
     6241 ggaaccccta tttgtttatt tttctaaata cattcaaata tgtatccgct catgagacaa
     6301 taaccctgat aaatgcttca ataatattga aaaaggaaga gtatgagtat tcaacatttc
     6361 cgtgtcgccc ttattccctt ttttgcggca ttttgccttc ctgtttttgc tcacccagaa
     6421 acgctggtga aagtaaaaga tgctgaagat cagttgggtg cacgagtggg ttacatcgaa
     6481 ctggatctca acagcggtaa gatccttgag agttttcgcc ccgaagaacg ttttccaatg
     6541 atgagcactt ttaaagttct gctatgtggc gcggtattat cccgtattga cgccgggcaa
     6601 gagcaactcg gtcgccgcat acactattct cagaatgact tggttgagta ctcaccagtc
     6661 acagaaaagc atcttacgga tggcatgaca gtaagagaat tatgcagtgc tgccataacc
     6721 atgagtgata acactgcggc caacttactt ctgacaacga tcggaggacc gaaggagcta
     6781 accgcttttt tgcacaacat gggggatcat gtaactcgcc ttgatcgttg ggaaccggag
     6841 ctgaatgaag ccataccaaa cgacgagcgt gacaccacga tgcctgtagc aatggcaaca
     6901 acgttgcgca aactattaac tggcgaacta cttactctag cttcccggca acaattaata
     6961 gactggatgg aggcggataa agttgcagga ccacttctgc gctcggccct tccggctggc
     7021 tggtttattg ctgataaatc tggagccggt gagcgtgggt ctggcggtat cattgcagca
     7081 ctggggccag atggtaagcc ctcccgtatc gtagttatct acacgacggg gagtcaggca
     7141 actatggatg aacgaaatag acagatcgct gagataggtg cctcactgat taagcattgg
     7201 taactgtcag accaagttta ctcatatata ctttagattg atttaaaact tcatttttaa
     7261 tttaaaagga tctaggtgaa gatccttttt gataatctca tgaccaaaat cccttaacgt
     7321 gagttttcgt tccactgagc gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat
     7381 cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct accagcggtg
     7441 gtttgtttgc cggatcaaga gctaccaact ctttttccga aggtaactgg cttcagcaga
     7501 gcgcagatac caaatactgt tcttctagtg tagccgtagt taggccacca cttcaagaac
     7561 tctgtagcac cgcctacata cctcgctctg ctaatcctgt taccagtggc tgctgccagt
     7621 ggcgataagt cgtgtcttac cgggttggac tcaagacgat agttaccgga taaggcgcag
     7681 cggtcgggct gaacgggggg ttcgtgcaca cagcccagct tggagcgaa
//
