LOCUS       sequence-358634-        6894 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Express Fluorescent RADARS in bicistronic vector with mCherry as 
            control gene.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6894)
  AUTHORS   Jiang K, Koob J, Chen XD, Krajeski RN, Zhang Y, Volf V, Zhou W, 
            Sgrizzi SR, Villiger L, Gootenberg JS, Chen F, Abudayyeh OO
  TITLE     Programmable eukaryotic protein synthesis with RNA sensors by 
            harnessing ADAR.
  JOURNAL   Nat Biotechnol. 2022 Oct 27. pii: 10.1038/s41587-022-01534-5. doi: 
            10.1038/s41587-022-01534-5.
  PUBMED    36302988
REFERENCE   2  (bases 1 to 6894)
  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 
            Biotechnol. 2022 Oct 27. pii: 10.1038/s41587-022-01534-5. doi: 
            10.1038/s41587-022-01534-5."
COMMENT     Sequence Label: pDY1243-Fluorescent RADARS
FEATURES             Location/Qualifiers
     source          1..6894
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        181..560
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        561..764
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     715..735
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     809..828
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        809..827
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             863..1570
                     /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(1008..1026)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     1276..1295
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     CDS             1622..1675
                     /codon_start=1
                     /product="2A peptide from Thosea asigna virus capsid 
                     protein"
                     /label=T2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="EGRGSLLTCGDVEENPGP"
     primer_bind     complement(2431..2448)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2437..2661
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      2707..3135
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2794..2813)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3004..3025
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3144..3164)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3149..3478
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3329..3464
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     3391..3410
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             3545..4339
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(3599..3618)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4209..4228
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    4513..4634
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4550..4569)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4604..4623
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4683..4699)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4683..4699)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4696..4718)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4707..4723
                     /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(4731..4737,4738..4755,4756..4761))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4776..4797
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4914..4931)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5085..5673)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5165..5184)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5844..6635,6636..6704))
                     /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     6467..6486
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6705..6809)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(join(6884..6894,1..9))
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 ctgatgccgc atagttaagc cagtatctgc tccctgcttg tgtgttggag gtcgctgagt
       61 agtgcgcgag caaaatttaa gctacaacaa ggcaaggctt gaccgacaat tgcatgaaga
      121 atctgcttag ggttaggcgt tttgcgctgc ttcgcgatgt acgggccaga tatacgcgtt
      181 gacattgatt attgactagt tattaatagt aatcaattac ggggtcatta gttcatagcc
      241 catatatgga gttccgcgtt acataactta cggtaaatgg cccgcctggc tgaccgccca
      301 acgacccccg cccattgacg tcaataatga cgtatgttcc catagtaacg ccaataggga
      361 ctttccattg acgtcaatgg gtggagtatt tacggtaaac tgcccacttg gcagtacatc
      421 aagtgtatca tatgccaagt acgcccccta ttgacgtcaa tgacggtaaa tggcccgcct
      481 ggcattatgc ccagtacatg accttatggg actttcctac ttggcagtac atctacgtat
      541 tagtcatcgc tattaccatg gtgatgcggt tttggcagta catcaatggg cgtggatagc
      601 ggtttgactc acggggattt ccaagtctcc accccattga cgtcaatggg agtttgtttt
      661 ggcaccaaaa tcaacgggac tttccaaaat gtcgtaacaa ctccgcccca ttgacgcaaa
      721 tgggcggtag gcgtgtacgg tgggaggtct atataagcag agctctctgg ctaactagag
      781 aacccactgc ttactggctt atcgaaatta atacgactca ctatagggag acccaagctg
      841 gctagcgttt aaacttaagc ttatggtgag caagggcgag gaggataaca tggccatcat
      901 caaggagttc atgcgcttca aggtgcacat ggagggctcc gtgaacggcc acgagttcga
      961 gatcgagggc gagggcgagg gccgccccta cgagggcacc cagaccgcca agctgaaggt
     1021 gaccaagggt ggccccctgc ccttcgcctg ggacatcctg tcccctcagt tcatgtacgg
     1081 ctccaaggcc tacgtgaagc accccgccga catccccgac tacttgaagc tgtccttccc
     1141 cgagggcttc aagtgggagc gcgtgatgaa cttcgaggac ggcggcgtgg tgaccgtgac
     1201 ccaggactcc tccctgcagg acggcgagtt catctacaag gtgaagctgc gcggcaccaa
     1261 cttcccctcc gacggccccg taatgcagaa gaagaccatg ggctgggagg cctcctccga
     1321 gcggatgtac cccgaggacg gcgccctgaa gggcgagatc aagcagaggc tgaagctgaa
     1381 ggacggcggc cactacgacg ctgaggtcaa gaccacctac aaggccaaga agcccgtgca
     1441 gctgcccggc gcctacaacg tcaacatcaa gttggacatc acctcccaca acgaggacta
     1501 caccatcgtg gaacagtacg aacgcgccga gggccgccac tccaccggcg gcatggacga
     1561 gctgtacaag cccggtgcag atggtaccgg cagctgagac gctgggaccg tctcaggcag
     1621 cgagggcaga ggaagtcttc taacatgcgg tgacgtggag gagaatcccg gccctatggt
     1681 cagcaaaggc gaagaagaca acatggcatc actgccagcg acacacgaac ttcatatttt
     1741 cggctccatc aatggtgtgg acttcgatat ggttggacag ggtactggga accctaacga
     1801 tggatatgag gagttgaact tgaagagtac caagggcgat cttcagtttt caccttggat
     1861 tctcgtacct catataggtt atggctttca tcaatatctg ccctatcctg acggtatgtc
     1921 acctttccaa gcagctatgg ttgacgggag cggttatcag gttcaccgga caatgcagtt
     1981 tgaagatgga gcctctttga ccgtaaatta tcggtatact tatgagggca gccatattaa
     2041 aggcgaggct caagttaagg ggactgggtt ccccgcggat ggtcccgtaa tgacaaacag
     2101 tctgacggct gcagactggt gccgctctaa gaagacatat ccaaatgaca agacgattat
     2161 tagcactttc aagtggtcat acactacagg aaatggtaaa cgctatagga gcactgcccg
     2221 gaccacgtat actttcgcga agccgatggc ggcaaattat ttgaaaaacc aaccgatgta
     2281 tgtgttcaga aagacagaac tcaagcattc taaaactgaa ctgaacttca aagagtggca
     2341 gaaagccttt actgatgtaa tgggcatgga cgaactctat aagtaagcgg ccgctcgagt
     2401 ctagagggcc cgtttaaacc cgctgatcag cctcgactgt gccttctagt tgccagccat
     2461 ctgttgtttg cccctccccc gtgccttcct tgaccctgga aggtgccact cccactgtcc
     2521 tttcctaata aaatgaggaa attgcatcgc attgtctgag taggtgtcat tctattctgg
     2581 ggggtggggt ggggcaggac agcaaggggg aggattggga agacaatagc aggcatgctg
     2641 gggatgcggt gggctctatg gcttctgagg cggaaagaac cagctggggc tctagggggt
     2701 atccccacgc gccctgtagc ggcgcattaa gcgcggcggg tgtggtggtt acgcgcagcg
     2761 tgaccgctac acttgccagc gccctagcgc ccgctccttt cgctttcttc ccttcctttc
     2821 tcgccacgtt cgccggcttt ccccgtcaag ctctaaatcg ggggctccct ttagggttcc
     2881 gatttagtgc tttacggcac ctcgacccca aaaaacttga ttagggtgat ggttcacgta
     2941 gtgggccatc gccctgatag acggtttttc gccctttgac gttggagtcc acgttcttta
     3001 atagtggact cttgttccaa actggaacaa cactcaaccc tatctcggtc tattcttttg
     3061 atttataagg gattttgccg atttcggcct attggttaaa aaatgagctg atttaacaaa
     3121 aatttaacgc gaattaattc tgtggaatgt gtgtcagtta gggtgtggaa agtccccagg
     3181 ctccccagca ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa ccaggtgtgg
     3241 aaagtcccca ggctccccag caggcagaag tatgcaaagc atgcatctca attagtcagc
     3301 aaccatagtc ccgcccctaa ctccgcccat cccgccccta actccgccca gttccgccca
     3361 ttctccgccc catggctgac taattttttt tatttatgca gaggccgagg ccgcctctgc
     3421 ctctgagcta ttccagaagt agtgaggagg cttttttgga ggcctaggct tttgcaaaaa
     3481 gctcccggga gcttgtatat ccattttcgg atctgatcaa gagacaggat gaggatcgtt
     3541 tcgcatgatt gaacaagatg gattgcacgc aggttctccg gccgcttggg tggagaggct
     3601 attcggctat gactgggcac aacagacaat cggctgctct gatgccgccg tgttccggct
     3661 gtcagcgcag gggcgcccgg ttctttttgt caagaccgac ctgtccggtg ccctgaatga
     3721 actgcaggac gaggcagcgc ggctatcgtg gctggccacg acgggcgttc cttgcgcagc
     3781 tgtgctcgac gttgtcactg aagcgggaag ggactggctg ctattgggcg aagtgccggg
     3841 gcaggatctc ctgtcatctc accttgctcc tgccgagaaa gtatccatca tggctgatgc
     3901 aatgcggcgg ctgcatacgc ttgatccggc tacctgccca ttcgaccacc aagcgaaaca
     3961 tcgcatcgag cgagcacgta ctcggatgga agccggtctt gtcgatcagg atgatctgga
     4021 cgaagagcat caggggctcg cgccagccga actgttcgcc aggctcaagg cgcgcatgcc
     4081 cgacggcgag gatctcgtcg tgacccatgg cgatgcctgc ttgccgaata tcatggtgga
     4141 aaatggccgc ttttctggat tcatcgactg tggccggctg ggtgtggcgg accgctatca
     4201 ggacatagcg ttggctaccc gtgatattgc tgaagagctt ggcggcgaat gggctgaccg
     4261 cttcctcgtg ctttacggta tcgccgctcc cgattcgcag cgcatcgcct tctatcgcct
     4321 tcttgacgag ttcttctgag cgggactctg gggttcgaaa tgaccgacca agcgacgccc
     4381 aacctgccat cacgagattt cgattccacc gccgccttct atgaaaggtt gggcttcgga
     4441 atcgttttcc gggacgccgg ctggatgatc ctccagcgcg gggatctcat gctggagttc
     4501 ttcgcccacc ccaacttgtt tattgcagct tataatggtt acaaataaag caatagcatc
     4561 acaaatttca caaataaagc atttttttca ctgcattcta gttgtggttt gtccaaactc
     4621 atcaatgtat cttatcatgt ctgtataccg tcgacctcta gctagagctt ggcgtaatca
     4681 tggtcatagc tgtttcctgt gtgaaattgt tatccgctca caattccaca caacatacga
     4741 gccggaagca taaagtgtaa agcctggggt gcctaatgag tgagctaact cacattaatt
     4801 gcgttgcgct cactgcccgc tttccagtcg ggaaacctgt cgtgccagct gcattaatga
     4861 atcggccaac gcgcggggag aggcggtttg cgtattgggc gctcttccgc ttcctcgctc
     4921 actgactcgc tgcgctcggt cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg
     4981 gtaatacggt tatccacaga atcaggggat aacgcaggaa agaacatgtg agcaaaaggc
     5041 cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca taggctccgc
     5101 ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga
     5161 ctataaagat accaggcgtt tccccctgga agctccctcg tgcgctctcc tgttccgacc
     5221 ctgccgctta ccggatacct gtccgccttt ctcccttcgg gaagcgtggc gctttctcat
     5281 agctcacgct gtaggtatct cagttcggtg taggtcgttc gctccaagct gggctgtgtg
     5341 cacgaacccc ccgttcagcc cgaccgctgc gccttatccg gtaactatcg tcttgagtcc
     5401 aacccggtaa gacacgactt atcgccactg gcagcagcca ctggtaacag gattagcaga
     5461 gcgaggtatg taggcggtgc tacagagttc ttgaagtggt ggcctaacta cggctacact
     5521 agaagaacag tatttggtat ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt
     5581 ggtagctctt gatccggcaa acaaaccacc gctggtagcg gtggtttttt tgtttgcaag
     5641 cagcagatta cgcgcagaaa aaaaggatct caagaagatc ctttgatctt ttctacgggg
     5701 tctgacgctc agtggaacga aaactcacgt taagggattt tggtcatgag attatcaaaa
     5761 aggatcttca cctagatcct tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata
     5821 tatgagtaaa cttggtctga cagttaccaa tgcttaatca gtgaggcacc tatctcagcg
     5881 atctgtctat ttcgttcatc catagttgcc tgactccccg tcgtgtagat aactacgata
     5941 cgggagggct taccatctgg ccccagtgct gcaatgatac cgcgagaccc acgctcaccg
     6001 gctccagatt tatcagcaat aaaccagcca gccggaaggg ccgagcgcag aagtggtcct
     6061 gcaactttat ccgcctccat ccagtctatt aattgttgcc gggaagctag agtaagtagt
     6121 tcgccagtta atagtttgcg caacgttgtt gccattgcta caggcatcgt ggtgtcacgc
     6181 tcgtcgtttg gtatggcttc attcagctcc ggttcccaac gatcaaggcg agttacatga
     6241 tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt tgtcagaagt
     6301 aagttggccg cagtgttatc actcatggtt atggcagcac tgcataattc tcttactgtc
     6361 atgccatccg taagatgctt ttctgtgact ggtgagtact caaccaagtc attctgagaa
     6421 tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca
     6481 catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg aaaactctca
     6541 aggatcttac cgctgttgag atccagttcg atgtaaccca ctcgtgcacc caactgatct
     6601 tcagcatctt ttactttcac cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc
     6661 gcaaaaaagg gaataagggc gacacggaaa tgttgaatac tcatactctt cctttttcaa
     6721 tattattgaa gcatttatca gggttattgt ctcatgagcg gatacatatt tgaatgtatt
     6781 tagaaaaata aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc acctgacgtc
     6841 gacggatcgg gagatctccc gatcccctat ggtgcactct cagtacaatc tgct
//
