LOCUS       sequence-396808-        6789 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Entry vector for closed-loop ADAR-based RNA sensors (DART VADAR). 
            Expresses TagBFP constitutively. Expresses NanoLuc and MCP-ADAR upon
            target detection.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6789)
  AUTHORS   Gayet RV, Ilia K, Razavi S, Tippens ND, Lalwani MA, Zhang K, Chen 
            JX, Chen JC, Vargas-Asencio J, Collins JJ
  TITLE     Autocatalytic base editing for RNA-responsive translational control.
  JOURNAL   Nat Commun. 2023 Mar 11;14(1):1339. doi: 10.1038/s41467-023-36851-z.
  PUBMED    36906659
REFERENCE   2  (bases 1 to 6789)
  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 
            Commun."; date: "2023-03-11"; volume: "14(1):1339. doi"; pages: " 
            10.1038/s41467-023-36851-z"
COMMENT     Sequence Label: 
            pDV21_feedback_entry_TagBFP-2A-MCS-2A-NLuc-2A-MCP-ADAR2dd(E488Q)-NES
FEATURES             Location/Qualifiers
     source          1..6789
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        348..559
                     /label=EF-1-alpha core promoter
                     /note="core promoter for human elongation factor 
                     EF-1-alpha"
     intron          560..1498
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
     primer_bind     1455..1475
                     /label=EF1a-F
                     /note="Human elongation factor-1a promoter, forward primer"
     CDS             2246..2302
                     /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             2366..2419
                     /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"
     CDS             2429..2941
                     /codon_start=1
                     /product="NanoLuc(R) luciferase"
                     /label=Nluc
                     /note="human codon-optimized"
                     /translation="MVFTLEDFVGDWRQTAGYNLDQVLEQGGVSSLFQNLGVSVTPIQR
                     IVLSGENGLKIDIHVIIPYEGLSGDQMGQIEKIFKVVYPVDDHHFKVILHYGTLVIDGV
                     TPNMIDYFGRPYEGIAVFDGKKITVTGTLWNGNKIIDERLINPDGSLLFRVTINGVTGW
                     RLCERILA"
     CDS             2951..3010
                     /codon_start=1
                     /product="2A peptide from equine rhinitis A virus 
                     polyprotein"
                     /label=E2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="QCTNYALLKLAGDVESNPGP"
     CDS             4616..4642
                     /codon_start=1
                     /product="nuclear export signal from the HIV Rev protein 
                     (Fischer et al., 1995)"
                     /label=NES
                     /translation="LPPLERLTL"
     polyA_signal    4697..4745
                     /note="synthetic polyadenylation signal"
     rep_origin      complement(5022..5610)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5102..5121)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5781..6572,6573..6641))
                     /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     6404..6423
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6642..6746)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 tgcgttttta tgcttgtagt attgtataat gtttttaaga tccggagaga gcgagattcc
       61 gtctcaaaga aaaaaaaagt aatgaaatga ataaaatgag tcctagagcc agtaaatgtc
      121 gtaaatgtct cagctagtca ggtagtaaaa ggtctcaact aggcagtggc agagcaggat
      181 tcaaattcag ggctgttgtg atgcctccgc agactctgag cgccacctgg tggtaatttg
      241 tctgtgcctc ttctgacgtg gaagaacagc aactaacaca ctaacacggc atttactatg
      301 ggccagccat tgttctaata ctttaattgg ctccggtgcc cgtcagtggg cagagcgcac
      361 atcgcccaca gtccccgaga agttgtgggg aggggtcggc aattgaaccg gtgcctagag
      421 aaggtggcgc ggggtaaact gggaaagtga tgtcgtgtac tggctccgcc tttttcccga
      481 gggtggggga gaaccgtata taagtgcagt agtcgccgtg aacgttcttt ttcgcaacgg
      541 gtttgccgcc agaacacagg taagtgccgt gtgtggttcc cgcgggcctg gcctctttac
      601 gggttatggc ccttgcgtgc cttgaattac ttccacctgg ctgcagtacg tgattcttga
      661 tcccgagctt cgggttggaa gtgggtggga gagttcgagg ccttgcgctt aaggagcccc
      721 ttcgcctcgt gcttgagttg aggcctggcc tgggcgctgg ggccgccgcg tgcgaatctg
      781 gtggcacctt cgcgcctgtc tcgctgcttt cgataagtct ctagccattt aaaatttttg
      841 atgacctgct gcgacgcttt ttttctggca agatagtctt gtaaatgcgg gccaagatct
      901 gcacactggt atttcggttt ttggggccgc gggcggcgac ggggcccgtg cgtcccagcg
      961 cacatgttcg gcgaggcggg gcctgcgagc gcggccaccg agaatcggac gggggtagtc
     1021 tcaagctggc cggcctgctc tggtgcctgg cctcgcgccg ccgtgtatcg ccccgccctg
     1081 ggcggcaagg ctggcccggt cggcaccagt tgcgtgagcg gaaagatggc cgcttcccgg
     1141 ccctgctgca gggagctcaa aatggaggac gcggcgctcg ggagagcggg cgggtgagtc
     1201 acccacacaa aggaaaaggg cctttccgtc ctcagccgtc gcttcatgtg actccacgga
     1261 gtaccgggcg ccgtccaggc acctcgatta gttctcgagc ttttggagta cgtcgtcttt
     1321 aggttggggg gaggggtttt atgcgatgga gtttccccac actgagtggg tggagactga
     1381 agttaggcca gcttggcact tgatgtaatt ctccttggaa tttgcccttt ttgagtttgg
     1441 atcttggttc attctcaagc ctcagacagt ggttcaaagt ttttttcttc catttcaggt
     1501 ggatcccaag ggcagataag tttgtacaaa aaagagaggt gccaccatga gcgagctgat
     1561 taaggagaac atgcacatga agctgtacat ggagggcacc gtggacaacc atcacttcaa
     1621 gtgcacatcc gagggcgaag gcaagcccta cgagggcacc cagaccatga gaatcaaggt
     1681 ggtcgagggc ggccctctcc ccttcgcctt cgacatcctg gctactagct tcctctacgg
     1741 cagcaagacc ttcatcaacc acacccaggg catccccgac ttcttcaagc agtccttccc
     1801 tgagggcttc acatgggaga gagtcaccac atacgaagac gggggcgtgc tgaccgctac
     1861 ccaggacacc agcctccagg acggctgcct catctacaac gtcaagatca gaggggtgaa
     1921 cttcacatcc aacggccctg tgatgcagaa gaaaacactc ggctgggagg ccttcaccga
     1981 gacgctgtac cccgctgacg gcggcctgga aggcagaaac gacatggccc tgaagctcgt
     2041 gggcgggagc catctgatcg caaacatcaa gaccacatat agatccaaga aacccgctaa
     2101 gaacctcaag atgcctggcg tctactatgt ggactacaga ctggaaagaa tcaaggaggc
     2161 caacaacgag acatacgtcg agcagcacga ggtggcagtg gccagatact gcgacctccc
     2221 tagcaaactg gggcacggta gcggcgcaac aaacttctct cttctgaaac aggcaggaga
     2281 tgtggaagaa aatccaggcc caggatccgg tttaattaag ggcctgcagg gtggaaagct
     2341 ttctactcga ggggctagcc agggagaggg cagaggaagt ctgcttacat gcggtgatgt
     2401 cgaagagaat cctggcccag gtagtggaat ggtcttcaca ctcgaagatt tcgttgggga
     2461 ctggcgacag acagccggct acaacctgga ccaagtcctt gaacagggag gtgtgtccag
     2521 tttgtttcag aatctcgggg tgtccgtaac tccgatccaa aggattgtcc tgagcggtga
     2581 aaatgggctg aagatcgaca tccatgtcat catcccgtat gaaggtctga gcggcgacca
     2641 aatgggccag atcgaaaaaa tttttaaggt ggtgtaccct gtggatgatc atcactttaa
     2701 ggtgatcctg cactatggca cactggtaat cgacggggtt acgccgaaca tgatcgacta
     2761 tttcggacgg ccgtatgaag gcatcgccgt gttcgacggc aaaaagatca ctgtaacagg
     2821 gaccctgtgg aacggcaaca aaattatcga cgagcgcctg atcaaccccg acggctccct
     2881 gctgttccga gtaaccatca acggagtgac cggctggcgg ctgtgcgaac gcattctggc
     2941 gggctctggg cagtgtacta attatgctct cttgaaattg gctggagatg ttgagagcaa
     3001 cccaggtccc ggctctggaa tggcgtccaa tttcactcag tttgtgctgg ttgacaacgg
     3061 cgggaccggg gacgttacgg tagccccctc aaactttgcc aacggtatag cggagtggat
     3121 aagcagcaat tctaggagtc aagcatacaa agttacatgc agcgtgcgcc aatctagcgc
     3181 tcagaatcgc aagtacacca ttaaagtaga ggtccccaag ggagcctgga gaagctatct
     3241 taacatggag ttgaccatac caatcttcgc taccaactct gactgtgaac tcattgtgaa
     3301 agccatgcaa ggtctgctca aggatggtaa cccaattccg tccgctatcg ctgccaactc
     3361 tgggatttac gggggcagtg ggagcggtgc aggatctggt agtccagctg ggggaggagc
     3421 accgggtagc ggtggggggt ctcagctgca cctgccccag gttctcgcag acgccgtatc
     3481 ccgccttgta ctgggcaagt ttggtgatct tactgacaat ttttcatctc ctcatgcgag
     3541 gcggaaagta ctcgcaggcg tcgtcatgac gaccggaact gacgtgaaag acgccaaagt
     3601 catctctgtc tccacgggca caaagtgcat aaacggggag tacatgagcg accgggggct
     3661 ggcactgaat gattgtcacg ctgaaataat atctaggcga tctctgctta gatttctcta
     3721 cactcaactc gaattgtacc ttaacaacaa agatgaccag aaacgcagta tatttcagaa
     3781 atcagaacgc ggcggatttc gacttaagga aaacgttcag ttccacttgt atatcagcac
     3841 atccccttgc ggtgacgccc gaatcttttc cccgcacgag ccgatattgg aggagcccgc
     3901 ggacagacat cctaatagga aggctagagg ccaacttcgg acgaagattg aaagtggcca
     3961 gggtactatc ccggtgcggt ccaacgctag tattcaaacg tgggacggag tccttcaagg
     4021 tgaacggctg ttgacaatga gctgctcaga caaaatcgcg cgctggaatg tagtgggaat
     4081 ccaaggcagc ctcttgagca tattcgtaga acccatatat ttctcatcca ttattttggg
     4141 ctctctgtat catggtgacc atctgtcaag ggctatgtac caacgaattt ctaatatcga
     4201 ggatcttcct ccactctata cactcaataa gcctctcttg tccgggatat caaacgctga
     4261 ggcccgccag ccagggaaag ctcctaactt cagtgttaac tggaccgttg gtgattctgc
     4321 gatagaggtc atcaacgcca cgacaggtaa ggatgagctc ggtagagcct cacgcctgtg
     4381 taaacacgcg ttgtattgta gatggatgag agtacatggg aaggtcccat ctcacttgct
     4441 ccgaagcaag atcactaagc ctaatgtgta tcatgagtca aaactcgcgg ctaaagaata
     4501 ccaggcagcc aaagctcgac tttttacagc ttttattaag gcagggctcg gggcatgggt
     4561 cgagaagccg accgagcagg accaattctc tctgacgggg agcggatccg gctctctgcc
     4621 accattggaa aggcttacgc tgtaacttcc actggattgt acaattacca actgatctag
     4681 aaccggtcat ggccgcaata aaatatcttt attttcatta catctgtgtg ttggtttttt
     4741 gtgtgttcga actagatgct gtcgaccgat gcgctggtct aatacccaat ctctcgtctt
     4801 atccagatgt tttatacgcc ttaattaata gggataacag ggtaatatct tgccctgcac
     4861 gaataccagc gaccccttgc ccaaatactt gccgtgggcc tcggcctgag agccaaaaca
     4921 cttgatgcgg aagaagtcgg tgcgctcctg cttgtcgccg gcatcgttgc gccacatcta
     4981 ggatctgcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag gctccgcccc
     5041 cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc gacaggacta
     5101 taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg
     5161 ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc
     5221 tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac
     5281 gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac
     5341 ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat tagcagagcg
     5401 aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg ctacactaga
     5461 agaacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa aagagttggt
     5521 agctcttgat ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag
     5581 cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc tacggggtct
     5641 gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt atcaaaaagg
     5701 atcttcacct agatcctttt aaattaaaaa tgaagtttta aatcaatcta aagtatatat
     5761 gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc
     5821 tgtctatttc gttcatccat agttgcctga ctccccgtcg tgtagataac tacgatacgg
     5881 gagggcttac catctggccc cagtgctgca atgataccgc gagaaccacg ctcaccggct
     5941 ccagatttat cagcaataaa ccagccagcc ggaagggccg agcgcagaag tggtcctgca
     6001 actttatccg cctccatcca gtctattaat tgttgccggg aagctagagt aagtagttcg
     6061 ccagttaata gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt gtcacgctcg
     6121 tcgtttggta tggcttcatt cagctccggt tcccaacgat caaggcgagt tacatgatcc
     6181 cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag
     6241 ttggccgcag tgttatcact catggttatg gcagcactgc ataattctct tactgtcatg
     6301 ccatccgtaa gatgcttttc tgtgactggt gagtactcaa ccaagtcatt ctgagaatag
     6361 tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac gggataatac cgcgccacat
     6421 agcagaactt taaaagtgct catcattgga aaacgttctt cggggcgaaa actctcaagg
     6481 atcttaccgc tgttgagatc cagttcgatg taacccactc gtgcacccaa ctgatcttca
     6541 gcatctttta ctttcaccag cgtttctggg tgagcaaaaa caggaaggca aaatgccgca
     6601 aaaaagggaa taagggcgac acggaaatgt tgaatactca tactcttcct ttttcaatat
     6661 tattgaagca tttatcaagg ttattgtctc atgagcggat acatatttga atgtatttag
     6721 aaaaataaac aaataggggt tccgcgcacg aattggccag cgctgcatta ccctgttatc
     6781 cctaaccgg
//
