LOCUS       sequence-339830-        7163 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses ADAR2 Deaminase Domain (DD) hyperactive mutant E488Q fused
            with 4 repeats of lambda N peptide (4LN) with a Nuclear Localization
            Signal (NLS).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7163)
  AUTHORS   Vallecillo-Viejo IC, Liscovitch-Brauer N, Montiel-Gonzalez MF, 
            Eisenberg E, Rosenthal JJC
  TITLE     Abundant off-target edits from site-directed RNA editing can be 
            reduced by nuclear localization of the editing enzyme.
  JOURNAL   RNA Biol. 2018 Jan 2;15(1):104-114. doi: 
            10.1080/15476286.2017.1387711. Epub 2017 Nov 13.
  PUBMED    29099293
REFERENCE   2  (bases 1 to 7163)
  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"; doi: 
            "10.1080/15476286.2017.1387711"; journalName: "RNA Biol"; date: 
            "2018-01-2- 2"; volume: "15"; issue: "1"; pages: "104-114"
COMMENT     Sequence Label: NLS-4LNDDE488Q-pCDNA3.1(+)
FEATURES             Location/Qualifiers
     source          1..7163
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        1..380
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        381..584
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     535..555
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     629..648
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        629..647
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      724..733
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             733..756
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             766..786
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             790..810
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             814..834
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             2455..2472
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(2520..2537)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2526..2750
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      2796..3224
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2883..2902)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3093..3114
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3233..3253)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3238..3567
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3418..3553
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     3480..3499
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             3634..4428
                     /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(3688..3707)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4298..4317
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    4602..4723
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4639..4658)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4693..4712
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4772..4788)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4772..4788)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4785..4807)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4796..4812
                     /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(4820..4826,4827..4844,4845..4850))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4865..4886
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5003..5020)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5174..5762)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5254..5273)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5933..6724,6725..6793))
                     /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     6556..6575
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6794..6898)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(6973..6992)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 gacattgatt attgactagt tattaatagt aatcaattac ggggtcatta gttcatagcc
       61 catatatgga gttccgcgtt acataactta cggtaaatgg cccgcctggc tgaccgccca
      121 acgacccccg cccattgacg tcaataatga cgtatgttcc catagtaacg ccaataggga
      181 ctttccattg acgtcaatgg gtggagtatt tacggtaaac tgcccacttg gcagtacatc
      241 aagtgtatca tatgccaagt acgcccccta ttgacgtcaa tgacggtaaa tggcccgcct
      301 ggcattatgc ccagtacatg accttatggg actttcctac ttggcagtac atctacgtat
      361 tagtcatcgc tattaccatg gtgatgcggt tttggcagta catcaatggg cgtggatagc
      421 ggtttgactc acggggattt ccaagtctcc accccattga cgtcaatggg agtttgtttt
      481 ggcaccaaaa tcaacgggac tttccaaaat gtcgtaacaa ctccgcccca ttgacgcaaa
      541 tgggcggtag gcgtgtacgg tgggaggtct atataagcag agctctctgg ctaactagag
      601 aacccactgc ttactggctt atcgaaatta atacgactca ctatagggag acccaagctg
      661 gctagcgttt aaacttaagc ttggtaccga gctcggatcc actagtccag tgtggtggaa
      721 ttcgccacca tggactacaa ggacgacgat gacaaactag tagctccaaa aaagaagaga
      781 aaggtagatc caaaaaagaa gagaaaggta gatccaaaaa agaagagaaa ggtaggatcc
      841 acctccggag taaatgcccg tacgcgccgg cgcgaacggc gagcagaaaa gcaggcacag
      901 tggaaggcag ctaatggcgg agggggaagc gggggcggtg gaagtggggg tggaggcagc
      961 aatgcgcgga cccgccgacg tgaacggcga gcagaaaaac aggcgcaatg gaaagcagcc
     1021 aacgacggcg gtggaagtgg aggtggtgga agcggaggcg gtggtagtaa cgccagaacc
     1081 cgacgcagag agaggagggc agagaaacag gcccagtgga aagccgctaa cggaggaggg
     1141 ggatccggtg gaggcggcag tggaggcggg ggatctaacg cacgaacacg acgacgtgag
     1201 cgtcgcgctg agaaacaagc tcaatggaaa gctgcaaact tgcacttgga tcagacgcca
     1261 tctcgccagc ctattcccag tgagggtctt cagctgcatt taccgcaggt tttagctgac
     1321 gctgtctcac gcctggtcct gggtaagttt ggtgacctga ccgacaactt ctcctcccct
     1381 cacgctcgca gaaaagtgct ggctggagtc gtcatgacaa caggcacaga tgttaaagat
     1441 gccaaggtga taagtgtttc tacaggaaca aaatgtatta atggtgaata catgagtgat
     1501 cgtggccttg cattaaatga ctgccatgca gaaataatat ctcggagatc cttgctcaga
     1561 tttctttata cacaacttga gctttactta aataacaaag atgatcaaaa aagatccatc
     1621 tttcagaaat cagagcgagg ggggtttagg ctgaaggaga atgtccagtt tcatctgtac
     1681 atcagcacct ctccctgtgg agatgccaga atcttctcac cacatgagcc aatcctggaa
     1741 gaaccagcag atagacaccc aaatcgtaaa gcaagaggac agctacggac caaaatagag
     1801 tctggtcagg ggacgattcc agtgcgctcc aatgcgagca tccaaacgtg ggacggggtg
     1861 ctgcaagggg agcggctgct caccatgtcc tgcagtgaca agattgcacg ctggaacgtg
     1921 gtgggcatcc agggatccct gctcagcatt ttcgtggagc ccatttactt ctcgagcatc
     1981 atcctgggca gcctttacca cggggaccac ctttccaggg ccatgtacca gcggatctcc
     2041 aacatagagg acctgccacc tctctacacc ctcaacaagc ctttgctcag tggcatcagc
     2101 aatgcagaag cacggcagcc agggaaggcc cccaacttca gtgtcaactg gacggtaggc
     2161 gactccgcta ttgaggtcat caacgccacg actgggaagg atgagctggg ccgcgcgtcc
     2221 cgcctgtgta agcacgcgtt gtactgtcgc tggatgcgtg tgcacggcaa ggttccctcc
     2281 cacttactac gctccaagat taccaaaccc aacgtgtacc atgagtccaa gctggcggca
     2341 aaggagtacc aggccgccaa ggcgcgtctg ttcacagcct tcatcaaggc ggggctgggg
     2401 gcctgggtgg agaagcccac cgagcaggac cagttctcac tcacgccact agtccaccat
     2461 caccatcacc attgagcggc cgctcgagtc tagagggccc gtttaaaccc gctgatcagc
     2521 ctcgactgtg ccttctagtt gccagccatc tgttgtttgc ccctcccccg tgccttcctt
     2581 gaccctggaa ggtgccactc ccactgtcct ttcctaataa aatgaggaaa ttgcatcgca
     2641 ttgtctgagt aggtgtcatt ctattctggg gggtggggtg gggcaggaca gcaaggggga
     2701 ggattgggaa gacaatagca ggcatgctgg ggatgcggtg ggctctatgg cttctgaggc
     2761 ggaaagaacc agctggggct ctagggggta tccccacgcg ccctgtagcg gcgcattaag
     2821 cgcggcgggt gtggtggtta cgcgcagcgt gaccgctaca cttgccagcg ccctagcgcc
     2881 cgctcctttc gctttcttcc cttcctttct cgccacgttc gccggctttc cccgtcaagc
     2941 tctaaatcgg gggctccctt tagggttccg atttagtgct ttacggcacc tcgaccccaa
     3001 aaaacttgat tagggtgatg gttcacgtag tgggccatcg ccctgataga cggtttttcg
     3061 ccctttgacg ttggagtcca cgttctttaa tagtggactc ttgttccaaa ctggaacaac
     3121 actcaaccct atctcggtct attcttttga tttataaggg attttgccga tttcggccta
     3181 ttggttaaaa aatgagctga tttaacaaaa atttaacgcg aattaattct gtggaatgtg
     3241 tgtcagttag ggtgtggaaa gtccccaggc tccccagcag gcagaagtat gcaaagcatg
     3301 catctcaatt agtcagcaac caggtgtgga aagtccccag gctccccagc aggcagaagt
     3361 atgcaaagca tgcatctcaa ttagtcagca accatagtcc cgcccctaac tccgcccatc
     3421 ccgcccctaa ctccgcccag ttccgcccat tctccgcccc atggctgact aatttttttt
     3481 atttatgcag aggccgaggc cgcctctgcc tctgagctat tccagaagta gtgaggaggc
     3541 ttttttggag gcctaggctt ttgcaaaaag ctcccgggag cttgtatatc cattttcgga
     3601 tctgatcaag agacaggatg aggatcgttt cgcatgattg aacaagatgg attgcacgca
     3661 ggttctccgg ccgcttgggt ggagaggcta ttcggctatg actgggcaca acagacaatc
     3721 ggctgctctg atgccgccgt gttccggctg tcagcgcagg ggcgcccggt tctttttgtc
     3781 aagaccgacc tgtccggtgc cctgaatgaa ctgcaggacg aggcagcgcg gctatcgtgg
     3841 ctggccacga cgggcgttcc ttgcgcagct gtgctcgacg ttgtcactga agcgggaagg
     3901 gactggctgc tattgggcga agtgccgggg caggatctcc tgtcatctca ccttgctcct
     3961 gccgagaaag tatccatcat ggctgatgca atgcggcggc tgcatacgct tgatccggct
     4021 acctgcccat tcgaccacca agcgaaacat cgcatcgagc gagcacgtac tcggatggaa
     4081 gccggtcttg tcgatcagga tgatctggac gaagagcatc aggggctcgc gccagccgaa
     4141 ctgttcgcca ggctcaaggc gcgcatgccc gacggcgagg atctcgtcgt gacccatggc
     4201 gatgcctgct tgccgaatat catggtggaa aatggccgct tttctggatt catcgactgt
     4261 ggccggctgg gtgtggcgga ccgctatcag gacatagcgt tggctacccg tgatattgct
     4321 gaagagcttg gcggcgaatg ggctgaccgc ttcctcgtgc tttacggtat cgccgctccc
     4381 gattcgcagc gcatcgcctt ctatcgcctt cttgacgagt tcttctgagc gggactctgg
     4441 ggttcgaaat gaccgaccaa gcgacgccca acctgccatc acgagatttc gattccaccg
     4501 ccgccttcta tgaaaggttg ggcttcggaa tcgttttccg ggacgccggc tggatgatcc
     4561 tccagcgcgg ggatctcatg ctggagttct tcgcccaccc caacttgttt attgcagctt
     4621 ataatggtta caaataaagc aatagcatca caaatttcac aaataaagca tttttttcac
     4681 tgcattctag ttgtggtttg tccaaactca tcaatgtatc ttatcatgtc tgtataccgt
     4741 cgacctctag ctagagcttg gcgtaatcat ggtcatagct gtttcctgtg tgaaattgtt
     4801 atccgctcac aattccacac aacatacgag ccggaagcat aaagtgtaaa gcctggggtg
     4861 cctaatgagt gagctaactc acattaattg cgttgcgctc actgcccgct ttccagtcgg
     4921 gaaacctgtc gtgccagctg cattaatgaa tcggccaacg cgcggggaga ggcggtttgc
     4981 gtattgggcg ctcttccgct tcctcgctca ctgactcgct gcgctcggtc gttcggctgc
     5041 ggcgagcggt atcagctcac tcaaaggcgg taatacggtt atccacagaa tcaggggata
     5101 acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg
     5161 cgttgctggc gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct
     5221 caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa
     5281 gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc
     5341 tcccttcggg aagcgtggcg ctttctcata gctcacgctg taggtatctc agttcggtgt
     5401 aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg
     5461 ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg
     5521 cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct
     5581 tgaagtggtg gcctaactac ggctacacta gaagaacagt atttggtatc tgcgctctgc
     5641 tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg
     5701 ctggtagcgg tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc
     5761 aagaagatcc tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt
     5821 aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa
     5881 aatgaagttt taaatcaatc taaagtatat atgagtaaac ttggtctgac agttaccaat
     5941 gcttaatcag tgaggcacct atctcagcga tctgtctatt tcgttcatcc atagttgcct
     6001 gactccccgt cgtgtagata actacgatac gggagggctt accatctggc cccagtgctg
     6061 caatgatacc gcgagaccca cgctcaccgg ctccagattt atcagcaata aaccagccag
     6121 ccggaagggc cgagcgcaga agtggtcctg caactttatc cgcctccatc cagtctatta
     6181 attgttgccg ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg
     6241 ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg
     6301 gttcccaacg atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa gcggttagct
     6361 ccttcggtcc tccgatcgtt gtcagaagta agttggccgc agtgttatca ctcatggtta
     6421 tggcagcact gcataattct cttactgtca tgccatccgt aagatgcttt tctgtgactg
     6481 gtgagtactc aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc
     6541 cggcgtcaat acgggataat accgcgccac atagcagaac tttaaaagtg ctcatcattg
     6601 gaaaacgttc ttcggggcga aaactctcaa ggatcttacc gctgttgaga tccagttcga
     6661 tgtaacccac tcgtgcaccc aactgatctt cagcatcttt tactttcacc agcgtttctg
     6721 ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg aataagggcg acacggaaat
     6781 gttgaatact catactcttc ctttttcaat attattgaag catttatcag ggttattgtc
     6841 tcatgagcgg atacatattt gaatgtattt agaaaaataa acaaataggg gttccgcgca
     6901 catttccccg aaaagtgcca cctgacgtcg acggatcggg agatctcccg atcccctatg
     6961 gtgcactctc agtacaatct gctctgatgc cgcatagtta agccagtatc tgctccctgc
     7021 ttgtgtgttg gaggtcgctg agtagtgcgc gagcaaaatt taagctacaa caaggcaagg
     7081 cttgaccgac aattgcatga agaatctgct tagggttagg cgttttgcgc tgcttcgcga
     7141 tgtacgggcc agatatacgc gtt
//
