LOCUS       sequence-339585-        7163 bp    DNA     circular SYN 14-APR-2025
DEFINITION  Expresses ADAR2 Deaminase Domain (DD) fused with 4 repeats of lambda
            N peptide (4LN) with a Nuclear Localization Signal (NLS) in 
            mammalian cells.
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 Apr 14, 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-4LNDD-pcDNA3.1(+)
FEATURES             Location/Qualifiers
     source          1..7163
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     regulatory      1..10
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             10..33
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             43..63
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             67..87
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             91..111
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             1732..1749
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(1797..1814)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    1803..2027
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      2073..2501
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2160..2179)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2370..2391
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(2510..2530)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        2515..2844
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      2695..2830
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     2757..2776
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             2911..3705
                     /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(2965..2984)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3575..3594
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    3879..4000
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3916..3935)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3970..3989
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4049..4065)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4049..4065)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4062..4084)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4073..4089
                     /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(4097..4103,4104..4121,4122..4127))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4142..4163
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4280..4297)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4451..5039)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4531..4550)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5210..6001,6002..6070))
                     /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     5833..5852
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6071..6175)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(6250..6269)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        6441..6820
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        6821..7024
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     6975..6995
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     7069..7088
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        7069..7087
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
ORIGIN
        1 gccaccatgg actacaagga cgacgatgac aaactagtag ctccaaaaaa gaagagaaag
       61 gtagatccaa aaaagaagag aaaggtagat ccaaaaaaga agagaaaggt aggatccacc
      121 tccggagtaa atgcccgtac gcgccggcgc gaacggcgag cagaaaagca ggcacagtgg
      181 aaggcagcta atggcggagg gggaagcggg ggcggtggaa gtgggggtgg aggcagcaat
      241 gcgcggaccc gccgacgtga acggcgagca gaaaaacagg cgcaatggaa agcagccaac
      301 ggcggcggtg gaagtggagg tggtggaagc ggaggcggtg gtagtaacgc cagaacccga
      361 cgcagagaga ggagggcaga gaaacaggcc cagtggaaag ccgctaacgg aggaggggga
      421 tccggtggag gcggcagtgg aggcggggga tctaacgcac gaacacgacg acgtgagcgt
      481 cgcgctgaga aacaagctca atggaaagct gcaaacttgc acttggatca gacgccatct
      541 cgccagccta ttcccagtga gggtcttcag ctgcatttac cgcaggtttt agctgacgct
      601 gtctcacgcc tggtcctggg taagtttggt gacctgaccg acaacttctc ctcccctcac
      661 gctcgcagaa aagtgctggc tggagtcgtc atgacaacag gcacagatgt taaagatgcc
      721 aaggtgataa gtgtttctac aggaacaaaa tgtattaatg gtgaatacat gagtgatcgt
      781 ggccttgcat taaatgactg ccatgcagaa ataatatctc ggagatcctt gctcagattt
      841 ctttatacac aacttgagct ttacttaaat aacaaagatg atcaaaaaag atccatcttt
      901 cagaaatcag agcgaggggg gtttaggctg aaggagaatg tccagtttca tctgtacatc
      961 agcacctctc cctgtggaga tgccagaatc ttctcaccac atgagccaat cctggaagaa
     1021 ccagcagata gacacccaaa tcgtaaagca agaggacagc tacggaccaa aatagagtct
     1081 ggtgagggga cgattccagt gcgctccaat gcgagcatcc aaacgtggga cggggtgctg
     1141 caaggggagc ggctgctcac catgtcctgc agtgacaaga ttgcacgctg gaacgtggtg
     1201 ggcatccagg gatccctgct cagcattttc gtggagccca tttacttctc gagcatcatc
     1261 ctgggcagcc tttaccacgg ggaccacctt tccagggcca tgtaccagcg gatctccaac
     1321 atagaggacc tgccacctct ctacaccctc aacaagcctt tgctcagtgg catcagcaat
     1381 gcagaagcac ggcagccagg gaaggccccc aacttcagtg tcaactggac ggtaggcgac
     1441 tccgctattg aggtcatcaa cgccacgact gggaaggatg agctgggccg cgcgtcccgc
     1501 ctgtgtaagc acgcgttgta ctgtcgctgg atgcgtgtgc acggcaaggt tccctcccac
     1561 ttactacgct ccaagattac caaacccaac gtgtaccatg agtccaagct ggcggcaaag
     1621 gagtaccagg ccgccaaggc gcgtctgttc acagccttca tcaaggcggg gctgggggcc
     1681 tgggtggaga agcccaccga gcaggaccag ttctcactca cgccactagt ccaccatcac
     1741 catcaccatt gagcggccgc tcgagtctag agggcccgtt taaacccgct gatcagcctc
     1801 gactgtgcct tctagttgcc agccatctgt tgtttgcccc tcccccgtgc cttccttgac
     1861 cctggaaggt gccactccca ctgtcctttc ctaataaaat gaggaaattg catcgcattg
     1921 tctgagtagg tgtcattcta ttctgggggg tggggtgggg caggacagca agggggagga
     1981 ttgggaagac aatagcaggc atgctgggga tgcggtgggc tctatggctt ctgaggcgga
     2041 aagaaccagc tggggctcta gggggtatcc ccacgcgccc tgtagcggcg cattaagcgc
     2101 ggcgggtgtg gtggttacgc gcagcgtgac cgctacactt gccagcgccc tagcgcccgc
     2161 tcctttcgct ttcttccctt cctttctcgc cacgttcgcc ggctttcccc gtcaagctct
     2221 aaatcggggg ctccctttag ggttccgatt tagtgcttta cggcacctcg accccaaaaa
     2281 acttgattag ggtgatggtt cacgtagtgg gccatcgccc tgatagacgg tttttcgccc
     2341 tttgacgttg gagtccacgt tctttaatag tggactcttg ttccaaactg gaacaacact
     2401 caaccctatc tcggtctatt cttttgattt ataagggatt ttgccgattt cggcctattg
     2461 gttaaaaaat gagctgattt aacaaaaatt taacgcgaat taattctgtg gaatgtgtgt
     2521 cagttagggt gtggaaagtc cccaggctcc ccagcaggca gaagtatgca aagcatgcat
     2581 ctcaattagt cagcaaccag gtgtggaaag tccccaggct ccccagcagg cagaagtatg
     2641 caaagcatgc atctcaatta gtcagcaacc atagtcccgc ccctaactcc gcccatcccg
     2701 cccctaactc cgcccagttc cgcccattct ccgccccatg gctgactaat tttttttatt
     2761 tatgcagagg ccgaggccgc ctctgcctct gagctattcc agaagtagtg aggaggcttt
     2821 tttggaggcc taggcttttg caaaaagctc ccgggagctt gtatatccat tttcggatct
     2881 gatcaagaga caggatgagg atcgtttcgc atgattgaac aagatggatt gcacgcaggt
     2941 tctccggccg cttgggtgga gaggctattc ggctatgact gggcacaaca gacaatcggc
     3001 tgctctgatg ccgccgtgtt ccggctgtca gcgcaggggc gcccggttct ttttgtcaag
     3061 accgacctgt ccggtgccct gaatgaactg caggacgagg cagcgcggct atcgtggctg
     3121 gccacgacgg gcgttccttg cgcagctgtg ctcgacgttg tcactgaagc gggaagggac
     3181 tggctgctat tgggcgaagt gccggggcag gatctcctgt catctcacct tgctcctgcc
     3241 gagaaagtat ccatcatggc tgatgcaatg cggcggctgc atacgcttga tccggctacc
     3301 tgcccattcg accaccaagc gaaacatcgc atcgagcgag cacgtactcg gatggaagcc
     3361 ggtcttgtcg atcaggatga tctggacgaa gagcatcagg ggctcgcgcc agccgaactg
     3421 ttcgccaggc tcaaggcgcg catgcccgac ggcgaggatc tcgtcgtgac ccatggcgat
     3481 gcctgcttgc cgaatatcat ggtggaaaat ggccgctttt ctggattcat cgactgtggc
     3541 cggctgggtg tggcggaccg ctatcaggac atagcgttgg ctacccgtga tattgctgaa
     3601 gagcttggcg gcgaatgggc tgaccgcttc ctcgtgcttt acggtatcgc cgctcccgat
     3661 tcgcagcgca tcgccttcta tcgccttctt gacgagttct tctgagcggg actctggggt
     3721 tcgaaatgac cgaccaagcg acgcccaacc tgccatcacg agatttcgat tccaccgccg
     3781 ccttctatga aaggttgggc ttcggaatcg ttttccggga cgccggctgg atgatcctcc
     3841 agcgcgggga tctcatgctg gagttcttcg cccaccccaa cttgtttatt gcagcttata
     3901 atggttacaa ataaagcaat agcatcacaa atttcacaaa taaagcattt ttttcactgc
     3961 attctagttg tggtttgtcc aaactcatca atgtatctta tcatgtctgt ataccgtcga
     4021 cctctagcta gagcttggcg taatcatggt catagctgtt tcctgtgtga aattgttatc
     4081 cgctcacaat tccacacaac atacgagccg gaagcataaa gtgtaaagcc tggggtgcct
     4141 aatgagtgag ctaactcaca ttaattgcgt tgcgctcact gcccgctttc cagtcgggaa
     4201 acctgtcgtg ccagctgcat taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta
     4261 ttgggcgctc ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc
     4321 gagcggtatc agctcactca aaggcggtaa tacggttatc cacagaatca ggggataacg
     4381 caggaaagaa catgtgagca aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt
     4441 tgctggcgtt tttccatagg ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa
     4501 gtcagaggtg gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggaagct
     4561 ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc
     4621 cttcgggaag cgtggcgctt tctcatagct cacgctgtag gtatctcagt tcggtgtagg
     4681 tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct
     4741 tatccggtaa ctatcgtctt gagtccaacc cggtaagaca cgacttatcg ccactggcag
     4801 cagccactgg taacaggatt agcagagcga ggtatgtagg cggtgctaca gagttcttga
     4861 agtggtggcc taactacggc tacactagaa gaacagtatt tggtatctgc gctctgctga
     4921 agccagttac cttcggaaaa agagttggta gctcttgatc cggcaaacaa accaccgctg
     4981 gtagcggtgg tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag
     5041 aagatccttt gatcttttct acggggtctg acgctcagtg gaacgaaaac tcacgttaag
     5101 ggattttggt catgagatta tcaaaaagga tcttcaccta gatcctttta aattaaaaat
     5161 gaagttttaa atcaatctaa agtatatatg agtaaacttg gtctgacagt taccaatgct
     5221 taatcagtga ggcacctatc tcagcgatct gtctatttcg ttcatccata gttgcctgac
     5281 tccccgtcgt gtagataact acgatacggg agggcttacc atctggcccc agtgctgcaa
     5341 tgataccgcg agacccacgc tcaccggctc cagatttatc agcaataaac cagccagccg
     5401 gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag tctattaatt
     5461 gttgccggga agctagagta agtagttcgc cagttaatag tttgcgcaac gttgttgcca
     5521 ttgctacagg catcgtggtg tcacgctcgt cgtttggtat ggcttcattc agctccggtt
     5581 cccaacgatc aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg gttagctcct
     5641 tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt gttatcactc atggttatgg
     5701 cagcactgca taattctctt actgtcatgc catccgtaag atgcttttct gtgactggtg
     5761 agtactcaac caagtcattc tgagaatagt gtatgcggcg accgagttgc tcttgcccgg
     5821 cgtcaatacg ggataatacc gcgccacata gcagaacttt aaaagtgctc atcattggaa
     5881 aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc agttcgatgt
     5941 aacccactcg tgcacccaac tgatcttcag catcttttac tttcaccagc gtttctgggt
     6001 gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt
     6061 gaatactcat actcttcctt tttcaatatt attgaagcat ttatcagggt tattgtctca
     6121 tgagcggata catatttgaa tgtatttaga aaaataaaca aataggggtt ccgcgcacat
     6181 ttccccgaaa agtgccacct gacgtcgacg gatcgggaga tctcccgatc ccctatggtg
     6241 cactctcagt acaatctgct ctgatgccgc atagttaagc cagtatctgc tccctgcttg
     6301 tgtgttggag gtcgctgagt agtgcgcgag caaaatttaa gctacaacaa ggcaaggctt
     6361 gaccgacaat tgcatgaaga atctgcttag ggttaggcgt tttgcgctgc ttcgcgatgt
     6421 acgggccaga tatacgcgtt gacattgatt attgactagt tattaatagt aatcaattac
     6481 ggggtcatta gttcatagcc catatatgga gttccgcgtt acataactta cggtaaatgg
     6541 cccgcctggc tgaccgccca acgacccccg cccattgacg tcaataatga cgtatgttcc
     6601 catagtaacg ccaataggga ctttccattg acgtcaatgg gtggagtatt tacggtaaac
     6661 tgcccacttg gcagtacatc aagtgtatca tatgccaagt acgcccccta ttgacgtcaa
     6721 tgacggtaaa tggcccgcct ggcattatgc ccagtacatg accttatggg actttcctac
     6781 ttggcagtac atctacgtat tagtcatcgc tattaccatg gtgatgcggt tttggcagta
     6841 catcaatggg cgtggatagc ggtttgactc acggggattt ccaagtctcc accccattga
     6901 cgtcaatggg agtttgtttt ggcaccaaaa tcaacgggac tttccaaaat gtcgtaacaa
     6961 ctccgcccca ttgacgcaaa tgggcggtag gcgtgtacgg tgggaggtct atataagcag
     7021 agctctctgg ctaactagag aacccactgc ttactggctt atcgaaatta atacgactca
     7081 ctatagggag acccaagctg gctagcgttt aaacttaagc ttggtaccga gctcggatcc
     7141 actagtccag tgtggtggaa ttc
//
