LOCUS       sequence-339581-        7073 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) in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7073)
  AUTHORS   Montiel-Gonzalez MF, Vallecillo-Viejo IC, Rosenthal JJ
  TITLE     An efficient system for selectively altering genetic information 
            within mRNAs.
  JOURNAL   Nucleic Acids Res. 2016 Dec 1;44(21):e157. doi: 10.1093/nar/gkw738. 
            Epub 2016 Aug 23.
  PUBMED    27557710
REFERENCE   2  (bases 1 to 7073)
  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: "Nucleic 
            Acids Res."; date: "2016-12-1"; volume: "44(21):e157. doi"; pages: "
            10.1093/nar/gkw738. Epub 2016 Aug 23"
COMMENT     Sequence Label: 4LNDDE488Q-pCDNA3.1(+)
FEATURES             Location/Qualifiers
     source          1..7073
                     /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             1642..1659
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(1707..1724)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    1713..1937
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      1983..2411
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2070..2089)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2280..2301
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(2420..2440)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        2425..2754
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      2605..2740
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     2667..2686
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             2821..3615
                     /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(2875..2894)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3485..3504
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    3789..3910
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3826..3845)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3880..3899
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(3959..3975)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3959..3975)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3972..3994)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3983..3999
                     /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(4007..4013,4014..4031,4032..4037))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4052..4073
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4190..4207)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4361..4949)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4441..4460)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5120..5911,5912..5980))
                     /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     5743..5762
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5981..6085)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(6160..6179)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        6351..6730
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        6731..6934
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     6885..6905
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     6979..6998
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        6979..6997
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
ORIGIN
        1 gccaccatgg actacaagga cgacgatgac aaactagtaa atgcccgtac gcgccggcgc
       61 gaacggcgag cagaaaagca ggcacagtgg aaggcagcta atggcggagg gggaagcggg
      121 ggcggtggaa gtgggggtgg aggcagcaat gcgcggaccc gccgacgtga acggcgagca
      181 gaaaaacagg cgcaatggaa agcagccaac ggcggcggtg gaagtggagg tggtggaagc
      241 ggaggcggtg gtagtaacgc cagaacccga cgcagagaga ggagggcaga gaaacaggcc
      301 cagtggaaag ccgctaacgg aggaggggga tccggtggag gcggcagtgg aggcggggga
      361 tctaacgcac gaacacgacg acgtgagcgt cgcgctgaga aacaagctca atggaaagct
      421 gcaaacttgc acttggatca gacgccatct cgccagccta ttcccagtga gggtcttcag
      481 ctgcatttac cgcaggtttt agctgacgct gtctcacgcc tggtcctggg taagtttggt
      541 gacctgaccg acaacttctc ctcccctcac gctcgcagaa aagtgctggc tggagtcgtc
      601 atgacaacag gcacagatgt taaagatgcc aaggtgataa gtgtttctac aggaacaaaa
      661 tgtattaatg gtgaatacat gagtgatcgt ggccttgcat taaatgactg ccatgcagaa
      721 ataatatctc ggagatcctt gctcagattt ctttatacac aacttgagct ttacttaaat
      781 aacaaagatg atcaaaaaag atccatcttt cagaaatcag agcgaggggg gtttaggctg
      841 aaggagaatg tccagtttca tctgtacatc agcacctctc cctgtggaga tgccagaatc
      901 ttctcaccac atgagccaat cctggaagaa ccagcagata gacacccaaa tcgtaaagca
      961 agaggacagc tacggaccaa aatagagtct ggtcagggga cgattccagt gcgctccaat
     1021 gcgagcatcc aaacgtggga cggggtgctg caaggggagc ggctgctcac catgtcctgc
     1081 agtgacaaga ttgcacgctg gaacgtggtg ggcatccagg gatccctgct cagcattttc
     1141 gtggagccca tttacttctc gagcatcatc ctgggcagcc tttaccacgg ggaccacctt
     1201 tccagggcca tgtaccagcg gatctccaac atagaggacc tgccacctct ctacaccctc
     1261 aacaagcctt tgctcagtgg catcagcaat gcagaagcac ggcagccagg gaaggccccc
     1321 aacttcagtg tcaactggac ggtaggcgac tccgctattg aggtcatcaa cgccacgact
     1381 gggaaggatg agctgggccg cgcgtcccgc ctgtgtaagc acgcgttgta ctgtcgctgg
     1441 atgcgtgtgc acggcaaggt tccctcccac ttactacgct ccaagattac caaacccaac
     1501 gtgtaccatg agtccaagct ggcggcaaag gagtaccagg ccgccaaggc gcgtctgttc
     1561 acagccttca tcaaggcggg gctgggggcc tgggtggaga agcccaccga gcaggaccag
     1621 ttctcactca cgccactagt ccaccatcac catcaccatt gagcggccgc tcgagtctag
     1681 agggcccgtt taaacccgct gatcagcctc gactgtgcct tctagttgcc agccatctgt
     1741 tgtttgcccc tcccccgtgc cttccttgac cctggaaggt gccactccca ctgtcctttc
     1801 ctaataaaat gaggaaattg catcgcattg tctgagtagg tgtcattcta ttctgggggg
     1861 tggggtgggg caggacagca agggggagga ttgggaagac aatagcaggc atgctgggga
     1921 tgcggtgggc tctatggctt ctgaggcgga aagaaccagc tggggctcta gggggtatcc
     1981 ccacgcgccc tgtagcggcg cattaagcgc ggcgggtgtg gtggttacgc gcagcgtgac
     2041 cgctacactt gccagcgccc tagcgcccgc tcctttcgct ttcttccctt cctttctcgc
     2101 cacgttcgcc ggctttcccc gtcaagctct aaatcggggg ctccctttag ggttccgatt
     2161 tagtgcttta cggcacctcg accccaaaaa acttgattag ggtgatggtt cacgtagtgg
     2221 gccatcgccc tgatagacgg tttttcgccc tttgacgttg gagtccacgt tctttaatag
     2281 tggactcttg ttccaaactg gaacaacact caaccctatc tcggtctatt cttttgattt
     2341 ataagggatt ttgccgattt cggcctattg gttaaaaaat gagctgattt aacaaaaatt
     2401 taacgcgaat taattctgtg gaatgtgtgt cagttagggt gtggaaagtc cccaggctcc
     2461 ccagcaggca gaagtatgca aagcatgcat ctcaattagt cagcaaccag gtgtggaaag
     2521 tccccaggct ccccagcagg cagaagtatg caaagcatgc atctcaatta gtcagcaacc
     2581 atagtcccgc ccctaactcc gcccatcccg cccctaactc cgcccagttc cgcccattct
     2641 ccgccccatg gctgactaat tttttttatt tatgcagagg ccgaggccgc ctctgcctct
     2701 gagctattcc agaagtagtg aggaggcttt tttggaggcc taggcttttg caaaaagctc
     2761 ccgggagctt gtatatccat tttcggatct gatcaagaga caggatgagg atcgtttcgc
     2821 atgattgaac aagatggatt gcacgcaggt tctccggccg cttgggtgga gaggctattc
     2881 ggctatgact gggcacaaca gacaatcggc tgctctgatg ccgccgtgtt ccggctgtca
     2941 gcgcaggggc gcccggttct ttttgtcaag accgacctgt ccggtgccct gaatgaactg
     3001 caggacgagg cagcgcggct atcgtggctg gccacgacgg gcgttccttg cgcagctgtg
     3061 ctcgacgttg tcactgaagc gggaagggac tggctgctat tgggcgaagt gccggggcag
     3121 gatctcctgt catctcacct tgctcctgcc gagaaagtat ccatcatggc tgatgcaatg
     3181 cggcggctgc atacgcttga tccggctacc tgcccattcg accaccaagc gaaacatcgc
     3241 atcgagcgag cacgtactcg gatggaagcc ggtcttgtcg atcaggatga tctggacgaa
     3301 gagcatcagg ggctcgcgcc agccgaactg ttcgccaggc tcaaggcgcg catgcccgac
     3361 ggcgaggatc tcgtcgtgac ccatggcgat gcctgcttgc cgaatatcat ggtggaaaat
     3421 ggccgctttt ctggattcat cgactgtggc cggctgggtg tggcggaccg ctatcaggac
     3481 atagcgttgg ctacccgtga tattgctgaa gagcttggcg gcgaatgggc tgaccgcttc
     3541 ctcgtgcttt acggtatcgc cgctcccgat tcgcagcgca tcgccttcta tcgccttctt
     3601 gacgagttct tctgagcggg actctggggt tcgaaatgac cgaccaagcg acgcccaacc
     3661 tgccatcacg agatttcgat tccaccgccg ccttctatga aaggttgggc ttcggaatcg
     3721 ttttccggga cgccggctgg atgatcctcc agcgcgggga tctcatgctg gagttcttcg
     3781 cccaccccaa cttgtttatt gcagcttata atggttacaa ataaagcaat agcatcacaa
     3841 atttcacaaa taaagcattt ttttcactgc attctagttg tggtttgtcc aaactcatca
     3901 atgtatctta tcatgtctgt ataccgtcga cctctagcta gagcttggcg taatcatggt
     3961 catagctgtt tcctgtgtga aattgttatc cgctcacaat tccacacaac atacgagccg
     4021 gaagcataaa gtgtaaagcc tggggtgcct aatgagtgag ctaactcaca ttaattgcgt
     4081 tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg ccagctgcat taatgaatcg
     4141 gccaacgcgc ggggagaggc ggtttgcgta ttgggcgctc ttccgcttcc tcgctcactg
     4201 actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc agctcactca aaggcggtaa
     4261 tacggttatc cacagaatca ggggataacg caggaaagaa catgtgagca aaaggccagc
     4321 aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg ctccgccccc
     4381 ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg acaggactat
     4441 aaagatacca ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc
     4501 cgcttaccgg atacctgtcc gcctttctcc cttcgggaag cgtggcgctt tctcatagct
     4561 cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg
     4621 aaccccccgt tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc
     4681 cggtaagaca cgacttatcg ccactggcag cagccactgg taacaggatt agcagagcga
     4741 ggtatgtagg cggtgctaca gagttcttga agtggtggcc taactacggc tacactagaa
     4801 gaacagtatt tggtatctgc gctctgctga agccagttac cttcggaaaa agagttggta
     4861 gctcttgatc cggcaaacaa accaccgctg gtagcggtgg tttttttgtt tgcaagcagc
     4921 agattacgcg cagaaaaaaa ggatctcaag aagatccttt gatcttttct acggggtctg
     4981 acgctcagtg gaacgaaaac tcacgttaag ggattttggt catgagatta tcaaaaagga
     5041 tcttcaccta gatcctttta aattaaaaat gaagttttaa atcaatctaa agtatatatg
     5101 agtaaacttg gtctgacagt taccaatgct taatcagtga ggcacctatc tcagcgatct
     5161 gtctatttcg ttcatccata gttgcctgac tccccgtcgt gtagataact acgatacggg
     5221 agggcttacc atctggcccc agtgctgcaa tgataccgcg agacccacgc tcaccggctc
     5281 cagatttatc agcaataaac cagccagccg gaagggccga gcgcagaagt ggtcctgcaa
     5341 ctttatccgc ctccatccag tctattaatt gttgccggga agctagagta agtagttcgc
     5401 cagttaatag tttgcgcaac gttgttgcca ttgctacagg catcgtggtg tcacgctcgt
     5461 cgtttggtat ggcttcattc agctccggtt cccaacgatc aaggcgagtt acatgatccc
     5521 ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc agaagtaagt
     5581 tggccgcagt gttatcactc atggttatgg cagcactgca taattctctt actgtcatgc
     5641 catccgtaag atgcttttct gtgactggtg agtactcaac caagtcattc tgagaatagt
     5701 gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg ggataatacc gcgccacata
     5761 gcagaacttt aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa ctctcaagga
     5821 tcttaccgct gttgagatcc agttcgatgt aacccactcg tgcacccaac tgatcttcag
     5881 catcttttac tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa
     5941 aaaagggaat aagggcgaca cggaaatgtt gaatactcat actcttcctt tttcaatatt
     6001 attgaagcat ttatcagggt tattgtctca tgagcggata catatttgaa tgtatttaga
     6061 aaaataaaca aataggggtt ccgcgcacat ttccccgaaa agtgccacct gacgtcgacg
     6121 gatcgggaga tctcccgatc ccctatggtg cactctcagt acaatctgct ctgatgccgc
     6181 atagttaagc cagtatctgc tccctgcttg tgtgttggag gtcgctgagt agtgcgcgag
     6241 caaaatttaa gctacaacaa ggcaaggctt gaccgacaat tgcatgaaga atctgcttag
     6301 ggttaggcgt tttgcgctgc ttcgcgatgt acgggccaga tatacgcgtt gacattgatt
     6361 attgactagt tattaatagt aatcaattac ggggtcatta gttcatagcc catatatgga
     6421 gttccgcgtt acataactta cggtaaatgg cccgcctggc tgaccgccca acgacccccg
     6481 cccattgacg tcaataatga cgtatgttcc catagtaacg ccaataggga ctttccattg
     6541 acgtcaatgg gtggagtatt tacggtaaac tgcccacttg gcagtacatc aagtgtatca
     6601 tatgccaagt acgcccccta ttgacgtcaa tgacggtaaa tggcccgcct ggcattatgc
     6661 ccagtacatg accttatggg actttcctac ttggcagtac atctacgtat tagtcatcgc
     6721 tattaccatg gtgatgcggt tttggcagta catcaatggg cgtggatagc ggtttgactc
     6781 acggggattt ccaagtctcc accccattga cgtcaatggg agtttgtttt ggcaccaaaa
     6841 tcaacgggac tttccaaaat gtcgtaacaa ctccgcccca ttgacgcaaa tgggcggtag
     6901 gcgtgtacgg tgggaggtct atataagcag agctctctgg ctaactagag aacccactgc
     6961 ttactggctt atcgaaatta atacgactca ctatagggag acccaagctg gctagcgttt
     7021 aaacttaagc ttggtaccga gctcggatcc actagtccag tgtggtggaa ttc
//
