LOCUS       sequence-396805-        6973 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses a compact, engineered version of ADAR2 constitutively in 
            mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6973)
  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 6973)
  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: pDV17_pCMV-MCP-ADAR2dd(E488Q)-NES
FEATURES             Location/Qualifiers
     source          1..6973
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        162..541
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        542..745
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     696..716
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     790..809
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        790..808
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      828..837
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             2430..2456
                     /codon_start=1
                     /product="nuclear export signal from the HIV Rev protein 
                     (Fischer et al., 1995)"
                     /label=NES
                     /translation="LPPLERLTL"
     primer_bind     complement(2491..2508)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2497..2721
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      2767..3195
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2854..2873)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3064..3085
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3204..3224)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3209..3538
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3389..3524
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     3451..3470
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             3605..4399
                     /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(3659..3678)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4269..4288
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    4573..4694
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4610..4629)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4664..4683
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4743..4759)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4743..4759)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4756..4778)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4767..4783
                     /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(4791..4797,4798..4815,4816..4821))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4836..4857
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4974..4991)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5145..5733)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5225..5244)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5904..6695,6696..6764))
                     /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     6527..6546
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6765..6869)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(6944..6963)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 ccagtatctg ctccctgctt gtgtgttgga ggtcgctgag tagtgcgcga gcaaaattta
       61 agctacaaca aggcaaggct tgaccgacaa ttgcatgaag aatctgctta gggttaggcg
      121 ttttgcgctg cttcgcgatg tacgggccag atatacgcgt tgacattgat tattgactag
      181 ttattaatag taatcaatta cggggtcatt agttcatagc ccatatatgg agttccgcgt
      241 tacataactt acggtaaatg gcccgcctgg ctgaccgccc aacgaccccc gcccattgac
      301 gtcaataatg acgtatgttc ccatagtaac gccaataggg actttccatt gacgtcaatg
      361 ggtggagtat ttacggtaaa ctgcccactt ggcagtacat caagtgtatc atatgccaag
      421 tacgccccct attgacgtca atgacggtaa atggcccgcc tggcattatg cccagtacat
      481 gaccttatgg gactttccta cttggcagta catctacgta ttagtcatcg ctattaccat
      541 ggtgatgcgg ttttggcagt acatcaatgg gcgtggatag cggtttgact cacggggatt
      601 tccaagtctc caccccattg acgtcaatgg gagtttgttt tggcaccaaa atcaacggga
      661 ctttccaaaa tgtcgtaaca actccgcccc attgacgcaa atgggcggta ggcgtgtacg
      721 gtgggaggtc tatataagca gagctctctg gctaactaga gaacccactg cttactggct
      781 tatcgaaatt aatacgactc actataggga gacccaagct ggctagcgcc accatggcgt
      841 ccaatttcac tcagtttgtg ctggttgaca acggcgggac cggggacgtt acggtagccc
      901 cctcaaactt tgccaacggt atagcggagt ggataagcag caattctagg agtcaagcat
      961 acaaagttac atgcagcgtg cgccaatcta gcgctcagaa tcgcaagtac accattaaag
     1021 tagaggtccc caagggagcc tggagaagct atcttaacat ggagttgacc ataccaatct
     1081 tcgctaccaa ctctgactgt gaactcattg tgaaagccat gcaaggtctg ctcaaggatg
     1141 gtaacccaat tccgtccgct atcgctgcca actctgggat ttacgggggc agtgggagcg
     1201 gtgcaggatc tggtagtcca gctgggggag gagcaccggg tagcggtggg gggtctcagc
     1261 tgcacctgcc ccaggttctc gcagacgccg tatcccgcct tgtactgggc aagtttggtg
     1321 atcttactga caatttttca tctcctcatg cgaggcggaa agtactcgca ggcgtcgtca
     1381 tgacgaccgg aactgacgtg aaagacgcca aagtcatctc tgtctccacg ggcacaaagt
     1441 gcataaacgg ggagtacatg agcgaccggg ggctggcact gaatgattgt cacgctgaaa
     1501 taatatctag gcgatctctg cttagatttc tctacactca actcgaattg taccttaaca
     1561 acaaagatga ccagaaacgc agtatatttc agaaatcaga acgcggcgga tttcgactta
     1621 aggaaaacgt tcagttccac ttgtatatca gcacatcccc ttgcggtgac gcccgaatct
     1681 tttccccgca cgagccgata ttggaggagc ccgcggacag acatcctaat aggaaggcta
     1741 gaggccaact tcggacgaag attgaaagtg gccagggtac tatcccggtg cggtccaacg
     1801 ctagtattca aacgtgggac ggagtccttc aaggtgaacg gctgttgaca atgagctgct
     1861 cagacaaaat cgcgcgctgg aatgtagtgg gaatccaagg cagcctcttg agcatattcg
     1921 tagaacccat atatttctca tccattattt tgggctctct gtatcatggt gaccatctgt
     1981 caagggctat gtaccaacga atttctaata tcgaggatct tcctccactc tatacactca
     2041 ataagcctct cttgtccggg atatcaaacg ctgaggcccg ccagccaggg aaagctccta
     2101 acttcagtgt taactggacc gttggtgatt ctgcgataga ggtcatcaac gccacgacag
     2161 gtaaggatga gctcggtaga gcctcacgcc tgtgtaaaca cgcgttgtat tgtagatgga
     2221 tgagagtaca tgggaaggtc ccatctcact tgctccgaag caagatcact aagcctaatg
     2281 tgtatcatga gtcaaaactc gcggctaaag aataccaggc agccaaagct cgacttttta
     2341 cagcttttat taaggcaggg ctcggggcat gggtcgagaa gccgaccgag caggaccaat
     2401 tctctctgac ggggagcgga tccggctctc tgccaccatt ggaaaggctt acgctgtaat
     2461 ctagagggcc cgtttaaacc cgctgatcag cctcgactgt gccttctagt tgccagccat
     2521 ctgttgtttg cccctccccc gtgccttcct tgaccctgga aggtgccact cccactgtcc
     2581 tttcctaata aaatgaggaa attgcatcgc attgtctgag taggtgtcat tctattctgg
     2641 ggggtggggt ggggcaggac agcaaggggg aggattggga agacaatagc aggcatgctg
     2701 gggatgcggt gggctctatg gcttctgagg cggaaagaac cagctggggc tctagggggt
     2761 atccccacgc gccctgtagc ggcgcattaa gcgcggcggg tgtggtggtt acgcgcagcg
     2821 tgaccgctac acttgccagc gccctagcgc ccgctccttt cgctttcttc ccttcctttc
     2881 tcgccacgtt cgccggcttt ccccgtcaag ctctaaatcg ggggctccct ttagggttcc
     2941 gatttagtgc tttacggcac ctcgacccca aaaaacttga ttagggtgat ggttcacgta
     3001 gtgggccatc gccctgatag acggtttttc gccctttgac gttggagtcc acgttcttta
     3061 atagtggact cttgttccaa actggaacaa cactcaaccc tatctcggtc tattcttttg
     3121 atttataagg gattttgccg atttcggcct attggttaaa aaatgagctg atttaacaaa
     3181 aatttaacgc gaattaattc tgtggaatgt gtgtcagtta gggtgtggaa agtccccagg
     3241 ctccccagca ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa ccaggtgtgg
     3301 aaagtcccca ggctccccag caggcagaag tatgcaaagc atgcatctca attagtcagc
     3361 aaccatagtc ccgcccctaa ctccgcccat cccgccccta actccgccca gttccgccca
     3421 ttctccgccc catggctgac taattttttt tatttatgca gaggccgagg ccgcctctgc
     3481 ctctgagcta ttccagaagt agtgaggagg cttttttgga ggcctaggct tttgcaaaaa
     3541 gctcccggga gcttgtatat ccattttcgg atctgatcaa gagacaggat gaggatcgtt
     3601 tcgcatgatt gaacaagatg gattgcacgc aggttctccg gccgcttggg tggagaggct
     3661 attcggctat gactgggcac aacagacaat cggctgctct gatgccgccg tgttccggct
     3721 gtcagcgcag gggcgcccgg ttctttttgt caagaccgac ctgtccggtg ccctgaatga
     3781 actgcaggac gaggcagcgc ggctatcgtg gctggccacg acgggcgttc cttgcgcagc
     3841 tgtgctcgac gttgtcactg aagcgggaag ggactggctg ctattgggcg aagtgccggg
     3901 gcaggatctc ctgtcatctc accttgctcc tgccgagaaa gtatccatca tggctgatgc
     3961 aatgcggcgg ctgcatacgc ttgatccggc tacctgccca ttcgaccacc aagcgaaaca
     4021 tcgcatcgag cgagcacgta ctcggatgga agccggtctt gtcgatcagg atgatctgga
     4081 cgaagagcat caggggctcg cgccagccga actgttcgcc aggctcaagg cgcgcatgcc
     4141 cgacggcgag gatctcgtcg tgacccatgg cgatgcctgc ttgccgaata tcatggtgga
     4201 aaatggccgc ttttctggat tcatcgactg tggccggctg ggtgtggcgg accgctatca
     4261 ggacatagcg ttggctaccc gtgatattgc tgaagagctt ggcggcgaat gggctgaccg
     4321 cttcctcgtg ctttacggta tcgccgctcc cgattcgcag cgcatcgcct tctatcgcct
     4381 tcttgacgag ttcttctgag cgggactctg gggttcgaaa tgaccgacca agcgacgccc
     4441 aacctgccat cacgagattt cgattccacc gccgccttct atgaaaggtt gggcttcgga
     4501 atcgttttcc gggacgccgg ctggatgatc ctccagcgcg gggatctcat gctggagttc
     4561 ttcgcccacc ccaacttgtt tattgcagct tataatggtt acaaataaag caatagcatc
     4621 acaaatttca caaataaagc atttttttca ctgcattcta gttgtggttt gtccaaactc
     4681 atcaatgtat cttatcatgt ctgtataccg tcgacctcta gctagagctt ggcgtaatca
     4741 tggtcatagc tgtttcctgt gtgaaattgt tatccgctca caattccaca caacatacga
     4801 gccggaagca taaagtgtaa agcctggggt gcctaatgag tgagctaact cacattaatt
     4861 gcgttgcgct cactgcccgc tttccagtcg ggaaacctgt cgtgccagct gcattaatga
     4921 atcggccaac gcgcggggag aggcggtttg cgtattgggc gctcttccgc ttcctcgctc
     4981 actgactcgc tgcgctcggt cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg
     5041 gtaatacggt tatccacaga atcaggggat aacgcaggaa agaacatgtg agcaaaaggc
     5101 cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca taggctccgc
     5161 ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga
     5221 ctataaagat accaggcgtt tccccctgga agctccctcg tgcgctctcc tgttccgacc
     5281 ctgccgctta ccggatacct gtccgccttt ctcccttcgg gaagcgtggc gctttctcat
     5341 agctcacgct gtaggtatct cagttcggtg taggtcgttc gctccaagct gggctgtgtg
     5401 cacgaacccc ccgttcagcc cgaccgctgc gccttatccg gtaactatcg tcttgagtcc
     5461 aacccggtaa gacacgactt atcgccactg gcagcagcca ctggtaacag gattagcaga
     5521 gcgaggtatg taggcggtgc tacagagttc ttgaagtggt ggcctaacta cggctacact
     5581 agaagaacag tatttggtat ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt
     5641 ggtagctctt gatccggcaa acaaaccacc gctggtagcg gtggtttttt tgtttgcaag
     5701 cagcagatta cgcgcagaaa aaaaggatct caagaagatc ctttgatctt ttctacgggg
     5761 tctgacgctc agtggaacga aaactcacgt taagggattt tggtcatgag attatcaaaa
     5821 aggatcttca cctagatcct tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata
     5881 tatgagtaaa cttggtctga cagttaccaa tgcttaatca gtgaggcacc tatctcagcg
     5941 atctgtctat ttcgttcatc catagttgcc tgactccccg tcgtgtagat aactacgata
     6001 cgggagggct taccatctgg ccccagtgct gcaatgatac cgcgagaccc acgctcaccg
     6061 gctccagatt tatcagcaat aaaccagcca gccggaaggg ccgagcgcag aagtggtcct
     6121 gcaactttat ccgcctccat ccagtctatt aattgttgcc gggaagctag agtaagtagt
     6181 tcgccagtta atagtttgcg caacgttgtt gccattgcta caggcatcgt ggtgtcacgc
     6241 tcgtcgtttg gtatggcttc attcagctcc ggttcccaac gatcaaggcg agttacatga
     6301 tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt tgtcagaagt
     6361 aagttggccg cagtgttatc actcatggtt atggcagcac tgcataattc tcttactgtc
     6421 atgccatccg taagatgctt ttctgtgact ggtgagtact caaccaagtc attctgagaa
     6481 tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca
     6541 catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg aaaactctca
     6601 aggatcttac cgctgttgag atccagttcg atgtaaccca ctcgtgcacc caactgatct
     6661 tcagcatctt ttactttcac cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc
     6721 gcaaaaaagg gaataagggc gacacggaaa tgttgaatac tcatactctt cctttttcaa
     6781 tattattgaa gcatttatca gggttattgt ctcatgagcg gatacatatt tgaatgtatt
     6841 tagaaaaata aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc acctgacgtc
     6901 gacggatcgg gagatctccc gatcccctat ggtgcactct cagtacaatc tgctctgatg
     6961 ccgcatagtt aag
//
