LOCUS       sequence-325957-        8480 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  CMV-bpNLS-dCas13d-bpNLS-ADARdd-mCherry, ADARdd embeded in loop3.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8480)
  AUTHORS   Liu Y, Mao S, Huang S, Li Y, Chen Y, Di M, Huang X, Lv J, Wang X, Ge
            J, Shen S, Zhang X, Liu D, Huang X, Chi T
  TITLE     REPAIRx, a specific yet highly efficient programmable A > I RNA base
            editor.
  JOURNAL   EMBO J. 2020 Nov 16;39(22):e104748. doi: 10.15252/embj.2020104748. 
            Epub 2020 Oct 15.
  PUBMED    33058207
REFERENCE   2  (bases 1 to 8480)
  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: "EMBO J."; 
            date: "2020-11-16"; volume: "39(22):e104748. doi"; pages: " 
            10.15252/embj.2020104748. Epub 2020 Oct 15"
COMMENT     Sequence Label: TC1316
FEATURES             Location/Qualifiers
     source          1..8480
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      complement(57..645)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(137..156)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(816..1607,1608..1676))
                     /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     1439..1458
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(1677..1781)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(1856..1875)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        2047..2426
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        2427..2630
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     2581..2601
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     2675..2694
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        2675..2693
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             2771..2791
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             4511..4531
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             5732..5752
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             6938..6958
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             6965..7030
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             7055..7111
                     /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             7118..7828
                     /codon_start=1
                     /product="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
                     /label=mCherry
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEG
                     TQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNF
                     EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK
                     GEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERA
                     EGRHSTGGMDELYK"
     primer_bind     complement(7263..7281)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     7531..7550
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     primer_bind     complement(7867..7884)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    7873..8097
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(8135..8151)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(8135..8151)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(8148..8170)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    8159..8175
                     /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(8183..8189,8190..8207,8208..8213))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    8228..8249
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(8366..8383)
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc
       61 cataggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagtca gaggtggcga
      121 aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct cgtgcgctct
      181 cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc gggaagcgtg
      241 gcgctttctc atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag
      301 ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc cggtaactat
      361 cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc cactggtaac
      421 aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg gtggcctaac
      481 tacggctaca ctagaagaac agtatttggt atctgcgctc tgctgaagcc agttaccttc
      541 ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag cggtggtttt
      601 tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc
      661 ttttctacgg ggtctgacgc tcagtggaac gaaaactcac gttaagggat tttggtcatg
      721 agattatcaa aaaggatctt cacctagatc cttttaaatt aaaaatgaag ttttaaatca
      781 atctaaagta tatatgagta aacttggtct gacagttacc aatgcttaat cagtgaggca
      841 cctatctcag cgatctgtct atttcgttca tccatagttg cctgactccc cgtcgtgtag
      901 ataactacga tacgggaggg cttaccatct ggccccagtg ctgcaatgat accgcgagac
      961 ccacgctcac cggctccaga tttatcagca ataaaccagc cagccggaag ggccgagcgc
     1021 agaagtggtc ctgcaacttt atccgcctcc atccagtcta ttaattgttg ccgggaagct
     1081 agagtaagta gttcgccagt taatagtttg cgcaacgttg ttgccattgc tacaggcatc
     1141 gtggtgtcac gctcgtcgtt tggtatggct tcattcagct ccggttccca acgatcaagg
     1201 cgagttacat gatcccccat gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc
     1261 gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg ttatggcagc actgcataat
     1321 tctcttactg tcatgccatc cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag
     1381 tcattctgag aatagtgtat gcggcgaccg agttgctctt gcccggcgtc aatacgggat
     1441 aataccgcgc cacatagcag aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg
     1501 cgaaaactct caaggatctt accgctgttg agatccagtt cgatgtaacc cactcgtgca
     1561 cccaactgat cttcagcatc ttttactttc accagcgttt ctgggtgagc aaaaacagga
     1621 aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat actcatactc
     1681 ttcctttttc aatattattg aagcatttat cagggttatt gtctcatgag cggatacata
     1741 tttgaatgta tttagaaaaa taaacaaata ggggttccgc gcacatttcc ccgaaaagtg
     1801 ccacctgacg tcgacggatc gggagatctc ccgatcccct atggtgcact ctcagtacaa
     1861 tctgctctga tgccgcatag ttaagccagt atctgctccc tgcttgtgtg ttggaggtcg
     1921 ctgagtagtg cgcgagcaaa atttaagcta caacaaggca aggcttgacc gacaattgca
     1981 tgaagaatct gcttagggtt aggcgttttg cgctgcttcg cgatgtacgg gccagatata
     2041 cgcgttgaca ttgattattg actagttatt aatagtaatc aattacgggg tcattagttc
     2101 atagcccata tatggagttc cgcgttacat aacttacggt aaatggcccg cctggctgac
     2161 cgcccaacga cccccgccca ttgacgtcaa taatgacgta tgttcccata gtaacgccaa
     2221 tagggacttt ccattgacgt caatgggtgg agtatttacg gtaaactgcc cacttggcag
     2281 tacatcaagt gtatcatatg ccaagtacgc cccctattga cgtcaatgac ggtaaatggc
     2341 ccgcctggca ttatgcccag tacatgacct tatgggactt tcctacttgg cagtacatct
     2401 acgtattagt catcgctatt accatggtga tgcggttttg gcagtacatc aatgggcgtg
     2461 gatagcggtt tgactcacgg ggatttccaa gtctccaccc cattgacgtc aatgggagtt
     2521 tgttttggca ccaaaatcaa cgggactttc caaaatgtcg taacaactcc gccccattga
     2581 cgcaaatggg cggtaggcgt gtacggtggg aggtctatat aagcagagct ctctggctaa
     2641 ctagagaacc cactgcttac tggcttatcg aaattaatac gactcactat agggagaccc
     2701 aagctggcta gcgtttaaac ttaagcttgc caccatgaaa cggactgctg acggcagcga
     2761 atttgagagc cccaagaaga agaggaaagt cggatccatc gagaagaaga agagcttcgc
     2821 caagggcatg ggagtgaaga gcaccctggt gtccggctct aaggtgtaca tgaccacatt
     2881 tgctgaggga agcgacgcca ggctggagaa gatcgtggag ggcgatagca tcagatccgt
     2941 gaacgaggga gaggctttca gcgccgagat ggctgacaag aacgctggct acaagatcgg
     3001 aaacgccaag ttttcccacc caaagggcta cgccgtggtg gctaacaacc cactgtacac
     3061 cggaccagtg cagcaggaca tgctgggact gaaggagaca ctggagaaga ggtacttcgg
     3121 cgagtccgcc gacggaaacg ataacatctg catccaggtc atccacaaca tcctggatat
     3181 cgagaagatc ctggctgagt acatcacaaa cgccgcttac gccgtgaaca acatctccgg
     3241 cctggacaag gatatcatcg gcttcggaaa gttttctacc gtgtacacat acgacgagtt
     3301 caaggatcca gagcaccacc gggccgcttt taacaacaac gacaagctga tcaacgccat
     3361 caaggctcag tacgacgagt tcgataactt tctggataac cccaggctgg gctacttcgg
     3421 acaggctttc ttttctaagg agggcagaaa ctacatcatc aactacggaa acgagtgtta
     3481 cgacatcctg gccctgctga gcggactggc ccactgggtg gtggccaaca acgaggagga
     3541 gtctcggatc agccgcacct ggctgtacaa cctggacaag aacctggata acgagtacat
     3601 ctccacactg aactacctgt acgacaggat caccaacgag ctgacaaaca gcttctccaa
     3661 gaactctgcc gctaacgtga actacatcgc tgagaccctg ggcatcaacc cagctgagtt
     3721 cgctgagcag tacttcagat tttccatcat gaaggagcag aagaacctgg gcttcaacat
     3781 cacaaagctg agagaagtga tgctggacag aaaggatatg tccgagatca ggaagaacca
     3841 caaggtgttc gattctatca gaaccaaggt gtacacaatg atggactttg tgatctacag
     3901 gtactacatc gaggaggatg ccaaggtggc cgctgccaac aagagcctgc ccgacaacga
     3961 gaagtctctg agcgagaagg atatcttcgt gatcaacctg agaggctcct ttaacgacga
     4021 tcagaaggac gctctgtact acgatgaggc caacaggatc tggagaaagc tggagaacat
     4081 catgcacaac atcaaggagt tccggggaaa caagacccgc gagtacaaga agaaggacgc
     4141 tccaaggctg cctaggatcc tgcctgctgg aagggacgtg agcgccttca gcaagctgat
     4201 gtacgccctg acaatgtttc tggacggaaa ggagatcaac gatctgctga ccacactgat
     4261 caacaagttc gacaacatcc agtcttttct gaaagtgatg cctctgatcg gcgtgaacgc
     4321 taagttcgtg gaggagtacg ccttctttaa ggacagcgcc aagatcgctg atgagctgcg
     4381 gctgatcaag tcctttgcca ggatgggaga gccaatcgct gacgctagga gagctatgta
     4441 catcgatgcc atccggatcc tgggaaccaa cggttctaag cgcaccgccg atggatccga
     4501 gttcgaatcc cctaaaaaga aaagaaaggt gggatcccag ctgcatttac cgcaggtttt
     4561 agctgacgct gtctcacgcc tggtcctggg taagtttggt gacctgaccg acaacttctc
     4621 ctcccctcac gctcgcagaa aagtgctggc tggagtcgtc atgacaacag gcacagatgt
     4681 taaagatgcc aaggtgataa gtgtttctac aggaacaaaa tgtattaatg gtgaatacat
     4741 gagtgatcgt ggccttgcat taaatgactg ccatgcagaa ataatatctc ggagatcctt
     4801 gctcagattt ctttatacac aacttgagct ttacttaaat aacaaagatg atcaaaaaag
     4861 atccatcttt cagaaatcag agcgaggggg gtttaggctg aaggagaatg tccagtttca
     4921 tctgtacatc agcacctctc cctgtggaga tgccagaatc ttctcaccac atgagccaat
     4981 cctggaagaa ccagcagata gacacccaaa tcgtaaagca agaggacagc tacggaccaa
     5041 aatagagtct ggtcagggga cgattccagt gcgctccaat gcgagcatcc aaacgtggga
     5101 cggggtgctg caaggggagc ggctgctcac catgtcctgc agtgacaaga ttgcacgctg
     5161 gaacgtggtg ggcatccagg gatcactgct cagcattttc gtggagccca tttacttctc
     5221 gagcatcatc ctgggcagcc tttaccacgg ggaccacctt tccagggcca tgtaccagcg
     5281 gatctccaac atagaggacc tgccacctct ctacaccctc aacaagcctt tgctcagtgg
     5341 catcagcaat gcagaagcac ggcagccagg gaaggccccc aacttcagtg tcaactggac
     5401 ggtaggcgac tccgctattg aggtcatcaa cgccacgact gggaaggatg agctgggccg
     5461 cgcgtcccgc ctgtgtaagc acgcgttgta ctgtcgctgg atgcgtgtgc acggcaaggt
     5521 tccctcccac ttactacgct ccaagattac caagcccaac gtgtaccatg agtccaagct
     5581 ggcggcaaag gagtaccagg ccgccaaggc gcgtctgttc acagccttca tcaaggcggg
     5641 gctgggggcc tgggtggaga agcccaccga gcaggaccag ttctcactca cgggatccaa
     5701 aagaactgcg gatggatctg agtttgagag tccaaaaaag aagaggaagg tcggttctat
     5761 gcgcaacttc atcatcaaca acgtgatcag caacaagcgg tttcactacc tgatcagata
     5821 cggcgaccca gctcacctgc acgagatcgc taagaacgag gccgtggtga agttcgtgct
     5881 gggacggatc gccgatatcc agaagaagca gggccagaac ggaaagaacc agatcgaccg
     5941 ctactacgag acctgcatcg gcaaggataa gggaaagtcc gtgtctgaga aggtggacgc
     6001 tctgaccaag atcatcacag gcatgaacta cgaccagttc gataagaaga gatctgtgat
     6061 cgaggacacc ggaagggaga acgccgagag agagaagttt aagaagatca tcagcctgta
     6121 cctgacagtg atctaccaca tcctgaagaa catcgtgaac atcaacgcta gatacgtgat
     6181 cggcttccac tgcgtggagc gcgatgccca gctgtacaag gagaagggat acgacatcaa
     6241 cctgaagaag ctggaggaga agggctttag ctccgtgacc aagctgtgcg ctggaatcga
     6301 cgagacagcc cccgacaaga ggaaggatgt ggagaaggag atggccgaga gagctaagga
     6361 gagcatcgac tccctggagt ctgctaaccc taagctgtac gccaactaca tcaagtactc
     6421 cgatgagaag aaggccgagg agttcaccag gcagatcaac agagagaagg ccaagaccgc
     6481 tctgaacgcc tacctgagga acacaaagtg gaacgtgatc atccgggagg acctgctgcg
     6541 catcgataac aagacctgta cactgttcgc taacaaggct gtggccctgg aggtggctcg
     6601 ctacgtgcac gcctacatca acgacatcgc cgaggtgaac tcctactttc agctgtacca
     6661 ctacatcatg cagaggatca tcatgaacga gagatacgag aagtctagcg gcaaggtgtc
     6721 tgagtacttc gacgccgtga acgatgagaa gaagtacaac gatagactgc tgaagctgct
     6781 gtgcgtgcct ttcggatact gtatcccacg gtttaagaac ctgagcatcg aggccctgtt
     6841 cgaccgcaac gaggctgcca agtttgataa ggagaagaag aaggtgagcg gcaactccgg
     6901 ttctaagcgc accgccgatg gatccgagtt cgaatcccct aaaaagaaaa gaaaggtggg
     6961 atccgactac aaagaccatg acggtgatta taaagatcat gatatcgatt acaaggatga
     7021 cgatgacaag ggatccaaaa gaactggaag cggagctact aacttcagcc tgctgaagca
     7081 ggctggagac gtggaggaga accctggacc tgttaacatg gtgagcaagg gcgaggagga
     7141 taacatggcc atcatcaagg agttcatgcg cttcaaggtg cacatggagg gctccgtgaa
     7201 cggccacgag ttcgagatcg agggcgaggg cgagggccgc ccctacgagg gcacccagac
     7261 cgccaagctg aaggtgacca agggtggccc cctgcccttc gcctgggaca tcctgtcccc
     7321 tcagttcatg tacggctcca aggcctacgt gaagcacccc gccgacatcc ccgactactt
     7381 gaagctgtcc ttccccgagg gcttcaagtg ggagcgcgtg atgaacttcg aggacggcgg
     7441 cgtggtgacc gtgacccagg actcctccct gcaggacggc gagttcatct acaaggtgaa
     7501 gctgcgcggc accaacttcc cctccgacgg ccccgtaatg cagaagaaga ccatgggctg
     7561 ggaggcctcc tccgagcgga tgtaccccga ggacggcgcc ctgaagggcg agatcaagca
     7621 gaggctgaag ctgaaggacg gcggccacta cgacgctgag gtcaagacca cctacaaggc
     7681 caagaagccc gtgcagctgc ccggcgccta caacgtcaac atcaagttgg acatcacctc
     7741 ccacaacgag gactacacca tcgtggaaca gtacgaacgc gccgagggcc gccactccac
     7801 cggcggcatg gacgagctgt acaagtaagc tcgagcctag agggcccgtt taaacccgct
     7861 gatcagcctc gactgtgcct tctagttgcc agccatctgt tgtttgcccc tcccccgtgc
     7921 cttccttgac cctggaaggt gccactccca ctgtcctttc ctaataaaat gaggaaattg
     7981 catcgcattg tctgagtagg tgtcattcta ttctgggggg tggggtgggg caggacagca
     8041 agggggagga ttgggaagac aatagcaggc atgctgggga tgcggtgggc tctatggtac
     8101 cgtcgacctc tagctagagc ttggcgtaat catggtcata gctgtttcct gtgtgaaatt
     8161 gttatccgct cacaattcca cacaacatac gagccggaag cataaagtgt aaagcctggg
     8221 gtgcctaatg agtgagctaa ctcacattaa ttgcgttgcg ctcactgccc gctttccagt
     8281 cgggaaacct gtcgtgccag ctgcattaat gaatcggcca acgcgcgggg agaggcggtt
     8341 tgcgtattgg gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc
     8401 tgcggcgagc ggtatcagct cactcaaagg cggtaatacg gttatccaca gaatcagggg
     8461 ataacgcagg aaagaacatg
//
