LOCUS       sequence-240561-        4551 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses the TadA-TadA* dimer in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4551)
  AUTHORS   Rees HA, Wilson C, Doman JL, Liu DR
  TITLE     Analysis and minimization of cellular RNA editing by DNA adenine 
            base editors.
  JOURNAL   Sci Adv. 2019 May 8;5(5):eaax5717. doi: 10.1126/sciadv.aax5717. 
            eCollection 2019 May.
  PUBMED    31086823
REFERENCE   2  (bases 1 to 4551)
  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: "Sci Adv."; 
            date: "2019-05-8"; volume: "5(5):eaax5717. doi"; pages: " 
            10.1126/sciadv.aax5717. eCollection 2019 May"
COMMENT     Sequence Label: pCMV-TadA-TadA*
FEATURES             Location/Qualifiers
     source          1..4551
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        132..335
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     286..306
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     377..396
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        377..395
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             445..465
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             1566..1583
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(1606..1623)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    1612..1836
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(1907..1923)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1907..1923)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1920..1942)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1931..1947
                     /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(1955..1961,1962..1979,1980..1985))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2000..2021
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2138..2155)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2309..2897)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2389..2408)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3068..3859,3860..3928))
                     /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     3691..3710
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3929..4033)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(4112..4131)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        join(4303..4551,1..131)
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
ORIGIN
        1 atatgccaag tacgccccct attgacgtca atgacggtaa atggcccgcc tggcattatg
       61 cccagtacat gaccttatgg gactttccta cttggcagta catctacgta ttagtcatcg
      121 ctattaccat ggtgatgcgg ttttggcagt acatcaatgg gcgtggatag cggtttgact
      181 cacggggatt tccaagtctc caccccattg acgtcaatgg gagtttgttt tggcaccaaa
      241 atcaacggga ctttccaaaa tgtcgtaaca actccgcccc attgacgcaa atgggcggta
      301 ggcgtgtacg gtgggaggtc tatataagca gagctggttt agtgaaccgt cagatccgct
      361 agagatccgc ggccgctaat acgactcact atagggagag ccgccaccat gaaacggaca
      421 gccgacggaa gcgagttcga gtcaccaaag aagaagcgga aagtctctga agtcgagttt
      481 agccacgagt attggatgag gcacgcactg accctggcaa agcgagcatg ggatgaaaga
      541 gaagtccccg tgggcgccgt gctggtgcac aacaatagag tgatcggaga gggatggaac
      601 aggccaatcg gccgccacga ccctaccgca cacgcagaga tcatggcact gaggcaggga
      661 ggcctggtca tgcagaatta ccgcctgatc gatgccaccc tgtatgtgac actggagcca
      721 tgcgtgatgt gcgcaggagc aatgatccac agcaggatcg gaagagtggt gttcggagca
      781 cgggacgcca agaccggcgc agcaggctcc ctgatggatg tgctgcacca ccccggcatg
      841 aaccaccggg tggagatcac agagggaatc ctggcagacg agtgcgccgc cctgctgagc
      901 gatttcttta gaatgcggag acaggagatc aaggcccaga agaaggcaca gagctccacc
      961 gactctggag gatctagcgg aggatcctct ggaagcgaga caccaggcac aagcgagtcc
     1021 gccacaccag agagctccgg cggctcctcc ggaggatcct ctgaggtgga gttttcccac
     1081 gagtactgga tgagacatgc cctgaccctg gccaagaggg cacgcgatga gagggaggtg
     1141 cctgtgggag ccgtgctggt gctgaacaat agagtgatcg gcgagggctg gaacagagcc
     1201 atcggcctgc acgacccaac agcccatgcc gaaattatgg ccctgagaca gggcggcctg
     1261 gtcatgcaga actacagact gattgacgcc accctgtacg tgacattcga gccttgcgtg
     1321 atgtgcgccg gcgccatgat ccactctagg atcggccgcg tggtgtttgg cgtgaggaac
     1381 gcaaaaaccg gcgccgcagg ctccctgatg gacgtgctgc actaccccgg catgaatcac
     1441 cgcgtcgaaa ttaccgaggg aatcctggca gatgaatgtg ccgccctgct gtgctatttc
     1501 tttcggatgc ctagacaggt gttcaatgct cagaagaagg cccagagctc caccgactaa
     1561 ccggtcatca tcaccatcac cattgagttt aaacccgctg atcagcctcg actgtgcctt
     1621 ctagttgcca gccatctgtt gtttgcccct cccccgtgcc ttccttgacc ctggaaggtg
     1681 ccactcccac tgtcctttcc taataaaatg aggaaattgc atcgcattgt ctgagtaggt
     1741 gtcattctat tctggggggt ggggtggggc aggacagcaa gggggaggat tgggaagaca
     1801 atagcaggca tgctggggat gcggtgggct ctatggcttc tgaggcggaa agaaccagct
     1861 ggggctcgat accgtcgacc tctagctaga gcttggcgta atcatggtca tagctgtttc
     1921 ctgtgtgaaa ttgttatccg ctcacaattc cacacaacat acgagccgga agcataaagt
     1981 gtaaagccta gggtgcctaa tgagtgagct aactcacatt aattgcgttg cgctcactgc
     2041 ccgctttcca gtcgggaaac ctgtcgtgcc agctgcatta atgaatcggc caacgcgcgg
     2101 ggagaggcgg tttgcgtatt gggcgctctt ccgcttcctc gctcactgac tcgctgcgct
     2161 cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa ggcggtaata cggttatcca
     2221 cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga
     2281 accgtaaaaa ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc
     2341 acaaaaatcg acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg
     2401 cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat
     2461 acctgtccgc ctttctccct tcgggaagcg tggcgctttc tcatagctca cgctgtaggt
     2521 atctcagttc ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc
     2581 agcccgaccg ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg
     2641 acttatcgcc actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg
     2701 gtgctacaga gttcttgaag tggtggccta actacggcta cactagaaga acagtatttg
     2761 gtatctgcgc tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg
     2821 gcaaacaaac caccgctggt agcggtggtt tttttgtttg caagcagcag attacgcgca
     2881 gaaaaaaagg atctcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga
     2941 acgaaaactc acgttaaggg attttggtca tgagattatc aaaaaggatc ttcacctaga
     3001 tccttttaaa ttaaaaatga agttttaaat caatctaaag tatatatgag taaacttggt
     3061 ctgacagtta ccaatgctta atcagtgagg cacctatctc agcgatctgt ctatttcgtt
     3121 catccatagt tgcctgactc cccgtcgtgt agataactac gatacgggag ggcttaccat
     3181 ctggccccag tgctgcaatg ataccgcgag acccacgctc accggctcca gatttatcag
     3241 caataaacca gccagccgga agggccgagc gcagaagtgg tcctgcaact ttatccgcct
     3301 ccatccagtc tattaattgt tgccgggaag ctagagtaag tagttcgcca gttaatagtt
     3361 tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg
     3421 cttcattcag ctccggttcc caacgatcaa ggcgagttac atgatccccc atgttgtgca
     3481 aaaaagcggt tagctccttc ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt
     3541 tatcactcat ggttatggca gcactgcata attctcttac tgtcatgcca tccgtaagat
     3601 gcttttctgt gactggtgag tactcaacca agtcattctg agaatagtgt atgcggcgac
     3661 cgagttgctc ttgcccggcg tcaatacggg ataataccgc gccacatagc agaactttaa
     3721 aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt
     3781 tgagatccag ttcgatgtaa cccactcgtg cacccaactg atcttcagca tcttttactt
     3841 tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa
     3901 gggcgacacg gaaatgttga atactcatac tcttcctttt tcaatattat tgaagcattt
     3961 atcagggtta ttgtctcatg agcggataca tatttgaatg tatttagaaa aataaacaaa
     4021 taggggttcc gcgcacattt ccccgaaaag tgccacctga cgtcgacgga tcgggagatc
     4081 gatctcccga tcccctaggg tcgactctca gtacaatctg ctctgatgcc gcatagttaa
     4141 gccagtatct gctccctgct tgtgtgttgg aggtcgctga gtagtgcgcg agcaaaattt
     4201 aagctacaac aaggcaaggc ttgaccgaca attgcatgaa gaatctgctt agggttaggc
     4261 gttttgcgct gcttcgcgat gtacgggcca gatatacgcg ttgacattga ttattgacta
     4321 gttattaata gtaatcaatt acggggtcat tagttcatag cccatatatg gagttccgcg
     4381 ttacataact tacggtaaat ggcccgcctg gctgaccgcc caacgacccc cgcccattga
     4441 cgtcaataat gacgtatgtt cccatagtaa cgccaatagg gactttccat tgacgtcaat
     4501 gggtggagta tttacggtaa actgcccact tggcagtaca tcaagtgtat c
//
