LOCUS       sequence-244349-        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"
     enhancer        74..453
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        454..657
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     608..628
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     699..718
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        699..717
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             767..787
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             1888..1905
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(1928..1945)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    1934..2158
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(2229..2245)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2229..2245)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2242..2264)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2253..2269
                     /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(2277..2283,2284..2301,2302..2307))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2322..2343
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2460..2477)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2631..3219)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2711..2730)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3390..4181,4182..4250))
                     /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     4013..4032
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4251..4355)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(4434..4453)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 aattgcatga agaatctgct tagggttagg cgttttgcgc tgcttcgcga tgtacgggcc
       61 agatatacgc gttgacattg attattgact agttattaat agtaatcaat tacggggtca
      121 ttagttcata gcccatatat ggagttccgc gttacataac ttacggtaaa tggcccgcct
      181 ggctgaccgc ccaacgaccc ccgcccattg acgtcaataa tgacgtatgt tcccatagta
      241 acgccaatag ggactttcca ttgacgtcaa tgggtggagt atttacggta aactgcccac
      301 ttggcagtac atcaagtgta tcatatgcca agtacgcccc ctattgacgt caatgacggt
      361 aaatggcccg cctggcatta tgcccagtac atgaccttat gggactttcc tacttggcag
      421 tacatctacg tattagtcat cgctattacc atggtgatgc ggttttggca gtacatcaat
      481 gggcgtggat agcggtttga ctcacgggga tttccaagtc tccaccccat tgacgtcaat
      541 gggagtttgt tttggcacca aaatcaacgg gactttccaa aatgtcgtaa caactccgcc
      601 ccattgacgc aaatgggcgg taggcgtgta cggtgggagg tctatataag cagagctggt
      661 ttagtgaacc gtcagatccg ctagagatcc gcggccgcta atacgactca ctatagggag
      721 agccgccacc atgaaacgga cagccgacgg aagcgagttc gagtcaccaa agaagaagcg
      781 gaaagtctct gaagtcgagt ttagccacga gtattggatg aggcacgcac tgaccctggc
      841 aaagcgagca tgggatgaaa gagaagtccc cgtgggcgcc gtgctggtgc acaacaatag
      901 agtgatcgga gagggatgga acaggccaat cggccgccac gaccctaccg cacacgcaga
      961 gatcatggca ctgaggcagg gaggcctggt catgcagaat taccgcctga tcgatgccac
     1021 cctgtatgtg acactggagc catgcgtgat gtgcgcagga gcaatgatcc acagcaggat
     1081 cggaagagtg gtgttcggag cacgggacgc caagaccggc gcagcaggct ccctgatgga
     1141 tgtgctgcac caccccggca tgaaccaccg ggtggagatc acagagggaa tcctggcaga
     1201 cgagtgcgcc gccctgctga gcgatttctt tagaatgcgg agacaggaga tcaaggccca
     1261 gaagaaggca cagagctcca ccgactctgg aggatctagc ggaggatcct ctggaagcga
     1321 gacaccaggc acaagcgagt ccgccacacc agagagctcc ggcggctcct ccggaggatc
     1381 ctctgaggtg gagttttccc acgagtactg gatgagacat gccctgaccc tggccaagag
     1441 ggcacgcgat gagagggagg tgcctgtggg agccgtgctg gtgctgaaca atagagtgat
     1501 cggcgagggc tggaacagag ccatcggcct gcacgaccca acagcccatg ccgaaattat
     1561 ggccctgaga cagggcggcc tggtcatgca gaactacaga ctgattgacg ccaccctgta
     1621 cgtgacattc gagccttgcg tgatgtgcgc cggcgccatg atccactcta ggatcggccg
     1681 cgtggtgttt ggcgtgagga acgcaaaaac cggcgccgca ggctccctga tggacgtgct
     1741 gcactacccc ggcatgaatc accgcgtcga aattaccgag ggaatcctgg cagatgaatg
     1801 tgccgccctg ctgtgctatt tctttcggat gcctagacag gtgttcaatg ctcagaagaa
     1861 ggcccagagc tccaccgact aaccggtcat catcaccatc accattgagt ttaaacccgc
     1921 tgatcagcct cgactgtgcc ttctagttgc cagccatctg ttgtttgccc ctcccccgtg
     1981 ccttccttga ccctggaagg tgccactccc actgtccttt cctaataaaa tgaggaaatt
     2041 gcatcgcatt gtctgagtag gtgtcattct attctggggg gtggggtggg gcaggacagc
     2101 aagggggagg attgggaaga caatagcagg catgctgggg atgcggtggg ctctatggct
     2161 tctgaggcgg aaagaaccag ctggggctcg ataccgtcga cctctagcta gagcttggcg
     2221 taatcatggt catagctgtt tcctgtgtga aattgttatc cgctcacaat tccacacaac
     2281 atacgagccg gaagcataaa gtgtaaagcc tagggtgcct aatgagtgag ctaactcaca
     2341 ttaattgcgt tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg ccagctgcat
     2401 taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta ttgggcgctc ttccgcttcc
     2461 tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc agctcactca
     2521 aaggcggtaa tacggttatc cacagaatca ggggataacg caggaaagaa catgtgagca
     2581 aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg
     2641 ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg
     2701 acaggactat aaagatacca ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt
     2761 ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag cgtggcgctt
     2821 tctcatagct cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc caagctgggc
     2881 tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt
     2941 gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccactgg taacaggatt
     3001 agcagagcga ggtatgtagg cggtgctaca gagttcttga agtggtggcc taactacggc
     3061 tacactagaa gaacagtatt tggtatctgc gctctgctga agccagttac cttcggaaaa
     3121 agagttggta gctcttgatc cggcaaacaa accaccgctg gtagcggtgg tttttttgtt
     3181 tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag aagatccttt gatcttttct
     3241 acggggtctg acgctcagtg gaacgaaaac tcacgttaag ggattttggt catgagatta
     3301 tcaaaaagga tcttcaccta gatcctttta aattaaaaat gaagttttaa atcaatctaa
     3361 agtatatatg agtaaacttg gtctgacagt taccaatgct taatcagtga ggcacctatc
     3421 tcagcgatct gtctatttcg ttcatccata gttgcctgac tccccgtcgt gtagataact
     3481 acgatacggg agggcttacc atctggcccc agtgctgcaa tgataccgcg agacccacgc
     3541 tcaccggctc cagatttatc agcaataaac cagccagccg gaagggccga gcgcagaagt
     3601 ggtcctgcaa ctttatccgc ctccatccag tctattaatt gttgccggga agctagagta
     3661 agtagttcgc cagttaatag tttgcgcaac gttgttgcca ttgctacagg catcgtggtg
     3721 tcacgctcgt cgtttggtat ggcttcattc agctccggtt cccaacgatc aaggcgagtt
     3781 acatgatccc ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc
     3841 agaagtaagt tggccgcagt gttatcactc atggttatgg cagcactgca taattctctt
     3901 actgtcatgc catccgtaag atgcttttct gtgactggtg agtactcaac caagtcattc
     3961 tgagaatagt gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg ggataatacc
     4021 gcgccacata gcagaacttt aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa
     4081 ctctcaagga tcttaccgct gttgagatcc agttcgatgt aacccactcg tgcacccaac
     4141 tgatcttcag catcttttac tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa
     4201 aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt gaatactcat actcttcctt
     4261 tttcaatatt attgaagcat ttatcagggt tattgtctca tgagcggata catatttgaa
     4321 tgtatttaga aaaataaaca aataggggtt ccgcgcacat ttccccgaaa agtgccacct
     4381 gacgtcgacg gatcgggaga tcgatctccc gatcccctag ggtcgactct cagtacaatc
     4441 tgctctgatg ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct
     4501 gagtagtgcg cgagcaaaat ttaagctaca acaaggcaag gcttgaccga c
//
