LOCUS       sequence-437107-        5366 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Express HACE Editor as a fusion of TadA-8e adenosine deaminase and 
            Ns3h helicase..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5366)
  AUTHORS   Chen XD, Chen Z, Wythes G, Zhang Y, Orr BC, Sun G, Chao YK, Navarro 
            Torres A, Thao K, Vallurupalli M, Sun J, Borji M, Tkacik E, Chen H, 
            Bernstein BE, Chen F
  TITLE     Helicase-assisted continuous editing for programmable mutagenesis of
            endogenous genomes.
  JOURNAL   Science. 2024 Oct 11;386(6718):eadn5876. doi: 
            10.1126/science.adn5876. Epub 2024 Oct 11.
  PUBMED    39388570
REFERENCE   2  (bases 1 to 5366)
  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: "Science."; 
            date: "2024-10-11"; volume: "386(6718):eadn5876. doi"; pages: " 
            10.1126/science.adn5876. Epub 2024 Oct 11"
COMMENT     Sequence Label: pCMV-TadA8e-Ns3h
FEATURES             Location/Qualifiers
     source          1..5366
                     /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"
     CDS             2352..2372
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             2381..2398
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(2421..2438)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2427..2651
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(2722..2738)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2722..2738)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2735..2757)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2746..2762
                     /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(2770..2776,2777..2794,2795..2800))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2815..2836
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2953..2970)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3124..3712)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3204..3223)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3883..4674,4675..4743))
                     /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     4506..4525
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4744..4848)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(4927..4946)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        join(5118..5366,1..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 ggccgcgaga gccgccacca tgagttctga ggtggagttt tcccacgagt
      421 actggatgag acatgccctg accctggcca agagggcacg ggatgagagg gaggtgcctg
      481 tgggagccgt gctggtgctg aacaatagag tgatcggcga gggctggaac agagccatcg
      541 gcctgcacga cccaacagcc catgccgaaa ttatggccct gagacagggc ggcctggtca
      601 tgcagaacta cagactgatt gacgccaccc tgtacgtgac attcgagcct tgcgtgatgt
      661 gcgccggcgc catgatccac tctaggatcg gccgcgtggt gtttggcgtg aggaactcaa
      721 aaagaggcgc cgcaggctcc ctgatgaacg tgctgaacta ccccggcatg aatcaccgcg
      781 tcgaaattac cgagggaatc ctggcagatg aatgtgccgc cctgctgtgc gatttctatc
      841 ggatgcctag acaggtgttc aatgctcaga agaaggccca gagctccatc aacagcggca
      901 gcgagactcc cgggacctca gagtccgcca cacccgaaag tatggtggac ttcatacccg
      961 ttgagtctat ggaaactacc atgcggtctc cggtcttcac agacaactca acccccccgg
     1021 ctgtaccgca gacattccaa gtggcacatc tgcacgctcc tactggcagc ggcaagagca
     1081 ccaaagtgcc ggctgcgtat gcagcccaag ggtacaaggt gctcgtcctg aacccgtccg
     1141 ttgccgccac cttagggttt ggggcgtata tgtccaaggc acacggtatc gaccctaaca
     1201 tcagaactgg ggtaaggacc attaccacgg gcggctccat tacgtactcc acctatggca
     1261 agttccttgc cgacggtggc tgttctgggg gcgcctatga catcataata tgtgatgagt
     1321 gccactcaac tgactcgact accatcttgg gcatcggcac agtcctggac caagcggaga
     1381 cggctggagc gcggctcgtc gtgctcgcca ccgctacacc tccgggatcg gttaccgtgc
     1441 cacaccccaa tatcgaggaa ataggcctgt ccaacaatgg agagatcccc ttctatggca
     1501 aagccatccc cattgaggcc atcaaggggg ggaggcatct cattttctgc cattccaaga
     1561 agaaatgtga cgagctcgcc gcaaagctga caggcctcgg actgaacgct gtagcatatt
     1621 accggggcct tgatgtgtcc gtcataccgc ctatcggaga cgtcgttgtc gtggcaacag
     1681 acgctctaat gacgggtttc accggcgatt ttgactcagt gatcgactgc aatacatgtg
     1741 tcacccagac agtcgacttc agcttggatc ccaccttcac cattgagacg acgaccgtgc
     1801 cccaagacgc ggtgtcgcgc tcgcaacggc gaggtagaac tggcaggggt aggagtggca
     1861 tctacaggtt tgtgactcca ggagaacggc cctcgggcat gttcgattct tcggtcctgt
     1921 gtgagtgcta tgacgcgggc tgtgcttggt atgagctcac gcccgctgag acctcggtta
     1981 ggttgcgggc ttacctaaat acaccagggt tgcccgtctg ccaggaccat ctggagttct
     2041 gggagagcgt cttcacaggc ctcacccaca tagatgccca cttcctgtcc cagactaaac
     2101 aggcaggaga caactttcct tacctggtgg catatcaagc tacagtgtgc gccagggctc
     2161 aagctccacc tccatcgtgg gaccaaatgt ggaagtgtct catacggctg aaacctacac
     2221 tgcacgggcc aacacccctg ctgtataggc taggagccgt ccaaaatgag gtcatcctca
     2281 cacaccccat aactaaatac atcatggcat gcatgtcggc tgacctggag gtcgtcactt
     2341 ctggtggttc tcccaagaag aagaggaaag tctaaccggt catcatcacc atcaccattg
     2401 agtttaaacc cgctgatcag cctcgactgt gccttctagt tgccagccat ctgttgtttg
     2461 cccctccccc gtgccttcct tgaccctgga aggtgccact cccactgtcc tttcctaata
     2521 aaatgaggaa attgcatcgc attgtctgag taggtgtcat tctattctgg ggggtggggt
     2581 ggggcaggac agcaaggggg aggattggga agacaatagc aggcatgctg gggatgcggt
     2641 gggctctatg gcttctgagg cggaaagaac cagctggggc tcgataccgt cgacctctag
     2701 ctagagcttg gcgtaatcat ggtcatagct gtttcctgtg tgaaattgtt atccgctcac
     2761 aattccacac aacatacgag ccggaagcat aaagtgtaaa gcctagggtg cctaatgagt
     2821 gagctaactc acattaattg cgttgcgctc actgcccgct ttccagtcgg gaaacctgtc
     2881 gtgccagctg cattaatgaa tcggccaacg cgcggggaga ggcggtttgc gtattgggcg
     2941 ctcttccgct tcctcgctca ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt
     3001 atcagctcac tcaaaggcgg taatacggtt atccacagaa tcaggggata acgcaggaaa
     3061 gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc
     3121 gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag
     3181 gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt
     3241 gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg
     3301 aagcgtggcg ctttctcata gctcacgctg taggtatctc agttcggtgt aggtcgttcg
     3361 ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg
     3421 taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac
     3481 tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg
     3541 gcctaactac ggctacacta gaagaacagt atttggtatc tgcgctctgc tgaagccagt
     3601 taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg
     3661 tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc
     3721 tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt
     3781 ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa aatgaagttt
     3841 taaatcaatc taaagtatat atgagtaaac ttggtctgac agttaccaat gcttaatcag
     3901 tgaggcacct atctcagcga tctgtctatt tcgttcatcc atagttgcct gactccccgt
     3961 cgtgtagata actacgatac gggagggctt accatctggc cccagtgctg caatgatacc
     4021 gcgagaccca cgctcaccgg ctccagattt atcagcaata aaccagccag ccggaagggc
     4081 cgagcgcaga agtggtcctg caactttatc cgcctccatc cagtctatta attgttgccg
     4141 ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg ccattgctac
     4201 aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg gttcccaacg
     4261 atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc
     4321 tccgatcgtt gtcagaagta agttggccgc agtgttatca ctcatggtta tggcagcact
     4381 gcataattct cttactgtca tgccatccgt aagatgcttt tctgtgactg gtgagtactc
     4441 aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat
     4501 acgggataat accgcgccac atagcagaac tttaaaagtg ctcatcattg gaaaacgttc
     4561 ttcggggcga aaactctcaa ggatcttacc gctgttgaga tccagttcga tgtaacccac
     4621 tcgtgcaccc aactgatctt cagcatcttt tactttcacc agcgtttctg ggtgagcaaa
     4681 aacaggaagg caaaatgccg caaaaaaggg aataagggcg acacggaaat gttgaatact
     4741 catactcttc ctttttcaat attattgaag catttatcag ggttattgtc tcatgagcgg
     4801 atacatattt gaatgtattt agaaaaataa acaaataggg gttccgcgca catttccccg
     4861 aaaagtgcca cctgacgtcg acggatcggg agatcgatct cccgatcccc tagggtcgac
     4921 tctcagtaca atctgctctg atgccgcata gttaagccag tatctgctcc ctgcttgtgt
     4981 gttggaggtc gctgagtagt gcgcgagcaa aatttaagct acaacaaggc aaggcttgac
     5041 cgacaattgc atgaagaatc tgcttagggt taggcgtttt gcgctgcttc gcgatgtacg
     5101 ggccagatat acgcgttgac attgattatt gactagttat taatagtaat caattacggg
     5161 gtcattagtt catagcccat atatggagtt ccgcgttaca taacttacgg taaatggccc
     5221 gcctggctga ccgcccaacg acccccgccc attgacgtca ataatgacgt atgttcccat
     5281 agtaacgcca atagggactt tccattgacg tcaatgggtg gagtatttac ggtaaactgc
     5341 ccacttggca gtacatcaag tgtatc
//
