LOCUS       sequence-437117-        6401 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Express HACE Editor as a fusion of the engineered dual base editor 
            TadE that can perform citosine and adenine deamination and PcrA 
            helicase..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6401)
  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 6401)
  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-tadDE-PcrA M6-UGI
FEATURES             Location/Qualifiers
     source          1..6401
                     /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             3126..3374
                     /codon_start=1
                     /product="uracil-DNA glycosylase inhibitor from a Bacillus 
                     subtilis bacteriophage (Mol et al., 1995)"
                     /label=UGI
                     /translation="TNLSDIIEKETGKQLVIQESILMLPEEVEEVIGNKPESDILVHTA
                     YDESTDENVMLLTSDAPEYKPWALVIQDSNGENKIKML"
     CDS             3387..3407
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             3416..3433
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     polyA_signal    3462..3686
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(3757..3773)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3757..3773)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3770..3792)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3781..3797
                     /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(3805..3811,3812..3829,3830..3835))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3850..3871
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3988..4005)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4159..4747)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4239..4258)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4918..5709,5710..5778))
                     /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     5541..5560
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5779..5883)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(5962..5981)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        join(6153..6401,1..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 ggatgaggga gaggcgcctg
      481 tgggagccgt gctggtgctg aacaatagag tgatcggcga gggctggaac agacgtatcg
      541 gcctgcacga cccaacagcc catgccgaaa ttatggccct gagacagggc ggcctggtca
      601 tgcagaactc cagactgatt gacgccaccc tgtacgtgac attcgagcct tgcgtgatgt
      661 gcgccggcgc catgatcaac 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 tatgaacttc ctgtctgagc
      961 aactgctggc acacctgaac aaggagcagc aagaagctgt gcggaccacc gagggacctc
     1021 tgctgatcat ggccggcgct ggaagcggaa aaacaagagt gctgacacac cgcatcgcct
     1081 acctgatggc tgagaagcac gtggcccctt ggaacatcct ggccattacc tttacaaaca
     1141 aggccgctag agagatgaga gaaagagtgc agagcctcct gggaggcgcc gccgaggacg
     1201 tgtggatcag caccttcgcc agcatggccg tgcggatcct gagaagagat atcgacagaa
     1261 tcggcatcaa ccggaacttc agcatcctgg acccaacaga ccagctgagc gtgatgaaaa
     1321 ccatcctgaa agaaaagaac atcgacccca agaaattcga gcctagaaca atcctgggca
     1381 caatcagcgc cgccaagaat gaactgctgc ctccagaaca gtttgcgaag cgggcctcca
     1441 cctactatga gaaagtggtg tctgacgtct accaggagta tcagcagcgg ctgctcaggt
     1501 gtcacagcct tgacttcgat gatctgatca tgaccacaat ccagctgttt gaccgggtcc
     1561 ccgacgtgct ccactactac caatacaagt ttcagtacat ccacatcgat gagtaccagg
     1621 acacaaacag agcccaatac accctggtga aaaagctggc tgagcggttc cagaacatcg
     1681 ccgccgtggg cgacgccgat cagtctatct acagatggcg gggcgccgac atccagaaca
     1741 tcctgagctt cgaaagagat taccccaacg ccaaagtgat cctgctggag caaaattacc
     1801 ggagcacgaa gcgcatcctg caggccgcaa acgaggtgat cgagcacaac gtgaacagaa
     1861 agcctaagcg gatctggacc gagaatcctg agggcaagcc catcctgtac tacgaggcca
     1921 tgaacgaagc cgacgaggcc cagttcgtgg ccggcagaat cagagaggcc gtggagcgcg
     1981 gcgagagaag ataccgagac ttcgccgtgc tgtacagaac caatgcccag tccagagtca
     2041 tggaagagat gctgctgaag gccaacatcc cttaccagat agtgggcggc gtgaagttct
     2101 acgacagaaa ggaaatcaag gacatcctgg cttatctgcg ggtcatcgca aacccagacg
     2161 acgactgcag cctgctgaga attatcaacg ttcctaaaag aggcatcgga gcctctacca
     2221 tcgacaagct ggtccggtac gccgccgacc acgagctgag cctgttcgag gccctgggcg
     2281 agctggagat gatcggtctt ggcgccaagg ccgctggggc cctggccgcc ttcagaagcc
     2341 agctggaaca gtggacacag ctgcaggagt acgtgagcgt gaccgaactg gtggaagagg
     2401 tgctggacaa gtctggctac cgggaaatgc tgaaggccga aagaaccatc gaagcccaga
     2461 gcagactgga gaacctggat gagttcctga gcgtgaccaa gcatttcgaa aacgtgtccg
     2521 acgacaagag cctgatcgcc ttcctgaccg atctggctct gatcagcgac ctggacgagc
     2581 tggatggcac cgagcaggcc gccgagggcg acgccgtgat gctgatgacc ctgcacgccg
     2641 ccaagggcct ggaattcccc gtggtctttc tgataggcat ggaagaaggc atcttccctc
     2701 acaatagaag cctggaggat gacgatgaga tggaagagga gagaagactg gcctacgtgg
     2761 gcatcaccag agctgaagag gaactggtgc tgaccagcgc ccagatgcgg accctgttcg
     2821 gcaacattca gatggaccct cctagccggt tccttaacga gatccccgcc catctgcttg
     2881 agacagccag cagacggcag gccggagctt ccaggcctgc cgtgagcaga ccccaggcct
     2941 ctggcgctgt gggcagctgg aaggtgggcg atcgggcaaa tcacagaaag tggggcatcg
     3001 gaaccgtggt ttccgtgcgg ggcggcggag atgatcagga actggacatc gcttttccat
     3061 ctcctatcgg cattaagaga ctgctagcta aattcgcccc tatcgagaag gtgtctggtg
     3121 gttctactaa tctgtcagat attattgaaa aggagaccgg taagcaactg gttatccagg
     3181 aatccatcct catgctccca gaggaggtgg aagaagtcat tgggaacaag ccggaaagcg
     3241 atatactcgt gcacaccgcc tacgacgaga gcaccgacga gaatgtcatg cttctgacta
     3301 gcgacgcccc tgaatacaag ccttgggctc tggtcataca ggatagcaac ggtgagaaca
     3361 agattaagat gctctctggt ggttctccca agaagaagag gaaagtctaa ccggtcatca
     3421 tcaccatcac cattgagttt aaacccgctg atcagcctcg actgtgcttt ctagttgcca
     3481 gccatctgtt gtttgcccct cccccgtgcc ttccttgacc ctggaaggtg ccactcccac
     3541 tgtcctttcc taataaaatg aggaaattgc atcgcattgt ctgagtaggt gtcattctat
     3601 tctggggggt ggggtggggc aggacagcaa gggggaggat tgggaagaca atagcaggca
     3661 tgctggggat gcggtgggct ctatggcttc tgaggcggaa agaaccagct ggggctcgat
     3721 accgtcgacc tctagctaga gcttggcgta atcatggtca tagctgtttc ctgtgtgaaa
     3781 ttgttatccg ctcacaattc cacacaacat acgagccgga agcataaagt gtaaagccta
     3841 gggtgcctaa tgagtgagct aactcacatt aattgcgttg cgctcactgc ccgctttcca
     3901 gtcgggaaac ctgtcgtgcc agctgcatta atgaatcggc caacgcgcgg ggagaggcgg
     3961 tttgcgtatt gggcgctctt ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg
     4021 gctgcggcga gcggtatcag ctcactcaaa ggcggtaata cggttatcca cagaatcagg
     4081 ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa
     4141 ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc acaaaaatcg
     4201 acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg cgtttccccc
     4261 tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat acctgtccgc
     4321 ctttctccct tcgggaagcg tggcgctttc tcatagctca cgctgtaggt atctcagttc
     4381 ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc agcccgaccg
     4441 ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg acttatcgcc
     4501 actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg gtgctacaga
     4561 gttcttgaag tggtggccta actacggcta cactagaaga acagtatttg gtatctgcgc
     4621 tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg gcaaacaaac
     4681 caccgctggt agcggtggtt tttttgtttg caagcagcag attacgcgca gaaaaaaagg
     4741 atctcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga acgaaaactc
     4801 acgttaaggg attttggtca tgagattatc aaaaaggatc ttcacctaga tccttttaaa
     4861 ttaaaaatga agttttaaat caatctaaag tatatatgag taaacttggt ctgacagtta
     4921 ccaatgctta atcagtgagg cacctatctc agcgatctgt ctatttcgtt catccatagt
     4981 tgcctgactc cccgtcgtgt agataactac gatacgggag ggcttaccat ctggccccag
     5041 tgctgcaatg ataccgcgag acccacgctc accggctcca gatttatcag caataaacca
     5101 gccagccgga agggccgagc gcagaagtgg tcctgcaact ttatccgcct ccatccagtc
     5161 tattaattgt tgccgggaag ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt
     5221 tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag
     5281 ctccggttcc caacgatcaa ggcgagttac atgatccccc atgttgtgca aaaaagcggt
     5341 tagctccttc ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt tatcactcat
     5401 ggttatggca gcactgcata attctcttac tgtcatgcca tccgtaagat gcttttctgt
     5461 gactggtgag tactcaacca agtcattctg agaatagtgt atgcggcgac cgagttgctc
     5521 ttgcccggcg tcaatacggg ataataccgc gccacatagc agaactttaa aagtgctcat
     5581 cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag
     5641 ttcgatgtaa cccactcgtg cacccaactg atcttcagca tcttttactt tcaccagcgt
     5701 ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg
     5761 gaaatgttga atactcatac tcttcctttt tcaatattat tgaagcattt atcagggtta
     5821 ttgtctcatg agcggataca tatttgaatg tatttagaaa aataaacaaa taggggttcc
     5881 gcgcacattt ccccgaaaag tgccacctga cgtcgacgga tcgggagatc gatctcccga
     5941 tcccctaggg tcgactctca gtacaatctg ctctgatgcc gcatagttaa gccagtatct
     6001 gctccctgct tgtgtgttgg aggtcgctga gtagtgcgcg agcaaaattt aagctacaac
     6061 aaggcaaggc ttgaccgaca attgcatgaa gaatctgctt agggttaggc gttttgcgct
     6121 gcttcgcgat gtacgggcca gatatacgcg ttgacattga ttattgacta gttattaata
     6181 gtaatcaatt acggggtcat tagttcatag cccatatatg gagttccgcg ttacataact
     6241 tacggtaaat ggcccgcctg gctgaccgcc caacgacccc cgcccattga cgtcaataat
     6301 gacgtatgtt cccatagtaa cgccaatagg gactttccat tgacgtcaat gggtggagta
     6361 tttacggtaa actgcccact tggcagtaca tcaagtgtat c
//
