LOCUS       sequence-461180-        6170 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Express HACE Editor as a fusion of TadA-8e adenosine deaminase and 
            an optimized PcrA M6 helicase..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6170)
  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 6170)
  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-PcrA M6
FEATURES             Location/Qualifiers
     source          1..6170
                     /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             3156..3176
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             3185..3202
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(3225..3242)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3231..3455
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(3526..3542)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3526..3542)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3539..3561)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3550..3566
                     /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(3574..3580,3581..3598,3599..3604))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3619..3640
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3757..3774)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3928..4516)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4008..4027)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4687..5478,5479..5547))
                     /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     5310..5329
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5548..5652)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(5731..5750)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        join(5922..6170,1..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 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 gttctaaaag aaccgccgac ggcagcgaat tcgagcccaa gaagaagagg aaagtctaac
     3181 cggtcatcat caccatcacc attgagttta aacccgctga tcagcctcga ctgtgccttc
     3241 tagttgccag ccatctgttg tttgcccctc ccccgtgcct tccttgaccc tggaaggtgc
     3301 cactcccact gtcctttcct aataaaatga ggaaattgca tcgcattgtc tgagtaggtg
     3361 tcattctatt ctggggggtg gggtggggca ggacagcaag ggggaggatt gggaagacaa
     3421 tagcaggcat gctggggatg cggtgggctc tatggcttct gaggcggaaa gaaccagctg
     3481 gggctcgata ccgtcgacct ctagctagag cttggcgtaa tcatggtcat agctgtttcc
     3541 tgtgtgaaat tgttatccgc tcacaattcc acacaacata cgagccggaa gcataaagtg
     3601 taaagcctag ggtgcctaat gagtgagcta actcacatta attgcgttgc gctcactgcc
     3661 cgctttccag tcgggaaacc tgtcgtgcca gctgcattaa tgaatcggcc aacgcgcggg
     3721 gagaggcggt ttgcgtattg ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc
     3781 ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag gcggtaatac ggttatccac
     3841 agaatcaggg gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa
     3901 ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc cgcccccctg acgagcatca
     3961 caaaaatcga cgctcaagtc agaggtggcg aaacccgaca ggactataaa gataccaggc
     4021 gtttccccct ggaagctccc tcgtgcgctc tcctgttccg accctgccgc ttaccggata
     4081 cctgtccgcc tttctccctt cgggaagcgt ggcgctttct catagctcac gctgtaggta
     4141 tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac cccccgttca
     4201 gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag tccaacccgg taagacacga
     4261 cttatcgcca ctggcagcag ccactggtaa caggattagc agagcgaggt atgtaggcgg
     4321 tgctacagag ttcttgaagt ggtggcctaa ctacggctac actagaagaa cagtatttgg
     4381 tatctgcgct ctgctgaagc cagttacctt cggaaaaaga gttggtagct cttgatccgg
     4441 caaacaaacc accgctggta gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag
     4501 aaaaaaagga tctcaagaag atcctttgat cttttctacg gggtctgacg ctcagtggaa
     4561 cgaaaactca cgttaaggga ttttggtcat gagattatca aaaaggatct tcacctagat
     4621 ccttttaaat taaaaatgaa gttttaaatc aatctaaagt atatatgagt aaacttggtc
     4681 tgacagttac caatgcttaa tcagtgaggc acctatctca gcgatctgtc tatttcgttc
     4741 atccatagtt gcctgactcc ccgtcgtgta gataactacg atacgggagg gcttaccatc
     4801 tggccccagt gctgcaatga taccgcgaga cccacgctca ccggctccag atttatcagc
     4861 aataaaccag ccagccggaa gggccgagcg cagaagtggt cctgcaactt tatccgcctc
     4921 catccagtct attaattgtt gccgggaagc tagagtaagt agttcgccag ttaatagttt
     4981 gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc
     5041 ttcattcagc tccggttccc aacgatcaag gcgagttaca tgatccccca tgttgtgcaa
     5101 aaaagcggtt agctccttcg gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt
     5161 atcactcatg gttatggcag cactgcataa ttctcttact gtcatgccat ccgtaagatg
     5221 cttttctgtg actggtgagt actcaaccaa gtcattctga gaatagtgta tgcggcgacc
     5281 gagttgctct tgcccggcgt caatacggga taataccgcg ccacatagca gaactttaaa
     5341 agtgctcatc attggaaaac gttcttcggg gcgaaaactc tcaaggatct taccgctgtt
     5401 gagatccagt tcgatgtaac ccactcgtgc acccaactga tcttcagcat cttttacttt
     5461 caccagcgtt tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa agggaataag
     5521 ggcgacacgg aaatgttgaa tactcatact cttccttttt caatattatt gaagcattta
     5581 tcagggttat tgtctcatga gcggatacat atttgaatgt atttagaaaa ataaacaaat
     5641 aggggttccg cgcacatttc cccgaaaagt gccacctgac gtcgacggat cgggagatcg
     5701 atctcccgat cccctagggt cgactctcag tacaatctgc tctgatgccg catagttaag
     5761 ccagtatctg ctccctgctt gtgtgttgga ggtcgctgag tagtgcgcga gcaaaattta
     5821 agctacaaca aggcaaggct tgaccgacaa ttgcatgaag aatctgctta gggttaggcg
     5881 ttttgcgctg cttcgcgatg tacgggccag atatacgcgt tgacattgat tattgactag
     5941 ttattaatag taatcaatta cggggtcatt agttcatagc ccatatatgg agttccgcgt
     6001 tacataactt acggtaaatg gcccgcctgg ctgaccgccc aacgaccccc gcccattgac
     6061 gtcaataatg acgtatgttc ccatagtaac gccaataggg actttccatt gacgtcaatg
     6121 ggtggagtat ttacggtaaa ctgcccactt ggcagtacat caagtgtatc
//
