LOCUS       sequence-437110-        6158 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Express HACE Editor as a fusion of TadA-8e adenosine deaminase and 
            PcrA helicase..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6158)
  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 6158)
  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
FEATURES             Location/Qualifiers
     source          1..6158
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     544..563
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        642..746
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(747..815,816..1607)
                     /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     complement(965..984)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      1778..2366
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2267..2286
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2520..2537
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    2654..2675
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(2690..2695,2696..2713,2714..2720)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2728..2744
                     /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)."
     primer_bind     2733..2755
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     2752..2768
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     2752..2768
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     polyA_signal    2839..3063
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     3052..3069
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     CDS             complement(3092..3109)
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             complement(3118..3138)
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     promoter        complement(5947..6150)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(5976..5996)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        join(6151..6158,1..372)
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
ORIGIN
        1 tagcgatgac taatacgtag atgtactgcc aagtaggaaa gtcccataag gtcatgtact
       61 gggcataatg ccaggcgggc catttaccgt cattgacgtc aatagggggc gtacttggca
      121 tatgatacac ttgatgtact gccaagtggg cagtttaccg taaatactcc acccattgac
      181 gtcaatggaa agtccctatt ggcgttacta tgggaacata cgtcattatt gacgtcaatg
      241 ggcgggggtc gttgggcggt cagccaggcg ggccatttac cgtaagttat gtaacgcgga
      301 actccatata tgggctatga actaatgacc ccgtaattga ttactattaa taactagtca
      361 ataatcaatg tcaacgcgta tatctggccc gtacatcgcg aagcagcgca aaacgcctaa
      421 ccctaagcag attcttcatg caattgtcgg tcaagccttg ccttgttgta gcttaaattt
      481 tgctcgcgca ctactcagcg acctccaaca cacaagcagg gagcagatac tggcttaact
      541 atgcggcatc agagcagatt gtactgagag tcgaccctag gggatcggga gatcgatctc
      601 ccgatccgtc gacgtcaggt ggcacttttc ggggaaatgt gcgcggaacc cctatttgtt
      661 tatttttcta aatacattca aatatgtatc cgctcatgag acaataaccc tgataaatgc
      721 ttcaataata ttgaaaaagg aagagtatga gtattcaaca tttccgtgtc gcccttattc
      781 ccttttttgc ggcattttgc cttcctgttt ttgctcaccc agaaacgctg gtgaaagtaa
      841 aagatgctga agatcagttg ggtgcacgag tgggttacat cgaactggat ctcaacagcg
      901 gtaagatcct tgagagtttt cgccccgaag aacgttttcc aatgatgagc acttttaaag
      961 ttctgctatg tggcgcggta ttatcccgta ttgacgccgg gcaagagcaa ctcggtcgcc
     1021 gcatacacta ttctcagaat gacttggttg agtactcacc agtcacagaa aagcatctta
     1081 cggatggcat gacagtaaga gaattatgca gtgctgccat aaccatgagt gataacactg
     1141 cggccaactt acttctgaca acgatcggag gaccgaagga gctaaccgct tttttgcaca
     1201 acatggggga tcatgtaact cgccttgatc gttgggaacc ggagctgaat gaagccatac
     1261 caaacgacga gcgtgacacc acgatgcctg tagcaatggc aacaacgttg cgcaaactat
     1321 taactggcga actacttact ctagcttccc ggcaacaatt aatagactgg atggaggcgg
     1381 ataaagttgc aggaccactt ctgcgctcgg cccttccggc tggctggttt attgctgata
     1441 aatctggagc cggtgagcgt gggtctcgcg gtatcattgc agcactgggg ccagatggta
     1501 agccctcccg tatcgtagtt atctacacga cggggagtca ggcaactatg gatgaacgaa
     1561 atagacagat cgctgagata ggtgcctcac tgattaagca ttggtaactg tcagaccaag
     1621 tttactcata tatactttag attgatttaa aacttcattt ttaatttaaa aggatctagg
     1681 tgaagatcct ttttgataat ctcatgacca aaatccctta acgtgagttt tcgttccact
     1741 gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg agatcctttt tttctgcgcg
     1801 taatctgctg cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc
     1861 aagagctacc aactcttttt ccgaaggtaa ctggcttcag cagagcgcag ataccaaata
     1921 ctgttcttct agtgtagccg tagttaggcc accacttcaa gaactctgta gcaccgccta
     1981 catacctcgc tctgctaatc ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc
     2041 ttaccgggtt ggactcaaga cgatagttac cggataaggc gcagcggtcg ggctgaacgg
     2101 ggggttcgtg cacacagccc agcttggagc gaacgaccta caccgaactg agatacctac
     2161 agcgtgagct atgagaaagc gccacgcttc ccgaagggag aaaggcggac aggtatccgg
     2221 taagcggcag ggtcggaaca ggagagcgca cgagggagct tccaggggga aacgcctggt
     2281 atctttatag tcctgtcggg tttcgccacc tctgacttga gcgtcgattt ttgtgatgct
     2341 cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc ggccttttta cggttcctgg
     2401 ccttttgctg gccttttgct cacatgttct ttcctgcgtt atcccctgat tctgtggata
     2461 accgtattac cgcctttgag tgagctgata ccgctcgccg cagccgaacg accgagcgca
     2521 gcgagtcagt gagcgaggaa gcggaagagc gcccaatacg caaaccgcct ctccccgcgc
     2581 gttggccgat tcattaatgc agctggcacg acaggtttcc cgactggaaa gcgggcagtg
     2641 agcgcaacgc aattaatgtg agttagctca ctcattaggc accctaggct ttacacttta
     2701 tgcttccggc tcgtatgttg tgtggaattg tgagcggata acaatttcac acaggaaaca
     2761 gctatgacca tgattacgcc aagctctagc tagaggtcga cggtatcgag ccccagctgg
     2821 ttctttccgc ctcagaagcc atagagccca ccgcatcccc agcatgcctg ctattgtctt
     2881 cccaatcctc ccccttgctg tcctgcccca ccccaccccc cagaatagaa tgacacctac
     2941 tcagacaatg cgatgcaatt tcctcatttt attaggaaag gacagtggga gtggcacctt
     3001 ccagggtcaa ggaaggcacg ggggaggggc aaacaacaga tggctggcaa ctagaaggca
     3061 cagtcgaggc tgatcagcgg gtttaaactc aatggtgatg gtgatgatga ccggttagac
     3121 tttcctcttc ttcttgggag aaccaccaga gtcttctttt ttctcgattg gagcgaactg
     3181 agcgagcagg cgctttgggc cctggctctt aaagatgatg tcgagctcta tggatccatt
     3241 tttctcattc acattggaaa ccatgccctc tccccaagct ttgtgcataa ccttatcgcc
     3301 aacattccaa tcgctggaga gcacctgctt cttcgtgcta gtcgtgcgct gtgagaagcc
     3361 gcgtttggcg aacggcttag cttttggttg tatggtctgc cgcttacctg agctatggtt
     3421 ctccagcagg ctctcgggta tttccttaag gaacctagag ggcatgtttg actgtgggcg
     3481 tccaaagagc atccgagacg ttgcgtgtgt aatataaaga acctcctccg ctcgtgtaat
     3541 cgcaacgtaa cagattctgc gctcctcttg catctcatga tcatcctcgg atttgattgc
     3601 ccgaatgtgt ggaaagaggg actcctccat acccatgatg aagactatgg gaaactccaa
     3661 tcccttagca ctgtgcatcg tcatcaatgt taccccattc tcagtgtcag cttcgtcaat
     3721 atcggcaacg agtgacaggt cagtaaggaa attgatcaat gactgctctt caaggggcgt
     3781 attctcctca tagtctttgg gcactgacat gaactcatct atgttctcga gtcttgacct
     3841 actttcgagc gtgttctccc tttccaacat ttcccgatag ccggattttt gcagaacttc
     3901 atctacgatc tcgtggatct caaggaattc ctgttccttt atcaggcttt gtatcagttc
     3961 gtaaaagtta agacactcct gggttacttt ctttgacaag ccgataaaat cagcttctcc
     4021 aagagcgtca aacatggaaa tattgttctg caacgcatag ttttgaactt tctcaactga
     4081 tgagggtcca accccgcgtt tcggtacgtt aataatccgt tgaagtgaga tgtcgtcgtt
     4141 actgttggca atgattcgga ggtagctcag caggtctttg atctctttcc tatcatagaa
     4201 tttttggcct ccaaccatcg tgtatggcat attgctcttc ataaaggttt cttcgagaac
     4261 tctggattgc gcattggtcc tgtacagaat tgccatgtct tggtatttct tgccgttgcg
     4321 ttggtgtttc attatttccc ggatgacgaa ctccgcttcg tctcgttcag tcattgcttc
     4381 gtagtaatga atcttttcac catttgtgtt agctgtccac agacctttag gctttcgctc
     4441 actattattt tttattactt cgttagccgc gttcaaaata gtttttgtag atctataatt
     4501 ctgctcaagg aagattgtat tcgcctcagg ataatctttc tcgaaggaca agatattctg
     4561 tatgtcggcg ccgcgccaac cgtatatgct ctggtcacta tccccgacca cgcaaagatt
     4621 tttaaacttt gacgccagca gctttaccaa tgtatactgt gccttattag tatcttggta
     4681 ttcatcaacg tggatatatt gaaatttgtt ttggtaatat tccaacactt ccggtactct
     4741 ttcaaaaagg ttaatagtag tcatgatcag atcatcaaaa tccagtgctt cattccgact
     4801 cagctgtcgt tggtagccgg aataaacagt tgcgaccatc tgactgtggt agtctgttgc
     4861 ttccttttgt gcgtccgcag gtgttttgag ttcgttcttc aagtttgata tggcacctat
     4921 gaacattcgg ggttcaaatt ttttgctatc tatattctca tttttgagaa catcttttat
     4981 tacagacttt tgatctgtag gatctattat cgtgaagttc cgttcgatcc caatcctatc
     5041 cgcgtctcgc cgaagtatgc gaacgcacat agaatggaag gtagacatcc aaatgacctc
     5101 cgcttggtca ccgacaagtt tttggactct ttccttcatc tctctagcag ccttgtttgt
     5161 gaaggtaatg gcaagcacgt tatatggact cacgtccttt tcgtcaagca agtacgctat
     5221 tcggtgagta aggaccctcg ttttaccact cccagccccc gccatgatca acaaggggcc
     5281 ttctgtggtc tttaccgcct cggattgttc tgtattcata tggttcagca gggcattcat
     5341 actttcgggt gtggcggact ctgaggtccc gggagtctcg ctgccgctgt tgatggagct
     5401 ctgggccttc ttctgagcat tgaacacctg tctaggcatc cgatagaaat cgcacagcag
     5461 ggcggcacat tcatctgcca ggattccctc ggtaatttcg acgcggtgat tcatgccggg
     5521 gtagttcagc acgttcatca gggagcctgc ggcgcctctt tttgagttcc tcacgccaaa
     5581 caccacgcgg ccgatcctag agtggatcat ggcgccggcg cacatcacgc aaggctcgaa
     5641 tgtcacgtac agggtggcgt caatcagtct gtagttctgc atgaccaggc cgccctgtct
     5701 cagggccata atttcggcat gggctgttgg gtcgtgcagg ccgatggctc tgttccagcc
     5761 ctcgccgatc actctattgt tcagcaccag cacggctccc acaggcacct ccctctcatc
     5821 ccgtgccctc ttggccaggg tcagggcatg tctcatccag tactcgtggg aaaactccac
     5881 ctcagaactc atggtggcgg ctctcgcggc cgcggatctc tagcggatct gacggttcac
     5941 taaaccagct ctgcttatat agacctccca ccgtacacgc ctaccgccca tttgcgtcaa
     6001 tggggcggag ttgttacgac attttggaaa gtcccgttga ttttggtgcc aaaacaaact
     6061 cccattgacg tcaatggggt ggagacttgg aaatccccgt gagtcaaacc gctatccacg
     6121 cccattgatg tactgccaaa accgcatcac catggtaa
//
