LOCUS       sequence-437106-        8219 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Express HACE Editor as a fusion of TadA-8e adenosine deaminase and 
            BLM helicase..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8219)
  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 8219)
  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-BLM
FEATURES             Location/Qualifiers
     source          1..8219
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        complement(820..1023)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(849..869)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        1024..1403
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     primer_bind     1575..1594
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        1673..1777
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1778..1846,1847..2638)
                     /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(1996..2015)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2809..3397
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3298..3317
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3551..3568
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    3685..3706
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(3721..3726,3727..3744,3745..3751)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3759..3775
                     /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     3764..3786
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     3783..3799
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     3783..3799
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     polyA_signal    3870..4094
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     4083..4100
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     CDS             complement(4123..4140)
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             complement(4149..4169)
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
ORIGIN
        1 aaagtcttcg gtaacattaa catctttatt tcttaataca ggtgtttttg ctactgacac
       61 attagttaca gatacattgt tatctgaaga tgttttcttt ttaaaagtga aacctgaaaa
      121 ttttggtttt gaaagactta atttattatt aagtgttctg gctgagtgac gttctagttg
      181 ctcctgtaga ttattttgag gaacagcagc catactttcg ggtgtggcgg actctgaggt
      241 cccgggagtc tcgctgccgc tgttgatgga gctctgggcc ttcttctgag cattgaacac
      301 ctgtctaggc atccgataga aatcgcacag cagggcggca cattcatctg ccaggattcc
      361 ctcggtaatt tcgacgcggt gattcatgcc ggggtagttc agcacgttca tcagggagcc
      421 tgcggcgcct ctttttgagt tcctcacgcc aaacaccacg cggccgatcc tagagtggat
      481 catggcgccg gcgcacatca cgcaaggctc gaatgtcacg tacagggtgg cgtcaatcag
      541 tctgtagttc tgcatgacca ggccgccctg tctcagggcc ataatttcgg catgggctgt
      601 tgggtcgtgc aggccgatgg ctctgttcca gccctcgccg atcactctat tgttcagcac
      661 cagcacggct cccacaggca cctccctctc atcccgtgcc ctcttggcca gggtcagggc
      721 atgtctcatc cagtactcgt gggaaaactc cacctcagaa ctcatggtgg cggctctcgc
      781 ggccgcggat ctctagcgga tctgacggtt cactaaacca gctctgctta tatagacctc
      841 ccaccgtaca cgcctaccgc ccatttgcgt caatggggcg gagttgttac gacattttgg
      901 aaagtcccgt tgattttggt gccaaaacaa actcccattg acgtcaatgg ggtggagact
      961 tggaaatccc cgtgagtcaa accgctatcc acgcccattg atgtactgcc aaaaccgcat
     1021 caccatggta atagcgatga ctaatacgta gatgtactgc caagtaggaa agtcccataa
     1081 ggtcatgtac tgggcataat gccaggcggg ccatttaccg tcattgacgt caataggggg
     1141 cgtacttggc atatgataca cttgatgtac tgccaagtgg gcagtttacc gtaaatactc
     1201 cacccattga cgtcaatgga aagtccctat tggcgttact atgggaacat acgtcattat
     1261 tgacgtcaat gggcgggggt cgttgggcgg tcagccaggc gggccattta ccgtaagtta
     1321 tgtaacgcgg aactccatat atgggctatg aactaatgac cccgtaattg attactatta
     1381 ataactagtc aataatcaat gtcaacgcgt atatctggcc cgtacatcgc gaagcagcgc
     1441 aaaacgccta accctaagca gattcttcat gcaattgtcg gtcaagcctt gccttgttgt
     1501 agcttaaatt ttgctcgcgc actactcagc gacctccaac acacaagcag ggagcagata
     1561 ctggcttaac tatgcggcat cagagcagat tgtactgaga gtcgacccta ggggatcggg
     1621 agatcgatct cccgatccgt cgacgtcagg tggcactttt cggggaaatg tgcgcggaac
     1681 ccctatttgt ttatttttct aaatacattc aaatatgtat ccgctcatga gacaataacc
     1741 ctgataaatg cttcaataat attgaaaaag gaagagtatg agtattcaac atttccgtgt
     1801 cgcccttatt cccttttttg cggcattttg ccttcctgtt tttgctcacc cagaaacgct
     1861 ggtgaaagta aaagatgctg aagatcagtt gggtgcacga gtgggttaca tcgaactgga
     1921 tctcaacagc ggtaagatcc ttgagagttt tcgccccgaa gaacgttttc caatgatgag
     1981 cacttttaaa gttctgctat gtggcgcggt attatcccgt attgacgccg ggcaagagca
     2041 actcggtcgc cgcatacact attctcagaa tgacttggtt gagtactcac cagtcacaga
     2101 aaagcatctt acggatggca tgacagtaag agaattatgc agtgctgcca taaccatgag
     2161 tgataacact gcggccaact tacttctgac aacgatcgga ggaccgaagg agctaaccgc
     2221 ttttttgcac aacatggggg atcatgtaac tcgccttgat cgttgggaac cggagctgaa
     2281 tgaagccata ccaaacgacg agcgtgacac cacgatgcct gtagcaatgg caacaacgtt
     2341 gcgcaaacta ttaactggcg aactacttac tctagcttcc cggcaacaat taatagactg
     2401 gatggaggcg gataaagttg caggaccact tctgcgctcg gcccttccgg ctggctggtt
     2461 tattgctgat aaatctggag ccggtgagcg tgggtctcgc ggtatcattg cagcactggg
     2521 gccagatggt aagccctccc gtatcgtagt tatctacacg acggggagtc aggcaactat
     2581 ggatgaacga aatagacaga tcgctgagat aggtgcctca ctgattaagc attggtaact
     2641 gtcagaccaa gtttactcat atatacttta gattgattta aaacttcatt tttaatttaa
     2701 aaggatctag gtgaagatcc tttttgataa tctcatgacc aaaatccctt aacgtgagtt
     2761 ttcgttccac tgagcgtcag accccgtaga aaagatcaaa ggatcttctt gagatccttt
     2821 ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg
     2881 tttgccggat caagagctac caactctttt tccgaaggta actggcttca gcagagcgca
     2941 gataccaaat actgttcttc tagtgtagcc gtagttaggc caccacttca agaactctgt
     3001 agcaccgcct acatacctcg ctctgctaat cctgttacca gtggctgctg ccagtggcga
     3061 taagtcgtgt cttaccgggt tggactcaag acgatagtta ccggataagg cgcagcggtc
     3121 gggctgaacg gggggttcgt gcacacagcc cagcttggag cgaacgacct acaccgaact
     3181 gagataccta cagcgtgagc tatgagaaag cgccacgctt cccgaaggga gaaaggcgga
     3241 caggtatccg gtaagcggca gggtcggaac aggagagcgc acgagggagc ttccaggggg
     3301 aaacgcctgg tatctttata gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt
     3361 tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac gccagcaacg cggccttttt
     3421 acggttcctg gccttttgct ggccttttgc tcacatgttc tttcctgcgt tatcccctga
     3481 ttctgtggat aaccgtatta ccgcctttga gtgagctgat accgctcgcc gcagccgaac
     3541 gaccgagcgc agcgagtcag tgagcgagga agcggaagag cgcccaatac gcaaaccgcc
     3601 tctccccgcg cgttggccga ttcattaatg cagctggcac gacaggtttc ccgactggaa
     3661 agcgggcagt gagcgcaacg caattaatgt gagttagctc actcattagg caccctaggc
     3721 tttacacttt atgcttccgg ctcgtatgtt gtgtggaatt gtgagcggat aacaatttca
     3781 cacaggaaac agctatgacc atgattacgc caagctctag ctagaggtcg acggtatcga
     3841 gccccagctg gttctttccg cctcagaagc catagagccc accgcatccc cagcatgcct
     3901 gctattgtct tcccaatcct cccccttgct gtcctgcccc accccacccc ccagaataga
     3961 atgacaccta ctcagacaat gcgatgcaat ttcctcattt tattaggaaa ggacagtggg
     4021 agtggcacct tccagggtca aggaaggcac gggggagggg caaacaacag atggctggca
     4081 actagaaggc acagtcgagg ctgatcagcg ggtttaaact caatggtgat ggtgatgatg
     4141 accggttaga ctttcctctt cttcttggga gaaccaccag atgagaatgc atatgaaggc
     4201 ttaagaaacg gtctatttat aggcttcggt ggagccataa tccccaattt gctattggct
     4261 cctgatgtcg cttgagaagt atgtgaggct gaactggatc caatgatgct ggaggatttc
     4321 gttttggaag atatctttct acatgtggca gaccccccct ttgccttgga accactggaa
     4381 gcagtttttc tcctcttaga cctttgggag gctggcatct ttttcctctt cctttcattt
     4441 ctggttttac ttgcaaagta gtgggaagat acgggtattt cctcgtcaag ctcctcagcg
     4501 gcacttcttc cggggcctct gctgctggac aggcttatcc ctggggaact gtcttcagct
     4561 ggcgatgtcc attcagagta tttctgtaat actgaaatca cttccgcacc atatttttcc
     4621 agtttgtctt cagtaacacc atcaatttga agcaaaacct caggatcaga agataaagat
     4681 tctgcaagct tcttgagagt gacggtatta aaaatattga agtaatggac accaaaaact
     4741 ttccccagag atttgcagac ttctgtaagt tctccaagac attttttaac catctcttcc
     4801 ctctgagaca cttttgctac taacgctttt tgttttttca cactgctgga attttctgtt
     4861 tccataaagt ctacctttaa attgccattt agtacagttt gtgctttatt tccgagcatc
     4921 acataagcga tcgcctggtc attggcattg atatataagt cttcatccaa aatcttgtca
     4981 agtatcagct ttttaaaaag tctttcggca ttgtgtcgtg aataagcaga tccttttcca
     5041 aatatacctg actggatttt tgcactctta ctccccaaga aaatgtcgac cagcatattc
     5101 atagtaaatc ttccagaagg acctacatgt tttatatttc tcattccttg tgatgaacta
     5161 tgttcttgaa caaatcttac aatacttttc acatcgtcag tcacatctct tgttttataa
     5221 tcctttgttt tacagcaatt atcacaagaa acatctgggt gtttcttaca aaaatcagga
     5281 ttaaatccat tttcaccaaa gtaggccaaa agctgtattc tcctgcattc tgttatattt
     5341 tcacagtaat gtaccatgct atacaaatta ttgaagtgag tttctcttgt atgatggttt
     5401 ccatcttttt ccatcattat aagtcttttc agtctggtca catcatgata ggtatagaaa
     5461 agcaggcagt gagatatttc cccatctctt ccagctctgc cagattcttg gtagtaaccc
     5521 tccacagatt tagggagaga tgcatgaatc acaaatcgca cgtccggttt gtcaatcccc
     5581 attccaaatg caattgtagc acagataacc tgacagccat cctgattaat ccacttctgc
     5641 tgcacttcat ctctggcaga atcactgagg ccagcatggt aagcaagagc agcgagccca
     5701 tctctctgta acgtgtcagc catggtgtca cattctcgcc tggagaggca gtaaattatc
     5761 cctgaatcat atgggtggtg ctttctgatc cattctaggc aatcaaatgc caccttttta
     5821 ggctttttcg gtaatacata gtatttcaga ttatgtctgt taaagctcat gctaaacacc
     5881 tgaggtctga gaatcttcag ctgagtcagg atgtccttct gtaccctggg attagctgtg
     5941 gccgtaagag ccatcaccgg aacagaagga aacttctggc gaagcatatt cattcttttg
     6001 taatcttgac gaaaatcatg tccccactga ctgacacaat gtgcttcatc aataacaaaa
     6061 cgtgccaaga gcttcctctc atagagattc tccagagtag aaatgagtct gttacttgca
     6121 cagatctttt ctggagtgac atataggagt tttatgattg ggtctttttt tgataactgg
     6181 aggtaaatat ttgtagcttc tgagtcagtc ttatcacctg tcagatatgt agctggaata
     6241 tccaaggaag tcagcttttg gacttgatct acgataagtg atctcaaggg agaaatgaca
     6301 acagtgaccc caggagaaac acaggcaggg agctggtaac acaaactctt accacctcca
     6361 gtcggcatca ggataaaaca gtcttcacca agcagtgcag cattgatcgc ctctagctga
     6421 ttagttctaa aattatgcag gccaaatttt ttatgaaaaa tcttcatcat ttcctttgta
     6481 tgaggaaaac taatactttg gaaacgctca tgtttcagat ttctggatgc taaattttgt
     6541 gctgacttgt cagtataatt cttaattgac tctgagaagt ttatattttg agcagtagat
     6601 gacactggaa gacagtctgt cttggctgag gaaagtcttt ctgatactga tttaattggc
     6661 cgaccttcct tgatgggttg ataggcagct gtggaagatt tgctggctgc taaattatgc
     6721 attatgtctt cccagtcatc atcatcatca aagtcatcta tgtcaaaatt atcaatatca
     6781 taggaaggtt gggtttctct ttctaagtca tttattgaag cagtatgttt attctgatct
     6841 ttcacagcag tgcttgtgag aacatttcct gggaaataag agctttcatt tttttttcct
     6901 agtcttggtg tttcagccca gttgctactt acaaaggact tctgtaaatg ggtattgaat
     6961 aaaggccttt caaaaagatt cttcctggtg gcagagaatc ctgtctttcc tagggtggta
     7021 gtcagtaaac agtccccagg agaaacagaa tttgagggaa ggtgtgaaaa atttaactcc
     7081 ttcatagaat tccctgtagg gcaggaatca ccctccatag ggccatcaag tgaatcaggc
     7141 ctgtatctcc acaatgagcc aagaagactg gcatcacttt tattaaaatc tacttccgtt
     7201 agaagtttcc ttcttatgtt ccgctgctga agcagttcgt tcccacaatc caaaagtttc
     7261 agtttatcat caggaatagt atcaattaat ttacagatgt gctccatcac atgaataagc
     7321 tgctgctgta aacttatctg tctagcgtca cagtctgtgc tggtttctgg attcagctcc
     7381 tgcatactca ttttctcagg ttttgacaaa agatcttttg atgtgctaag aacatcctct
     7441 tttctgtcag aggtgtcaag gtcctttaac gtactaaggc attttgaaga ggaagaagaa
     7501 gcagaaataa tttcttctgg agaaggtgga acaaaatccg tatcataatc atcatcatca
     7561 aattcaatac atggaacttt ctcagttgaa tgtaattcag cttcttccaa attcttcttc
     7621 ttttcgctat tatctctttc atcttccaaa tgagttttca gagagtcact ttcctgagca
     7681 tcttcattta tatgcacttc agcaatgggg ccatcatcga tgcaaatcac atcgctgctt
     7741 aaccattctg agtcatcctt ctgttcctca gtcaaatcta tttgctcgct ttcagaggag
     7801 ggtggaggca agtcagtctt tactgtgttt gttgtataaa gctgtgcttt aaaaaagttt
     7861 ctcttaccct tttttgattt ctgagcagtg cttactctta caaagtgact ttggggtggt
     7921 gtaacaaatg attttgaagt ctcagaagta tcaaagtcat ccatatcatc ccaatcattg
     7981 atggtactta aagaatctgg tgaagaacta aattctaatt tcttgagagc agtatcccgg
     8041 gatttcttta cagttggtgt gttttgggta gtgcatacaa cttccttcgg agtctgcaag
     8101 aaatctggca ataatgattt tgatccacct ctctgtgttt cctgtcctgc tggagcattt
     8161 ttaaagaagt ccttgaccct ttgctgattt gtggtgttgg gtagaggttc actgaagga
//
