LOCUS       sequence-437074-        5708 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Express HACE Editor as a fusion of AID cytidine deaminase and Ns3h 
            helicase..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5708)
  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 5708)
  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-AID-Ns3h-UGI
FEATURES             Location/Qualifiers
     source          1..5708
                     /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"
     regulatory      384..393
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             2433..2681
                     /codon_start=1
                     /product="uracil-DNA glycosylase inhibitor from a Bacillus 
                     subtilis bacteriophage (Mol et al., 1995)"
                     /label=UGI
                     /translation="TNLSDIIEKETGKQLVIQESILMLPEEVEEVIGNKPESDILVHTA
                     YDESTDENVMLLTSDAPEYKPWALVIQDSNGENKIKML"
     CDS             2694..2714
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             2723..2740
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(2763..2780)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2769..2993
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(3064..3080)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3064..3080)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3077..3099)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3088..3104
                     /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(3112..3118,3119..3136,3137..3142))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3157..3178
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3295..3312)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3466..4054)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3546..3565)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4225..5016,5017..5085))
                     /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     4848..4867
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5086..5190)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(5269..5288)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        join(5460..5708,1..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 tggacagcct cttgatgaac cggagggagt
      421 ttctttacca attcaaaaat gtccgctggg ctaagggtcg gcgtgagacc tacctgtgct
      481 acgtagtgaa gaggcgtgac agtgctacat ccttttcact ggactttggt tatcttcgca
      541 ataagaacgg ctgccacgtg gaattgctct tcctccgcta catctcggac tgggacctag
      601 accctggccg ctgctaccgc gtcacctggt tcatctcctg gagcccctgc tacgactgtg
      661 cccgacatgt ggccgacttt ctgcgaggga accccaacct cagtctgagg atcttcaccg
      721 cgcgcctcta cttctgtgag gaccgcaagg ctgagcccga ggggctgcgg cggctgcacc
      781 gcgccggggt gcaaatagcc atcatgacct tcaaagatta tttttactgc tggaatactt
      841 ttgtagaaaa ccacggaaga actttcaaag cctgggaagg gctgcatgaa aattcagttc
      901 gtctctccag acagcttcgg cgcatccttt tgcccctgta tgaggttgat gacttacgag
      961 acgcatttcg tactagcggc agcgagactc ccgggacctc agagtccgcc acacccgaaa
     1021 gtatggtgga cttcataccc gttgagtcta tggaaactac catgcggtct ccggtcttca
     1081 cagacaactc aacccccccg gctgtaccgc agacattcca agtggcacat ctgcacgctc
     1141 ctactggcag cggcaagagc accaaagtgc cggctgcgta tgcagcccaa gggtacaagg
     1201 tgctcgtcct gaacccgtcc gttgccgcca ccttagggtt tggggcgtat atgtccaagg
     1261 cacacggtat cgaccctaac atcagaactg gggtaaggac cattaccacg ggcggctcca
     1321 ttacgtactc cacctatggc aagttccttg ccgacggtgg ctgttctggg ggcgcctatg
     1381 acatcataat atgtgatgag tgccactcaa ctgactcgac taccatcttg ggcatcggca
     1441 cagtcctgga ccaagcggag acggctggag cgcggctcgt cgtgctcgcc accgctacac
     1501 ctccgggatc ggttaccgtg ccacacccca atatcgagga aataggcctg tccaacaatg
     1561 gagagatccc cttctatggc aaagccatcc ccattgaggc catcaagggg gggaggcatc
     1621 tcattttctg ccattccaag aagaaatgtg acgagctcgc cgcaaagctg acaggcctcg
     1681 gactgaacgc tgtagcatat taccggggcc ttgatgtgtc cgtcataccg cctatcggag
     1741 acgtcgttgt cgtggcaaca gacgctctaa tgacgggttt caccggcgat tttgactcag
     1801 tgatcgactg caatacatgt gtcacccaga cagtcgactt cagcttggat cccaccttca
     1861 ccattgagac gacgaccgtg ccccaagacg cggtgtcgcg ctcgcaacgg cgaggtagaa
     1921 ctggcagggg taggagtggc atctacaggt ttgtgactcc aggagaacgg ccctcgggca
     1981 tgttcgattc ttcggtcctg tgtgagtgct atgacgcggg ctgtgcttgg tatgagctca
     2041 cgcccgctga gacctcggtt aggttgcggg cttacctaaa tacaccaggg ttgcccgtct
     2101 gccaggacca tctggagttc tgggagagcg tcttcacagg cctcacccac atagatgccc
     2161 acttcctgtc ccagactaaa caggcaggag acaactttcc ttacctggtg gcatatcaag
     2221 ctacagtgtg cgccagggct caagctccac ctccatcgtg ggaccaaatg tggaagtgtc
     2281 tcatacggct gaaacctaca ctgcacgggc caacacccct gctgtatagg ctaggagccg
     2341 tccaaaatga ggtcatcctc acacacccca taactaaata catcatggca tgcatgtcgg
     2401 ctgacctgga ggtcgtcact tctggtggtt ctactaatct gtcagatatt attgaaaagg
     2461 agaccggtaa gcaactggtt atccaggaat ccatcctcat gctcccagag gaggtggaag
     2521 aagtcattgg gaacaagccg gaaagcgata tactcgtgca caccgcctac gacgagagca
     2581 ccgacgagaa tgtcatgctt ctgactagcg acgcccctga atacaagcct tgggctctgg
     2641 tcatacagga tagcaacggt gagaacaaga ttaagatgct ctctggtggt tctcccaaga
     2701 agaagaggaa agtctaaccg gtcatcatca ccatcaccat tgagtttaaa cccgctgatc
     2761 agcctcgact gtgccttcta gttgccagcc atctgttgtt tgcccctccc ccgtgccttc
     2821 cttgaccctg gaaggtgcca ctcccactgt cctttcctaa taaaatgagg aaattgcatc
     2881 gcattgtctg agtaggtgtc attctattct ggggggtggg gtggggcagg acagcaaggg
     2941 ggaggattgg gaagacaata gcaggcatgc tggggatgcg gtgggctcta tggcttctga
     3001 ggcggaaaga accagctggg gctcgatacc gtcgacctct agctagagct tggcgtaatc
     3061 atggtcatag ctgtttcctg tgtgaaattg ttatccgctc acaattccac acaacatacg
     3121 agccggaagc ataaagtgta aagcctaggg tgcctaatga gtgagctaac tcacattaat
     3181 tgcgttgcgc tcactgcccg ctttccagtc gggaaacctg tcgtgccagc tgcattaatg
     3241 aatcggccaa cgcgcgggga gaggcggttt gcgtattggg cgctcttccg cttcctcgct
     3301 cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc actcaaaggc
     3361 ggtaatacgg ttatccacag aatcagggga taacgcagga aagaacatgt gagcaaaagg
     3421 ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg
     3481 cccccctgac gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg
     3541 actataaaga taccaggcgt ttccccctgg aagctccctc gtgcgctctc ctgttccgac
     3601 cctgccgctt accggatacc tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca
     3661 tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt
     3721 gtacgaaccc cccgttcagc ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc
     3781 caacccggta agacacgact tatcgccact ggcagcagcc actggtaaca ggattagcag
     3841 agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg tggcctaact acggctacac
     3901 tagaagaaca gtatttggta tctgcgctct gctgaagcca gttaccttcg gaaaaagagt
     3961 tggtagctct tgatccggca aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa
     4021 gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat cctttgatct tttctacggg
     4081 gtctgacgct cagtggaacg aaaactcacg ttaagggatt ttggtcatga gattatcaaa
     4141 aaggatcttc acctagatcc ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat
     4201 atatgagtaa acttggtctg acagttacca atgcttaatc agtgaggcac ctatctcagc
     4261 gatctgtcta tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga taactacgat
     4321 acgggagggc ttaccatctg gccccagtgc tgcaatgata ccgcgagacc cacgctcacc
     4381 ggctccagat ttatcagcaa taaaccagcc agccggaagg gccgagcgca gaagtggtcc
     4441 tgcaacttta tccgcctcca tccagtctat taattgttgc cgagaagcta gagtaagtag
     4501 ttcgccagtt aatagtttgc gcaacgttgt tgccattgct acaggcatcg tggtgtcacg
     4561 ctcgtcgttt ggtatggctt cattcagctc cggttcccaa cgatcaaggc gagttacatg
     4621 atcccccatg ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg ttgtcagaag
     4681 taagttggcc gcagtgttat cactcatggt tatggcagca ctgcataatt ctcttactgt
     4741 catgccatcc gtaagatgct tttctgtgac tggtgagtac tcaaccaagt cattctgaga
     4801 atagtgtatg cggcgaccga gttgctcttg cccggcgtca atacgggata ataccgcgcc
     4861 acatagcaga actttaaaag tgctcatcat tggaaaacgt tcttcggggc gaaaactctc
     4921 aaggatctta ccgctgttga gatccagttc gatgtaaccc actcgtgcac ccaactgatc
     4981 ttcagcatct tttactttca ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc
     5041 cgcaaaaaag ggaataaggg cgacacggaa atgttgaata ctcatactct tcctttttca
     5101 atattattga agcatttatc agggttattg tctcatgagc ggatacatat ttgaatgtat
     5161 ttagaaaaat aaacaaatag gggttccgcg cacatttccc cgaaaagtgc cacctgacgt
     5221 cgacggatcg ggagatcgat ctcccgatcc cctagggtcg actctcagta caatctgctc
     5281 tgatgccgca tagttaagcc agtatctgct ccctgcttgt gtgttggagg tcgctgagta
     5341 gtgcgcgagc aaaatttaag ctacaacaag gcaaggcttg accgacaatt gcatgaagaa
     5401 tctgcttagg gttaggcgtt ttgcgctgct tcgcgatgta cgggccagat atacgcgttg
     5461 acattgatta ttgactagtt attaatagta atcaattacg gggtcattag ttcatagccc
     5521 atatatggag ttccgcgtta cataacttac ggtaaatggc ccgcctggct gaccgcccaa
     5581 cgacccccgc ccattgacgt caataatgac gtatgttccc atagtaacgc caatagggac
     5641 tttccattga cgtcaatggg tggagtattt acggtaaact gcccacttgg cagtacatca
     5701 agtgtatc
//
