LOCUS       sequence-485458-        8801 bp    DNA     circular SYN 08-SEP-2025
DEFINITION  Expresses the QBEmax base editor in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8801)
  AUTHORS   Hu J, Guo M, Gao Q, Jia H, He M, Wang Z, Guo L, Liu G, Gao Q, Zhao 
            KT
  TITLE     QBEmax is a sequence-permuted and internally protected base editor.
  JOURNAL   Nat Biotechnol. 2025 Apr 21. doi: 10.1038/s41587-025-02641-9.
  PUBMED    40258958
REFERENCE   2  (bases 1 to 8801)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Sep 8, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Nat 
            Biotechnol. 2025 Apr 21. doi: 10.1038/s41587-025-02641-9."
COMMENT     Sequence Label: QBEmax
FEATURES             Location/Qualifiers
     source          1..8801
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(join(172..963,964..1032))
                     /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     795..814
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(1033..1137)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(1216..1235)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        1407..1786
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        1787..1990
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     1941..1961
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     CDS             2068..2088
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             3781..5496
                     /codon_start=1
                     /product="N-terminal portion of Streptococcus pyogenes Cas9
                     (Zetsche et al., 2015)"
                     /label=Cas9(N)
                     /translation="DKKYSIGLAIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKKN
                     LIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEESF
                     LVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIKF
                     RGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRLE
                     NLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQI
                     GDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVRQ
                     QLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLLR
                     KQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARGN
                     SRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEYF
                     TVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIE"
     CDS             6898..7146
                     /codon_start=1
                     /product="uracil-DNA glycosylase inhibitor from a Bacillus 
                     subtilis bacteriophage (Mol et al., 1995)"
                     /label=UGI
                     /translation="TNLSDIIEKETGKQLVIQESILMLPEEVEEVIGNKPESDILVHTA
                     YDESTDENVMLLTSDAPEYKPWALVIQDSNGENKIKML"
     CDS             7177..7425
                     /codon_start=1
                     /product="uracil-DNA glycosylase inhibitor from a Bacillus 
                     subtilis bacteriophage (Mol et al., 1995)"
                     /label=UGI
                     /translation="TNLSDIIEKETGKQLVIQESILMLPEEVEEVIGNKPESDILVHTA
                     YDESTDENVMLLTSDAPEYKPWALVIQDSNGENKIKML"
     CDS             7468..7488
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     primer_bind     complement(7511..7528)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    7517..7741
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(7812..7828)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(7812..7828)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(7825..7847)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    7836..7852
                     /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(7860..7866,7867..7884,7885..7890))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7905..7926
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(8043..8060)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(join(8214..8801,1))
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(8294..8313)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 agaagatcct ttgatctttt ctacggggtc tgacactcag tggaacgaaa actcacgtta
       61 agggattttg gtcatgagat tatcaaaaag gatcttcacc tagatccttt taaattaaaa
      121 atgaagtttt aaatcaatct aaagtatata tgagtaaact tggtctgaca gttaccaatg
      181 cttaatcagt gaggcaccta tctcagcgat ctgtctattt cgttcatcca tagttgcctg
      241 actccccgtc gtgtagataa ctacgatacg ggagggctta ccatctggcc ccagtgctgc
      301 aatgataccg cgagacccac gctcaccggc tccagattta tcagcaataa accagccagc
      361 cggaagggcc gagcgcagaa gtggtcctgc aactttatcc gcctccatcc agtctattaa
      421 ttgttgccgg gaagctagag taagtagttc gccagttaat agtttgcgca acgttgttgc
      481 cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat tcagctccgg
      541 ttcccaacga tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag cggttagctc
      601 cttcggtcct ccgatcgttg tcagaagtaa gttggccgca gtgttatcac tcatggttat
      661 ggcagcactg cataattctc ttactgtcat gccatccgta agatgctttt ctgtgactgg
      721 tgagtactca accaagtcat tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc
      781 ggcgtcaata cgggataata ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg
      841 aaaacgttct tcggggcgaa aactctcaag gatcttaccg ctgttgagat ccagttcgat
      901 gtaacccact cgtgcaccca actgatcttc agcatctttt actttcacca gcgtttctgg
      961 gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga ataagggcga cacggaaatg
     1021 ttgaatactc atactcttcc tttttcaata ttattgaagc atttatcagg gttattgtct
     1081 catgagcgga tacatatttg aatgtattta gaaaaataaa caaatagggg ttccgcgcac
     1141 atttccccga aaagtgccac ctgacgtcga cggatcggga gatcgatctc ccgatcccct
     1201 agggtcgact ctcagtacaa tctgctctga tgccgcatag ttaagccagt atctgctccc
     1261 tgcttgtgtg ttggaggtcg ctgagtagtg cgcgagcaaa atttaagcta caacaaggca
     1321 aggcttgacc gacaattgca tgaagaatct gcttagggtt aggcgttttg cgctgcttcg
     1381 cgatgtacgg gccagatata cgcgttgaca ttgattattg actagttatt aatagtaatc
     1441 aattacgggg tcattagttc atagcccata tattgagttc cgcgttacat aacttacggt
     1501 aaatggcccg cctggctgac cgcccaacga cccccgccca ttgacgtcaa taatgacgta
     1561 tgttcccata gtaacgccaa tagggacttt ccattgacgt caatgggtgg agtatttacg
     1621 gtaaactgcc cacttggcag tacatcaagt gtatcatatg ccaagtacgc cccctattga
     1681 cgtcaatgac ggtaaatggc ccgcctggca ttatgcccag tacatgacct tatgggactt
     1741 tcctacttgg cagtacatct acgtattagt catcgctatt accatggtga tgcggttttg
     1801 gcagtacatc aatgggcgtg gatagcggtt tgactcacgg ggatttccaa gtctccaccc
     1861 cattgacgtc aatgggagtt tgttttggca ccaaaatcaa cgggactttc caaaatgtcg
     1921 taacaactcc gccccattga cgcaaatggg cggtaggcgt gtacggtggg aggtctatat
     1981 aagcagagct ggtttagtga accgtcagat ccgctagaga tccgcggccg catgaaaaga
     2041 accgctgatg gcagcgagtt cgagagccct aagaagaagc ggaaggtgaa ggctaccgcc
     2101 aagtacttct tctacagcaa catcatgaac tttttcaaga ccgagattac cctggccaac
     2161 ggcgagatcc ggaagcggcc tctgatcgag acaaacggcg aaaccgggga gatcgtgtgg
     2221 gataagggcc gggattttgc caccgtgcgg aaagtgctga gcatgcccca agtgaatatc
     2281 gtgaaaaaga ccgaggtgca gacaggcggc ttcagcaaag agtctatcct gcccaagagg
     2341 aacagcgata agctgatcgc cagaaagaag gactgggacc ctaagaagta cggcggcttc
     2401 gacagcccca ccgtggccta ttctgtgctg gtggtggcca aagtggaaaa gggcaagtcc
     2461 aagaaactga agagtgtgaa agagctgctg gggatcacca tcatggaaag aagcagcttc
     2521 gagaagaatc ccatcgactt tctggaagcc aagggctaca aagaagtgaa aaaggacctg
     2581 atcatcaagc tgcctaagta ctccctgttc gagctggaaa acggccggaa gagaatgctg
     2641 gcctctgccg gcgaactgca gaagggaaac gaactggccc tgccctccaa atatgtgaac
     2701 ttcctgtacc tggccagcca ctatgagaag tctggaggat ctagcggagg atcctctgga
     2761 agcgagacac caggcacaag cgagtccgcc acaccagaga gctccggcgg ctcctccgga
     2821 ggatcctgtc ccatctgggt caagaacgcc ctgcaagagc tggtgggccg gaaggagaca
     2881 agcggcaagg tgtttgatgt ggatggaaag cccattggcc ctgacatcca aagcggctac
     2941 aaagaccggg aactgagaag aggtgtgtat gaaaccctca gaaagtcccc cttcttccag
     3001 aagcacttcc ctagcacagc cacctggtac gtgagcctcc acgtggaagc ccagtacgct
     3061 gtgtggatgc tgaggaacag aatcaaacat gctaccgtgg tgatcaacaa cacatacgtc
     3121 tgctctgaca tgaaccggct gcatgacaac tgcatgaccg ctgtcccaca cattctgcca
     3181 gagggctaca ccatgacagt ctggaaggct gacaggacag aggtaacact tcggggcaaa
     3241 gcccccaaag aatccggagg atctagcgga ggctcctctg gctctgagac acctggcaca
     3301 agcgagagcg caacacctga aagcagcggg ggcagcagcg gggggtcact gaagggctcc
     3361 cccgaggata atgagcagaa acagctgttt gtggaacagc acaagcacta cctggacgag
     3421 atcatcgagc agatcagcga gttctccaag agagtgatcc tggccgacgc taatctggac
     3481 aaagtgctgt ccgcctacaa caagcaccgg gataagccca tcagagagca ggccgagaat
     3541 atcatccacc tgtttaccct gaccaatctg ggagcccctg ccgccttcaa gtactttgac
     3601 accaccatcg accggaagag gtacaccagc accaaagagg tgctggacgc caccctgatc
     3661 caccagagca tcaccggcct gtacgagaca cggatcgacc tgtctcagct gggaggcgac
     3721 ggcggatcag gaggctctgg cggttcaggt ggatcaggcg gtagcggagg ttcaggtggt
     3781 gacaagaagt acagcatcgg cctggccatc ggcaccaact ctgtgggctg ggccgtgatc
     3841 accgacgagt acaaggtgcc cagcaagaaa ttcaaggtgc tgggcaacac cgaccggcac
     3901 agcatcaaga agaacctgat cggagccctg ctgttcgaca gcggcgaaac agccgaggcc
     3961 acccggctga agagaaccgc cagaagaaga tacaccagac ggaagaaccg gatctgctat
     4021 ctgcaagaga tcttcagcaa cgagatggcc aaggtggacg acagcttctt ccacagactg
     4081 gaagagtcct tcctggtgga agaggataag aagcacgagc ggcaccccat cttcggcaac
     4141 atcgtggacg aggtggccta ccacgagaag taccccacca tctaccacct gagaaagaaa
     4201 ctggtggaca gcaccgacaa ggccgacctg cggctgatct atctggccct ggcccacatg
     4261 atcaagttcc ggggccactt cctgatcgag ggcgacctga accccgacaa cagcgacgtg
     4321 gacaagctgt tcatccagct ggtgcagacc tacaaccagc tgttcgagga aaaccccatc
     4381 aacgccagcg gcgtggacgc caaggccatc ctgtctgcca gactgagcaa gagcagacgg
     4441 ctggaaaatc tgatcgccca gctgcccggc gagaagaaga atggcctgtt cggaaacctg
     4501 attgccctga gcctgggcct gacccccaac ttcaagagca acttcgacct ggccgaggat
     4561 gccaaactgc agctgagcaa ggacacctac gacgacgacc tggacaacct gctggcccag
     4621 atcggcgacc agtacgccga cctgtttctg gccgccaaga acctgtccga cgccatcctg
     4681 ctgagcgaca tcctgagagt gaacaccgag atcaccaagg cccccctgag cgcctctatg
     4741 atcaagagat acgacgagca ccaccaggac ctgaccctgc tgaaagctct cgtgcggcag
     4801 cagctgcctg agaagtacaa agagattttc ttcgaccaga gcaagaacgg ctacgccggc
     4861 tacattgacg gcggagccag ccaggaagag ttctacaagt tcatcaagcc catcctggaa
     4921 aagatggacg gcaccgagga actgctcgtg aagctgaaca gagaggacct gctgcggaag
     4981 cagcggacct tcgacaacgg cagcatcccc caccagatcc acctgggaga gctgcacgcc
     5041 attctgcggc ggcaggaaga tttttaccca ttcctgaagg acaaccggga aaagatcgag
     5101 aagatcctga ccttccgcat cccctactac gtgggccctc tggccagggg aaacagcaga
     5161 ttcgcctgga tgaccagaaa gagcgaggaa accatcaccc cctggaactt cgaggaagtg
     5221 gtggacaagg gcgcttccgc ccagagcttc atcgagcgga tgaccaactt cgataagaac
     5281 ctgcccaacg agaaggtgct gcccaagcac agcctgctgt acgagtactt caccgtgtat
     5341 aacgagctga ccaaagtgaa atacgtgacc gagggaatga gaaagcccgc cttcctgagc
     5401 ggcgagcaga aaaaggccat cgtggacctg ctgttcaaga ccaaccggaa agtgaccgtg
     5461 aagcagctga aagaggacta cttcaagaaa atcgagtgct tcgactccgt ggaaatctcc
     5521 ggcgtggaag atcggttcaa cgcctccctg ggcacatacc acgatctgct gaaaattatc
     5581 aaggacaagg acttcctgga caatgaggaa aacgaggaca ttctggaaga tatcgtgctg
     5641 accctgacac tgtttgagga cagagagatg atcgaggaac ggctgaaaac ctatgcccac
     5701 ctgttcgacg acaaagtgat gaagcagctg aagcggcgga gatacaccgg ctggggcagg
     5761 ctgagccgga agctgatcaa cggcatccgg gacaagcagt ccggcaagac aatcctggat
     5821 ttcctgaagt ccgacggctt cgccaacaga aacttcatgc agctgatcca cgacgacagc
     5881 ctgaccttta aagaggacat ccagaaagcc caggtgtccg gccagggcga tagcctgcac
     5941 gagcacattg ccaatctggc cggcagcccc gccattaaga agggcatcct gcagacagtg
     6001 aaggtggtgg acgagctcgt gaaagtgatg ggccggcaca agcccgagaa catcgtgatc
     6061 gaaatggcca gagagaacca gaccacccag aagggacaga agaacagccg cgagagaatg
     6121 aagcggatcg aagagggcat caaagagctg ggcagccaga tcctgaaaga acaccccgtg
     6181 gaaaacaccc agctgcagaa cgagaagctg tacctgtact acctgcagaa tgggcgggat
     6241 atgtacgtgg accaggaact ggacatcaac cggctgtccg actacgatgt ggaccatatc
     6301 gtgcctcaga gctttctgaa ggacgactcc atcgacaaca aggtgctgac cagaagcgac
     6361 aagaaccggg gcaagagcga caacgtgccc tccgaagagg tcgtgaagaa gatgaagaac
     6421 tactggcggc agctgctgaa cgccaagctg attacccaga gaaagttcga caatctgacc
     6481 aaggccgaga gaggcggcct gagcgaactg gataaggccg gcttcatcaa gagacagctg
     6541 gtggaaaccc ggcagatcac aaagcacgtg gcacagatcc tggactcccg gatgaacact
     6601 aagtacgacg agaatgacaa gctgatccgg gaagtgaaag tgatcaccct gaagtccaag
     6661 ctggtgtccg atttccggaa ggatttccag ttttacaaag tgcgcgagat caacaactac
     6721 caccacgccc acgacgccta cctgaacgcc gtcgtgggaa ccgccctgat caaaaagtac
     6781 cctaagctgg aaagcgagtt cgtgtacggc gactacaagg tgtacgacgt gcggaagatg
     6841 atcgccaaga gcgagcagga aatcggcagc ggcgggagcg gcgggagcgg ggggagcact
     6901 aatctgagcg acatcattga gaaggagact gggaaacagc tggtcattca ggagtccatc
     6961 ctgatgctgc ctgaggaggt ggaggaagtg atcggcaaca agccagagtc tgacatcctg
     7021 gtgcacaccg cctacgacga gtccacagat gagaatgtga tgctgctgac ctctgacgcc
     7081 cccgagtata agccttgggc cctggtcatc caggattcta acggcgagaa taagatcaag
     7141 atgctgagcg gaggatccgg aggatctgga ggcagcacca acctgtctga catcatcgag
     7201 aaggagacag gcaagcagct ggtcatccag gagagcatcc tgatgctgcc cgaagaagtc
     7261 gaagaagtga tcggaaacaa gcctgagagc gatatcctgg tccataccgc ctacgacgag
     7321 agtaccgacg aaaatgtgat gctgctgaca tccgacgccc cagagtataa gccctgggct
     7381 ctggtcatcc aggattccaa cggagagaac aaaatcaaaa tgctgagcgg cggcagcaaa
     7441 agaaccgccg acggctcaga gttcgagccc aaaaagaagc ggaaagtgta agtttaaacc
     7501 cgctgatcag cctcgactgt gccttctagt tgccagccat ctgttgtttg cccctccccc
     7561 gtgccttcct tgaccctgga aggtgccact cccactgtcc tttcctaata aaatgaggaa
     7621 attgcatcgc attgtctgag taggtgtcat tctattctgg ggggtggggt ggggcaggac
     7681 agcaaggggg aggattggga agacaatagc aggcatgctg gggatgcggt gggctctatg
     7741 gcttctgagg cggaaagaac cagctggggc tcgataccgt cgacctctag ctagagcttg
     7801 gcgtaatcat ggtcatagct gtttcctgtg tgaaattgtt atccgctcac aattccacac
     7861 aacatacgag ccggaagcat aaagtgtaaa gcctagggtg cctaatgagt gagctaactc
     7921 acattaattg cgttgcgctc actgcccgct ttccagtcgg gaaacctgtc gtgccagctg
     7981 cattaatgaa tcggccaacg cgcggggaga ggcggtttgc gtattgggcg ctcttccgct
     8041 tcctcgctca ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt atcagctcac
     8101 tcaaaggcgg taatacggtt atccacagaa tcaggggata acgcaggaaa gaacatgtga
     8161 gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc gtttttccat
     8221 aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag gtggcgaaac
     8281 ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt gcgctctcct
     8341 gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg aagcgtggcg
     8401 ctttctcata gctcacgctg taggtatctc agttcggtgt aggtcgttcg ctccaagctg
     8461 ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg taactatcgt
     8521 cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac tggtaacagg
     8581 attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg gcctaactac
     8641 ggctacacta gaagaacagt atttggtatc tgcgctctgc tgaagccagt taccttcgga
     8701 aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg tggttttttt
     8761 gtttgcaagc agcagattac gcgcagaaaa aaaggatctc a
//
