LOCUS       sequence-485708-        8801 bp    DNA     circular SYN 10-SEP-2025
DEFINITION  Expresses the QBEn 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 10, 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: QBEn
FEATURES             Location/Qualifiers
     source          1..8801
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(join(171..962,963..1031))
                     /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     794..813
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(1032..1136)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(1215..1234)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        1406..1785
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        1786..1989
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     1940..1960
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     CDS             2067..2087
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             6897..7145
                     /codon_start=1
                     /product="uracil-DNA glycosylase inhibitor from a Bacillus 
                     subtilis bacteriophage (Mol et al., 1995)"
                     /label=UGI
                     /translation="TNLSDIIEKETGKQLVIQESILMLPEEVEEVIGNKPESDILVHTA
                     YDESTDENVMLLTSDAPEYKPWALVIQDSNGENKIKML"
     CDS             7176..7424
                     /codon_start=1
                     /product="uracil-DNA glycosylase inhibitor from a Bacillus 
                     subtilis bacteriophage (Mol et al., 1995)"
                     /label=UGI
                     /translation="TNLSDIIEKETGKQLVIQESILMLPEEVEEVIGNKPESDILVHTA
                     YDESTDENVMLLTSDAPEYKPWALVIQDSNGENKIKML"
     CDS             7467..7487
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     primer_bind     complement(7510..7527)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    7516..7740
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(7811..7827)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(7811..7827)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(7824..7846)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    7835..7851
                     /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(7859..7865,7866..7883,7884..7889))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7904..7925
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(8042..8059)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(8213..8801)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(8293..8312)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 gaagatcctt tgatcttttc tacggggtct gacactcagt ggaacgaaaa ctcacgttaa
       61 gggattttgg tcatgagatt atcaaaaagg atcttcacct agatcctttt aaattaaaaa
      121 tgaagtttta aatcaatcta aagtatatat gagtaaactt ggtctgacag ttaccaatgc
      181 ttaatcagtg aggcacctat ctcagcgatc tgtctatttc gttcatccat agttgcctga
      241 ctccccgtcg tgtagataac tacgatacgg gagggcttac catctggccc cagtgctgca
      301 atgataccgc gagacccacg ctcaccggct ccagatttat cagcaataaa ccagccagcc
      361 ggaagggccg agcgcagaag tggtcctgca actttatccg cctccatcca gtctattaat
      421 tgttgccggg aagctagagt aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc
      481 attgctacag gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt cagctccggt
      541 tcccaacgat caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc
      601 ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag tgttatcact catggttatg
      661 gcagcactgc ataattctct tactgtcatg ccatccgtaa gatgcttttc tgtgactggt
      721 gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg
      781 gcgtcaatac gggataatac cgcgccacat agcagaactt taaaagtgct catcattgga
      841 aaacgttctt cggggcgaaa actctcaagg atcttaccgc tgttgagatc cagttcgatg
      901 taacccactc gtgcacccaa ctgatcttca gcatctttta ctttcaccag cgtttctggg
      961 tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa taagggcgac acggaaatgt
     1021 tgaatactca tactcttcct ttttcaatat tattgaagca tttatcaggg ttattgtctc
     1081 atgagcggat acatatttga atgtatttag aaaaataaac aaataggggt tccgcgcaca
     1141 tttccccgaa aagtgccacc tgacgtcgac ggatcgggag atcgatctcc cgatccccta
     1201 gggtcgactc tcagtacaat ctgctctgat gccgcatagt taagccagta tctgctccct
     1261 gcttgtgtgt tggaggtcgc tgagtagtgc gcgagcaaaa tttaagctac aacaaggcaa
     1321 ggcttgaccg acaattgcat gaagaatctg cttagggtta ggcgttttgc gctgcttcgc
     1381 gatgtacggg ccagatatac gcgttgacat tgattattga ctagttatta atagtaatca
     1441 attacggggt cattagttca tagcccatat attgagttcc gcgttacata acttacggta
     1501 aatggcccgc ctggctgacc gcccaacgac ccccgcccat tgacgtcaat aatgacgtat
     1561 gttcccatag taacgccaat agggactttc cattgacgtc aatgggtgga gtatttacgg
     1621 taaactgccc acttggcagt acatcaagtg tatcatatgc caagtacgcc ccctattgac
     1681 gtcaatgacg gtaaatggcc cgcctggcat tatgcccagt acatgacctt atgggacttt
     1741 cctacttggc agtacatcta cgtattagtc atcgctatta ccatggtgat gcggttttgg
     1801 cagtacatca atgggcgtgg atagcggttt gactcacggg gatttccaag tctccacccc
     1861 attgacgtca atgggagttt gttttggcac caaaatcaac gggactttcc aaaatgtcgt
     1921 aacaactccg ccccattgac gcaaatgggc ggtaggcgtg tacggtggga ggtctatata
     1981 agcagagctg gtttagtgaa ccgtcagatc cgctagagat ccgcggccgc atgaaaagaa
     2041 ccgctgatgg cagcgagttc gagagcccta agaagaagcg gaaggtggcc agactgagca
     2101 agagcagacg gctggaaaat ctgatcgccc agctgcccgg cgagaagaag aatggcctgt
     2161 tcggaaacct gattgccctg agcctgggcc tgacccccaa cttcaagagc aacttcgacc
     2221 tggccgagga tgccaaactg cagctgagca aggacaccta cgacgacgac ctggacaacc
     2281 tgctggccca gatcggcgac cagtacgccg acctgtttct ggccgccaag aacctgtccg
     2341 acgccatcct gctgagcgac atcctgagag tgaacaccga gatcaccaag gcccccctga
     2401 gcgcctctat gatcaagaga tacgacgagc accaccagga cctgaccctg ctgaaagctc
     2461 tcgtgcggca gcagctgcct gagaagtaca aagagatttt cttcgaccag agcaagaacg
     2521 gctacgccgg ctacattgac ggcggagcca gccaggaaga gttctacaag ttcatcaagc
     2581 ccatcctgga aaagatggac ggcaccgagg aactgctcgt gaagctgaac agagaggacc
     2641 tgctgcggaa gcagcggacc ttcgacaacg gcagcatccc ccaccagatc cacctgggag
     2701 agctgcacgc cattctgcgg cggcaggaag atttttaccc attcctgaag gacaaccggg
     2761 aaaagatcga gaagatcctg accttccgca tcccctacta cgtgggccct ctggccaggg
     2821 gaaacagcag attcgcctgg atgaccagaa agagcgagga aaccatcacc ccctggaact
     2881 tcgaggaagt ggtggacaag ggcgcttccg cccagagctt catcgagcgg atgaccaact
     2941 tcgataagaa cctgcccaac gagaaggtgc tgcccaagca cagcctgctg tacgagtact
     3001 tcaccgtgta taacgagctg accaaagtga aatacgtgac cgagggaatg agaaagcccg
     3061 ccttcctgag cggcgagcag aaaaaggcca tcgtggacct gctgttcaag accaaccgga
     3121 aagtgaccgt gaagcagctg aaagaggact acttcaagaa aatcgagtgc ttcgactccg
     3181 tggaaatctc cggcgtggaa gatcggttca acgcctccct gggcacatac cacgatctgc
     3241 tgaaaattat caaggacaag gacttcctgg acaatgagga aaacgaggac attctggaag
     3301 atatcgtgct gaccctgaca ctgtttgagg acagagagat gatcgaggaa cggctgaaaa
     3361 cctatgccca cctgttcgac gacaaagtga tgaagcagct gaagcggcgg agatacaccg
     3421 gctggggcag gctgagccgg aagctgatca acggcatccg ggacaagcag tccggcaaga
     3481 caatcctgga tttcctgaag tccgacggct tcgccaacag aaacttcatg cagctgatcc
     3541 acgacgacag cctgaccttt aaagaggaca tccagaaagc ccaggtgtcc ggccagggcg
     3601 atagcctgca cgagcacatt gccaatctgg ccggcagccc cgccattaag aagggcatcc
     3661 tgcagacagt gaaggtggtg gacgagctcg tgaaagtgat gggccggcac aagcccgaga
     3721 acatcgtgat cgaaatggcc agagagaacc agaccaccca gaagggacag aagaacagcc
     3781 gcgagagaat gaagcggatc gaagagggca tcaaagagct gggcagccag atcctgaaag
     3841 aacaccccgt ggaaaacacc cagctgcaga acgagaagct gtacctgtac tacctgcaga
     3901 atgggcggga tatgtacgtg gaccaggaac tggacatcaa ccggctgtcc gactacgatg
     3961 tggaccatat cgtgcctcag agctttctga aggacgactc catcgacaac aaggtgctga
     4021 ccagaagcga caagaaccgg ggcaagagcg acaacgtgcc ctccgaagag gtcgtgaaga
     4081 agatgaagaa ctactggcgg cagctgctga acgccaagct gattacccag agaaagttcg
     4141 acaatctgac caaggccgag agaggcggcc tgagcgaact ggataaggcc ggcttcatca
     4201 agagacagct ggtggaaacc cggcagatca caaagcacgt ggcacagatc ctggactccc
     4261 ggatgaacac taagtacgac gagaatgaca agctgatccg ggaagtgaaa gtgatcaccc
     4321 tgaagtccaa gctggtgtcc gatttccgga aggatttcca gttttacaaa gtgcgcgaga
     4381 tcaacaacta ccaccacgcc cacgacgcct acctgaacgc cgtcgtggga accgccctga
     4441 tcaaaaagta ccctaagctg gaaagcgagt tcgtgtacgg cgactacaag gtgtacgacg
     4501 tgcggaagat gatcgccaag agcgagtctg gaggatctag cggaggatcc tctggaagcg
     4561 agacaccagg cacaagcgag tccgccacac cagagagctc cggcggctcc tccggaggat
     4621 cctgtcccat ctgggtcaag aacgccctgc aagagctggt gggccggaag gagacaagcg
     4681 gcaaggtgtt tgatgtggat ggaaagccca ttggccctga catccaaagc ggctacaaag
     4741 accgggaact gagaagaggt gtgtatgaaa ccctcagaaa gtcccccttc ttccagaagc
     4801 acttccctag cacagccacc tggtacgtga gcctccacgt ggaagcccag tacgctgtgt
     4861 ggatgctgag gaacagaatc aaacatgcta ccgtggtgat caacaacaca tacgtctgct
     4921 ctgacatgaa ccggctgcat gacaactgca tgaccgctgt cccacacatt ctgccagagg
     4981 gctacaccat gacagtctgg aaggctgaca ggacagaggt aacacttcgg ggcaaagccc
     5041 ccaaagaatc cggaggatct agcggaggct cctctggctc tgagacacct ggcacaagcg
     5101 agagcgcaac acctgaaagc agcgggggca gcagcggggg gtcacaggaa atcggcaagg
     5161 ctaccgccaa gtacttcttc tacagcaaca tcatgaactt tttcaagacc gagattaccc
     5221 tggccaacgg cgagatccgg aagcggcctc tgatcgagac aaacggcgaa accggggaga
     5281 tcgtgtggga taagggccgg gattttgcca ccgtgcggaa agtgctgagc atgccccaag
     5341 tgaatatcgt gaaaaagacc gaggtgcaga caggcggctt cagcaaagag tctatcctgc
     5401 ccaagaggaa cagcgataag ctgatcgcca gaaagaagga ctgggaccct aagaagtacg
     5461 gcggcttcga cagccccacc gtggcctatt ctgtgctggt ggtggccaaa gtggaaaagg
     5521 gcaagtccaa gaaactgaag agtgtgaaag agctgctggg gatcaccatc atggaaagaa
     5581 gcagcttcga gaagaatccc atcgactttc tggaagccaa gggctacaaa gaagtgaaaa
     5641 aggacctgat catcaagctg cctaagtact ccctgttcga gctggaaaac ggccggaaga
     5701 gaatgctggc ctctgccggc gaactgcaga agggaaacga actggccctg ccctccaaat
     5761 atgtaaactt cctgtacctg gccagccact atgagaagct gaagggctcc cccgaggata
     5821 atgagcagaa acagctgttt gtggaacagc acaagcacta cctggacgag atcatcgagc
     5881 agatcagcga gttctccaag agagtgatcc tggccgacgc taatctggac aaagtgctgt
     5941 ccgcctacaa caagcaccgg gataagccca tcagagagca ggccgagaat atcatccacc
     6001 tgtttaccct gaccaatctg ggagcccctg ccgccttcaa gtactttgac accaccatcg
     6061 accggaagag gtacaccagc accaaagagg tgctggacgc caccctgatc caccagagca
     6121 tcaccggcct gtacgagaca cggatcgacc tgtctcagct gggaggcgac ggcggatcag
     6181 gaggctctgg cggttcaggt ggatcaggcg gtagcggagg ttcaggtggt gacaagaagt
     6241 acagcatcgg cctggccatc ggcaccaact ctgtgggctg ggccgtgatc accgacgagt
     6301 acaaggtgcc cagcaagaaa ttcaaggtgc tgggcaacac cgaccggcac agcatcaaga
     6361 agaacctgat cggagccctg ctgttcgaca gcggcgaaac agccgaggcc acccggctga
     6421 agagaaccgc cagaagaaga tacaccagac ggaagaaccg gatctgctat ctgcaagaga
     6481 tcttcagcaa cgagatggcc aaggtggacg acagcttctt ccacagactg gaagagtcct
     6541 tcctggtgga agaggataag aagcacgagc ggcaccccat cttcggcaac atcgtggacg
     6601 aggtggccta ccacgagaag taccccacca tctaccacct gagaaagaaa ctggtggaca
     6661 gcaccgacaa ggccgacctg cggctgatct atctggccct ggcccacatg atcaagttcc
     6721 ggggccactt cctgatcgag ggcgacctga accccgacaa cagcgacgtg gacaagctgt
     6781 tcatccagct ggtgcagacc tacaaccagc tgttcgagga aaaccccatc aacgccagcg
     6841 gcgtggacgc caaggccatc ctgtctagcg gcgggagcgg cgggagcggg gggagcacta
     6901 atctgagcga catcattgag aaggagactg ggaaacagct ggtcattcag gagtccatcc
     6961 tgatgctgcc tgaggaggtg gaggaagtga tcggcaacaa gccagagtct gacatcctgg
     7021 tgcacaccgc ctacgacgag tccacagatg agaatgtgat gctgctgacc tctgacgccc
     7081 ccgagtataa gccttgggcc ctggtcatcc aggattctaa cggcgagaat aagatcaaga
     7141 tgctgagcgg aggatccgga ggatctggag gcagcaccaa cctgtctgac atcatcgaga
     7201 aggagacagg caagcagctg gtcatccagg agagcatcct gatgctgccc gaagaagtcg
     7261 aagaagtgat cggaaacaag cctgagagcg atatcctggt ccataccgcc tacgacgaga
     7321 gtaccgacga aaatgtgatg ctgctgacat ccgacgcccc agagtataag ccctgggctc
     7381 tggtcatcca ggattccaac ggagagaaca aaatcaaaat gctgagcggc ggcagcaaaa
     7441 gaaccgccga cggctcagag ttcgagccca aaaagaagcg gaaagtgtaa gtttaaaccc
     7501 gctgatcagc ctcgactgtg ccttctagtt gccagccatc tgttgtttgc ccctcccccg
     7561 tgccttcctt gaccctggaa ggtgccactc ccactgtcct ttcctaataa aatgaggaaa
     7621 ttgcatcgca ttgtctgagt aggtgtcatt ctattctggg gggtggggtg gggcaggaca
     7681 gcaaggggga ggattgggaa gacaatagca ggcatgctgg ggatgcggtg ggctctatgg
     7741 cttctgaggc ggaaagaacc agctggggct cgataccgtc gacctctagc tagagcttag
     7801 cgtaatcatg gtcatagctg tttcctgtgt gaaattgtta tccgctcaca attccacaca
     7861 acatacgagc cggaagcata aagtgtaaag cctagggtgc ctaatgagtg agctaactca
     7921 cattaattgc gttgcgctca ctgcccgctt tccagtcggg aaacctgtcg tgccagctgc
     7981 attaatgaat cggccaacgc gcggggagag gcggtttgcg tattgggcgc tcttccgctt
     8041 cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta tcagctcact
     8101 caaaggcggt aatacggtta tccacagaat caggggataa cgcaggaaag aacatgtgag
     8161 caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg tttttccata
     8221 ggctccgccc ccctgacgag catcacaaaa atcgacgctc aagtcagagg tggcgaaacc
     8281 cgacaggact ataaagatac caggcgtttc cccctggaag ctccctcgtg cgctctcctg
     8341 ttccgaccct gccgcttacc ggatacctgt ccgcctttct cccttcggga agcgtggcgc
     8401 tttctcatag ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg
     8461 gctgtgtgca cgaacccccc gttcagcccg accgctgcgc cttatccggt aactatcgtc
     8521 ttgagtccaa cccggtaaga cacgacttat cgccactggc agcagccact ggtaacagga
     8581 ttagcagagc gaggtatgta ggcggtgcta cagagttctt gaagtggtgg cctaactacg
     8641 gctacactag aagaacagta tttggtatct gcgctctgct gaagccagtt accttcggaa
     8701 aaagagttgg tagctcttga tccggcaaac aaaccaccgc tggtagcggt ggtttttttg
     8761 tttgcaagca gcagattacg cgcagaaaaa aaggatctca a
//
