LOCUS       sequence-240463-        6357 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  BE PACE selection, express C-intein-Cas9-ugi fusion.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6357)
  AUTHORS   Thuronyi BW, Koblan LW, Levy JM, Yeh WH, Zheng C, Newby GA, Wilson 
            C, Bhaumik M, Shubina-Oleinik O, Holt JR, Liu DR
  TITLE     Continuous evolution of base editors with expanded target 
            compatibility and improved activity.
  JOURNAL   Nat Biotechnol. 2019 Jul 22. pii: 10.1038/s41587-019-0193-0. doi: 
            10.1038/s41587-019-0193-0.
  PUBMED    31332326
REFERENCE   2  (bases 1 to 6357)
  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: "Nat 
            Biotechnol. 2019 Jul 22. pii: 10.1038/s41587-019-0193-0. doi: 
            10.1038/s41587-019-0193-0."
COMMENT     Sequence Label: pBT44c
FEATURES             Location/Qualifiers
     source          1..6357
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             4483..4737
                     /codon_start=1
                     /product="uracil-DNA glycosylase inhibitor from a Bacillus 
                     subtilis bacteriophage (Mol et al., 1995)"
                     /label=UGI
                     /translation="MTNLSDIIEKETGKQLVIQESILMLPEEVEEVIGNKPESDILVHT
                     AYDESTDENVMLLTSDAPEYKPWALVIQDSNGENKIKML"
     CDS             complement(4807..5622)
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     CDS             complement(4807..5622)
                     /codon_start=1
                     /gene="aph(3')-Ia"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     5530..5549
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     5614..5633
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     rep_origin      complement(5763..6308)
                     /direction=LEFT
                     /label=p15A ori
                     /note="Plasmids containing the medium-copy-number p15A 
                     origin of replication can be propagated in E. coli cells 
                     that contain a second plasmid with the ColE1 origin.
                     "
     primer_bind     complement(5843..5862)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 caaataaaac gaaaggctca gtcgaaagac tgggcctttc gttttgctga ggagacttag
       61 ggactctcac agctaacacc acgtcgtccc tatctgctgc cctaggtcta tgagtggttg
      121 ctggataact ttacgggcat gcataaggct cgtatgatat attcagggag accacaacgg
      181 tttccctcta caaataattt tgtttaactt ttactagagt gggaccctac ctgcaggtgc
      241 agtaaggagg aaaaaaaaat gatcaagata gctacaagga agtatcttgg caaacaaaac
      301 gtttatgata ttggagtcga aagagatcac aactttgctc tgaagaacgg attcatagct
      361 tctaattgtt ttaacaaata ctcaataggc ttagctatcg gcacaaatag cgtcggatgg
      421 gcggtgatca ctgatgaata taaggttccg tctaaaaagt tcaaggttct gggaaataca
      481 gaccgccaca gtatcaaaaa aaatcttata ggggctcttt tatttgacag tggagagaca
      541 gcggaagcga ctcgtctcaa acggacagct cgtagaaggt atacacgtcg gaagaatcgt
      601 atttgttatc tacaggagat tttttcaaat gagatggcga aagtagatga tagtttcttt
      661 catcgacttg aagagtcttt tttggtggaa gaagacaaga agcatgaacg tcatcctatt
      721 tttggaaata tagtagatga agttgcttat catgagaaat atccaactat ctatcatctg
      781 cgaaaaaaat tggtagattc tactgataaa gcggatttgc gcttaatcta tttggcctta
      841 gcgcatatga ttaagtttcg tggtcatttt ttgattgagg gagatttaaa tcctgataat
      901 agtgatgtgg acaaactatt tatccagttg gtacaaacct acaatcaatt atttgaagaa
      961 aaccctatta acgcaagtgg agtagatgct aaagcgattc tttctgcacg attgagtaaa
     1021 tcaagacgat tagaaaatct cattgctcag ctccccggtg agaagaaaaa tggcttattt
     1081 gggaatctca ttgctttgtc attgggtttg acccctaatt ttaaatcaaa ttttgatttg
     1141 gcagaagatg ctaaattaca gctttcaaaa gatacttacg atgatgattt agataattta
     1201 ttggcgcaaa ttggagatca atatgctgat ttgtttttgg cagctaagaa tttatcagat
     1261 gctattttac tttcagatat cctaagagta aatactgaaa taactaaggc tcccctatca
     1321 gcttcaatga ttaaacgcta cgatgaacat catcaagact tgactctttt aaaagcttta
     1381 gttcgacaac aacttccaga aaagtataaa gaaatctttt ttgatcaatc aaaaaacgga
     1441 tatgcaggtt atattgatgg gggagctagc caagaagaat tttataaatt tatcaaacca
     1501 attttagaaa aaatggatgg tactgaggaa ttattggtga aactaaatcg tgaagatttg
     1561 ctgcgcaagc aacggacctt tgacaacggc tctattcccc atcaaattca cttgggtgag
     1621 ctgcatgcta ttttgagaag acaagaagac ttttatccat ttttaaaaga caatcgtgag
     1681 aagattgaaa aaatcttgac ttttcgaatt ccttattatg ttggtccatt ggcgcgtggc
     1741 aatagtcgtt ttgcatggat gactcggaag tctgaagaaa caattacccc atggaatttt
     1801 gaagaagttg tcgataaagg tgcttcagct caatcattta ttgaacgcat gacaaacttt
     1861 gataaaaatc ttccaaatga aaaagtacta ccaaaacata gtttgcttta tgagtatttt
     1921 acggtttata acgaattgac aaaggtcaaa tatgttactg aaggaatgcg aaaaccagca
     1981 tttctttcag gtgaacagaa gaaagccatt gttgatttac tcttcaaaac aaatcgaaaa
     2041 gtaaccgtta agcaattaaa agaagattat ttcaaaaaaa tagaatgttt tgatagtgtt
     2101 gaaatttcag gagttgaaga tagatttaat gcttcattag gtacctacca tgatttgcta
     2161 aaaattatta aagataaaga ttttttggat aatgaagaaa atgaagatat cttagaggat
     2221 attgttttaa cattgacctt atttgaagat agggagatga ttgaggaaag acttaaaaca
     2281 tatgctcacc tctttgatga taaggtgatg aaacagctta aacgtcgccg ttatactggt
     2341 tggggacgtt tgtctcgaaa attgattaat ggtattaggg ataagcaatc tggcaaaaca
     2401 atattagatt ttttgaaatc agatggtttt gccaatcgca attttatgca gctgatccat
     2461 gatgatagtt tgacatttaa agaagacatt caaaaagcac aagtgtctgg acaaggcgat
     2521 agtttacatg aacatattgc aaatttagct ggtagccctg ctattaaaaa aggtatttta
     2581 cagactgtaa aagttgttga tgaattggtc aaagtaatgg ggcggcataa gccagaaaat
     2641 atcgttattg aaatggcacg tgaaaatcag acaactcaaa agggccagaa aaattcgcga
     2701 gagcgtatga aacgaatcga agaaggtatc aaagaattag gaagtcagat tcttaaagag
     2761 catcctgttg aaaatactca attgcaaaat gaaaagctct atctctatta tctccaaaat
     2821 ggaagagaca tgtatgtgga ccaagaatta gatattaatc gtttaagtga ttatgatgtc
     2881 gatgccattg ttccacaaag tttccttaaa gacgattcaa tagacaataa ggtcttaacg
     2941 cgttctgata aaaatcgtgg taaatcggat aacgttccaa gtgaagaagt agtcaaaaag
     3001 atgaaaaact attggagaca acttctaaac gccaagttaa tcactcaacg taagtttgat
     3061 aatttaacga aagctgaacg tggaggtttg agtgaacttg ataaagctgg ttttatcaaa
     3121 cgccaattgg ttgaaactcg ccaaatcact aagcatgtgg cacaaatttt ggatagtcgc
     3181 atgaatacta aatacgatga aaatgataaa cttattcgag aggttaaagt gattacctta
     3241 aaatctaaat tagtttctga cttccgaaaa gatttccaat tctataaagt acgtgagatt
     3301 aacaattacc atcatgccca tgatgcgtat ctaaatgccg tcgttggaac tgctttgatt
     3361 aagaaatatc caaaacttga atcggagttt gtctatggtg attataaagt ttatgatgtt
     3421 cgtaaaatga ttgctaagtc tgagcaagaa ataggcaaag caaccgcaaa atatttcttt
     3481 tactctaata tcatgaactt cttcaaaaca gaaattacac ttgcaaatgg agagattcgc
     3541 aaacgccctc taatcgaaac taatggggaa actggagaaa ttgtctggga taaagggcga
     3601 gattttgcca cagtgcgcaa agtattgtcc atgccccaag tcaatattgt caagaaaaca
     3661 gaagtacaga caggcggatt ctccaaggag tcaattttac caaaaagaaa ttcggacaag
     3721 cttattgctc gtaaaaaaga ctgggatcca aaaaaatatg gtggttttga tagtccaacg
     3781 gtagcttatt cagtcctagt ggttgctaag gtggaaaaag ggaaatcgaa gaagttaaaa
     3841 tccgttaaag agttactagg gatcacaatt atggaaagaa gttcctttga aaaaaatccg
     3901 attgactttt tagaagctaa aggatataag gaagttaaaa aagacttaat cattaaacta
     3961 cctaaatata gtctttttga gttagaaaac ggtcgtaaac ggatgctggc tagtgccgga
     4021 gaattacaaa aaggaaatga gctggctctg ccaagcaaat atgtgaattt tttatattta
     4081 gctagtcatt atgaaaagtt gaagggtagt ccagaagata acgaacaaaa acaattgttt
     4141 gtggagcagc ataagcatta tttagatgag attattgagc aaatcagtga attttctaag
     4201 cgtgttattt tagcagatgc caatttagat aaagttctta gtgcatataa caaacataga
     4261 gacaaaccaa tacgtgaaca agcagaaaat attattcatt tatttacgtt gacgaatctt
     4321 ggagctcccg ctgcttttaa atattttgat acaacaattg atcgtaaacg atatacgtct
     4381 acaaaagaag ttttagatgc cactcttatc catcaatcca tcactggtct ttatgaaaca
     4441 cgcattgatt tgagtcagct aggaggtgac agcggcggtt caatgacaaa tctgtcggat
     4501 atcatcgaaa aagaaaccgg taaacagctg gttatccaag aaagtatttt aatgcttccc
     4561 gaagaagtcg aagaggtgat tggcaacaaa cccgaaagtg atattctcgt gcatactgcg
     4621 tatgatgagt cgaccgatga aaacgtcatg ctgctgacct ccgacgcgcc cgagtataaa
     4681 ccgtgggctt tggttatcca ggatagcaac ggtgaaaata agattaaaat gttataaggc
     4741 actcctcagc aaatataatg accctcttga taacccaaga gggcattttt taatgcccat
     4801 ggcgtcttag aaaaactcat cgagcatcaa atgaaactgc aatttattca tatcaggatt
     4861 atcaatacca tatttttgaa aaagccgttt ctgtaatgaa ggagaaaact caccgaggca
     4921 gttccatagg atggcaagat cctggtatcg gtctgcgatt ccgactcgtc caacatcaat
     4981 acaacctatt aatttcccct cgtcaaaaat aaggttatca agtgagaaat caccatgagt
     5041 gacgactgaa tccggtgaga atggcaaaag tttatgcatt tctttccaga cttgttcaac
     5101 aggccagcca ttacgctcgt catcaaaatc actcgcatca accaaaccgt tattcattcg
     5161 tgattgcgcc tgagcgagac gaaatacgcg atcgctgtta aaaggacaat tacaaacagg
     5221 aatcgaatgc aaccggcgca ggaacactgc cagcgcatca acaatatttt cacctgaatc
     5281 aggatattct tctaatacct ggaatgctgt tttcccgggg atcgcagtgg tgagtaacca
     5341 tgcatcatca ggagtacgga taaaatgctt gatggtcgga agaggcataa attccgtcag
     5401 ccagtttagt ctgaccatct catctgtaac atcattggca acgctacctt tgccatgttt
     5461 cagaaacaac tctggcgcat cgggcttccc atacaatcga tagattgtcg cacctgattg
     5521 cccgacatta tcgcgagccc atttataccc atataaatca gcatccatgt tggaatttaa
     5581 tcgcggccta gagcaagacg tttcccgttg aatatggctc ataacacccc ttgtattact
     5641 gtttatgtaa gcagacagtt ttattgttca tgaagagaaa cctacggatc ggggcggaga
     5701 tttcctggaa gatgccagga agatacttaa cagggaagtg agagggccgc ggcaaagccg
     5761 tttttccata ggctccgccc ccctgacaag catcacgaaa tctgacgctc aaatcagtgg
     5821 tggcgaaacc cgacaggact ataaagatac caggcgtttc cccctggcgg ctccctcgtg
     5881 cgctctcctg ttcctgcctt tcggtttacc ggtgtcattc cgctgttatg gccgcgtttg
     5941 tctcattcca cgcctgacac tcagttccgg gtaggcagtt cgctccaagc tggactgtat
     6001 gcacgaaccc cccgttcagt ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc
     6061 caacccggaa agacatgcaa aagcaccact ggcagcagcc actggtaatt gatttagagg
     6121 agttagtctt gaagtcatgc gccggttaag gctaaactga aaggacaagt tttggtgact
     6181 gcgctcctcc aagccagtta cctcggttca aagagttggt agctcagaga accttcgaaa
     6241 aaccgccctg caaggcggtt ttttcgtttt cagagcaaga tattacgcgc agaccaaaac
     6301 gatctcaaga agatcatctt attaaggagc cccatacgat ataagaggac atccggt
//
