LOCUS       sequence-238421-        5265 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  BE PACE selection, express T7 RNA polymerase with degron-GTCCA 
            target.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5265)
  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 5265)
  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: pBT2dR3-proB
FEATURES             Location/Qualifiers
     source          1..5265
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(3072..3863)
                     /codon_start=1
                     /gene="aadA"
                     /product="aminoglycoside adenylyltransferase (Murphy, 
                     1985)"
                     /label=SmR
                     /note="confers resistance to spectinomycin and 
                     streptomycin"
                     /translation="MREAVIAEVSTQLSEVVGVIERHLEPTLLAVHLYGSAVDGGLKPH
                     SDIDLLVTVTVRLDETTRRALINDLLETSASPGESEILRAVEVTIVVHDDIIPWRYPAK
                     RELQFGEWQRNDILAGIFEPATIDIDLAILLTKAREHSVALVGPAAEELFDPVPEQDLF
                     EALNETLTLWNSPPDWAGDERNVVLTLSRIWYSAVTGKIAPKDVAADWAMERLPAQYQP
                     VILEARQAYLGQEEDRLASRADQLEEFVHYVKGEITKVVGK"
     promoter        complement(3864..3935)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      3963..4551
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4452..4471
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4705..4722
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    4737..4877
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(4803..4822)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4963..4985
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(4979..5170)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
ORIGIN
        1 aagaggacat ccggtcaaat aaaacgaaag gctcagtcga aagactgggc ctttcgtttt
       61 gctgaggaga cttagggact ctcacagcta acaccacgtc gtccctatct gctgccctag
      121 gtctatgagt ggttgctgga taactttacg ggcatgcata aggctcgtaa tatatattca
      181 gggagaccac aacggtttcc ctctacaaat aattttgttt aacttttact agagtgggac
      241 cctacctgca ggtgcagtac tacatcatca ggcatgaaca cgattaacat cgctaagaac
      301 gacttctctg acatcgaact ggctgctatc ccgttcaaca ctctggctga ccattacggt
      361 gagcgtttag ctcgcgaaca gttggccctt gagcatgagt cttacgagat gggtgaagca
      421 cgcttccgca agatgtttga gcgtcaactt aaagctggtg aggttgcgga taacgctgcc
      481 gccaagcctc tcatcactac cctactccct aagatgattg cacgcatcaa cgactggttt
      541 gaggaagtga aagctaagcg cggcaagcgc ccgacagcct tccagttcct gcaagaaatc
      601 aagccggaag ccgtagcgta catcaccatt aagaccactc tggcttgcct aaccagtgct
      661 gacaatacaa ccgttcaggc tgtagcaagc gcaatcggtc gggccattga ggacgaggct
      721 cgcttcggtc gtatccgtga ccttaaagct aagcacttca agaaaaacgt tgaggaacaa
      781 ctcaacaagc gcgtagggca cgtctacaag aaagcattta tgcaagttgt cgaggctgac
      841 atgctctcta agggtctact cggtggcgag gcgtggtctt cgtggcataa ggaagactct
      901 attcatgtag gagtacgctg catcgagatg ctcattgagt caaccggaat ggttagctta
      961 caccgccaaa atgctggcgt agtaggtcaa gactctgaga ctatcgaact cgcacctgaa
     1021 tacgctgagg ctatcgcaac ccgtgcaggt gcgctggctg gcatctctcc gatgttccaa
     1081 ccttgcgtag ttcctcctaa gccgtggact ggcattactg gtggtggcta ttgggctaac
     1141 ggtcgtcgtc ctctggcgct ggtgcgtact cacagtaaga aagcactgat gcgctacgaa
     1201 gacgtttaca tgcctgaggt gtacaaagcg attaacattg cgcaaaacac cgcatggaaa
     1261 atcaacaaga aagtcctagc ggtcgccaac gtaatcacca agtggaagca ttgtccggtc
     1321 gaggacatcc ctgcgattga gcgtgaagaa ctcccgatga aaccggaaga catcgacatg
     1381 aatcctgagg ctctcaccgc gtggaaacgt gctgccgctg ctgtgtaccg caaggacaag
     1441 gctcgcaagt ctcgccgtat cagccttgag ttcatgcttg agcaagccaa taagtttgct
     1501 aaccataagg ccatctggtt cccttacaac atggactggc gcggtcgtgt ttacgctgtg
     1561 tcaatgttca acccgcaagg taacgatatg accaaaggac tgcttacgct ggcgaaaggt
     1621 aaaccaatcg gtaaggaagg ttactactgg ctgaaaatcc acggtgcaaa ctgtgcgggt
     1681 gtcgataagg ttccgttccc tgagcgcatc aagttcattg aggaaaacca cgagaacatc
     1741 atggcttgcg ctaagtctcc actggagaac acttggtggg ctgagcaaga ttctccgttc
     1801 tgcttccttg cgttctgctt tgagtacgct ggggtacagc accacggcct gagctataac
     1861 tgctcccttc cgctggcgtt tgacgggtct tgctctggca tccagcactt ctccgcgatg
     1921 ctccgagatg aggtaggtgg tcgcgcggtt aacttgcttc ctagtgaaac cgttcaggac
     1981 atctacggga ttgttgctaa gaaagtcaac gagattctac aagcagacgc aatcaatggg
     2041 accgataacg aagtagttac cgtgaccgat gagaacactg gtgaaatctc tgagaaagtc
     2101 aagctgggca ctaaggcact ggctggtcaa tggctggctt acggtgttac tcgcagtgtg
     2161 actaagcgtt cagtcatgac gctggcttac gggtccaaag agttcggctt ccgtcaacaa
     2221 gtgctggaag ataccattca gccagctatt gattccggca agggtctgat gttcactcag
     2281 ccgaatcagg ctgctggata catggctaag ctgatttggg aatctgtgag cgtgacggtg
     2341 gtagctgcgg ttgaagcaat gaactggctt aagtctgctg ctaagctgct ggctgctgag
     2401 gtcaaagata agaagactgg agagattctt cgcaagcgtt gcgctgtgca ttgggtaact
     2461 cctgatggtt tccctgtgtg gcaggaatac aagaagccta ttcagacgcg cttgaacctg
     2521 atgttcctcg gtcagttccg cttacagcct accattaaca ccaacaaaga tagcgagatt
     2581 gatgcacaca aacaggagtc tggtatcgct cctaactttg tacacagcca agacggtagc
     2641 caccttcgta agactgtagt gtgggcacac gagaagtacg gaatcgaatc ttttgcactg
     2701 attcacgact ccttcggtac cattccggct gacgctgcga acctgttcaa agcagtgcgc
     2761 gaaactatgg ttgacacata tgagtcttgt gatgtactgg ctgatttcta cgaccagttc
     2821 gctgaccagt tgcacgagtc tcaattggac aaaatgccag cacttccggc taaaggtaac
     2881 ttgaacctcc gtgacatctt agagtccgac ttcgcgttcg cctggaccag ggcagcaaac
     2941 gacgaaaact acaactacgc tttagcagct taataaactt aattaacggc actcctcagc
     3001 aaaggcactc ctcagcaaat ataatgaccc tcttgataac ccaagagggc attttttaat
     3061 gcccatggcg tttatttgcc gactaccttg gtgatctcgc ctttcacgta gtggacaaat
     3121 tcttccaact gatctgcgcg cgaggccaag cgatcttctt cttgtccaag ataagcctgt
     3181 ctagcttcaa gtatgacggg ctgatactgg gccggcaggc gctccattgc ccagtcggca
     3241 gcgacatcct tcggcgcgat tttgccggtt actgcgctgt accaaatgcg ggacaacgta
     3301 agcactacat ttcgctcatc gccagcccag tcgggcggcg agttccatag cgttaaggtt
     3361 tcatttagcg cctcaaatag atcctgttca ggaaccggat caaagagttc ctccgccgct
     3421 ggacctacca aggcaacgct atgttctctt gcttttgtca gcaagatagc cagatcaatg
     3481 tcgatcgtgg ctggctcgaa gatacctgca agaatgtcat tgcgctgcca ttctccaaat
     3541 tgcagttcgc gcttagctgg ataacgccac ggaatgatgt cgtcgtgcac aacaatggtg
     3601 acttctacag cgcggagaat ctcgctctct ccaggggaag ccgaagtttc caaaaggtcg
     3661 ttgatcaaag ctcgccgcgt tgtttcatca agccttacgg tcaccgtaac cagcaaatca
     3721 atatcactgt gtggcttcag gccgccatcc actgcggagc cgtacaaatg tacggccagc
     3781 aacgtcggtt cgagatggcg ctcgatgacg ccaactacct ctgatagttg agtcgatact
     3841 tcggcgatca ccgcttccct catactcttc ctttttcaat attattgaag catttatcag
     3901 ggttattgtc tcatgagcgt atacatattt gaatgtattt agaaaaatag gccaaatagg
     3961 gtttgagatc ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta
     4021 ccagcggtgg tttgtttgcc ggatcaagag ctaccacctc tttttccgaa ggtaactggc
     4081 ttcagcagag cgcagatacc aaatactgtc cttctagtgt agccgtagtt aggccaccac
     4141 ttcaagaact ctgtagcacc gcctacatac ctcgctctgc taatcctgtt accagtggct
     4201 gctgccagtg gcgataagtc gtgtcttacc gggttggact caagacgata gttaccggat
     4261 aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac agcccagctt ggagcgaacg
     4321 acctacaccg aactgagata cctacagcgt gagctatgag aaagcgccac gcttcccgaa
     4381 gggagaaagg cggacaggta tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg
     4441 gagcttccag ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg ccacctctga
     4501 cttgagcgtc gatttttgtg atgctcgtca ggggggcgga gcctatggaa aaacgccagc
     4561 aacgcggcct ttttacggtt cctggccttt tgctggcctt ttgctcacat gttctttcct
     4621 gcgttatccc ctgattctgt ggataaccgt attaccgcct ttgagtgagc tgataccgct
     4681 cgccgcagcc gaacgaccga gcgcagcgag tcagtgagcg aggaagcgga tgagcgcctg
     4741 atgcggtatt ttctccttac gcatctgtgc ggtatttcac accgcatatg gtgcactctc
     4801 agtacaatct gctctgatgc cgcatagtta agccagtata cactccgcta tcgctacgtg
     4861 actgggtcat ggctgcgccc cgacacccgc caacacccgc tgacgcgccc tgacgggctt
     4921 gtctgctccc ggcatccgct tacagacaag ctgtgaccgt ctccgggagc tgcatgtgtc
     4981 agaggttttc accgtcatca ccgaaacgcg cgaggcagct gcggtaaagc tcatcagcgt
     5041 ggtcgtgaag cgattcacag atgtctgcct gttcatccgc gtccagctcg ttgagtttct
     5101 ccagaagcgt taatgtctgg cttctgataa agcgggccat gttaagggcg gttttttcct
     5161 gtttggtcac tgatgcctcc gtgtaagggg gatttctgtt catgggggta atgataccga
     5221 tgaaacgaga gaggatgctc acgatacggg ttactgatga tgaac
//
