LOCUS       sequence-352077-        5264 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Complementary plasmid .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5264)
  AUTHORS   Mok BY, Kotrys AV, Raguram A, Huang TP, Mootha VK, Liu DR
  TITLE     CRISPR-free base editors with enhanced activity and expanded 
            targeting scope in mitochondrial and nuclear DNA.
  JOURNAL   Nat Biotechnol. 2022 Sep;40(9):1378-1387. doi: 
            10.1038/s41587-022-01256-8. Epub 2022 Apr 4.
  PUBMED    35379961
REFERENCE   2  (bases 1 to 5264)
  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"; doi: 
            "10.1038/s41587-022-01256-8"; journalName: "Nat Biotechnol"; date: 
            "2022-09"; volume: "40"; issue: "9"; pages: "1378-1387"
COMMENT     Sequence Label: CP2-TCC
FEATURES             Location/Qualifiers
     source          1..5264
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     96..113
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    128..268
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(194..213)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     354..376
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(370..561)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(3727..4518)
                     /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(4519..4590)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      4618..5206
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5107..5126
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 tgttctttcc tgcgttatcc cctgattctg tggataaccg tattaccgcc tttgagtgag
       61 ctgataccgc tcgccgcagc cgaacgaccg agcgcagcga gtcagtgagc gaggaagcgg
      121 atgagcgcct gatgcggtat tttctcctta cgcatctgtg cggtatttca caccgcatat
      181 ggtgcactct cagtacaatc tgctctgatg ccgcataatt aagccagtat acactccgct
      241 atcgctacgt gactgggtca tggctgcgcc ccgacacccg ccaacacccg ctgacgcgcc
      301 ctgacgggct tgtctgctcc cggcatccgc ttacagacaa gctgtgaccg tctccgggag
      361 ctgcatgtgt cagaggtttt caccgtcatc accgaaacgc gcgaggcagc tgcggtaaag
      421 ctcatcagcg tggtcgtgaa gcgattcaca gatgtctgcc tgttcatccg cgtccagctc
      481 gttgagtttc tccagaagcg ttaatgtctg gcttctgata aagcgggcca tgttaagggc
      541 ggttttttcc tgtttggtca ctgatgcctc cgtgtaaggg ggatttctgt tcatgggggt
      601 aatgataccg atgaaacgag agaggatgct cacgatacgg gttactgatg atgaacaaga
      661 ggacatccgg tcaaataaaa cgaaaggctc agtcgaaaga ctgggccttt cgttttgctg
      721 aggagactta gggactctca cagctaacac cacgtcgtcc ctatctgctg ccctaggtct
      781 atgagtggtt gctggataac tttacgggca tgcataaggc tcggtatcta tattcaggga
      841 gaccacaacg gtttccctct acaaataatt ttgtttaact tttactagag tgggacccta
      901 cctgcaggtg cagtaaaaaa aggaaaaatt gaacacgatt aacatcgcta agaacgactt
      961 ctctgacatc gaactggctg ctatcccgtt caacactctg gctgaccatt acggtgagcg
     1021 tttagctcgc gaacagttgg cccttgagca tgagtcttac gagatgggtg aagcacgctt
     1081 ccgcaagatg tttgagcgtc aacttaaagc tggtgaggtt gcggataacg ctgccgccaa
     1141 gcctctcatc actaccctac tccctaagat gattgcacgc atcaacgact ggtttgagga
     1201 agtgaaagct aagcgcggca agcgcccgac agccttccag ttcctgcaag aaatcaagcc
     1261 ggaagccgta gcgtacatca ccattaagac cactctggct tgcctaacca gtgctgacaa
     1321 tacaaccgtt caggctgtag caagcgcaat cggtcgggcc attgaggacg aggctcgctt
     1381 cggtcgtatc cgtgacctta aagctaagca cttcaagaaa aacgttgagg aacaactcaa
     1441 caagcgcgta gggcacgtct acaagaaagc atttatgcaa gttgtcgagg ctgacatgct
     1501 ctctaagggt ctactcggtg gcgaggcgtg gtcttcgtgg cataaggaag actctattca
     1561 tgtaggagta cgctgcatcg agatgctcat tgagtcaacc ggaatggtta gcttacaccg
     1621 ccaaaatgct ggcgtagtag gtcaagactc tgagactatc gaactcgcac ctgaatacgc
     1681 tgaggctatc gcaacccgtg caggtgcgct ggctggcatc tctccgatgt tccaaccttg
     1741 cgtagttcct cctaagccgt ggactggcat tactggtggt ggctattggg ctaacggtcg
     1801 tcgtcctctg gcgctggtgc gtactcacag taagaaagca ctgatgcgct acgaagacgt
     1861 ttacatgcct gaggtgtaca aagcgattaa cattgcgcaa aacaccgcat ggaaaatcaa
     1921 caagaaagtc ctagcggtcg ccaacgtaat caccaagtgg aagcattgtc cggtcgagga
     1981 catccctgcg attgagcgtg aagaactccc gatgaaaccg gaagacatcg acatgaatcc
     2041 tgaggctctc accgcgtgga aacgtgctgc cgctgctgtg taccgcaagg acaaggctcg
     2101 caagtctcgc cgtatcagcc ttgagttcat gcttgagcaa gccaataagt ttgctaacca
     2161 taaggccatc tggttccctt acaacatgga ctggcgcggt cgtgtttacg ctgtgtcaat
     2221 gttcaacccg caaggtaacg atatgaccaa aggactgctt acgctggcga aaggtaaacc
     2281 aatcggtaag gaaggttact actggctgaa aatccacggt gcaaactgtg cgggtgtcga
     2341 taaggttccg ttccctgagc gcatcaagtt cattgaggaa aaccacgaga acatcatggc
     2401 ttgcgctaag tctccactgg agaacacttg gtgggctgag caagattctc cgttctgctt
     2461 ccttgcgttc tgctttgagt acgctggggt acagcaccac ggcctgagct ataactgctc
     2521 ccttccgctg gcgtttgacg ggtcttgctc tggcatccag cacttctccg cgatgctccg
     2581 agatgaggta ggtggtcgcg cggttaactt gcttcctagt gaaaccgttc aggacatcta
     2641 cgggattgtt gctaagaaag tcaacgagat tctacaagca gacgcaatca atgggaccga
     2701 taacgaagta gttaccgtga ccgatgagaa cactggtgaa atctctgaga aagtcaagct
     2761 gggcactaag gcactggctg gtcaatggct ggcttacggt gttactcgca gtgtgactaa
     2821 gcgttcagtc atgacgctgg cttacgggtc caaagagttc ggcttccgtc aacaagtgct
     2881 ggaagatacc attcagccag ctattgattc cggcaagggt ctgatgttca ctcagccgaa
     2941 tcaggctgct ggatacatgg ctaagctgat ttgggaatct gtgagcgtga cggtggtagc
     3001 tgcggttgaa gcaatgaact ggcttaagtc tgctgctaag ctgctggctg ctgaggtcaa
     3061 agataagaag actggagaga ttcttcgcaa gcgttgcgct gtgcattggg taactcctga
     3121 tggtttccct gtgtggcagg aatacaagaa gcctattcag acgcgcttga acctgatgtt
     3181 cctcggtcag ttccgcttac agcctaccat taacaccaac aaagatagcg agattgatgc
     3241 acacaaacag gagtctggta tcgctcctaa ctttgtacac agccaagacg gtagccacct
     3301 tcgtaagact gtagtgtggg cacacgagaa gtacggaatc gaatcttttg cactgattca
     3361 cgactccttc ggtaccattc cggctgacgc tgcgaacctg ttcaaagcag tgcgcgaaac
     3421 tatggttgac acatatgagt cttgtgatgt actggctgat ttctacgacc agttcgctga
     3481 ccagttgcac gagtctcaat tggacaaaat gccagcactt ccggctaaag gtaacttgaa
     3541 cctccgtgac atcttagagt ccgacttcgc gttcgcctgg accagggcag caaacgacga
     3601 aaactacaac tacgctttag cagcttaata aacttaatta acggcactcc tcagcaaagg
     3661 cactcctcag caaatataat gaccctcttg ataacccaag agggcatttt ttaatgccca
     3721 tggcgtttat ttgccgacta ccttggtgat ctcgcctttc acgtagtgga caaattcttc
     3781 caactgatct gcgcgcgagg ccaagcgatc ttcttcttgt ccaagataag cctgtctagc
     3841 ttcaagtatg acgggctgat actgggccgg caggcgctcc attgcccagt cggcagcgac
     3901 atccttcggc gcgattttgc cggttactgc gctgtaccaa atgcgggaca acgtaagcac
     3961 tacatttcgc tcatcgccag cccagtcggg cggcgagttc catagcgtta aggtttcatt
     4021 tagcgcctca aatagatcct gttcaggaac cggatcaaag agttcctccg ccgctggacc
     4081 taccaaggca acgctatgtt ctcttgcttt tgtcagcaag atagccagat caatgtcgat
     4141 cgtggctggc tcgaagatac ctgcaagaat gtcattgcgc tgccattctc caaattgcag
     4201 ttcgcgctta gctggataac gccacggaat gatgtcgtcg tgcacaacaa tggtgacttc
     4261 tacagcgcgg agaatctcgc tctctccagg ggaagccgaa gtttccaaaa ggtcgttgat
     4321 caaagctcgc cgcgttgttt catcaagcct tacggtcacc gtaaccagca aatcaatatc
     4381 actgtgtggc ttcaggccgc catccactgc ggagccgtac aaatgtacgg ccagcaacgt
     4441 cggttcgaga tggcgctcga tgacgccaac tacctctgat agttgagtcg atacttcggc
     4501 gatcaccgct tccctcatac tcttcctttt tcaatattat tgaagcattt atcagggtta
     4561 ttgtctcatg agcgtataca tatttgaatg tatttagaaa aataggccaa atagggtttg
     4621 agatcctttt tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac cgctaccagc
     4681 ggtggtttgt ttgccggatc aagagctacc acctcttttt ccgaaggtaa ctggcttcag
     4741 cagagcgcag ataccaaata ctgtccttct agtgtagccg tagttaggcc accacttcaa
     4801 gaactctgta gcaccgccta catacctcgc tctgctaatc ctgttaccag tggctgctgc
     4861 cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac cggataaggc
     4921 gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc agcttggagc gaacgaccta
     4981 caccgaactg agatacctac agcgtgagct atgagaaagc gccacgcttc ccgaagggag
     5041 aaaggcggac aggtatccgg taagcggcag ggtcggaaca ggagagcgca cgagggagct
     5101 tccaggggga aacgcctggt atctttatag tcctgtcggg tttcgccacc tctgacttga
     5161 gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc
     5221 ggccttttta cggttcctgg ccttttgctg gccttttgct caca
//
