LOCUS       sequence-351810-        4690 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Accessory plasmids .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4690)
  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 4690)
  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: AP1
FEATURES             Location/Qualifiers
     source          1..4690
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     15..33
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     CDS             complement(54..1004)
                     /codon_start=1
                     /gene="rep101"
                     /product="RepA protein needed for replication with the 
                     pSC101 origin"
                     /label=Rep101
                     /translation="MSELVVFKANELAISRYDLTEHETKLILCCVALLNPTIENPTRKE
                     RTVSFTYNQYAQMMNISRENAYGVLAKATRELMTRTVEIRNPLVKGFEIFQWTNYAKFS
                     SEKLELVFSEEILPYLFQLKKFIKYNLEHVKSFENKYSMRIYEWLLKELTQKKTHKANI
                     EISLDEFKFMLMLENNYHEFKRLNQWVLKPISKDLNTYSNMKLVVDKRGRPTDTLIFQV
                     ELDRQMDLVTELENNQIKMNGDKIPTTITSDSYLRNGLRKTLHDALTAKIQLTSFEAKF
                     LSDMQSKHDLNGSFSWLTQKQRTTLENILAKYGRI"
     rep_origin      1052..1274
                     /label=pSC101 ori
                     /note="low-copy replication origin that requires the Rep101
                     protein"
     primer_bind     1838..1857
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1838..1856
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             1894..3168
                     /codon_start=1
                     /gene="III"
                     /product="pIII"
                     /label=M13 gene III
                     /note="minor coat protein"
                     /translation="MKKLLFAIPLVVPFYSHSAETVESCLAKPHTENSFTNVWKDDKTL
                     DRYANYEGCLWNATGVVVCTGDETQCYGTWVPIGLAIPENEGGGSEGGGSEGGGSEGGG
                     TKPPEYGDTPIPGYTYINPLDGTYPPGTEQNPANPNPSLEESQPLNTFMFQNNRFRNRQ
                     GALTVYTGTVTQGTDPVKTYYQYTPVSSKAMYDAYWNGKFRDCAFHSGFNEDPFVCEYQ
                     GQSSDLPQPPVNAGGGSGGGSGGGSEGGGSEGGGSEGGGSEGGGSGGGSGSGDFDYEKM
                     ANANKGAMTENADENALQSDAKGKLDSVATDYGAAIDGFIGDVSGLANGNGATGDFAGS
                     NSQMAQVGDGDNSPLMNNFRQYLPSLPQSVECRPFVFGAGKPYEFSIDCDKINLFRGVF
                     AFLLYVATFMYVFSTFANILRNKES"
     terminator      complement(3251..3278)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(3370..3456)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     3609..3626
                     /label=pBAD Reverse
                     /note="For vectors with E. coli araBAD promoter, reverse 
                     primer"
     CDS             complement(join(3672..4463,4464..4532))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQDQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     4295..4314
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4533..4637)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 cattaaccta taaaaatagg cgtatcacga ggcccttagg ccaaataggc cgttcagatc
       61 cttccgtatt tagccagtat gttctctagt gtggttcgtt gtttttgcgt gagccatgag
      121 aacgaaccat tgagatcatg cttactttgc atgtcactca aaaattttgc ctcaaaactg
      181 gtgagctgaa tttttgcagt taaagcatcg tgtagtgttt ttcttagtcc gttacgtagg
      241 taggaatctg atgtaatggt tgttggtatt ttgtcaccat tcatttttat ctggttgttc
      301 tcaagttcgg ttacgagatc catttgtcta tctagttcaa cttggaaaat caacgtatca
      361 gtcgggcggc ctcgcttatc aaccaccaat ttcatattgc tgtaagtgtt taaatcttta
      421 cttattggtt tcaaaaccca ttggttaagc cttttaaact catggtagtt attttcaagc
      481 attaacatga acttaaattc atcaaggcta atctctatat ttgccttgtg agttttcttt
      541 tgtgttagtt cttttaataa ccactcataa atcctcatag agtatttgtt ttcaaaagac
      601 ttaacatgtt ccagattata ttttatgaat ttttttaact ggaaaagata aggcaatatc
      661 tcttcactaa aaactaattc taatttttcg cttgagaact tggcatagtt tgtccactgg
      721 aaaatctcaa agcctttaac caaaggattc ctgatttcca cagttctcgt catcagctct
      781 ctggttgctt tagctaatac accataagca ttttccctac tgatgttcat catctgagcg
      841 tattggttat aagtgaacga taccgtccgt tctttccttg tagggttttc aatcgtgggg
      901 ttgagtagtg ccacacagca taaaattagc ttggtttcat gctccgttaa gtcatagcga
      961 ctaatcgcta gttcatttgc tttgaaaaca actaattcag acatacatct caattggtct
     1021 aggtgatttt aatcactata ccaattgaga tgggctagtc aatgataatt actagtcctt
     1081 ttcctttgag ttgtgggtat ctgtaaattc tgctagacct ttgctggaaa acttgtaaat
     1141 tctgctagac cctctgtaaa ttccgctaga cctttgtgtg ttttttttgt ttatattcaa
     1201 gtggttataa tttatagaat aaagaaagaa taaaaaaaga taaaaagaat agatcccagc
     1261 cctgtgtata actcactact ttagtcagtt ccgcagtatt acaaaaggat gtcgcaaacg
     1321 ctgtttgctc ctctacaaaa cagaccttaa aaccctaaag gcttaagtag caccctcgca
     1381 agctcgggca aatcgctgaa tattcctttt gtctccgacc atcaggcacc tgagtcgctg
     1441 tctttttcgt gacattcagt tcgctgcgct cacggctctg gcagtgaatg ggggtaaatg
     1501 gcactacagg cgccttttat ggattcatgc aaggaaacta cccataatac aagaaaagcc
     1561 cgtcacgggc ttctcagggc gttttatggc gggtctgcta tgtggtgcta tctgactttt
     1621 tgctgttcag cagttcctgc cctctgattt tccagtctga ccacttcgga ttatcccgtg
     1681 acaggtcatt cagactggct aatgcaccca gtaaggcagc ggtatcatca acaggcttac
     1741 ccgtcttact gtcaagagga catccggtca aataaaacga aaggctcagt cgaaagactg
     1801 ggcctttcgt tttgctgagg agacttaggg accctactaa tacgactcac tatagggaga
     1861 aagacctgca ggtgcagtaa ggaggaaaaa aaaatgaaaa aattattatt cgcaattcct
     1921 ttagttgttc ctttctattc tcactccgct gaaactgttg aaagttgttt agcaaaaccc
     1981 catacagaaa attcatttac taacgtctgg aaagacgaca aaactttaga tcgttacgct
     2041 aactatgagg gctgtctgtg gaatgctaca ggcgttgtag tttgtactgg tgacgaaact
     2101 cagtgttacg gtacatgggt tcctattggg cttgctatcc ctgaaaatga gggtggtggc
     2161 tctgagggtg gcggttctga gggtggcggt tctgagggtg gcggtactaa acctcctgag
     2221 tacggtgata cacctattcc gggctatact tatatcaacc ctctcgacgg cacttatccg
     2281 cctggtactg agcaaaaccc cgctaatcct aatccttctc ttgaggagtc tcagcctctt
     2341 aatactttca tgtttcagaa taataggttc cgaaataggc agggggcatt aactgtttat
     2401 acgggcactg ttactcaagg cactgacccc gttaaaactt attaccagta cactcctgta
     2461 tcatcaaaag ccatgtatga cgcttactgg aacggtaaat tcagagactg cgctttccat
     2521 tctggcttta atgaggatcc attcgtttgt gaatatcaag gccaatcgtc tgacctgcct
     2581 caacctcctg tcaatgctgg cggcggctct ggtggtggtt ctggtggcgg ctctgagggt
     2641 ggtggctctg agggtggcgg ttctgagggt ggcggctctg agggaggcgg ttccggtggt
     2701 ggctctggtt ccggtgattt tgattatgaa aagatggcaa acgctaataa gggggctatg
     2761 accgaaaatg ccgatgaaaa cgcgctacag tctgacgcta aaggcaaact tgattctgtc
     2821 gctactgatt acggtgctgc tatcgatggt ttcattggtg acgtttccgg ccttgctaat
     2881 ggtaatggtg ctactggtga ttttgctggc tctaattccc aaatggctca agtcggtgac
     2941 ggtgataatt cacctttaat gaataatttc cgtcaatatt taccttccct ccctcaatcg
     3001 gttgaatgtc gcccttttgt ctttggcgct ggtaaacctt acgagttcag tatcgactgc
     3061 gataagatca acctgttccg cggtgtcttt gcgtttcttt tatatgttgc cacctttatg
     3121 tatgtatttt ctacgtttgc taacatactg cgtaataagg agtcttaaac ggagccaatg
     3181 tacgcaaaaa accccgcttc agcggggttt tttcgccctc tagaatcctc agcgagtttg
     3241 tagaaacgca aaaaggccat ccgtcaggat ggccttctgc ttaatttgat gcctggcagt
     3301 ttatggcggg cgtcctgccc gccaccctcc gggccgttgc ttcgcaacgt tcaaatccgc
     3361 tcccggcgga tttgtcctac tcaggagagc gttcaccgac aaacaacaga taaaacgaaa
     3421 ggcccagtct ttcgactgag cctttcgttt tatttgatgc ctggcagttc cctactctcg
     3481 catggggaga ccccacacta ccatcggcgc tacggcgttt cacttctgag ttcggcatgg
     3541 ggtcaggtgg gaccaccgcg ctactgccgc caggcaaatt ctgttttatc agaccgcttc
     3601 tgcgttctga tttaatctgt atcaggctga aaatcttctc tcatccgcca aaacagccat
     3661 gcccatggcg tttaccaatg cttaatcagt gaggcaccta tctcagcgat ctgtctattt
     3721 cgttcatcca tagttgcctg actccccgtc gtgtagataa ctacgatacg ggagggctta
     3781 ccatctggcc ccagtgctgc aatgataccg cgagacccac gctcaccggc tccagattta
     3841 tcagcaataa accagccagc cggaagggcc gagcgcagaa gtggtcctgc aactttatcc
     3901 gcctccatcc agtctattaa ttgttgccgg gaagctagag taagtagttc gccagttaat
     3961 agtttgcgca acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt
     4021 atggcttcat tcagctccgg ttcccaacga tcaaggcgag ttacatgatc ccccatgttg
     4081 tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg tcagaagtaa gttggccgca
     4141 gtgttatcac tcatggttat ggcagcactg cataattctc ttactgtcat gccatccgta
     4201 agatgctttt ctgtgactgg tgagtactca accaagtcat tctgagaata gtgtatgcgg
     4261 cgaccgagtt gatcttgccc ggcgtcaata cgggataata ccgcgccaca tagcagaact
     4321 ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa aactctcaag gatcttaccg
     4381 ctgttgagat ccagttcgat gtaacccact cgtgcaccca actgatcttc agcatctttt
     4441 actttcacca gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga
     4501 ataagggcga cacggaaatg ttgaatactc atactcttcc tttttcaata ttattgaagc
     4561 atttatcagg gttattgtct catgagcgga tacatatttg aatgtattta gaaaaataaa
     4621 caaatagggg ttccgcgcac atttccccga aaagtgccac ctgacgtcta agaaaccatt
     4681 attatcatga
//
