LOCUS       sequence-351799-        4135 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Target plasmid for context profiling.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4135)
  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 4135)
  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: pBM10a
FEATURES             Location/Qualifiers
     source          1..4135
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(join(129..920,921..989))
                     /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     752..771
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(990..1094)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     1162..1180
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     CDS             complement(1201..2151)
                     /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      2199..2421
                     /label=pSC101 ori
                     /note="low-copy replication origin that requires the Rep101
                     protein"
     terminator      2916..2961
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     CDS             complement(join(3276..3986,3987..3989))
                     /codon_start=1
                     /product="GFP variant that folds robustly even when fused 
                     to poorly folded proteins (Pedelacq et al., 2006)"
                     /label=superfolder GFP
                     /note="mammalian codon-optimized"
                     /translation="MSKGEELFTGVVPILVELDGDVNGHKFSVRGEGEGDATNGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKRHDFFKSAMPEGYVQERTISFKDDG
                     TYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNFNSHNVYITADKQKNGIKA
                     NFKIRHNVEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSVLSKDPNEKRDHMVLLE
                     FVTAAGITHGMDELYK"
ORIGIN
        1 ggacgacgtg gtgttagctg tgacttaatt aacggcactg cgaaaaaacc ccgccgaatc
       61 ggggtttttt gcgaaatata atgaccctct tgataaccca agagggcatt ttttaatgcc
      121 catggcgttt accaatgctt aatcagtgag gcacctatct cagcgatctg tctatttcgt
      181 tcatccatag ttgcctgact ccccgtcgtg tagataacta cgatacggga gggcttacca
      241 tctggcccca gtgctgcaat gataccgcga gacccacgct caccggctcc agatttatca
      301 gcaataaacc agccagccgg aagggccgag cgcagaagtg gtcctgcaac tttatccgcc
      361 tccatccagt ctattaattg ttgccgggaa gctagagtaa gtagttcgcc agttaatagt
      421 ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg
      481 gcttcattca gctccggttc ccaacgatca aggcgagtta catgatcccc catgttgtgc
      541 aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg
      601 ttatcactca tggttatggc agcactgcat aattctctta ctgtcatgcc atccgtaaga
      661 tgcttttctg tgactggtga gtactcaacc aagtcattct gagaatagtg tatgcggcga
      721 ccgagttgct cttgcccggc gtcaatacgg gataataccg cgccacatag cagaacttta
      781 aaagtgctca tcattggaaa acgttcttcg gggcgaaaac tctcaaggat cttaccgctg
      841 ttgagatcca gttcgatgta acccactcgt gcacccaact gatcttcagc atcttttact
      901 ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata
      961 agggcgacac ggaaatgttg aatactcata ctcttccttt ttcaatatta ttgaagcatt
     1021 tatcagggtt attgtctcat gagcggatac atatttgaat gtatttagaa aaataaacaa
     1081 ataggggttc cgcgcacatt tccccgaaaa gtgccacctg acgtctaaga aaccattatt
     1141 atcatgacat taacctataa aaataggcgt atcacgaggc ccttaggcca aataggccgt
     1201 tcagatcctt ccgtatttag ccagtatgtt ctctagtgtg gttcgttgtt tttgcgtgag
     1261 ccatgagaac gaaccattga gatcatgctt actttgcatg tcactcaaaa attttgcctc
     1321 aaaactggtg agctgaattt ttgcagttaa agcatcgtgt agtgtttttc ttagtccgtt
     1381 acgtaggtag gaatctgatg taatggttgt tggtattttg tcaccattca tttttatctg
     1441 gttgttctca agttcggtta cgagatccat ttgtctatct agttcaactt ggaaaattaa
     1501 cgtatcagtc gggcggcctc gcttatcaac caccaatttc atattgctgt aagtgtttaa
     1561 atctttactt attggtttca aaacccattg gttaagcctt ttaaactcat ggtagttatt
     1621 ttcaagcatt aacatgaact taaattcatc aaggctaatc tctatatttg ccttgtgagt
     1681 tttcttttgt gttagttctt ttaataacca ctcataaatc ctcatagagt atttgttttc
     1741 aaaagactta acatgttcca gattatattt tatgaatttt tttaactgga aaagataagg
     1801 caatatctct tcactaaaaa ctaattctaa tttttcgctt gagaacttgg catagtttgt
     1861 ccactggaaa atctcaaagc ctttaaccaa aggattcctg atttccacag ttctcgtcat
     1921 cagctctctg gttgctttag ctaatacacc ataagcattt tccctactga tgttcatcat
     1981 ctgagcgtat tggttataag tgaacgatac cgtccgttct ttccttgtag ggttttcaat
     2041 cgtggggttg agtagtgcca cacagcataa aattagcttg gtttcatgct ccgttaagtc
     2101 atagcgacta atcgctagtt catttgcttt gaaaacaact aattcagaca tacatctcaa
     2161 ttggtctagg tgattttaat cactatacca attgagatgg gctagtcaat gataattact
     2221 agtccttttc ctttgagttg tgggtatctg taaattctgc tagacctttg ctggaaaact
     2281 tgtaaattct gctagaccct ctgtaaattc cgctagacct ttgtatgttt tttttgttta
     2341 tattcaagtg gttataattt atagaataaa gaaagaataa aaaaagataa aaagaataga
     2401 tcccagccct gtgtataact cactacttta gtcagttccg cagtattaca aaaggatgtc
     2461 gcaaacgctg tttgctcctc tacaaaacag accttaaaac cctaaaggct taagtagcac
     2521 cctcgcaagc tcgggcaaat cgctgaatat tccttttgtc tccgaccatc aggcacctga
     2581 gtcgctgtct ttttcgtgac attcagttcg ctgcgctcac ggctctggca gtgaatgagg
     2641 gtaaatggca ctacaggcgc cttttatgga ttcatgcaag gaaactaccc ataatacaag
     2701 aaaagcccgt cacgggcttc tcagggcgtt ttatggcggg tctgctatgt ggtgctatct
     2761 gactttttgc tgttcagcag ttcctgccct ctgattttcc agtctgacca cttcggatta
     2821 tcccgtgaca ggtcattcag actggctaat gcacccagta aggcagcggt atcatcaaca
     2881 ggcttacccg tcttactgtc aagaggacat ccggtcaaat aaaacgaaag gctcagtcga
     2941 aagactgggc ctttcgtttt agactgagca cgtcaatacg caaaccgcct ctccccgcgc
     3001 gttggccgat tcattaatgc agctggcacg acaggtttcc cgactggagt ttgtagtcta
     3061 gtctgtccga cttcgcgttt tttttgttta tttcagcaaa cgacgaaccc gccaaacgcg
     3121 ccctgacggg attgtctgct cccggcatcc gcttacagac aagacggagc caatgtacgc
     3181 aaaaaacccc gcttcagcgg ggttttttcg ccaaaaaaaa ccccgccctg tcaggggcgg
     3241 ggtttttttt tagggcccta ctgactttat gagcctttat acagttcgtc cataccgtgc
     3301 gtgatgccgg ccgcggtcac gaattccagc aggaccatgt ggtcacgctt ttcgttcgga
     3361 tctttactca gcacagactg cgtgctcagg taatgattgt ccggcagcag gaccgggcca
     3421 tcaccaatcg gggtgttttg ctgatagtgg tcggccagtt gaacgctgcc atcttccaca
     3481 ttatggcgga tcttgaagtt agccttgata ccgttcttct gcttatctgc ggtgatgtac
     3541 acgttgtgcg aattgaagtt atattccagt ttatggccca gaatattacc gtcttcctta
     3601 aaatcgatgc ctttcagttc aatacggtta accagcgtat caccttcgaa tttcacttcg
     3661 gcgcgcgttt tgtaggtgcc gtcatcttta aatgagatgg tacgttcttg gacataacct
     3721 tccggcatcg cggacttgaa aaagtcgtgg cgtttcatgt gatccgggta acgactgaaa
     3781 cactgaacac catacgtcag cgtggtgacc agggtcggcc acggaaccgg cagtttaccc
     3841 gtggtgcaaa taaacttcag cgtcagttta ccattggttg catcgccttc accttcgccg
     3901 cgcacgctga atttgtgacc attgacatca ccatccagtt ccaccagaat cggcaccacg
     3961 cccgtaaaca gttcttcgcc cttgctcatt tttttttcct ccttactcta gtaaaagtta
     4021 aacaaaatta tttgtagagg gaaaccgttg tggtctccct gaatatagat accgagcctt
     4081 atgcatgccc gtaaagttat ccagcaacca ctcatagacc tagggcagca gatag
//
