LOCUS       sequence-349149-        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(107..898,899..967))
                     /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     730..749
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(968..1072)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     1140..1158
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     CDS             complement(1179..2129)
                     /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      2177..2399
                     /label=pSC101 ori
                     /note="low-copy replication origin that requires the Rep101
                     protein"
     terminator      2894..2939
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     CDS             complement(join(3254..3964,3965..3967))
                     /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 acttaattaa cggcactgcg aaaaaacccc gccgaagcgg ggttttttgc gaaatataat
       61 gaccctcttg ataacccaag agggcatttt ttaatgccca tggcgtttac caatgcttaa
      121 tcagtgaggc acctatctca gcgatctgtc tatttcgttc atccatagtt gcctgactcc
      181 ccgtcgtgta gataactacg atacgggagg gcttaccatc tggccccagt gctgcaatga
      241 taccgcgaga cccacgctca ccggctccag atttatcagc aataaaccag ccagccggaa
      301 gggccgagcg cagaagtggt cctgcaactt tatccgcctc catccagtct attaattgtt
      361 gccgggaagc tagagtaagt agttcgccag ttaatagttt gcgcaacgtt gttgccattg
      421 ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc tccggttccc
      481 aacgatcaag gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt agctccttcg
      541 gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg gttatggcag
      601 cactgcataa ttctcttact gtcatgccat ccgtaagatg cttttctgtg actggtgagt
      661 actcaaccaa gtcattctga gaatagtgta tgcggcgacc gagttgctct tgcccggcgt
      721 caatacggga taataccgcg ccacatagca gaactttaaa agtgctcatc attggaaaac
      781 gttcttcggg gcgaaaactc tcaaggatct taccgctgtt gagatccagt tcgatgtaac
      841 ccactcgtgc acccaactga tcttcagcat cttttacttt caccagcgtt tctgggtgag
      901 caaaaacagg aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa
      961 tactcatact cttccttttt caatattatt gaagcattta tcagggttat tgtctcatga
     1021 gcggatacat atttgaatgt atttagaaaa ataaacaaat aggggttccg cgcacatttc
     1081 cccgaaaagt gccacctgac gtctaagaaa ccattattat catgacatta acctataaaa
     1141 ataggcgtat cacgaggccc ttaggccaaa taggccgttc agatccttcc gtatttagcc
     1201 agtatgttct ctagtgtggt tcgttgtttt tgcgtgagcc atgagaacga accattgaga
     1261 tcatgcttac tttgcatgtc actcaaaaat tttgcctcaa aactggtgag ctgaattttt
     1321 gcagttaaag catcgtgtag tgtttttctt agtccgttac gtaggtagga atctgatgta
     1381 atggttgttg gtattttgtc accattcatt tttatctggt tgttctcaag ttcggttacg
     1441 agatccattt gtctatctag ttcaacttgg aaaatcaacg tatcagtcgg gcggcctcgc
     1501 ttatcaacca ccaatttcat attgctgtaa gtgtttaaat ctttacttat tggtttcaaa
     1561 acccattggt taagcctttt aaactcatgg tagttatttt caagcattaa catgaactta
     1621 aattcatcaa ggctaatctc tatatttgcc ttgtgagttt tcttttgtgt tagttctttt
     1681 aataaccact cataaatcct catagagtat ttgttttcaa aagacttaac atgttccaga
     1741 ttatatttta tgaatttttt taactggaaa agataaggca atatctcttc actaaaaact
     1801 aattctaatt tttcgcttga gaacttggca tagtttgtcc actggaaaat ctcaaagcct
     1861 ttaaccaaag gattcctgat ttccacagtt ctcgtcatca gctctctggt tgctttagct
     1921 aatacaccat aagcattttc cctactgatg ttcatcatct gagcgtattg gttataagtg
     1981 aacgataccg tccgttcttt ccttgtaggg ttttcaatcg tggggttgag tagtgccaca
     2041 cagcataaaa ttagcttggt ttcatgctcc gttaagtcat agcgactaat cgctagttca
     2101 tttgctttga aaacaactaa ttcagacata catctcaatt ggtctaggtg attttaatca
     2161 ctataccaat tgagatgggc tagtcaatga taattactag tccttttcct ttgagttgtg
     2221 ggtatctgta aattctgcta gacctttgct ggaaaacttg taaattctgc tagaccctct
     2281 gtaaattccg ctagaccttt gtgtgttttt tttgtttata ttcaagtggt tataatttat
     2341 agaataaaga aagaataaaa aaagataaaa agaatagatc ccagccctgt gtataactca
     2401 ctactttagt cagttccgca gtattacaaa aggatgtcgc aaacgctgtt tgctcctcta
     2461 caaaacagac cttaaaaccc taaaggctta agtagcaccc tcgcaagctc gggcaaatcg
     2521 ctgaatattc cttttgtctc cgaccatcag gcacctgagt cgctgtcttt ttcgtgacat
     2581 tcagttcgct gcgctcacgg ctctggcagt gaatgggggt aaatggcact acaggcgcct
     2641 tttatggatt catgcaagga aactacccat aatacaagaa aagcccgtca cgggcttctc
     2701 agggcgtttt atggcgggtc tgctatgtgg tgctatctga ctttttgctg ttcagcagtt
     2761 cctgccctct gattttccag tctgaccact tcggattatc ccgtgacagg tcattcagac
     2821 tggctaatgc acccagtaag gcagcggtat catcaacagg cttacccgtc ttactgtcaa
     2881 gaggacatcc ggtcaaataa aacgaaaggc tcagtcgaaa gactgggcct ttcgttttag
     2941 actgagcacg tcaatacgca aaccgcctct ccccgcgcgt tggccgattc attaatgcag
     3001 ctggcacgac aggtttcccg actggagttt gtagtctagt ctgtccgact tcgcgttttt
     3061 tttgtttatt tcagcaaacg acgaacccgc caaacgcgcc ctgacgggat tgtctgctcc
     3121 cggcatccgc ttacagacaa gacggagcca atgtacgcaa aaaaccccgc ttcagcgggg
     3181 ttttttcgcc aaaaaaaacc ccgccctgtc aggggcgggg ttttttttta gggccctact
     3241 gactttatga gcctttatac agttcgtcca taccgtgcgt gatgccggcc gcggtcacga
     3301 attccagcag gaccatgtgg tcacgctttt cgttcggatc tttactcagc acagactgcg
     3361 tgctcaggta atgattgtcc ggcagcagga ccgggccatc accaatcggg gtgttttgct
     3421 gatagtggtc ggccagttga acgctgccat cttccacatt atggcggatc ttgaagttag
     3481 ccttgatacc gttcttctgc ttatctgcgg tgatgtacac gttgtgcgaa ttgaagttat
     3541 attccagttt atggcccaga atattaccgt cttccttaaa atcgatgcct ttcagttcaa
     3601 tacggttaac cagcgtatca ccttcgaatt tcacttcggc gcgcgttttg taggtgccgt
     3661 catctttaaa tgagatggta cgttcttgga cataaccttc cggcatcgcg gacttgaaaa
     3721 agtcgtggcg tttcatgtga tccgggtaac gactgaaaca ctgaacacca tacgtcagcg
     3781 tggtgaccag ggtcggccac ggaaccggca gtttacccgt ggtgcaaata aacttcagcg
     3841 tcagtttacc attggttgca tcgccttcac cttcgccgcg cacgctgaat ttgtgaccat
     3901 tgacatcacc atccagttcc accagaatcg gcaccacgcc cgtaaacagt tcttcgccct
     3961 tgctcatttt tttttcctcc ttactctagt aaaagttaaa caaaattatt tgtagaggga
     4021 aaccgttgtg gtctccctga atatagatac cgagccttat gcatgcccgt aaagttatcc
     4081 agcaaccact catagaccta gggcagcaga tagggacgac gtggtgttag ctgtg
//
