LOCUS       sequence-348313-        3471 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  MoClo Level1 plasmid, allows transition into Cermak, et al., 2017 GG
            cloning system.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3471)
  AUTHORS   Cody JP, Maher MF, Nasti RA, Starker CG, Chamness JC, Voytas DF
  TITLE     Direct delivery and fast-treated Agrobacterium co-culture 
            (Fast-TrACC) plant transformation methods for Nicotiana benthamiana.
  JOURNAL   Nat Protoc. 2022 Oct 17. pii: 10.1038/s41596-022-00749-9. doi: 
            10.1038/s41596-022-00749-9.
  PUBMED    36253612
REFERENCE   2  (bases 1 to 3471)
  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 Protoc.
            2022 Oct 17. pii: 10.1038/s41596-022-00749-9. doi: 
            10.1038/s41596-022-00749-9."
COMMENT     Sequence Label: pJC225
FEATURES             Location/Qualifiers
     source          1..3471
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        join(57..62,63..80,81..87)
                     /label=lac UV5 promoter
                     /note="E. coli lac promoter with an ""up"" mutation"
     CDS             141..821
                     /codon_start=1
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQAGRNLE
                     DPAY"
     CDS             1141..1446
                     /codon_start=1
                     /gene="ccdB"
                     /product="CcdB, a bacterial toxin that poisons DNA gyrase"
                     /label=ccdB
                     /note="Plasmids containing the ccdB gene cannot be 
                     propagated in standard E. coli strains."
                     /translation="MQFKVYTYKRESRYRLFVDVQSDIIDTPGRRMVIPLASARLLSDK
                     VSRELYPVVHIGDESWRMMTTDMASVPVSVIGEEVADLSHRENDIKNAINLMFWGI"
     primer_bind     1325..1344
                     /label=ccdB-fwd
                     /note="ccdB gene, forward primer"
     promoter        1524..1628
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      2696..3284
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3185..3204
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3438..3455
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 cacctgcttt tggacggcgc gccggagtga gaccggccgc attaggcacc ccaggcttta
       61 cactttatgc ttccggctcg tataatgtgt ggattttgag ttaggatccg tcgagatttt
      121 caggagctaa ggaagctaaa atggagaaaa aaatcactgg atataccacc gttgatatat
      181 cccaatggca tcgtaaagaa cattttgagg catttcagtc agtcgctcaa tgtacctata
      241 accagaccgt tcagctggat attacggcct ttttaaagac cgtaaagaaa aataagcaca
      301 agttttatcc ggcctttatt cacattcttg cccgcctgat gaatgctcat ccggaattcc
      361 gtatggcaat gaaagacggt gagctggtga tatgggatag tgttcaccct tgttacaccg
      421 ttttccatga gcaaactgaa acgttttcat cgctctggag tgaataccac gacgatttcc
      481 ggcagtttct acacatatat tcgcaagatg tggcgtgtta cggtgaaaac ctggcctatt
      541 tccctaaagg gtttattgag aatatgtttt tcgtttcagc caatccctgg gtgagtttca
      601 ccagttttga tttaaacgtg gccaatatgg acaacttctt cgcccccgtt ttcaccatgg
      661 gcaaatatta tacgcaaggc gacaaggtgc tgatgccgct ggcgattcag gttcatcatg
      721 ccgtttgtga tggcttccat gtcggcagaa tgcttaatga attacaacag tactgcgatg
      781 agtggcaggc ggggcgtaat ctagaggatc cggcttacta aaagccagat aacagtatgc
      841 gtatttgcgc gctgattttt gcggtataag aatatatact gatatgtata cccgaagtat
      901 gtcaaaagga ggtatgctat gaagcagcgt attacagtga cagttgacag cgacagctat
      961 cagttgctca aggcatatat gatgtcaata tctccggtct ggtaagcaca accatgcaga
     1021 atgaagcccg tcgtctgcgt gccgaacgct ggaaagcgga aaatcaggaa gggatggctg
     1081 aggtcgcccg gtttattgaa atgaacggct cttttgctga cgagaacagg ggctggtgaa
     1141 atgcagttta aggtttacac ctataaaaga gagagccgtt atcgtctgtt tgtggatgta
     1201 cagagtgata ttattgacac gcccgggcga cggatggtga tccccctggc cagtgcacgt
     1261 ctgctgtcag ataaagtctc ccgtgaactt tacccggtgg tgcatatcgg ggatgaaagc
     1321 tggcgcatga tgaccaccga tatggccagt gtgccggttt ccgttatcgg ggaagaagtg
     1381 gctgatctca gccaccgcga aaatgacatc aaaaacgcca ttaacctgat gttctgggga
     1441 atataaggtc tctcgctgag ctcccggttt tgcaggtgtg atgaaaagcc ttagggtcag
     1501 gtggcacttt tcggggaaat gtgcgatgaa cccctatttg tttatttttc taaatacatt
     1561 caaatatgta tccgctaatg agacaataac cctgataaat gcttcaatta tattgaaaaa
     1621 ggaagagtat gagtatccaa catttccgtg tcgcccttat tccctttttt gcggccttct
     1681 gccttcctgt ttttgctcac ccagaaacgc tggtgaaagt aaaagatgct gaagatcagt
     1741 tgggtgcccg agtgggttac attgaactgg atctcaacag cggcaaaatc ttagagagtt
     1801 ttcgccccga agaacgcttc ccaatgatga gcactttcaa agttctgcta tgtggcgcgg
     1861 tgttatcccg tattgatgct gggcaagagc aactcggtcg ccgcatacac tattctaaga
     1921 atgacttggt tgagtattca ccagtaacag aaaagcatct tacggatggc atgacagtaa
     1981 gagaattatg tagtgctgcc ataaccatga gtgataacac tgcggccaac ttacttctga
     2041 caacgattgg aggaccgaag gagctaaccg cttttttgca caacatgggg gatcatgtaa
     2101 ctcgccttga tcgttgggaa ccggagctga atgaagccat accaaacgac gagcgtgaca
     2161 ccacgatgcc tgtagcaatg gcaacaactc tccgcaaact attaactggc gaactactta
     2221 ctcttgcttc acgccagcaa ctcattgact ggatggaggc ggacaaagtt gcaggaccac
     2281 ttctgcgctc ggcacttccg gctggctggt ttattgctga taaatctgga gccggtgagc
     2341 gtggctctcg cggtatcatt gcagcactgg ggccagatgg taagccctcc cgtatcgtag
     2401 ttatctacac gacggggagt caggcaacta tggatgaacg aaatagacag atcgctgaga
     2461 taggtgcctc actgattaag cattggtaac tgtcagacga agggaaataa atagtagccc
     2521 gcctgatgtg cgggcttttt tttggtggtg gttaccagtg tcgcttagct accgtaggtc
     2581 ctcttaggcg gtccgattag cgacatagtc atcaatgcct tgacggcaaa atcccttaac
     2641 gtgagttttc gttccactga gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag
     2701 atcctttttt tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg
     2761 tggtttgttt gccggatcaa gagctactaa ctctttttcc gaaggtaact ggcttcagca
     2821 gagcgcagat accaaatact gttcttctag tgtagccgta gttaggccac cacttcaaga
     2881 actctgtagc accgcctaca tacctcgctc tgctaatcct gttaccagtg gctgctgcca
     2941 gtggcgataa gtcgtgtctt accgggttgg actcaagacg atagttaccg gataaggcgc
     3001 agcggtcggg ctgaacgggg ggttcgtgct cacagcccag cttggagcga acgacctaca
     3061 ccgaactgag atacctacag cgtgagctat gagaaagcgc cacgcttccc gaagggagaa
     3121 aggcggacag gtatccggta agcggcaggg tcggaacagg agagcgcaca agggagcttc
     3181 cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc
     3241 gtcgattttt gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg
     3301 cctttttacg gttcctggcc ttttgctggc cttttgctca catgttcttt cctgcgttat
     3361 cccctgattc tgtggataac cgtattaccg cctttgagtg agctgatacc gctcgccgca
     3421 gccgaacgac cgagcgcagc gagtcagtga gcgaggaagc ggaagagcgc c
//
