LOCUS       sequence-390624-        2894 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Level 0 J1-J4 destination vector, BbsI entry.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 2894)
  AUTHORS   Chamness JC, Kumar J, Cruz AJ, Rhuby E, Holum MJ, Cody JP, Tibebu R,
            Gamo ME, Starker CG, Zhang F, Voytas DF
  TITLE     An extensible vector toolkit and parts library for advanced 
            engineering of plant genomes.
  JOURNAL   Plant Genome. 2023 Mar 9:e20312. doi: 10.1002/tpg2.20312.
  PUBMED    36896468
REFERENCE   2  (bases 1 to 2894)
  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: "Plant 
            Genome. 2023 Mar 9:e20312. doi: 10.1002/tpg2.20312."
COMMENT     Sequence Label: pMC-1-ccdB-4
FEATURES             Location/Qualifiers
     source          1..2894
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             421..726
                     /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     605..624
                     /label=ccdB-fwd
                     /note="ccdB gene, forward primer"
     primer_bind     complement(774..792)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        860..964
                     /gene="bla"
                     /label=AmpR promoter
     CDS             1186..1977
                     /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"
     rep_origin      2148..2736
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2637..2656
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     join(2890..2894,1..13)
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 gtcagtgagc gaggaagcgg aagagcgccc aatacgcaaa ccgcctctcc ccgcgcgttg
       61 gccgattcat taatcactct gtggtctcag gagttgtctt caagccagat aacagtatgc
      121 gtatttgcgc gctgattttt gcggtataag aatatatact gatatgtata cccgaagtat
      181 gtcaaaagga ggtatgctat gaagcagcgt attacagtga cagttgacag cgacagctat
      241 cagttgctca aggcatatat gatgtcaata tctccggtct ggtaagcaca accatgcaga
      301 atgaagcccg tcgtctgcgt gccgaacgct ggaaagcgga aaatcaggaa gggatggctg
      361 aggtcgcccg gtttattgaa atgaacggct cttttgctga cgagaacagg ggctggtgaa
      421 atgcagttta aggtttacac ctataaaaga gagagccgtt atcgtctgtt tgtggatgta
      481 cagagtgata ttattgacac gcccgggcga cggatggtga tccccctggc cagtgcacgt
      541 ctgctgtcag ataaagtctc ccgtgaactt tacccggtgg tgcatatcgg ggatgaaagc
      601 tggcgcatga tgaccaccga tatggccagt gtgccggttt ccgttatcgg ggaagaagtg
      661 gctgatctca gccaccgcga aaatgacatc aaaaacgcca ttaacctgat gttctgggga
      721 atataagaag acaatacttg agaccacgaa gtggctcttc agtggacgaa agggcctcgt
      781 gatacgccta tttttatagg ttaatgtcat gataataatg gtttcttaga cgtcaggtgg
      841 cacttttcgg ggaaatgtgc gcggaacccc tatttgttta tttttctaaa tacattcaaa
      901 tatgtatccg ctcatgagac aataaccctg ataaatgctt caataatatt gaaaaaggaa
      961 gagtatgcgc tcacgcaact ggtccagaac cttgaccgaa cgcagcggtg gtaacggcgc
     1021 agtggcggtt ttcatggctt gttatgactg tttttttttg gggtacagtc tatgcctcgg
     1081 gcatccaagc agcaagcgcg ttacgccgtg ggtcgatgtt tgatgttatg gagcagcaac
     1141 gatgttacgc agcagggcag tcgccctaaa acaaagttaa acatcatgag ggaagcggtg
     1201 atcgccgaag tatcgactca actatcagag gtagttggcg tcatcgagcg ccatctcgaa
     1261 ccgacgttgc tggccgtaca tttgtacggc tccgcagtgg atggcggcct gaagccacac
     1321 agcgatattg atttgctggt tacggtgacc gtaaggcttg atgaaacaac gcggcgagct
     1381 ttgatcaacg accttttgga aacttcggct tcccctggag agagcgagat tctccgcgct
     1441 gtagaagtca ccattgttgt gcacgacgac atcattccgt ggcgttatcc agctaagcgc
     1501 gaactgcaat ttggagaatg gcagcgcaat gacattcttg caggtatctt cgagccagcc
     1561 acgatcgaca ttgatctggc tatcttgctg acaaaagcaa gagaacatag cgttgccttg
     1621 gtaggtccag cggcggagga actctttgat ccggttcctg aacaggatct atttgaggcg
     1681 ctaaatgaaa ccttaacgct atggaactcg ccgcccgact gggctggcga tgagcgaaat
     1741 gtagtgctta cgttgtcccg catttggtac agcgcagtaa ccggcaaaat cgcgccgaag
     1801 gatgtcgctg ccgactgggc aatggagcgc ctgccggccc agtatcagcc cgtcatactt
     1861 gaagctagac aggcttatct tggacaagaa gaagatcgct tggcctcgcg cgcagatcag
     1921 ttggaagaat ttgtccatta cgtaaaaggc gagatcacca aggtagtcgg caaataactg
     1981 tcagaccaag tttactcata tatactttag attgatttaa aacttcattt ttaatttaaa
     2041 aggatctagg tgaagatcct ttttgataat ctcatgacca aaatccctta acgtgagttt
     2101 tcgttccact gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg agatcctttt
     2161 tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt
     2221 ttgccggatc aagagctacc aactcttttt ccgaaggtaa ctggcttcag cagagcgcag
     2281 ataccaaata ctgttcttct agtgtagccg tagttaggcc accacttcaa gaactctgta
     2341 gcaccgccta catacctcgc tctgctaatc ctgttaccag tggctgctgc cagtggcgat
     2401 aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac cggataaggc gcagcggtcg
     2461 ggctgaacgg ggggttcgtg cacacagccc agcttggagc gaacgaccta caccgaactg
     2521 agatacctac agcgtgagct atgagaaagc gccacgcttc ccgaagggag aaaggcggac
     2581 aggtatccgg taagcggcag ggtcggaaca ggagagcgca cgagggagct tccaggggga
     2641 aacgcctggt atctttatag tcctgtcggg tttcgccacc tctgacttga gcgtcgattt
     2701 ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc ggccttttta
     2761 cggttcctgg ccttttgctg gccttttgct cacatgttct ttcctgcgtt atcccctgat
     2821 tctgtggata accgtattac cgcctttgag tgagctgata ccgctcgccg cagccgaacg
     2881 accgagcgca gcga
//
