LOCUS       sequence-393135-        3167 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Level 2 destination vector: minimal (non-T-DNA) backbone.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3167)
  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 3167)
  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: pMIN-0
FEATURES             Location/Qualifiers
     source          1..3167
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     504..523
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        join(761..766,767..784,785..791)
                     /label=lac UV5 promoter
                     /note="E. coli lac promoter with an ""up"" mutation"
     CDS             845..1525
                     /codon_start=1
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQAGRNLE
                     DPAY"
     CDS             1845..2150
                     /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     2029..2048
                     /label=ccdB-fwd
                     /note="ccdB gene, forward primer"
     CDS             2281..3096
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     CDS             2281..3096
                     /codon_start=1
                     /gene="aph(3')-Ia"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     complement(2354..2373)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
ORIGIN
        1 atcaattgat cttcttgaaa tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa
       61 aaaccaccgc taccagcggt ggtttgtttg ccggatcaag agctaccaac tctttttccg
      121 aaggtaactg gcttcagcag agcgcagata ccaaatactg ttcttctagt gtagccgtag
      181 ttaggccacc acttcaagaa ctctgtagca ccgcctacat acctcgctct gctaatcctg
      241 ttaccagtgg ctgcttccag tggcgataag tcgtgtctta ccgggttgga ctcaagacga
      301 tagttaccgg ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac acagcccagc
      361 ttggagcgaa cgacctacac cgaactgaga tacctacagc gtgagctatg agaaagcgcc
      421 acgcttcccg aagggagaaa ggcggacagg tatccggtaa gcggcagggt cggaacagga
      481 gagcgcacga gggagcttcc agggggaaac gcctggtatc tttatagtcc tgtcgggttt
      541 cgccacctct gacttgagcg tcgatttttg tgatgctcgt caggggggcg gagcctatgg
      601 aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct tttgctggcc ttttgctcac
      661 atgttctttc ctgcgttatc ccctgattct gtggataacc gtattaccgc ctttgagtga
      721 tcaccgtttt gcaggtgcgg ccgcattagg caccccaggc tttacacttt atgcttccgg
      781 ctcgtataat gtgtggattt tgagttagga tccgtcgaga ttttcaggag ctaaggaagc
      841 taaaatggag aaaaaaatca ctggatatac caccgttgat atatcccaat ggcatcgtaa
      901 agaacatttt gaggcatttc agtcagtcgc tcaatgtacc tataaccaga ccgttcagct
      961 ggatattacg gcctttttaa agaccgtaaa gaaaaataag cacaagtttt atccggcctt
     1021 tattcacatt cttgcccgcc tgatgaatgc tcatccggaa ttccgtatgg caatgaaaga
     1081 cggtgagctg gtgatatggg atagtgttca cccttgttac accgttttcc atgagcaaac
     1141 tgaaacgttt tcatcgctct ggagtgaata ccacgacgat ttccggcagt ttctacacat
     1201 atattcgcaa gatgtggcgt gttacggtga aaacctggcc tatttcccta aagggtttat
     1261 tgagaatatg tttttcgttt cagccaatcc ctgggtgagt ttcaccagtt ttgatttaaa
     1321 cgtggccaat atggacaact tcttcgcccc cgttttcacc atgggcaaat attatacgca
     1381 aggcgacaag gtgctgatgc cgctggcgat tcaggttcat catgccgttt gtgatggctt
     1441 ccatgtcggc agaatgctta atgaattaca acagtactgc gatgagtggc aggcggggcg
     1501 taatctagag gatccggctt actaaaagcc agataacagt atgcgtattt gcgcgctgat
     1561 ttttgcggta taagaatata tactgatatg tatacccgaa gtatgtcaaa aggaggtatg
     1621 ctatgaagca gcgtattaca gtgacagttg acagcgacag ctatcagttg ctcaaggcat
     1681 atatgatgtc aatatctccg gtctggtaag cacaaccatg cagaatgaag cccgtcgtct
     1741 gcgtgccgaa cgctggaaag cggaaaatca ggaagggatg gctgaggtcg cccggtttat
     1801 tgaaatgaac ggctcttttg ctgacgagaa caggggctgg tgaaatgcag tttaaggttt
     1861 acacctataa aagagagagc cgttatcgtc tgtttgtgga tgtacagagt gatattattg
     1921 acacgcccgg gcgacggatg gtgatccccc tggccagtgc acgtctgctg tcagataaag
     1981 tctcccgtga actttacccg gtggtgcata tcggggatga aagctggcgc atgatgacca
     2041 ccgatatggc cagtgtgccg gtttccgtta tcggggaaga agtggctgat ctcagccacc
     2101 gcgaaaatga catcaaaaac gccattaacc tgatgttctg gggaatataa cacctgcaaa
     2161 aagtgcaatt agatccacgt tgtgtctcaa aatctctgat gttacattgc acaagataaa
     2221 aatatatcat catgaacaat aaaactgtct gcttacataa acagtaatac aatgggtgtt
     2281 atgagccata ttcaacggga aacgtcttgt tcgaggccgc gattaaattc caacatggat
     2341 gctgatttat atgggtataa atgggctcgc gataatgtcg ggcaatcagg tgcgacaatc
     2401 taccgattgt atgggaagcc cgatgcgcca gagttgtttc tgaaacatgg caaaggtagc
     2461 gttgccaatg atgttacaga tgagatggtc agactaaact ggctgacgga atttatgcct
     2521 cttccgacca tcaagcattt tatccgtact cctgatgatg catggttact caccactgcg
     2581 atcccaggga aaacagcatt ccaggtatta gaagaatatc ctgattcagg tgaaaatatt
     2641 gttgatgcgc tggcagtgtt cctgcgccgg ttgcattcga ttcctgtttg taattgtcct
     2701 tttaacagcg atcgcgtatt tcgtctggct caggcgcaat cacgaatgaa taacggtttg
     2761 gttgatgcga gtgattttga tgacgagcgt aatggctggc ctgttgaaca agtctggaaa
     2821 gaaatgcata aacttttgcc attctcaccg gattcagtcg tcactcatgg tgatttctca
     2881 cttgataacc ttatttttga cgaggggaaa ttaataggtt gtattgatgt tggacgagtc
     2941 ggaatcgcag accgatacca ggatcttgcc atcctatgga actgcctcgg tgagttttct
     3001 ccttcattac agaaacggct ttttcaaaaa tatggtattg ataatcctga tatgaataaa
     3061 ttgcagtttc atttgatgct cgatgagttt ttctaatcag aattggttaa ttggttgtaa
     3121 cactggcaga gcattacgct gacttgacgg gacggcggct ttgttga
//
