LOCUS       sequence-390529-        7508 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Level 2 destination vector: pMIN-Ri backbone.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7508)
  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 7508)
  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-Ri
FEATURES             Location/Qualifiers
     source          1..7508
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    1612..1636
                     /label=LB T-DNA repeat
                     /note="left border repeat from nopaline C58 T-DNA"
     promoter        join(1739..1744,1745..1762,1763..1769)
                     /label=lac UV5 promoter
                     /note="E. coli lac promoter with an ""up"" mutation"
     CDS             1823..2503
                     /codon_start=1
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQAGRNLE
                     DPAY"
     CDS             2823..3128
                     /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     3007..3026
                     /label=ccdB-fwd
                     /note="ccdB gene, forward primer"
     misc_feature    3165..3189
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     CDS             3325..4140
                     /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             3325..4140
                     /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(3398..3417)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      4226..4814
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4715..4734
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 caacgtcgcc gtcgcggaag cccttctgtc tctctagcgc gaacagaatc gcatttcctc
       61 gaatcctcgt caagagtttt tagcgccgtt ttggtgagct gatttccttt gcctgctgaa
      121 aggtgaaaga tgatgcagac aggaagtgta acgacgccat tcgggcggcg gccaatgacg
      181 cttgcgcttg tgcggcgcca gacggcgctg gccgatatca aacaaggcaa gacagcggac
      241 aagtggaagg tctttagaga tgcgtccgcg gctatggaac tacttggaat ccagtccaac
      301 agtcttgccg tccttgatgc gctattgagc tttcacccgg aaacggagtt gcgtcaggag
      361 gcacagctga tcgttttccc gtcgaatgct cagcttgccc ttcgggcgca tgggatggct
      421 ggcgcgactt tgcgtaggca catcgccatg ctcgtggagt caggcttgat cgtccggaag
      481 gatagcgcca acggaaagcg ttacgctcgt aaggatggcg ctggtcagat cgagcgcgcg
      541 tttggcttcg atttgtctcc gcttctcgcg cggtcggaag aactagcgat gatggcacag
      601 caggttatgg ccgatcgagc agcattcagg atggccaaag aaagtctgac gatttgccga
      661 cgggacgttc ggaagctaat tacggcagct atggaagagg gagcggaggg cgactggcaa
      721 gctgtcgagg aagtctatgt ggaacttgtg ggtagaattc cacgcgcccc gacgcttgct
      781 gatgtagagt caattctcga agagatgtgg atgctccagg aagagataat caaccggttg
      841 gaaattagag acaattcaga aaataatagc accaatgctg cccagagcga gcagcacata
      901 cagaattcaa aacccgaatc cgttaatgaa cttgaacctc gctctgaaaa ggagcagggc
      961 gctaagccga gtgaaataga ccgggcaagg agcgagccga taaaagcgtt ccccctcggg
     1021 atgatcctga aagcatgccc gaccattggc aactatgggc cgagcggtgc ggttgctagc
     1081 tggcgtgacc tcatgtcggc tgcggtggtg gttcggtcta tgctgggggt cagcccgtcg
     1141 gcttaccaag acgcgtgtga ggcaatggga ccggagaatg cggcagcagc gatggcgtgc
     1201 attttggagc gagcgaactt catcaattcg cccgggggct atctccgaga tctgacacgg
     1261 cggagcgagc ttgggaagtt ttcacttggc ccgatgataa tggcgctctt gaaggctagc
     1321 gggcagggga cgttgcggtt tggctagaat tagcgagtat ggagcaggat ggtctgtggt
     1381 cagctgacca cagacctaat aggttgaaaa catgagcgtt ttttggatga tcgacagacc
     1441 atccgattcc cggagtacca agcgtgctct gatgggagcg ataacattac tcaacaagca
     1501 cgaaggcccc atgccgatcg ttgatcgtga aggagagcct gctctacatg cggcggtatt
     1561 ttgccggccg aggcatgtag tcgcggagca ctgcctattt actgccctag gtggcaggat
     1621 atattgtggt gtaaacaaat tgacgcttag acaacttaat aacacattgc ggacgttttt
     1681 aatgtactga atgcccgggc accgttttgc aggtgcggcc gcattaggca ccccaggctt
     1741 tacactttat gcttccggct cgtataatgt gtggattttg agttaggatc cgtcgagatt
     1801 ttcaggagct aaggaagcta aaatggagaa aaaaatcact ggatatacca ccgttgatat
     1861 atcccaatgg catcgtaaag aacattttga ggcatttcag tcagtcgctc aatgtaccta
     1921 taaccagacc gttcagctgg atattacggc ctttttaaag accgtaaaga aaaataagca
     1981 caagttttat ccggccttta ttcacattct tgcccgcctg atgaatgctc atccggaatt
     2041 ccgtatggca atgaaagacg gtgagctggt gatatgggat agtgttcacc cttgttacac
     2101 cgttttccat gagcaaactg aaacgttttc atcgctctgg agtgaatacc acgacgattt
     2161 ccggcagttt ctacacatat attcgcaaga tgtggcgtgt tacggtgaaa acctggccta
     2221 tttccctaaa gggtttattg agaatatgtt tttcgtttca gccaatccct gggtgagttt
     2281 caccagtttt gatttaaacg tggccaatat ggacaacttc ttcgcccccg ttttcaccat
     2341 gggcaaatat tatacgcaag gcgacaaggt gctgatgccg ctggcgattc aggttcatca
     2401 tgccgtttgt gatggcttcc atgtcggcag aatgcttaat gaattacaac agtactgcga
     2461 tgagtggcag gcggggcgta atctagagga tccggcttac taaaagccag ataacagtat
     2521 gcgtatttgc gcgctgattt ttgcggtata agaatatata ctgatatgta tacccgaagt
     2581 atgtcaaaag gaggtatgct atgaagcagc gtattacagt gacagttgac agcgacagct
     2641 atcagttgct caaggcatat atgatgtcaa tatctccggt ctggtaagca caaccatgca
     2701 gaatgaagcc cgtcgtctgc gtgccgaacg ctggaaagcg gaaaatcagg aagggatggc
     2761 tgaggtcgcc cggtttattg aaatgaacgg ctcttttgct gacgagaaca ggggctggtg
     2821 aaatgcagtt taaggtttac acctataaaa gagagagccg ttatcgtctg tttgtggatg
     2881 tacagagtga tattattgac acgcccgggc gacggatggt gatccccctg gccagtgcac
     2941 gtctgctgtc agataaagtc tcccgtgaac tttacccggt ggtgcatatc ggggatgaaa
     3001 gctggcgcat gatgaccacc gatatggcca gtgtgccggt ttccgttatc ggggaagaag
     3061 tggctgatct cagccaccgc gaaaatgaca tcaaaaacgc cattaacctg atgttctggg
     3121 gaatataaca cctgcaaaaa gtgcccgggc aaactatcag tgtttgacag gatatattgg
     3181 cgggtaaacc taagagaaaa gagcgtttac aattagatcc acgttgtgtc tcaaaatctc
     3241 tgatgttaca ttgcacaaga taaaaatata tcatcatgaa caataaaact gtctgcttac
     3301 ataaacagta atacaatggg tgttatgagc catattcaac gggaaacgtc ttgttcgagg
     3361 ccgcgattaa attccaacat ggatgctgat ttatatgggt ataaatgggc tcgcgataat
     3421 gtcgggcaat caggtgcgac aatctaccga ttgtatggga agcccgatgc gccagagttg
     3481 tttctgaaac atggcaaagg tagcgttgcc aatgatgtta cagatgagat ggtcagacta
     3541 aactggctga cggaatttat gcctcttccg accatcaagc attttatccg tactcctgat
     3601 gatgcatggt tactcaccac tgcgatccca gggaaaacag cattccaggt attagaagaa
     3661 tatcctgatt caggtgaaaa tattgttgat gcgctggcag tgttcctgcg ccggttgcat
     3721 tcgattcctg tttgtaattg tccttttaac agcgatcgcg tatttcgtct ggctcaggcg
     3781 caatcacgaa tgaataacgg tttggttgat gcgagtgatt ttgatgacga gcgtaatggc
     3841 tggcctgttg aacaagtctg gaaagaaatg cataaacttt tgccattctc accggattca
     3901 gtcgtcactc atggtgattt ctcacttgat aaccttattt ttgacgaggg gaaattaata
     3961 ggttgtattg atgttggacg agtcggaatc gcagaccgat accaggatct tgccatccta
     4021 tggaactgcc tcggtgagtt ttctccttca ttacagaaac ggctttttca aaaatatggt
     4081 attgataatc ctgatatgaa taaattgcag tttcatttga tgctcgatga gtttttctaa
     4141 tcagaattgg ttaattggtt gtaacactgg cagagcatta cgctgacttg acgggacggc
     4201 ggctttgttg aatcaattga tcttcttgaa atcctttttt tctgcgcgta atctgctgct
     4261 tgcaaacaaa aaaaccaccg ctaccagcgg tggttttttt gccggatcaa gagctaccaa
     4321 ctctttttcc gaaggtaact ggcttcagca gagcgcagat accaaatact gttcttctag
     4381 tgtagccgta gttaggccac cacttcaaga actctgtagc accgcctaca tacctcgctc
     4441 tgctaatcct gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt accgggttgg
     4501 actcaagacg atagttaccg gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca
     4561 cacagcccag cttggagcga acgacctaca ccgaactgag atacctacag cgtgagctat
     4621 gagaaagcgc cacgcttccc gaagggagaa aggcggacag gtatccggta agcggcaggg
     4681 tcggaacagg agagcgcacg agggagcttc cagggggaaa cgcctggtat ctttatagtc
     4741 ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt gtgatgctcg tcaggggggc
     4801 ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc ttttgctggc
     4861 cttttgctca catgttcttt cctgcgttat cccctgattc tgtggataac cgtattaccg
     4921 cctttgagtg atcggcccag gtagaatccg cctgagtcgc aagggtgact tcgcctatat
     4981 tggacgacgg cgcgcagagg gcgacctctt tttgggttac gattgtagga ttatcactaa
     5041 aacaatacat gaacatattc aaatggcaat ctctctaagg cattggaaat aaatacaaat
     5101 aacagttggg tggagttttt cgacctgtgg gcgttaacct tctgttaacc taaaagctct
     5161 tgcccaaaca gcagaatcgg cgctaattgc cagcggcgga acttttccag tttcgcgaaa
     5221 aatatcgcca ctggcaagga atgggtttga gatggcgaag tctgtcctaa aagcagcgcc
     5281 tgtagttgta gggttgacgg ccttgatgga gcgtcatgcc gatgccctct cgagccaact
     5341 tcaagcacat catcttaagg ttttcccgcc gcattccgag aagggcattc gaacattcgg
     5401 gccatcggag gcgtccaagc tgctcggcgt tggcgagtca tatttacggc agaccgcgtc
     5461 tgagatgcca gagttgaatg ttagcatgag cccgggtggc aggcgaatgt tctcaattga
     5521 agatatccat gcgattcgga agtatatgga tcaggtcggc cgcgggaacc ggcgctacct
     5581 gccacatcgt cgaggcggcg agcagcttca ggttatctct gtgatgaatt tcaaaggtgg
     5641 gtcgggtaag accaccaccg ccgcgcatct ggcgcagtac ctcgctatgc gcggatatcg
     5701 agtcttggcc attgatctcg atcctcaagc gagcctttct gcactctttg ggagccaacc
     5761 ggaaacggac gttggcccga acgaaacgct ctacggcgct ataaggtatg atgatgagca
     5821 ggttgcaatc gaacgagtcg tccgagggac ttacattccc gacctccacc tgattcctgg
     5881 taatcttgag ctgatggagt ttgaacacga tacgccacgc gcgctgatga accgcaaaga
     5941 gggcgacacg ctcttttatg gtcgcatcag ccaagtaatt gaagatatcg cggataacta
     6001 tgacgtcgtg gtcatcgact gccctcccca gcttgggtat ctcacgctat ccgcattgac
     6061 tgcggcgacg tccattcttg tcacggtcca tccgcagatg ctggatgtga tgtcgatgaa
     6121 ccagtttctg gcaatgacat cgaacctttt gcgtgaaatc gagaatgctg gcgccaagtt
     6181 caagtttaat tggatgcgct atctgataac ccgtttcgaa ccgagcgacg gaccacagaa
     6241 ccaaatggta ggttatctgc ggtcgatttt tggcgaaaat gtcctcaatt ttccgatgct
     6301 taaaaccacc gcggtttcgg acgctggcct gacaaaccag actctattcg aagtggagcg
     6361 tggcctgttc acgcgctcga cctatgatcg agccttggag gcgatgaacg ccgtcaacga
     6421 cgagatcgaa acactgatca aaaaagcatg gggtaggccc acatgagccg gaagcacatc
     6481 cttggcgtca caactgacgc ccctgagtcg tcgcccgccg acaataggac ggcaaagaac
     6541 cgctccatgc cgctcctcgg cgtaacaagg aaggagcgcg atccggcaac gaagctcaca
     6601 gcgaacattg gtaacgcact gcgagagcaa aacgatcgtc ttagccgtgc cgaagagatc
     6661 gagcggcgcc tcgctgaagg tcaggcagtg atagagttgg atgcctcgtc aatagaaccg
     6721 tctttcgtgc aggatcgtat gcgaggggac attgacgggc tccttacttc gatccgggaa
     6781 caaggacagc aagtcccaat ccttgtgcga ccgcatccga gccagccggg ccgatatcag
     6841 gttgccttcg gccaccgccg gctacgcgcc gtttcagaac tcggacttcc ggtcagagcg
     6901 gtcgttcgcg aactgacgga cgagcaagtg gtcgtagcac agggtcagga aaacaatgag
     6961 cgcgaagatc ttaccttcat cgaaaaggcg cgcttcgcac atcgcctgaa caggcagttt
     7021 tctcgagaga ttgtcatcgc cgcgatgtcg atcgacaaga gcaatttgtc caagatgctt
     7081 ctgctcgttg acgccctccc ctctgaactg accgatgcta ttggtgccgc tcctggtgtt
     7141 ggacggccga gttggcaaca acttgccgag ctgattgaga aagtttcttc accggccgac
     7201 gtggctaaat atgctatgtc ggaggaagtt caagcgctgc catcggcaga acgattcaag
     7261 gcggtgatcg ctagtctgaa gcccagtcgg gttgcgcgtg gacttcccga ggtcatggcc
     7321 accccagacg gcaccagaat tgcacaggtg acgcagagca aggccaaact ggaaatcacg
     7381 attgacagga aggcgacgcc cgattttgcg accttcgtgc tcgatcatgt gccagcgctg
     7441 tatcaagcgt accacgctga gaaccaacgg aaacggggag agtaaaccgc aaaagaaaag
     7501 agccccct
//
