LOCUS       sequence-487263-        6411 bp    DNA     circular SYN 26-SEP-2025
DEFINITION  T-DNA plasmid, Level 2 destination vector, replaces pMIN-VS1.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6411)
  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 6411)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Sep 26, 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: pEM790
FEATURES             Location/Qualifiers
     source          1..6411
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    32..56
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     CDS             365..994
                     /codon_start=1
                     /product="stability protein from the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
                     /label=pVS1 StaA
                     /translation="MKVIAVLNQKGGSGKTTIATHLARALQLAGADVLLVDSDPQGSAR
                     DWAAVREDQPLTVVGIDRPTIDRDVKAIGRRDFVVIDGAPQAADLAVSAIKAADFVLIP
                     VQPSPYDIWATADLVELVKQRIEVTDGRLQAAFVVSRAIKGTRIGGEVAEALAGYELPI
                     LESRITQRVSYPGTAAAGTTVLESEPEGDAAREVQALAAEIKSKLI"
     CDS             1423..2496
                     /codon_start=1
                     /product="replication protein from the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
                     /label=pVS1 RepA
                     /translation="MSGRKPSGPVQIGAALGDDLVEKLKAAQAAQRQRIEAEARPGESW
                     QAAADRIRKESRQPPAAGASSIRKPPKGDEQPDFFVPMLYDVGTRDSRSIMDVAVFRLS
                     KRDRRAGEVIRYELPDGHVEVSAGPAGMASVWDYDLVLMAVSHLTESMNRYREGKGDKP
                     GRVFRPHVADVLKFCRRADGGKQKDDLVETCIRLNTTHVAMQRTKKAKNGRLVTVSEGE
                     ALISRYKIVKSETGRPEYIEIELADWMYREITEGKNPDVLTVHPDYFLIDPGIGRFLYR
                     LARRAAGKAEARWLFKTIYERSGSAGEFKKFCFTVRKLIGSNDLPEYDLKEEAGQAGPI
                     LVMRYRNLIEGEASAGS"
     rep_origin      2562..2756
                     /label=pVS1 oriV
                     /note="origin of replication for the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
     promoter        2818..2922
                     /gene="bla"
                     /label=AmpR promoter
     CDS             2923..3717
                     /codon_start=1
                     /gene="aphA-3"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin"
                     /translation="MAKMRISPELKKLIEKYRCVKDTEGMSPAKVYKLVGENENLYLKM
                     TDSRYKGTTYDVEREKDMMLWLEGKLPVPKVLHFERHDGWSNLLMSEADGVLCSEEYED
                     EQSPEKIIELYAECIRLFHSIDISDCPYTNSLDSRLAELDYLLNNDLADVDCENWEEDT
                     PFKDPRELYDFLKTEKPEEELVFSHGDLGDSNIFVKDGKVSGFIDLGRSGRADKWYDIA
                     FCVRSLREDIGEEQYVELFFDLLGIKPDWEKIKYYILLDELF"
     rep_origin      3888..4476
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4377..4396
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     misc_feature    4612..4636
                     /label=LB T-DNA repeat
                     /note="left border repeat from nopaline C58 T-DNA"
     promoter        join(4747..4752,4753..4770,4771..4777)
                     /label=lac UV5 promoter
                     /note="E. coli lac promoter with an ""up"" mutation"
     CDS             4831..5511
                     /codon_start=1
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQAGRNLE
                     DPAY"
     CDS             5831..6136
                     /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     6015..6034
                     /label=ccdB-fwd
                     /note="ccdB gene, forward primer"
ORIGIN
        1 cacgaagtga tccgtttaaa ctatcagtgt ttgacaggat atattggcgg gtaaacctaa
       61 gagaaaagag cgtttattag aataatcgga tatttaaaag ggcgtgaaaa ggtttatccg
      121 ttcgtccatt tgtatgtgca tgccaaccac agggttcccc tcgggagtca gccgtgcggc
      181 tgcatgaaat cctggccggt ttgtctgatg ccaagctggc ggcctggccg gccagcttgg
      241 ccgctgaaga aaccgagcgc cgccgtctaa aaaggtgatg tgtatttgag taaaacagct
      301 tgcgtcatgc ggtcgctgcg tatatgatgc gatgagtaaa taaacaaata cgcaagggga
      361 acgcatgaag gttatcgctg tacttaacca gaaaggcggg tcaggcaaga cgaccatcgc
      421 aacccatcta gcccgcgccc tgcaactcgc cggggccgat gttctgttag tcgattccga
      481 tccccagggc agtgcccgcg attgggcggc cgtgcgggaa gatcaaccgc taaccgttgt
      541 cggcatcgac cgcccgacga ttgaccgcga cgtgaaggcc atcggccggc gcgacttcgt
      601 agtgatcgac ggagcgcccc aggcggcgga cttggctgtg tccgcgatca aggcagccga
      661 cttcgtgctg attccggtgc agccaagccc ttacgacata tgggccaccg ccgacctggt
      721 ggagctggtt aagcagcgca ttgaggtcac ggatggaagg ctacaagcgg cctttgtcgt
      781 gtcgcgggcg atcaaaggca cgcgcatcgg cggtgaggtt gccgaggcgc tggccgggta
      841 cgagctgccc attcttgagt cccgtatcac gcagcgcgtg agctacccag gcactgccgc
      901 cgccggcaca accgttcttg aatcagaacc cgagggcgac gctgcccgcg aggtccaggc
      961 gctggccgct gaaattaaat caaaactcat ttgagttaat gaggtaaaga gaaaatgagc
     1021 aaaagcacaa acacgctaag tgccggccgt ccgagcgcac gcagcagcaa ggctgcaacg
     1081 ttggccagcc tggcagacac gccagccatg aagcgggtca actttcagtt gccggcggag
     1141 gatcacacca agctgaagat gtacgcggta cgccaaggca agaccattac cgagctgcta
     1201 tctgaataca tcgcgcagct accagagtaa atgagcaaat gaataaatga gtagatgaat
     1261 tttagcggct aaaggaggcg gcatggaaaa tcaagaacaa ccaggcaccg acgccgtgga
     1321 atgccccatg tgtggaggaa cgggcggttg gccaggcgta agcggctggg ttgtctgccg
     1381 gccctgcaat ggcactggaa cccccaagcc cgaggaatcg gcgtgagcgg tcgcaaacca
     1441 tccggcccgg tacaaatcgg cgcggcgctg ggtgatgacc tggtggagaa gttgaaggcc
     1501 gcgcaggccg cccagcggca acgcatcgag gcagaagcac gccccggtga atcgtggcaa
     1561 gcggccgctg atcgaatccg caaagaatcc cggcaaccgc cggcagccgg tgcgtcgtcg
     1621 attaggaagc cgcccaaggg cgacgagcaa ccagattttt tcgttccgat gctctatgac
     1681 gtgggcaccc gcgatagtcg cagcatcatg gacgtggccg ttttccgtct gtcgaagcgt
     1741 gaccgacgag ctggcgaggt gatccgctac gagcttccag acgggcacgt agaggtttcc
     1801 gcagggccgg ccggcatggc cagtgtgtgg gattacgacc tggtactgat ggcggtttcc
     1861 catctaaccg aatccatgaa ccgataccgg gaagggaagg gagacaagcc cggccgcgtg
     1921 ttccgtccac acgttgcgga cgtactcaag ttctgccggc gagccgatgg cggaaagcag
     1981 aaagacgacc tggtagaaac ctgcattcgg ttaaacacca cgcacgttgc catgcagcgt
     2041 acgaagaagg ccaagaacgg ccgcctggtg acggtatccg agggtgaagc cttgattagc
     2101 cgctacaaga tcgtaaagag cgaaaccggg cggccggagt acatcgagat cgagctagct
     2161 gattggatgt accgcgagat tacagaaggc aagaacccgg acgtgctgac ggttcacccc
     2221 gattactttt tgatcgatcc cggcatcggc cgttttctct accgcctggc acgccgcgcc
     2281 gcaggcaagg cagaagccag atggttgttc aagacgatct acgaacgcag tggcagcgcc
     2341 ggagagttca agaagttctg tttcaccgtg cgcaagctga tcgggtcaaa tgacctgccg
     2401 gagtacgatt tgaaggagga ggcggggcag gctggcccga tcctagtcat gcgctaccgc
     2461 aacctgatcg agggcgaagc atccgccggt tcctaatgta cggagcagat gctagggcaa
     2521 attgccctag caggggaaaa aggtcgaaaa agcttctttc ctgtggatag cacgtacatt
     2581 gggaacccaa agccgtacat tgggaaccgg aacccgtaca ttgggaaccc aaagccgtac
     2641 attgggaacc ggtcacacat gtaagtgact gatataaaag agaaaaaagg cgatttttcc
     2701 gcctaaaact ctttaaaact tattaaaact cttaaaaccc gcctggcctg tgcataactg
     2761 tctggccagc gcacagccga acagctgcaa aaagcgccca catcaaggct ccgagtgcgc
     2821 ggaaccccta tttgtttatt tttctaaata cattcaaata tgtatccgct catgagacaa
     2881 taaccctgat aaatgcttca ataatattga aaaaggaaga gtatggctaa aatgagaata
     2941 tcaccggaat tgaaaaaact gatcgaaaaa taccgctgcg taaaagatac ggaaggaatg
     3001 tctcctgcta aggtatataa gctggtggga gaaaatgaaa acctatattt aaaaatgacg
     3061 gacagccggt ataaagggac cacctatgat gtggaacggg aaaaggacat gatgctatgg
     3121 ctggaaggaa agctgcctgt tccaaaggtc ctgcactttg aacggcatga tggctggagc
     3181 aatctgctca tgagtgaggc cgatggcgtc ctttgctcgg aagagtatga agatgaacaa
     3241 agccctgaaa agattatcga gctgtatgcg gagtgcatca ggctctttca ctccatcgac
     3301 atatcggatt gtccctatac gaatagctta gacagccgct tagccgaatt ggattactta
     3361 ctgaataacg atctggccga tgtggattgc gaaaactggg aagaggacac tccatttaaa
     3421 gatccgcgcg agctgtatga ttttttaaag acggaaaagc ccgaagagga acttgtcttt
     3481 tcccacggcg acctgggaga cagcaacatc tttgtgaaag atggcaaagt aagtggcttt
     3541 attgatcttg ggagaagcgg cagggcggac aagtggtatg acattgcctt ctgcgtccgg
     3601 tcgctcaggg aggatatcgg ggaagaacag tatgtcgagc tattttttga cttactgggg
     3661 atcaagcctg attgggagaa aataaaatat tatattttac tggatgaatt gttttagctg
     3721 tcagaccaag tttactcata tatactttag attgatttaa aacttcattt ttaatttaaa
     3781 aggatctagg tgaagatcct ttttgataat ctcatgacca aaatccctta acgtgagttt
     3841 tcgttccact gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg agatcctttt
     3901 tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt
     3961 ttgccggatc aagagctacc aactcttttt ccgaaggtaa ctggcttcag cagagcgcag
     4021 ataccaaata ctgttcttct agtgtagccg tagttaggcc accacttcaa gaactctgta
     4081 gcaccgccta catacctcgc tctgctaatc ctgttaccag tggctgctgc cagtggcgat
     4141 aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac cggataaggc gcagcggtcg
     4201 ggctgaacgg ggggttcgtg cacacagccc agcttggagc gaacgaccta caccgaactg
     4261 agatacctac agcgtgagct atgagaaagc gccacgcttc ccgaagggag aaaggcggac
     4321 aggtatccgg taagcggcag ggtcggaaca ggagagcgca cgagggagct tccaggggga
     4381 aacgcctggt atctttatag tcctgtcggg tttcgccacc tctgacttga gcgtcgattt
     4441 ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc ggccttttta
     4501 cggttcctgc tcggatctgt tggaccggac agtagtcatg gttgatgggc tgcctgtatc
     4561 gagtggtgat tttgtgccga gctgccggtc ggggagctgt tggctggctg gtggcaggat
     4621 atattgtggt gtaaacaaat tgacgcttag acaacttaat aacacattgc ggacgttttt
     4681 aatgtactgg ggttgaacac tctgtgccac cgttttgcag gtgcggccgc attaggcacc
     4741 ccaggcttta cactttatgc ttccggctcg tataatgtgt ggattttgag ttaggatccg
     4801 tcgagatttt caggagctaa ggaagctaaa atggagaaaa aaatcactgg atataccacc
     4861 gttgatatat cccaatggca tcgtaaagaa cattttgagg catttcagtc agtcgctcaa
     4921 tgtacctata accagaccgt tcagctggat attacggcct ttttaaagac cgtaaagaaa
     4981 aataagcaca agttttatcc ggcctttatt cacattcttg cccgcctgat gaatgctcat
     5041 ccggaattcc gtatggcaat gaaagacggt gagctggtga tatgggatag tgttcaccct
     5101 tgttacaccg ttttccatga gcaaactgaa acgttttcat cgctctggag tgaataccac
     5161 gacgatttcc ggcagtttct acacatatat tcgcaagatg tggcgtgtta cggtgaaaac
     5221 ctggcctatt tccctaaagg gtttattgag aatatgtttt tcgtttcagc caatccctgg
     5281 gtgagtttca ccagttttga tttaaacgtg gccaatatgg acaacttctt cgcccccgtt
     5341 ttcaccatgg gcaaatatta tacgcaaggc gacaaggtgc tgatgccgct ggcgattcag
     5401 gttcatcatg ccgtttgtga tggcttccat gtcggcagaa tgcttaatga attacaacag
     5461 tactgcgatg agtggcaggc ggggcgtaat ctagaggatc cggcttacta aaagccagat
     5521 aacagtatgc gtatttgcgc gctgattttt gcggtataag aatatatact gatatgtata
     5581 cccgaagtat gtcaaaagga ggtatgctat gaagcagcgt attacagtga cagttgacag
     5641 cgacagctat cagttgctca aggcatatat gatgtcaata tctccggtct ggtaagcaca
     5701 accatgcaga atgaagcccg tcgtctgcgt gccgaacgct ggaaagcgga aaatcaggaa
     5761 gggatggctg aggtcgcccg gtttattgaa atgaacggct cttttgctga cgagaacagg
     5821 ggctggtgaa atgcagttta aggtttacac ctataaaaga gagagccgtt atcgtctgtt
     5881 tgtggatgta cagagtgata ttattgacac gcccgggcga cggatggtga tccccctggc
     5941 cagtgcacgt ctgctgtcag ataaagtctc ccgtgaactt tacccggtgg tgcatatcgg
     6001 ggatgaaagc tggcgcatga tgaccaccga tatggccagt gtgccggttt ccgttatcgg
     6061 ggaagaagtg gctgatctca gccaccgcga aaatgacatc aaaaacgcca ttaacctgat
     6121 gttctgggga atataagccg gtaaatcgac gctggtcaat ctgttaggcc cgagggcggt
     6181 tgaagaacgt ctgagacaac atcttcagct tgccagtgag catctctctg cggcctgcca
     6241 acacgggcac gccactcaac attttattca ggcctggttt gacaaaaaac tcgctgccgt
     6301 cagttaagct tgcaagaatt caagcttgga gtacgttaat tagttgtctg ttttaagccc
     6361 cgtgctctcc gccggggctt ttttattaaa aacacctgca aaaagtgggg a
//
