LOCUS       sequence-404799-        5994 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Level 2 destination vector: pMIN-VS1 backbone.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5994)
  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 5994)
  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-VS1
FEATURES             Location/Qualifiers
     source          1..5994
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    1..25
                     /label=LB T-DNA repeat
                     /note="left border repeat from nopaline C58 T-DNA"
     promoter        join(128..133,134..151,152..158)
                     /label=lac UV5 promoter
                     /note="E. coli lac promoter with an ""up"" mutation"
     CDS             212..892
                     /codon_start=1
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQAGRNLE
                     DPAY"
     CDS             1212..1517
                     /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     1396..1415
                     /label=ccdB-fwd
                     /note="ccdB gene, forward primer"
     misc_feature    1554..1578
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     CDS             1714..2529
                     /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             1714..2529
                     /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(1787..1806)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      2615..3203
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3104..3123
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             3444..4073
                     /codon_start=1
                     /product="stability protein from the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
                     /label=pVS1 StaA
                     /translation="MKVIAVLNQKGGSGKTTIATHLARALQLAGADVLLVDSDPQGSAR
                     DWAAVREDQPLTVVGIDRPTIDRDVKAIGRRDFVVIDGAPQAADLAVSAIKAADFVLIP
                     VQPSPYDIWATADLVELVKQRIEVTDGRLQAAFVVSRAIKGTRIGGEVAEALAGYELPI
                     LESRITQRVSYPGTAAAGTTVLESEPEGDAAREVQALAAEIKSKLI"
     CDS             4502..5575
                     /codon_start=1
                     /product="replication protein from the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
                     /label=pVS1 RepA
                     /translation="MSGRKPSGPVQIGAALGDDLVEKLKAAQAAQRQRIEAEARPGESW
                     QAAADRIRKESRQPPAAGAPSIRKPPKGDEQPDFFVPMLYDVGTRDSRSIMDVAVFRLS
                     KRDRRAGEVIRYELPDGHVEVSAGPAGMASVWDYDLVLMAVSHLTESMNRYREGKGDKP
                     GRVFRPHVADVLKFCRRADGGKQKDDLVETCIRLNTTHVAMQRTKKAKNGRLVTVSEGE
                     ALISRYKIVKSETGRPEYIEIELADWMYREITEGKNPDVLTVHPDYFLIDPGIGRFLYR
                     LARRAAGKAEARWLFKTIYERSGSAGEFKKFCFTVRKLIGSNDLPEYDLKEEAGQAGPI
                     LVMRYRNLIEGEASAGS"
     rep_origin      5641..5835
                     /label=pVS1 oriV
                     /note="origin of replication for the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
ORIGIN
        1 tggcaggata tattgtggtg taaacaaatt gacgcttaga caacttaata acacattgcg
       61 gacgttttta atgtactgaa tgcccgggca ccgttttgca ggtgcggccg cattaggcac
      121 cccaggcttt acactttatg cttccggctc gtataatgtg tggattttga gttaggatcc
      181 gtcgagattt tcaggagcta aggaagctaa aatggagaaa aaaatcactg gatataccac
      241 cgttgatata tcccaatggc atcgtaaaga acattttgag gcatttcagt cagtcgctca
      301 atgtacctat aaccagaccg ttcagctgga tattacggcc tttttaaaga ccgtaaagaa
      361 aaataagcac aagttttatc cggcctttat tcacattctt gcccgcctga tgaatgctca
      421 tccggaattc cgtatggcaa tgaaagacgg tgagctggtg atatgggata gtgttcaccc
      481 ttgttacacc gttttccatg agcaaactga aacgttttca tcgctctgga gtgaatacca
      541 cgacgatttc cggcagtttc tacacatata ttcgcaagat gtggcgtgtt acggtgaaaa
      601 cctggcctat ttccctaaag ggtttattga gaatatgttt ttcgtttcag ccaatccctg
      661 ggtgagtttc accagttttg atttaaacgt ggccaatatg gacaacttct tcgcccccgt
      721 tttcaccatg ggcaaatatt atacgcaagg cgacaaggtg ctgatgccgc tggcgattca
      781 ggttcatcat gccgtttgtg atggcttcca tgtcggcaga atgcttaatg aattacaaca
      841 gtactgcgat gagtggcagg cggggcgtaa tctagaggat ccggcttact aaaagccaga
      901 taacagtatg cgtatttgcg cgctgatttt tgcggtataa gaatatatac tgatatgtat
      961 acccgaagta tgtcaaaagg aggtatgcta tgaagcagcg tattacagtg acagttgaca
     1021 gcgacagcta tcagttgctc aaggcatata tgatgtcaat atctccggtc tggtaagcac
     1081 aaccatgcag aatgaagccc gtcgtctgcg tgccgaacgc tggaaagcgg aaaatcagga
     1141 agggatggct gaggtcgccc ggtttattga aatgaacggc tcttttgctg acgagaacag
     1201 gggctggtga aatgcagttt aaggtttaca cctataaaag agagagccgt tatcgtctgt
     1261 ttgtggatgt acagagtgat attattgaca cgcccgggcg acggatggtg atccccctgg
     1321 ccagtgcacg tctgctgtca gataaagtct cccgtgaact ttacccggtg gtgcatatcg
     1381 gggatgaaag ctggcgcatg atgaccaccg atatggccag tgtgccggtt tccgttatcg
     1441 gggaagaagt ggctgatctc agccaccgcg aaaatgacat caaaaacgcc attaacctga
     1501 tgttctgggg aatataacac ctgcaaaaag tgcccgggca aactatcagt gtttgacagg
     1561 atatattggc gggtaaacct aagagaaaag agcgtttaca attagatcca cgttgtgtct
     1621 caaaatctct gatgttacat tgcacaagat aaaaatatat catcatgaac aataaaactg
     1681 tctgcttaca taaacagtaa tacaatgggt gttatgagcc atattcaacg ggaaacgtct
     1741 tgttcgaggc cgcgattaaa ttccaacatg gatgctgatt tatatgggta taaatgggct
     1801 cgcgataatg tcgggcaatc aggtgcgaca atctaccgat tgtatgggaa gcccgatgcg
     1861 ccagagttgt ttctgaaaca tggcaaaggt agcgttgcca atgatgttac agatgagatg
     1921 gtcagactaa actggctgac ggaatttatg cctcttccga ccatcaagca ttttatccgt
     1981 actcctgatg atgcatggtt actcaccact gcgatcccag ggaaaacagc attccaggta
     2041 ttagaagaat atcctgattc aggtgaaaat attgttgatg cgctggcagt gttcctgcgc
     2101 cggttgcatt cgattcctgt ttgtaattgt ccttttaaca gcgatcgcgt atttcgtctg
     2161 gctcaggcgc aatcacgaat gaataacggt ttggttgatg cgagtgattt tgatgacgag
     2221 cgtaatggct ggcctgttga acaagtctgg aaagaaatgc ataaactttt gccattctca
     2281 ccggattcag tcgtcactca tggtgatttc tcacttgata accttatttt tgacgagggg
     2341 aaattaatag gttgtattga tgttggacga gtcggaatcg cagaccgata ccaggatctt
     2401 gccatcctat ggaactgcct cggtgagttt tctccttcat tacagaaacg gctttttcaa
     2461 aaatatggta ttgataatcc tgatatgaat aaattgcagt ttcatttgat gctcgatgag
     2521 tttttctaat cagaattggt taattggttg taacactggc agagcattac gctgacttga
     2581 cgggacggcg gctttgttga atcaattgat cttcttgaaa tccttttttt ctgcgcgtaa
     2641 tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg ccggatcaag
     2701 agctaccaac tctttttccg aaggtaactg gcttcagcag agcgcagata ccaaatactg
     2761 ttcttctagt gtagccgtag ttaggccacc acttcaagaa ctctgtagca ccgcctacat
     2821 acctcgctct gctaatcctg ttaccagtgg ctgctgccag tggcgataag tcgtgtctta
     2881 ccgggttgga ctcaagacga tagttaccgg ataaggcgca gcggtcgggc tgaacggggg
     2941 gttcgtgcac acagcccagc ttggagcgaa cgacctacac cgaactgaga tacctacagc
     3001 gtgagctatg agaaagcgcc acgcttcccg aagggagaaa ggcggacagg tatccggtaa
     3061 gcggcagggt cggaacagga gagcgcacga gggagcttcc agggggaaac gcctggtatc
     3121 tttatagtcc tgtcgggttt cgccacctct gacttgagcg tcgatttttg tgatgctcgt
     3181 caggggggcg gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct
     3241 tttgctggcc ttttgctcac atgttctttc ctgcgttatc ccctgattct gtggataacc
     3301 gtattaccgc ctttgagtga tcctgaagaa accgagcgcc gccgtctaaa aaggtgatgt
     3361 gtatttgagt aaaacagctt gcgtcatgcg gtcgctgcgt atatgatgcg atgagtaaat
     3421 aaacaaatac gcaaggggaa cgcatgaagg ttatcgctgt acttaaccag aaaggcgggt
     3481 caggcaagac gaccatcgca acccatctag cccgcgccct gcaactcgcc ggggccgatg
     3541 ttctgttagt cgattccgat ccccagggca gtgcccgcga ttgggcggcc gtgcgggaag
     3601 atcaaccgct aaccgttgtc ggcatcgacc gcccgacgat tgaccgcgac gtgaaggcca
     3661 tcggccggcg cgacttcgta gtgatcgacg gagcgcccca ggcggcggac ttggctgtgt
     3721 ccgcgatcaa ggcagccgac ttcgtgctga ttccggtgca gccaagccct tacgacatat
     3781 gggccaccgc cgacctggtg gagctggtta agcagcgcat tgaggtcacg gatggaaggc
     3841 tacaagcggc ctttgtcgtg tcgcgggcga tcaaaggcac gcgcatcggc ggtgaggttg
     3901 ccgaggcgct ggccgggtac gagctgccca ttcttgagtc ccgtatcacg cagcgcgtga
     3961 gctacccagg cactgccgcc gccggcacaa ccgttcttga atcagaaccc gagggcgacg
     4021 ctgcccgcga ggtccaggcg ctggccgctg aaattaaatc aaaactcatt tgagttaatg
     4081 aggtaaagag aaaatgagca aaagcacaaa cacgctaagt gccggccgtc cgagcgcacg
     4141 cagcagcaag gctgcaacgt tggccagcct ggcagacacg ccagccatga agcgggtcaa
     4201 ctttcagttg ccggcggagg atcacaccaa gctgaagatg tacgcggtac gccaaggcaa
     4261 gaccattacc gagctgctat ctgaatacat cgcgcagcta ccagagtaaa tgagcaaatg
     4321 aataaatgag tagatgaatt ttagcggcta aaggaggcgg catggaaaat caagaacaac
     4381 caggcaccga cgccgtggaa tgccccatgt gtggaggaac gggcggttgg ccaggcgtaa
     4441 gcggctgggt tgcctgccgg ccctgcaatg gcactggaac ccccaagccc gaggaatcgg
     4501 cgtgagcggt cgcaaaccat ccggcccggt acaaatcggc gcggcgctgg gtgatgacct
     4561 ggtggagaag ttgaaggccg cgcaggccgc ccagcggcaa cgcatcgagg cagaagcacg
     4621 ccccggtgaa tcgtggcaag cggccgctga tcgaatccgc aaagaatccc ggcaaccgcc
     4681 ggcagccggt gcgccgtcga ttaggaagcc gcccaagggc gacgagcaac cagatttttt
     4741 cgttccgatg ctctatgacg tgggcacccg cgatagtcgc agcatcatgg acgtggccgt
     4801 tttccgtctg tcgaagcgtg accgacgagc tggcgaggtg atccgctacg agcttccaga
     4861 cgggcacgta gaggtttccg cagggccggc cggcatggcc agtgtgtggg attacgacct
     4921 ggtactgatg gcggtttccc atctaaccga atccatgaac cgataccggg aagggaaggg
     4981 agacaagccc ggccgcgtgt tccgtccaca cgttgcggac gtactcaagt tctgccggcg
     5041 agccgatggc ggaaagcaga aagacgacct ggtagaaacc tgcattcggt taaacaccac
     5101 gcacgttgcc atgcagcgta cgaagaaggc caagaacggc cgcctggtga cggtatccga
     5161 gggtgaagcc ttgattagcc gctacaagat cgtaaagagc gaaaccgggc ggccggagta
     5221 catcgagatc gagctagctg attggatgta ccgcgagatc acagaaggca agaacccgga
     5281 cgtgctgacg gttcaccccg attacttttt gatcgatccc ggcatcggcc gttttctcta
     5341 ccgcctggca cgccgcgccg caggcaaggc agaagccaga tggttgttca agacgatcta
     5401 cgaacgcagt ggcagcgccg gagagttcaa gaagttctgt ttcaccgtgc gcaagctgat
     5461 cgggtcaaat gacctgccgg agtacgattt gaaggaggag gcggggcagg ctggcccgat
     5521 cctagtcatg cgctaccgca acctgatcga gggcgaagca tccgccggtt cctaatgtac
     5581 ggagcagatg ctagggcaaa ttgccctagc aggggaaaaa ggtcgaaaag gcctctttcc
     5641 tgtggatagc acgtacattg ggaacccaaa gccgtacatt gggaaccgga acccgtacat
     5701 tgggaaccca aagccgtaca ttgggaaccg gtcacacatg taagtgactg atataaaaga
     5761 gaaaaaaggc gatttttccg cctaaaactc tttaaaactt attaaaactc ttaaaacccg
     5821 cctggcctgt gcataactgt ctggccagcg cacagcccaa gagctgcaaa aagcgcctac
     5881 ccttcggtcg ctgcgctccc tacgccccgc cgcttcgcgt cggcctatcg cggccgctgg
     5941 ccgctcaaaa atggctggcc tacggccagg caatctacca gggcgcggac aagc
//
