LOCUS       sequence-243112-        6010 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  destination vector for GreenGate cloning method, ""Effector line"", 
            crossed with a ""Driver line"" the GOI, here VND7, will be expressed
            upon Dex-induction .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6010)
  AUTHORS   Schurholz AK, Lopez-Salmeron V, Li Z, Forner J, Wenzl C, Gaillochet 
            C, Augustin S, Vilches-Barro A, Fuchs M, Gebert M, Lohmann JU, Greb 
            T, Wolf S
  TITLE     A Comprehensive Toolkit for Inducible, Cell Type-Specific Gene 
            Expression in Arabidopsis.
  JOURNAL   Plant Physiol. 2018 Jul 19. pii: pp.18.00463. doi: 
            10.1104/pp.18.00463.
  PUBMED    30026289
REFERENCE   2  (bases 1 to 6010)
  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 
            Physiol. 2018 Jul 19. pii: pp.18.00463. doi: 10.1104/pp.18.00463."
COMMENT     Sequence Label: pVL94 - pOp4:VND7
FEATURES             Location/Qualifiers
     source          1..6010
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     269..288
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     misc_feature    459..483
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     protein_bind    620..639
                     /label=lac operator (symmetric)
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG). The 
                     symmetric lac operator was optimized for tight binding of 
                     lac repressor."
     protein_bind    672..691
                     /label=lac operator (symmetric)
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG). The 
                     symmetric lac operator was optimized for tight binding of 
                     lac repressor."
     protein_bind    724..743
                     /label=lac operator (symmetric)
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG). The 
                     symmetric lac operator was optimized for tight binding of 
                     lac repressor."
     promoter        767..813
                     /label=minimal CaMV 35S promoter
                     /note="minimal 35S promoter from cauliflower mosaic virus"
     CDS             1831..2064
                     /codon_start=1
                     /gene="UL48"
                     /product="transcriptional activation domain of herpes 
                     simplex virus protein VP16 (Triezenberg et al., 1988; 
                     Cousens et al., 1989)"
                     /label=VP16 AD
                     /translation="APPTDVSLGDELHLDGEDVAMAHADALDDFDLDMLGDGDSPGPGF
                     TPHDSAPYGALDMADFEFEQMFTDALGIDEYGG"
     promoter        2706..3086
                     /label=MAS promoter
                     /note="mannopine synthase promoter (Velten et al., 1984)"
     CDS             3097..3648
                     /codon_start=1
                     /gene="Streptomyces hygroscopicus bar"
                     /product="phosphinothricin acetyltransferase"
                     /label=BlpR
                     /note="confers resistance to bialophos or phosphinothricin"
                     /translation="MSPERRPADIRRATEADMPAVCTIVNHYIETSTVNFRTEPQEPQE
                     WTDDLVRLRERYPWLVAEVDGEVAGIAYAGPWKARNAYDWTAESTVYVSPRHQRTGLGS
                     TLYTHLLKSLEAQGFKSVVAVIGLPNDPSVRMHEALGYAPRGMLRAAGFKHGNWHDVGF
                     WQLDFSLPVPPRPVLPVTEI"
     terminator      3654..3906
                     /label=MAS terminator
                     /note="mannopine synthase terminator"
     misc_feature    3956..3978
                     /label=LB T-DNA repeat
                     /note="left border repeat from nopaline C58 T-DNA 
                     (truncated)"
     rep_origin      4105..4540
                     /label=pSa ori
                     /note="origin of replication from bacterial plasmid pSa"
     CDS             4831..5619
                     /codon_start=1
                     /gene="aadA"
                     /product="aminoglycoside adenylyltransferase (Murphy, 
                     1985)"
                     /label=SmR
                     /note="confers resistance to spectinomycin and 
                     streptomycin"
                     /translation="MREAVIAEVSTQLSEVVGVIERHLEPTLLAVHLYGSAVDGGLKPH
                     SDIDLLVTVTVRLDETTRRALINDLLETSASPGESEILRAVEVTIVVHDDIIPWRYPAK
                     RELQFGEWQRNDILAGIFEPATIDIDLAILLTKAREHSVALVGPAAEELFDPVPEQDLF
                     EALNETLTLWNSPPDWAGDERNVVLTLSRIWYSAVTGKIAPKDVAADWAMERLPAQYQP
                     VILEARQAYLGQEDRLASRADQLEEFVHYVKGEITKVVGK"
     rep_origin      join(5790..6010,1..368)
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
ORIGIN
        1 tcctgttacc agtggctgct gccagtggcg ataagtcgtg tcttaccggg ttggactcaa
       61 gacgatagtt accggataag gcgcagcggt cgggctgaac ggggggttcg tgcacacagc
      121 ccagcttgga gcgaacgacc tacaccgaac tgagatacct acagcgtgag ctatgagaaa
      181 gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa
      241 caggagagcg cacgagggag cttccagggg gaaacgcctg gtatctttat agtcctgtcg
      301 ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc
      361 tatggaaaaa cgccagcaac gcggcctttt tacggttcct ggccttttgc tggccttttg
      421 ctcacatgag atctcaaaca aacacataca gcgacttagt ttacccgcca atatatcctg
      481 tcaaggctcg aactaggctc ggacgaagta agcggccgca accttaatta aacacggcgc
      541 gccaacagcg atcgcaccag gtaccaccta tgcatatgtc gagctcaaga aatctagaaa
      601 gaagaaaggg aagagaaaga attgtgagcg ctcacaattg aaagactaga aagaagaaag
      661 ggaagagaaa gaattgtgag cgctcacaat tgaaagacta gaaagaagaa agggaagaga
      721 aagaattgtg agcgctcaca attgaaagac tagtggatcg atcttcgcaa gacccttcct
      781 ctatataagg aagttcattt catttggaga ggacgaacag tattcagtcg actggtacca
      841 acaggctcaa tggataatat aatgcaatcg tcaatgccac cgggattccg atttcatccg
      901 acagaggaag agcttgtggg ttattaccta gataggaaga tcaattcaat gaagagtgct
      961 ttagatgtca ttgtagagat tgatctctac aaaatggagc catgggatat acaagcgagg
     1021 tgtaaactag ggtatgaaga gcaaaacgag tggtacttct ttagtcataa ggacaggaag
     1081 taccctaccg ggactaggac caaccgagcc actgcggctg ggttctggaa agccacgggt
     1141 agagacaagg cggtactatc aaaaaacagt gtcatcggaa tgcggaagac acttgtctac
     1201 tacaagggtc gagctcctaa tggaagaaag tccgattgga tcatgcacga ataccgtctc
     1261 caaaactccg agcttgcccc ggttcaggag gaaggctggg tggtgtgtcg agcatttagg
     1321 aagccaattc caaaccagag gccattaggg tacgagccat ggcagaacca gctctaccac
     1381 gtcgaaagta gtaacaacta ctcatcttca gtgacaatga acacgagtca tcatatcggt
     1441 gcatcttcat caagtcataa ccttaatcaa atgctcatga gcaataacca ctacaatcct
     1501 aataatacat cctcatcgat gcatcaatat ggcaacattg agctcccgca gttggacagc
     1561 ccgagcttgt cgcctagttt agggacgaat aaagatcaga acgagagttt cgagcaagaa
     1621 gaagagaaga gctttaactg tgtggattgg agaacactag ataccttgct tgagacacaa
     1681 gtcatacatc cgcataaccc taatattctt atgttcgaaa cgcagtcgta taatccggcg
     1741 ccaagcttcc cttccatgca tcaaagctat aatgaggtcg aagctaatat tcatcattct
     1801 cttggatgct tccctgactc gaaatcagca gcccccccga ccgatgtcag cctgggggac
     1861 gagctccact tagacggcga ggacgtggcg atggcgcatg ccgacgcgct agacgatttc
     1921 gatctggaca tgttggggga cggggattcc ccgggtccgg gatttacccc ccacgactcc
     1981 gccccctacg gcgctctgga tatggccgac ttcgagtttg agcagatgtt taccgatgcc
     2041 cttggaattg acgagtacgg tgggtaactg catctcgtct ctgttatgct taagaagttc
     2101 aatgtttcgt ttcatgtaaa actttggtgg tttgtgtttt ggggccttgt ataatccctg
     2161 atgaataagt gttctactat gtttccgttc ctgttatctc tttctttcta atgacaagtc
     2221 gaacttcttc tttatcatcg cttcgttttt attatctgtg cttcttttgt ttaatacgcc
     2281 tgcaaagtga ctcgactctg tttagtgcag ttctgcgaaa cttgtaaata gtccaattgt
     2341 tggcctctag taatagatgt agcgaaagtg ttgagctgtt gggttctaag gatggcttga
     2401 acatgttaat cttttaggtt ctgagtatga tgaacattcg ttgttgctaa gaaatgcctg
     2461 taatgtccca caaatgtaga aaatggttcg tacctttgtc caagcattga tatgtctgat
     2521 gagaggaaac tgcaagatac tgagcttggt ttaacgaagg agaggcagtt tcttccttcc
     2581 aaagcatttc atttgacaat gccttgatca tcttaagtag agtttctgtt gtggaaagtt
     2641 tgaaactttg aagaaacgac tctcaagtaa attgatgatc acaagtgaaa gtgtatgtta
     2701 ctagtttttc aaatcagtgc gcaagacgtg acgtaagtat ccgagtcagt ttttattttt
     2761 ctactaattt ggtcgtttat ttcggcgtgt aggacatggc aaccgggcct gaatttcgcg
     2821 ggtattctgt ttctattcca actttttctt gatccgcagc cattaacgac ttttgaatag
     2881 atacgctgac acgccaagcc tcgctagtca aaagtgtacc aaacaacgct ttacagcaag
     2941 aacggaatgc gcgtgacgct cgcggtgacg ccatttcgcc ttttcagaaa tggataaata
     3001 gccttgcttc ctattatatc ttcccaaatt accaatacat tacactagca tctgaatttc
     3061 ataaccaatc tcgatacacc aaatcgggta ccaacaatga gcccagaacg acgcccggcc
     3121 gacatccgcc gtgccaccga ggcggacatg ccggcggtct gcaccatcgt caaccactac
     3181 atcgagacaa gcacggtcaa cttccgtacc gagccgcagg aaccgcagga gtggacggac
     3241 gacctcgtcc gtctgcggga gcgctatccc tggctcgtcg ccgaggtgga cggcgaggtc
     3301 gccggcatcg cctacgcggg tccctggaag gcacgcaacg cctacgactg gacggccgag
     3361 tccaccgtgt acgtctcccc ccgccaccag cggacgggac tgggctccac gctctacacc
     3421 cacctgctga agtccctgga ggcacagggc ttcaagagcg tggtcgctgt catcgggctg
     3481 cccaacgacc cgagcgtgcg catgcacgag gcgctcggat atgccccccg cggcatgctg
     3541 cgggcggccg gcttcaagca cgggaactgg catgacgtgg gtttctggca gctggacttc
     3601 agcctgccgg tgccgccccg tccggtcctg cccgtcaccg aaatctgaga gctcttggac
     3661 tcccatgttg gcaaaggcaa ccaaacaaac aatgaatgat ccgctcctgc atatggggcg
     3721 gtttgagtat ttcaactgcc atttgggctg aattgaagac atgctcctgt cagaaattcc
     3781 gtgatcttac tcaatattca gtaatctcgg ccaatatcct aaatgtgcgt ggctttatct
     3841 gtctttgtat tgtttcatca attcatgtaa cgtttgcttt tcttatgaat tttcaaataa
     3901 attatcgtat tctagaccaa gcatgcacag ctggcgttct cagccagatc caacgttaca
     3961 ccacaatata tcctgccaag atctaattcc ggggatcgga aatccagaag cccgagaggt
     4021 tgccgccttt cgggcttttt ctttttcaaa aaaaaaaatt tataaaacga tctgttgcgg
     4081 ccggccgccg ggttgtgggc aaaggcgctc gacggtgggc aaccgcttgc ggttgtccac
     4141 gggcggagcc ggtgcgcgta gcgcattgtc cacaagccaa gggcgaccaa taattgatat
     4201 atatattcat aattgaaaag ctaattgaac atactacttg ctgtaactac ttgccggagc
     4261 gaggggtgtt tgcaagctgt tgatctgaaa gggctattag cgttctcacg tgcctttttg
     4321 attagcgatt tcacgtgacc ttattagcga tttcacgtac tccgattagc gatttcacgt
     4381 accctgatta gcgatttcac gtggatagtt tttggagcgg gccggaaagc cccgtgaatc
     4441 aaggctttgc ggggcattag cggtttcacg tggataacta ccctctatcc acaggcttcc
     4501 ggggataaaa aagcccgctc gacggcgggc tgttggatgg ggatcgcctg aatcgcccca
     4561 tcatccagcc agaaagtgag ggagccacgg ttgatgagag ctttgttgta ggtggaccag
     4621 ttggtgattt tgaacttttg ctttgccacg gaacggtctg cgttgtcggg aagatgcgtg
     4681 atctgatcct tcaactcagc aaaagttcga tttattcaac aaagccacgt tgtgtctcaa
     4741 aatctctgat gttacattgc acaagataaa aatatatcat catgaacaat aaaactgtct
     4801 gcttacataa acagtaatac aaggggtgtt atgagggaag cggtgatcgc cgaagtatcg
     4861 actcaactat cagaggtagt tggcgtcatc gagcgccatc tcgaaccgac gttgctggcc
     4921 gtacatttgt acggctccgc agtggatggc ggcctgaagc cacacagtga tattgatttg
     4981 ctggttacgg tgaccgtaag gcttgatgaa acaacgcggc gagctttgat caacgacctt
     5041 ttggaaactt cggcttcccc tggagagagc gagattctcc gcgctgtaga agtcaccatt
     5101 gttgtgcacg acgacatcat tccgtggcgt tatccagcta agcgcgaact gcaatttgga
     5161 gaatggcagc gcaatgacat tcttgcaggt atcttcgagc cagccacgat cgacattgat
     5221 ctggctatct tgctgacaaa agcaagagaa catagcgttg ccttggtagg tccagcggcg
     5281 gaggaactct ttgatccggt tcctgaacag gatctatttg aggcgctaaa tgaaacctta
     5341 acgctatgga actcgccgcc cgactgggct ggcgatgagc gaaatgtagt gcttacgttg
     5401 tcccgcattt ggtacagcgc agtaaccggc aaaatcgcgc cgaaggatgt cgctgccgac
     5461 tgggcaatgg agcgcctgcc ggcccagtat cagcccgtca tacttgaagc taggcaggct
     5521 tatcttggac aagaagatcg cttggcctcg cgcgcagatc agttggaaga atttgttcac
     5581 tacgtgaaag gcgagatcac caaggtagtc ggcaaataat cactagacca atgttacaca
     5641 tatatacttt agattgattt aaaacttcat ttttaattta aaaggatcta ggtgaagatc
     5701 ctttttgata atctcatgac caaaatccct taacgtgagt tttcgttcca ctgagcgtca
     5761 gaccccgtag aaaagatcaa aggatcttct tgagatcctt tttttctgcg cgtaatctgc
     5821 tgcttgcaaa caaaaaaacc accgctacca gcggtggttt gtttgccgga tcaagagcta
     5881 ccaactcttc ttccgaaggt aactggcttc agcagagcgc agataccaaa tactgttctt
     5941 ctagtgtagc cgtagttagg ccaccacttc aagaactctg tagcaccgcc tacatacctc
     6001 gctctgctaa
//
