LOCUS       sequence-227518-        6107 bp    DNA     circular SYN 15-APR-2025
DEFINITION  destination vector for GreenGate cloning method, contains 2x a set 
            of modules from A-F, ""Driver line"", tissues-specific promoter, 
            Dex-inducible mTurquoise2 expression.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6107)
  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 6107)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 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: pSW554 - pOp5:SP-mVenus-HDEL
FEATURES             Location/Qualifiers
     source          1..6107
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    534..556
                     /label=LB T-DNA repeat
                     /note="left border repeat from nopaline C58 T-DNA 
                     (truncated)"
     protein_bind    694..713
                     /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    746..765
                     /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    798..817
                     /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    850..869
                     /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    902..921
                     /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        945..991
                     /label=minimal CaMV 35S promoter
                     /note="minimal 35S promoter from cauliflower mosaic virus"
     CDS             join(1073..1075,1076..1078,1079..1789)
                     /codon_start=1
                     /product="yellow fluorescent protein (YFP) with fast and 
                     efficient maturation (Nagai et al., 2002)"
                     /label=Venus
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KLICTTGKLPVPWPTLVTTLGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYITADKQKNGIK
                     ANFKIRHNIEDGGVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(1118..1139)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1379..1398)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1726..1747
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     promoter        2453..2833
                     /label=MAS promoter
                     /note="mannopine synthase promoter (Velten et al., 1984)"
     misc_feature    4126..4150
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     rep_origin      complement(4241..4829)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4321..4340)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(5000..5788)
                     /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(6079..6107,1..407)
                     /label=pSa ori
                     /note="origin of replication from bacterial plasmid pSa"
ORIGIN
        1 tttttatccc cggaagcctg tggatagagg gtagttatcc acgtgaaacc gctaatgccc
       61 cgcaaagcct tgattcacgg ggctttccgg cccgctccaa aaactatcca cgtgaaatcg
      121 ctaatcaggg tacgtgaaat cgctaatcgg agtacgtgaa atcgctaata aggtcacgtg
      181 aaatcgctaa tcaaaaaggc acgtgagaac gctaatagcc ctttcagatc aacagcttgc
      241 aaacacccct cgctccggca agtagttaca gcaagtagta tgttcaatta gcttttcaat
      301 tatgaatata tatatcaatt attggtcgcc cttggcttgt ggacaatgcg ctacgcgcac
      361 cggctccgcc cgtggacaac cgcaagcggt tgcccaccgt cgagcgcctt tgcccacaac
      421 ccggcggccg gccgcaacag atcgttttat aaattttttt ttttgaaaaa gaaaaagccc
      481 gaaaggcggc aacctctcgg gcttctggat ttccgatccc cggaattaga tcttggcagg
      541 atatattgtg gtgtaacgtt ctcgaactag gctcggacga agtaagcggc cgcaacctta
      601 attaaacacg gcgcgccaac agcgatcgca ccaggtacca cctatgcata tgtcgagctc
      661 aagaaatcta gaaagaagaa agggaagaga aagaattgtg agcgctcaca attgaaagac
      721 tagaaagaag aaagggaaga gaaagaattg tgagcgctca caattgaaag actagaaaga
      781 agaaagggaa gagaaagaat tgtgagcgct cacaattgaa agactagaaa gaagaaaggg
      841 aagagaaaga attgtgagcg ctcacaattg aaagactaga aagaagaaag ggaagagaaa
      901 gaattgtgag cgctcacaat tgaaagacta gtggatcgat cttcgcaaga cccttcctct
      961 atataaggaa gttcatttca tttggagagg acgaacaatg aaagccttca cactcgctct
     1021 cttcttagct ctttccctct atctcctgcc caatccagcc actagtggct ccatggtgag
     1081 caagggcgag gagctgttca ccggggtggt gcccatcctg gtcgagctgg acggcgacgt
     1141 aaacggccac aagttcagcg tgtccggcga gggcgagggc gatgccacct acggcaagct
     1201 gaccctgaag ctgatctgca ccaccggcaa gctgcccgtg ccctggccca ccctcgtgac
     1261 caccctgggc tacggcctgc agtgcttcgc ccgctacccc gaccacatga agcagcacga
     1321 cttcttcaag tccgccatgc ccgaaggcta cgtccaggag cgcaccatct tcttcaagga
     1381 cgacggcaac tacaagaccc gcgccgaggt gaagttcgag ggcgacaccc tggtgaaccg
     1441 catcgagctg aagggcatcg acttcaagga ggacggcaac atcctggggc acaagctgga
     1501 gtacaactac aacagccaca acgtctatat caccgccgac aagcagaaga acggcatcaa
     1561 ggccaacttc aagatccgcc acaacatcga ggacggcggc gtgcagctcg ccgaccacta
     1621 ccagcagaac acccccatcg gcgacggccc cgtgctgctg cccgacaacc actacctgag
     1681 ctaccagtcc gccctgagca aagaccccaa cgagaagcgc gatcacatgg tcctgctgga
     1741 gttcgtgacc gccgccggga tcactctcgg catggacgag ctgtacaagt caggtcatga
     1801 tgagctttag ctgcatctcg tctctgttat gcttaagaag ttcaatgttt cgtttcatgt
     1861 aaaactttgg tggtttgtgt tttggggcct tgtataatcc ctgatgaata agtgttctac
     1921 tatgtttccg ttcctgttat ctctttcttt ctaatgacaa gtcgaacttc ttctttatca
     1981 tcgcttcgtt tttattatct gtgcttcttt tgtttaatac gcctgcaaag tgactcgact
     2041 ctgtttagtg cagttctgcg aaacttgtaa atagtccaat tgttggcctc tagtaataga
     2101 tgtagcgaaa gtgttgagct gttgggttct aaggatggct tgaacatgtt aatcttttag
     2161 gttctgagta tgatgaacat tcgttgttgc taagaaatgc ctgtaatgtc ccacaaatgt
     2221 agaaaatggt tcgtaccttt gtccaagcat tgatatgtct gatgagagga aactgcaaga
     2281 tactgagctt ggtttaacga aggagaggca gtttcttcct tccaaagcat ttcatttgac
     2341 aatgccttga tcatcttaag tagagtttct gttgtggaaa gtttgaaact ttgaagaaac
     2401 gactctcaag taaattgatg atcacaagtg aaagtgtatg ttactaggat ccttttcaaa
     2461 tcagtgcgca agacgtgacg taagtatccg agtcagtttt tatttttcta ctaatttggt
     2521 cgtttatttc ggcgtgtagg acatggcaac cgggcctgaa tttcgcgggt attctgtttc
     2581 tattccaact ttttcttgat ccgcagccat taacgacttt tgaatagata cgctgacacg
     2641 ccaagcctcg ctagtcaaaa gtgtaccaaa caacgcttta cagcaagaac ggaatgcgcg
     2701 tgacgctcgc ggtgacgcca tttcgccttt tcagaaatgg ataaatagcc ttgcttccta
     2761 ttatatcttc ccaaattacc aatacattac actagcatct gaatttcata accaatctcg
     2821 atacaccaaa tcgccatggc ttctatgata tcctcttcag ctgtgactac agtcagccgt
     2881 gcttctacgg tgcaatcggc cgcggtggct ccattcggcg gcctcaaatc catgactgga
     2941 ttcccagtta agaaggtcaa cactgacatt acttccatta caagcaatgg tggaagagta
     3001 aagtgcatgg tgacggtgtt cggcattctg aatctcaccg aggactcctt cttcgatgag
     3061 agccggcggc tagaccccgc cggcgctgtc accgcggcga tcgaaatgct gcgagtcgga
     3121 tcagacgtcg tggatgtcgg accggccgcc agccatccgg acgcgaggcc tgtatcgccg
     3181 gccgatgaga tcagacgtat tgcgccgctc ttagacgccc tgtccgatca gatgcaccgt
     3241 gtttcaatcg acagcttcca accggaaacc cagcgctatg cgctcaagcg cggcgtgggc
     3301 tacctgaacg atatccaagg atttcctgac cctgcgctct atcccgatat tgctgaggcg
     3361 gactgcaggc tggtggttat gcactcagcg cagcgggatg gcatcgccac ccgcaccggt
     3421 caccttcgac ccgaagacgc gctcgacgag attgtgcggt tcttcgaggc gcgggtttcc
     3481 gccttgcgac ggagcggggt cgctgccgac cggctcatcc tcgatccggg gatgggattt
     3541 ttcttgagcc ccgcaccgga aacatcgctg cacgtgctgt cgaaccttca aaagctgaag
     3601 tcggcgttgg ggcttccgct acttgtctcg gtgtcgcgga aatccttctt gggcgccacc
     3661 gttggccttc ctgtaaagga tctgggtcca gcgagccttg cggcggaact tcacgcgatc
     3721 ggcaatggcg ctgactacgt ccgcacccac gcgcctggag atctgcgaag cgcaatcacc
     3781 atctcggaaa ccctcgcgaa atttcgcagt cgcgacgcca gagacagagg gttagatcat
     3841 gcctagcttg gctgcaggtc gacctgcagg catgcccgct gaaatcacca gtctctctct
     3901 acaaatctat ctctctctat aataatgtgt gagtagttcc cagataaggg aattagggtt
     3961 cttatagggt ttcgctcatg tgttgagcat ataagaaacc cttagtatgt atttgtattt
     4021 gtaaaatact tctatcaata aaatttctaa ttcctaaaac caaaatccag gggtaccgta
     4081 ttctagacca agcatgcaca gctggcgttc tcagccagat ccccttgaca ggatatattg
     4141 gcgggtaaac taagtcgctg tatgtgtttg tttgagatct catgtgagca aaaggccagc
     4201 aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg ctccgccccc
     4261 ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg acaggactat
     4321 aaagatacca ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc
     4381 cgcttaccgg atacctgtcc gcctttctcc cttcgggaag cgtggcgctt tctcatagct
     4441 cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg
     4501 aaccccccgt tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc
     4561 cggtaagaca cgacttatcg ccactggcag cagccactgg taacaggatt agcagagcga
     4621 ggtatgtagg cggtgctaca gagttcttga agtggtggcc taactacggc tacactagaa
     4681 gaacagtatt tggtatctgc gctctgctga agccagttac cttcggaaga agagttggta
     4741 gctcttgatc cggcaaacaa accaccgctg gtagcggtgg tttttttgtt tgcaagcagc
     4801 agattacgcg cagaaaaaaa ggatctcaag aagatccttt gatcttttct acggggtctg
     4861 acgctcagtg gaacgaaaac tcacgttaag ggattttggt catgagatta tcaaaaagga
     4921 tcttcaccta gatcctttta aattaaaaat gaagttttaa atcaatctaa agtatatatg
     4981 tgtaacattg gtctagtgat tatttgccga ctaccttggt gatctcgcct ttcacgtagt
     5041 gaacaaattc ttccaactga tctgcgcgcg aggccaagcg atcttcttgt ccaagataag
     5101 cctgcctagc ttcaagtatg acgggctgat actgggccgg caggcgctcc attgcccagt
     5161 cggcagcgac atccttcggc gcgattttgc cggttactgc gctgtaccaa atgcgggaca
     5221 acgtaagcac tacatttcgc tcatcgccag cccagtcggg cggcgagttc catagcgtta
     5281 aggtttcatt tagcgcctca aatagatcct gttcaggaac cggatcaaag agttcctccg
     5341 ccgctggacc taccaaggca acgctatgtt ctcttgcttt tgtcagcaag atagccagat
     5401 caatgtcgat cgtggctggc tcgaagatac ctgcaagaat gtcattgcgc tgccattctc
     5461 caaattgcag ttcgcgctta gctggataac gccacggaat gatgtcgtcg tgcacaacaa
     5521 tggtgacttc tacagcgcgg agaatctcgc tctctccagg ggaagccgaa gtttccaaaa
     5581 ggtcgttgat caaagctcgc cgcgttgttt catcaagcct tacggtcacc gtaaccagca
     5641 aatcaatatc actgtgtggc ttcaggccgc catccactgc ggagccgtac aaatgtacgg
     5701 ccagcaacgt cggttcgaga tggcgctcga tgacgccaac tacctctgat agttgagtcg
     5761 atacttcggc gatcaccgct tccctcataa caccccttgt attactgttt atgtaagcag
     5821 acagttttat tgttcatgat gatatatttt tatcttgtgc aatgtaacat cagagatttt
     5881 gagacacaac gtggctttgt tgaataaatc gaacttttgc tgagttgaag gatcagatca
     5941 cgcatcttcc cgacaacgca gaccgttccg tggcaaagca aaagttcaaa atcaccaact
     6001 ggtccaccta caacaaagct ctcatcaacc gtggctccct cactttctgg ctggatgatg
     6061 gggcgattca ggcgatcccc atccaacagc ccgccgtcga gcgggct
//
