LOCUS       sequence-228173-        6261 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  entry vector for GreenGate cloning method, promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6261)
  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 6261)
  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: pVL23 - pVND7
FEATURES             Location/Qualifiers
     source          1..6261
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(10..28)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        96..200
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(201..269,270..1061)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(419..438)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      1232..1820
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1721..1740
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     1974..1991
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    2108..2129
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(2144..2149,2150..2167,2168..2174)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2182..2198
                     /label=lac operator
                     /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)."
     promoter        2212..2230
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     2212..2229
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(5847..5866)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(5848..5866)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(5876..5898)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     complement(6092..6111)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     6211..6233
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
ORIGIN
        1 gacgaaaggg cctcgtgata cgcctatttt tataggttaa tgtcatgata ataatggttt
       61 cttagacgtc aggtggcact tttcggggaa atgtgcgcgg aacccctatt tgtttatttt
      121 tctaaataca ttcaaatatg tatccgctca tgagacaata accctgataa atgcttcaat
      181 aatattgaaa aaggaagagt atgagtattc aacatttccg tgtcgccctt attccctttt
      241 ttgcggcatt ttgccttcct gtttttgctc acccagaaac gctggtgaaa gtaaaagatg
      301 ctgaagatca gttgggtgca cgagtgggtt acatcgaact ggatctcaac agcggtaaga
      361 tccttgagag ttttcgcccc gaagaacgtt ttccaatgat gagcactttt aaagttctgc
      421 tatgtggcgc ggtattatcc cgtattgacg ccgggcaaga gcaactcggt cgccgcatac
      481 actattctca gaatgacttg gttgagtact caccagtcac agaaaagcat cttacggatg
      541 gcatgacagt aagagaatta tgcagtgctg ccataaccat gagtgataac actgcggcca
      601 acttacttct gacaacgatc ggaggaccga aggagctaac cgcttttttg cacaacatgg
      661 gggatcatgt aactcgcctt gatcgttggg aaccggagct gaatgaagcc ataccaaacg
      721 acgagcgtga caccacgatg cctgtagcaa tggcaacaac gttgcgcaaa ctattaactg
      781 gcgaactact tactctagct tcccggcaac aattaataga ctggatggag gcggataaag
      841 ttgcaggacc acttctgcgc tcggcccttc cggctggctg gtttattgct gataaatctg
      901 gagccggtga gcgtggctct cgaggtatca ttgcagcact ggggccagat ggtaagccct
      961 cccgtatcgt agttatctac acgacgggga gtcaggcaac tatggatgaa cgaaatagac
     1021 agatcgctga gataggtgcc tcactgatta agcattggta actgtcagac caagtttact
     1081 catatatact ttagattgat ttaaaacttc atttttaatt taaaaggatc taggtgaaga
     1141 tcctttttga taatctcatg accaaaatcc cttaacgtga gttttcgttc cactgagcgt
     1201 cagaccccgt agaaaagatc aaaggatctt cttgagatcc tttttttctg cgcgtaatct
     1261 gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg gatcaagagc
     1321 taccaactct ttttccgaag gtaactggct tcagcagagc gcagatacca aatactgttc
     1381 ttctagtgta gccgtagtta ggccaccact tcaagaactc tgtagcaccg cctacatacc
     1441 tcgctctgct aatcctgtta ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg
     1501 ggttggactc aagacgatag ttaccggata aggcgcagcg gtcgggctga acggggggtt
     1561 cgtgcacaca gcccagcttg gagcgaacga cctacaccga actgagatac ctacagcgtg
     1621 agctatgaga aagcgccacg cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg
     1681 gcagggtcgg aacaggagag cgcacgaggg agcttccagg gggaaacgcc tggtatcttt
     1741 atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg atttttgtga tgctcgtcag
     1801 gggggcggag cctatggaaa aacgccagca acgcggcctt tttacggttc ctggcctttt
     1861 gctggccttt tgctcacatg ttctttcctg cgttatcccc tgattctgtg gataaccgta
     1921 ttaccgcctt tgagtgagct gataccgctc gccgcagccg aacgaccgag cgcagcgagt
     1981 cagtgagcga ggaagcggaa gagcgcccaa tacgcaaacc gcctctcccc gcgcgttggc
     2041 cgattcatta atgcagctgg cacgacaggt ttcccgactg gaaagcgggc agtgagcgca
     2101 acgcaattaa tgtgagttag ctcactcatt aggcacccca ggctttacac tttatgcttc
     2161 cggctcgtat gttgtgtgga attgtgagcg gataacaatt tcacacgagc tatttaggtg
     2221 acactataga agtgaagctt ggtctcaacc ttagcacgtt gacgtatgct gagtgaaaaa
     2281 agtcaaaaat taaattgcta aacccactaa ttacagttac cgctaattat aattagtgga
     2341 gagtggcaag tacacacgca ctatgatttg tgagttgtga ccatccatat atgtagttca
     2401 atgcatcata tgtatagata ataaaactga taaaagtata gttcaaaaag cttcgataat
     2461 ctgtaaataa gaaaaacaca tgatttgcta tctcatgaca cgacttagta aaatcaatct
     2521 atacgtaaca tatatagcca attcttttaa taaaaatata tatatgaaat cattttatct
     2581 ttttggagaa acaaaaggca tgcttagata tgctttggtg cttcatgata tatccaaaat
     2641 tagtagtatc aaactaaacc gaaactgaaa gaaaaagaaa aaaagcgtag tcattctaca
     2701 tggtgaacgt aactcgtata tgccatattg ccatacataa aagagtccat aaatatggtt
     2761 cgactgcaaa gaattagggg ttatattgtt atatacgtgt gtgtgtataa cgagacttag
     2821 agacactatg attagagaaa ggaaaattag ggatcgaggg acatttatat aacgaaaaaa
     2881 gtgaatggtg tgaaagggtt aatggataga tggtcttggg atttgaacaa aaggctctct
     2941 ctccctcgcg cagtagacaa ataggcatgc atgtgaatct gactccaaat caagtgctac
     3001 ctcttttgcc ttttctattt ttggtaccca atatttgtta taattttcat tttttctgtt
     3061 ttatatatcc attctcctcg aattctgtgt ctaccttgtc atatgttaag tattttggat
     3121 ttttctttgg actagggtca aagagttact gagatgacgt caccttaagc cgaaaggtga
     3181 tgaggtaaaa gaaacgacag ccaacggctt tgggtggacc cgtgcaagcg gataaatcag
     3241 atgaggaaca aatgattagt tatgtgatag aaacaggtct gaatcttacg tgtcttacat
     3301 tcgtaatgac acgtgttaag gacggcgcca ccaaaccaga caatgggagc tggattgctg
     3361 gagaaagact taagaacata tgaaacatat tcctgtgttt tactcttctc cttatttttt
     3421 ctttcataat ataatgttca tatatacacg attcctgatg ttttaatggc caatgttcgt
     3481 tcctttaggt ttactataaa tctgagttta gtttaatcat ttaagaaaaa gttgagattg
     3541 tggtttcgag tggatctagc tagactcaca ttataaaatt agctgcggtg gtatattgag
     3601 tgagatttac gtagtaatcc ataaatgtac atgcgtgtaa atcctttcga cgttgagatg
     3661 tttatatact taataagtta atagtaatat ctctttgaca aaaaagttaa tagtaatatc
     3721 tcaacaactt tgaggtatta cgtgtcttat ctatgggtta ataagacata ctaaaactaa
     3781 gacacatttc atcagtacct gatccagcaa acatatagga atatatttct gtagttcttc
     3841 ttcgggttta caaatccagt cctttttaat ccttgaattg tgaatttcca acaagccact
     3901 cacaatttat aactagtttt tattactgaa aatatcttac atggtataag aatttatggt
     3961 gtgtagctga acatagtgtt ttaagaaaat agattaatta tattcaaaac gtaatttttt
     4021 tgtcatctga ttttattgat aatacatgga atcgtgttat gaaatattat ccaatcaaac
     4081 tgaagttttt ggatatattc ggaacatcca tttgacctaa ttacgccaca taaaaaccgt
     4141 taatctaaag gatttgtctt gtgtgatgta ataataacta atgagcaata gacaaataaa
     4201 aaaaaggttt ggcttgtgca gtaactgccg gtaaagtgga gaagattgga aaaactttaa
     4261 acaaaaaatt gcggtaagag agagagagag acgccgacca aaaacaaacc gtagatgtct
     4321 atcccacgcg caatcaaacc tttccttcat ctccttcttc ttcttcttct ccttcttctt
     4381 atatcatcat tttgcaccca catgtcacac acttaccata tacatataat taagcatact
     4441 cgcataaacc tatatataca ctctaagatt tctagctggt ccatatgcat gtgatctaat
     4501 caaaagccaa actctccttg tctttgctat attaacaatc atttgatcca caccaccaga
     4561 gatttcaatc ttcctctcct tcttcctttt gctactctct tttagatcgt tcttgtgaaa
     4621 atcgcttgca aataacaagc ccttcgtcca atctttatta gctcgtaagt ttcaccccat
     4681 tgctcctttt agggtttttt tgttttgttg ctgtactcac ttcatgcacg aaccagatga
     4741 tgatacatcg gcgtttgtgt tttacgtttt tattattcaa aaatacatac atattacata
     4801 tagtatcgtt ttgtatcgtt tcatatcttg aatagtatac atgtgtgtgg tcctgttgag
     4861 attgacccac aaatattaga gtaaactggg gaattgaaat caatcatggt ggggatttcg
     4921 gaatctatgt gtgtttctaa gatattattg tttcttggaa attaattgtt gttgcaccat
     4981 aatataattt atattatata gatagtctag gaaaataact caagacaaag agtgtgtaac
     5041 acactccaac ctcagtgact tccacccgtc aagcttttcg gctacatagc ctttatatac
     5101 gattatttta catatagtta cagatatata cagtactgct tgtatatttt tggtcttata
     5161 atactacctt atgtcattat atatagtaca tattatacaa agtttcaatc ttttcattta
     5221 taagcatcgt tcggtatgta gaaggcgaaa caaaccctta aacctaatat ttttttctgg
     5281 tccaaaaact ttctctatca agcttttgca ttttttcttt taaaatagta acagctcctc
     5341 cattgttggg tggtcccata aaaatacatc ataaccatca tcagcaacat ataatagcga
     5401 tagccttaag cttaaagatc caagcttttg gatataatat ctagttacat gtatatatat
     5461 atatatgtgt ttgtatattt ataggtatat agtatattat cgtgtgcgtc cgtgtgtaac
     5521 atcactatca ttacccaaca tacagctttt ggaatttcat tggacccaag agccattcga
     5581 tccctttctt ataattaaat ataacgaagc ttgtaatagg ctttggtttt ggtcccaaat
     5641 acaaaacttt aaggttgtgc cttgtttgtc taaggttttc gcaatagctt ccaaaaagac
     5701 acgcttatgt ttatatacat ttcattttcc ttcatatcat tctcttctct gccacttctc
     5761 catcttgttt cttactcatt tctctaacaa ttttccaaat taaatacgtt tataggatca
     5821 tcgtggaaca tgagaccgaa ttctcgccct atagtgagtc gtattacctg gccgtcgttt
     5881 tacaacgtcg tgactgggaa aaccctggcg ttacccaact taatcgcctt gcagcacatc
     5941 cccctttcgc cagctggcgt aatagcgaag aggcccgcac cgatcgccct tcccaacagt
     6001 tgcgcagcct gaatggcgaa tggcgcctga tgcggtattt tctccttacg catctgtgcg
     6061 gtatttcaca ccgcatatgg tgcactctca gtacaatctg ctctgatgcc gcatagttaa
     6121 gccagccccg acacccgcca acacccgctg acgcgccctg acgggcttgt ctgctcccgg
     6181 catccgctta cagacaagct gtgaccgtct ccgggagctg catgtgtcag aggttttcac
     6241 cgtcatcacc gaaacgcgcg a
//
