LOCUS       sequence-228245-        2992 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  entry vector for GreenGate cloning method, terminator.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 2992)
  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 2992)
  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: pVL12 - tAt4g24550
FEATURES             Location/Qualifiers
     source          1..2992
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(331..350)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(332..350)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(360..382)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     complement(576..595)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     695..717
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(755..773)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        841..945
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(946..1014,1015..1806)
                     /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(1164..1183)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      1977..2565
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2466..2485
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2719..2736
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    2853..2874
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(2889..2894,2895..2912,2913..2919)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2927..2943
                     /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        2957..2975
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     2957..2974
                     /label=SP6
                     /note="SP6 promoter, forward primer"
ORIGIN
        1 ctgccctgtt ttcagataat gtttatcctt cacttgtatg tctaagtact ttctcttttc
       61 attttcggat cacacttgct tcacagtact gtgttgaatg ttgtctcttc tatttggata
      121 ctttcttgtt tcggtatcaa catatatggc tatatgcttt tattttgttt gcccctgaaa
      181 acatcatatg atctcttttc ttggaactgg caaaattgca tcttttggga tttttcttga
      241 atttctgaac tgaactgaat ccaaagattc aaagttaagc aaattgcaaa agaaataaag
      301 gtgcaacgat actatgagac cgaattctcg ccctatagtg agtcgtatta cctggccgtc
      361 gttttacaac gtcgtgactg ggaaaaccct ggcgttaccc aacttaatcg ccttgcagca
      421 catccccctt tcgccagctg gcgtaatagc gaagaggccc gcaccgatcg cccttcccaa
      481 cagttgcgca gcctgaatgg cgaatggcgc ctgatgcggt attttctcct tacgcatctg
      541 tgcggtattt cacaccgcat atggtgcact ctcagtacaa tctgctctga tgccgcatag
      601 ttaagccagc cccgacaccc gccaacaccc gctgacgcgc cctgacgggc ttgtctgctc
      661 ccggcatccg cttacagaca agctgtgacc gtctccggga gctgcatgtg tcagaggttt
      721 tcaccgtcat caccgaaacg cgcgagacga aagggcctcg tgatacgcct atttttatag
      781 gttaatgtca tgataataat ggtttcttag acgtcaggtg gcacttttcg gggaaatgtg
      841 cgcggaaccc ctatttgttt atttttctaa atacattcaa atatgtatcc gctcatgaga
      901 caataaccct gataaatgct tcaataatat tgaaaaagga agagtatgag tattcaacat
      961 ttccgtgtcg cccttattcc cttttttgcg gcattttgcc ttcctgtttt tgctcaccca
     1021 gaaacgctgg tgaaagtaaa agatgctgaa gatcagttgg gtgcacgagt gggttacatc
     1081 gaactggatc tcaacagcgg taagatcctt gagagttttc gccccgaaga acgttttcca
     1141 atgatgagca cttttaaagt tctgctatgt ggcgcggtat tatcccgtat tgacgccggg
     1201 caagagcaac tcggtcgccg catacactat tctcagaatg acttggttga gtactcacca
     1261 gtcacagaaa agcatcttac ggatggcatg acagtaagag aattatgcag tgctgccata
     1321 accatgagtg ataacactgc ggccaactta cttctgacaa cgatcggagg accgaaggag
     1381 ctaaccgctt ttttgcacaa catgggggat catgtaactc gccttgatcg ttgggaaccg
     1441 gagctgaatg aagccatacc aaacgacgag cgtgacacca cgatgcctgt agcaatggca
     1501 acaacgttgc gcaaactatt aactggcgaa ctacttactc tagcttcccg gcaacaatta
     1561 atagactgga tggaggcgga taaagttgca ggaccacttc tgcgctcggc ccttccggct
     1621 ggctggttta ttgctgataa atctggagcc ggtgagcgtg gctctcgagg tatcattgca
     1681 gcactggggc cagatggtaa gccctcccgt atcgtagtta tctacacgac ggggagtcag
     1741 gcaactatgg atgaacgaaa tagacagatc gctgagatag gtgcctcact gattaagcat
     1801 tggtaactgt cagaccaagt ttactcatat atactttaga ttgatttaaa acttcatttt
     1861 taatttaaaa ggatctaggt gaagatcctt tttgataatc tcatgaccaa aatcccttaa
     1921 cgtgagtttt cgttccactg agcgtcagac cccgtagaaa agatcaaagg atcttcttga
     1981 gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg
     2041 gtggtttgtt tgccggatca agagctacca actctttttc cgaaggtaac tggcttcagc
     2101 agagcgcaga taccaaatac tgttcttcta gtgtagccgt agttaggcca ccacttcaag
     2161 aactctgtag caccgcctac atacctcgct ctgctaatcc tgttaccagt ggctgctgcc
     2221 agtggcgata agtcgtgtct taccgggttg gactcaagac gatagttacc ggataaggcg
     2281 cagcggtcgg gctgaacggg gggttcgtgc acacagccca gcttggagcg aacgacctac
     2341 accgaactga gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga
     2401 aaggcggaca ggtatccggt aagcggcagg gtcggaacag gagagcgcac gagggagctt
     2461 ccagggggaa acgcctggta tctttatagt cctgtcgggt ttcgccacct ctgacttgag
     2521 cgtcgatttt tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg
     2581 gcctttttac ggttcctggc cttttgctgg ccttttgctc acatgttctt tcctgcgtta
     2641 tcccctgatt ctgtggataa ccgtattacc gcctttgagt gagctgatac cgctcgccgc
     2701 agccgaacga ccgagcgcag cgagtcagtg agcgaggaag cggaagagcg cccaatacgc
     2761 aaaccgcctc tccccgcgcg ttggccgatt cattaatgca gctggcacga caggtttccc
     2821 gactggaaag cgggcagtga gcgcaacgca attaatgtga gttagctcac tcattaggca
     2881 ccccaggctt tacactttat gcttccggct cgtatgttgt gtggaattgt gagcggataa
     2941 caatttcaca cgagctattt aggtgacact atagaagtga agcttggtct ca
//
