LOCUS       sequence-239396-        3335 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  G9a SET domain with GreenGate D-E flanking sequences.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3335)
  AUTHORS   Lee JE, Neumann M, Duro DI, Schmid M
  TITLE     CRISPR-based tools for targeted transcriptional and epigenetic 
            regulation in plants.
  JOURNAL   PLoS One. 2019 Sep 26;14(9):e0222778. doi: 
            10.1371/journal.pone.0222778. eCollection 2019.
  PUBMED    31557222
REFERENCE   2  (bases 1 to 3335)
  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: "PLoS One.";
            date: "2019-09-26"; volume: "14(9):e0222778. doi"; pages: " 
            10.1371/journal.pone.0222778. eCollection 2019"
COMMENT     Sequence Label: pJL035
FEATURES             Location/Qualifiers
     source          1..3335
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(11..33)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     133..152
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(1109..1131)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1120..1136
                     /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        complement(join(1144..1150,1151..1168,1169..1174))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1189..1210
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(1327..1344)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1498..2086)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1578..1597)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2257..3048,3049..3117))
                     /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     2880..2899
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3118..3222)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     3290..3308
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
ORIGIN
        1 acggtgaaaa cctctgacac atgcagctcc cggagacggt cacagcttgt ctgtaagcgg
       61 atgccgggag cagacaagcc cgtcagggcg cgtcagcggg tgttggcggg tgtcggggct
      121 ggcttaacta tgcggcatca gagcagattg tactgagagt gcaccaattg ggtaccgagc
      181 tcgcggccgc aagcggtctc atcaggtatg gctggtgaga atgtgccaat cccttgtgtc
      241 aatggggtag atggagaacc ttgtcctgaa gattacaagt acatatcgga aaattgcgag
      301 acaagcacga tgaatatcga tcggaacatt actcatctcc aacactgtac atgtgttgat
      361 gactgctcat cttcaaactg tctttgcggt caattgtcaa tcagatgttg gtacgataaa
      421 gacggaagat tgcttcagga gtttaacaaa atcgaaccac ctctcatctt tgagtgtaat
      481 caagcctgca gttgttggag atcttgcaag aatcgagttg tccaaagtgg catcaaagtt
      541 cgtttacagc tctatcgtac tgctaaaatg ggatggggtg taagagcact tcagaccatt
      601 ccacaaggta ccttcatttg cgagtatgtg ggagagctaa tttccgacgc tgaagcagac
      661 gttagagagg atgatagcta cttgtttgac ctcgataaca aggatggcga agtctactgc
      721 atagatgctc gatattatgg gaacatttct aggttcataa accatctgtg tgatcccaat
      781 atcattccag tgagagtttt catgttacac caggatttac ggtttcctag gattgccttc
      841 ttttctagca gagatatacg tacaggagag gaattggggt ttgactatgg tgacagattc
      901 tgggatatta agtccaagta tttcacttgt cagtgtggat ctgagaaatg caagcatagt
      961 gctgaagcga ttgcgttgga acaatctagg ctagcaaggc ttgatccgca tcctgaactg
     1021 cttcccgatc tttcgtcact accaccgata aacacgactg gttagtaact gcagagacca
     1081 cctgcttttg ctcgcttgga tccgaattcc tgtgtgaaat tgttatccgc tcacaattcc
     1141 acacaacata cgagccggaa gcataaagtg taaagcctgg ggtgcctaat gagtgagcta
     1201 actcacatta attgcgttgc gctcactgcc cgctttccag tcgggaaacc tgtcgtgcca
     1261 gctgcattaa tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg ggcgctcttc
     1321 cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc
     1381 tcactcaaag gcggtaatac ggttatccac agaatcaggg gataacgcag gaaagaacat
     1441 gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt
     1501 ccataggctc cgcccccctg acgagcatca caaaaatcga cgctcaagtc agaggtggcg
     1561 aaacccgaca ggactataaa gataccaggc gtttccccct ggaagctccc tcgtgcgctc
     1621 tcctgttccg accctgccgc ttaccggata cctgtccgcc tttctccctt cgggaagcgt
     1681 ggcgctttct catagctcac gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa
     1741 gctgggctgt gtgcacgaac cccccgttca gcccgaccgc tgcgccttat ccggtaacta
     1801 tcgtcttgag tccaacccgg taagacacga cttatcgcca ctggcagcag ccactggtaa
     1861 caggattagc agagcgaggt atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa
     1921 ctacggctac actagaagaa cagtatttgg tatctgcgct ctgctgaagc cagttacctt
     1981 cggaaaaaga gttggtagct cttgatccgg caaacaaacc accgctggta gcggtggttt
     2041 ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag atcctttgat
     2101 cttttctacg gggtctgacg ctcagtggaa cgaaaactca cgttaaggga ttttggtcat
     2161 gagattatca aaaaggatct tcacctagat ccttttaaat taaaaatgaa gttttaaatc
     2221 aatctaaagt atatatgagt aaacttggtc tgacagttac caatgcttaa tcagtgaggc
     2281 acctatctca gcgatctgtc tatttcgttc atccatagtt gcctgactcc ccgtcgtgta
     2341 gataactacg atacgggagg gcttaccatc tggccccagt gctgcaatga taccgcgact
     2401 cccacgctca ccggctccag atttatcagc aataaaccag ccagccggaa gggccgagcg
     2461 cagaagtggt cctgcaactt tatccgcctc catccagtct attaattgtt gccgggaagc
     2521 tagagtaagt agttcgccag ttaatagttt gcgcaacgtt gttgccattg ctacaggcat
     2581 cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc tccggttccc aacgatcaag
     2641 gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat
     2701 cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg gttatggcag cactgcataa
     2761 ttctcttact gtcatgccat ccgtaagatg cttttctgtg actggtgagt actcaaccaa
     2821 gtcattctga gaatagtgta tgcggcgacc gagttgctct tgcccggcgt caatacggga
     2881 taataccgcg ccacatagca gaactttaaa agtgctcatc attggaaaac gttcttcggg
     2941 gcgaaaactc tcaaggatct taccgctgtt gagatccagt tcgatgtaac ccactcgtgc
     3001 acccaactga tcttcagcat cttttacttt caccagcgtt tctgggtgag caaaaacagg
     3061 aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa tactcatact
     3121 cttccttttt caatattatt gaagcattta tcagggttat tgtctcatga gcggatacat
     3181 atttgaatgt atttagaaaa ataaacaaat aggggttccg cgcacatttc cccgaaaagt
     3241 gccacctgac gtctaagaaa ccattattat catgacatta acctataaaa ataggcgtat
     3301 cacgaggccc tttcgtctcg cgcgtttcgg tgatg
//
