LOCUS       sequence-239373-        2733 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  SRDX repressor domain with GreenGate D-E flanking sequences.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 2733)
  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 2733)
  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: pDG058
FEATURES             Location/Qualifiers
     source          1..2733
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     209..228
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     462..479
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    596..617
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(632..637,638..655,656..662)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    670..686
                     /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        700..718
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     700..717
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(807..826)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(808..826)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(836..858)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     complement(1052..1071)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     1171..1193
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(1231..1249)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        1317..1421
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1422..1490,1491..2282)
                     /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(1640..1659)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      join(2453..2733,1..308)
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
ORIGIN
        1 gacgatagtt accggataag gcgcagcggt cgggctgaac ggggggttcg tgcacacagc
       61 ccagcttgga gcgaacgacc tacaccgaac tgagatacct acagcgtgag ctatgagaaa
      121 gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa
      181 caggagagcg cacgagggag cttccagggg gaaacgcctg gtatctttat agtcctgtcg
      241 ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc
      301 tatggaaaaa cgccagcaac gcggcctttt tacggttcct ggccttttgc tggccttttg
      361 ctcacatgtt ctttcctgcg ttatcccctg attctgtgga taaccgtatt accgcctttg
      421 agtgagctga taccgctcgc cgcagccgaa cgaccgagcg cagcgagtca gtgagcgagg
      481 aagcggaaga gcgcccaata cgcaaaccgc ctctccccgc gcgttggccg attcattaat
      541 gcagctggca cgacaggttt cccgactgga aagcgggcag tgagcgcaac gcaattaatg
      601 tgagttagct cactcattag gcaccccagg ctttacactt tatgcttccg gctcgtatgt
      661 tgtgtggaat tgtgagcgga taacaatttc acacgagcta tttaggtgac actatagaag
      721 tgaagcttgg tctcatcagg tatgcttgat ctcgacttgg agcttagact cggattcgct
      781 taatagctgc tgagaccgaa ttctcgccct atagtgagtc gtattacctg gccgtcgttt
      841 tacaacgtcg tgactgggaa aaccctggcg ttacccaact taatcgcctt gcagcacatc
      901 cccctttcgc cagctggcgt aatagcgaag aggcccgcac cgatcgccct tcccaacagt
      961 tgcgcagcct gaatggcgaa tggcgcctga tgcggtattt tctccttacg catctgtgcg
     1021 gtatttcaca ccgcatatgg tgcactctca gtacaatctg ctctgatgcc gcatagttaa
     1081 gccagccccg acacccgcca acacccgctg acgcgccctg acgggcttgt ctgctcccgg
     1141 catccgctta cagacaagct gtgaccgtct ccgggagctg catgtgtcag aggttttcac
     1201 cgtcatcacc gaaacgcgcg agacgaaagg gcctcgtgat acgcctattt ttataggtta
     1261 atgtcatgat aataatggtt tcttagacgt caggtggcac ttttcgggga aatgtgcgcg
     1321 gaacccctat ttgtttattt ttctaaatac attcaaatat gtatccgctc atgagacaat
     1381 aaccctgata aatgcttcaa taatattgaa aaaggaagag tatgagtatt caacatttcc
     1441 gtgtcgccct tattcccttt tttgcggcat tttgccttcc tgtttttgct cacccagaaa
     1501 cgctggtgaa agtaaaagat gctgaagatc agttgggtgc acgagtgggt tacatcgaac
     1561 tggatctcaa cagcggtaag atccttgaga gttttcgccc cgaagaacgt tttccaatga
     1621 tgagcacttt taaagttctg ctatgtggcg cggtattatc ccgtattgac gccgggcaag
     1681 agcaactcgg tcgccgcata cactattctc agaatgactt ggttgagtac tcaccagtca
     1741 cagaaaagca tcttacggat ggcatgacag taagagaatt atgcagtgct gccataacca
     1801 tgagtgataa cactgcggcc aacttacttc tgacaacgat cggaggaccg aaggagctaa
     1861 ccgctttttt gcacaacatg ggggatcatg taactcgcct tgatcgttgg gaaccggagc
     1921 tgaatgaagc cataccaaac gacgagcgtg acaccacgat gcctgtagca atggcaacaa
     1981 cgttgcgcaa actattaact ggcgaactac ttactctagc ttcccggcaa caattaatag
     2041 actggatgga ggcggataaa gttgcaggac cacttctgcg ctcggccctt ccggctggct
     2101 ggtttattgc tgataaatct ggagccggtg agcgtggctc tcgaggtatc attgcagcac
     2161 tggggccaga tggtaagccc tcccgtatcg tagttatcta cacgacgggg agtcaggcaa
     2221 ctatggatga acgaaataga cagatcgctg agataggtgc ctcactgatt aagcattggt
     2281 aactgtcaga ccaagtttac tcatatatac tttagattga tttaaaactt catttttaat
     2341 ttaaaaggat ctaggtgaag atcctttttg ataatctcat gaccaaaatc ccttaacgtg
     2401 agttttcgtt ccactgagcg tcagaccccg tagaaaagat caaaggatct tcttgagatc
     2461 ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg
     2521 tttgtttgcc ggatcaagag ctaccaactc tttttccgaa ggtaactggc ttcagcagag
     2581 cgcagatacc aaatactgtt cttctagtgt agccgtagtt aggccaccac ttcaagaact
     2641 ctgtagcacc gcctacatac ctcgctctgc taatcctgtt accagtggct gctgccagtg
     2701 gcgataagtc gtgtcttacc gggttggact caa
//
