LOCUS       sequence-239372-        2572 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  VP64 activator domain with GreenGate D-E flanking sequences.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 2572)
  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 2572)
  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: pMH185
FEATURES             Location/Qualifiers
     source          1..2572
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     175..197
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     189..206
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     190..206
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     217..236
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        217..235
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             309..458
                     /codon_start=1
                     /product="tetrameric repeat of the minimal activation 
                     domain of herpes simplex virus VP16 (Beerli et al., 1998)"
                     /label=VP64
                     /translation="DALDDFDLDMLGSDALDDFDLDMLGSDALDDFDLDMLGSDALDDF
                     DLDML"
     primer_bind     complement(612..629)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(722..1310)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(802..821)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(1481..2272,2273..2341))
                     /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     2104..2123
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(2342..2446)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 tcaaaagaat agaccgagat agggttgagt ggccgctaca gggcgctccc attcgccatt
       61 caggctgcgc aactgttggg aagggcgttt cggtgcgggc ctcttcgcta ttacgccagc
      121 tggcgaaagg gggatgtgct gcaaggcgat taagttgggt aacgccaggg ttttcccagt
      181 cacgacgttg taaaacgacg gccagtgagc gcgacgtaat acgactcact atagggcgaa
      241 ttggcggaag gccgtcaagg ccgcatcaac gagctcagtg aagcttggtc tcatcaggtg
      301 gaagagctga tgctttggat gattttgatt tggatatgtt gggatctgat gctttggatg
      361 attttgattt ggatatgttg ggatctgatg ctttggatga ttttgatttg gatatgttgg
      421 gatctgatgc tttggatgat tttgatttgg atatgttgat taattaatag ctgctgagac
      481 cgaattctcg gtaccctctg ggcctcatgg gccttccgct cactgcccgc tttccagtcg
      541 ggaaacctgt cgtgccagct gcattaacat ggtcatagct gtttccttgc gtattgggcg
      601 ctctccgctt cctcgctcac tgactcgctg cgctcggtcg ttcgggtaaa gcctggggtg
      661 cctaatgagc aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt
      721 ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt
      781 ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc
      841 gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa
      901 gcgtggcgct ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct
      961 ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta
     1021 actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg
     1081 gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc
     1141 ctaactacgg ctacactaga agaacagtat ttggtatctg cgctctgctg aagccagtta
     1201 ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg
     1261 gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt
     1321 tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg
     1381 tcatgagatt atcaaaaagg atcttcacct agatcctttt aaattaaaaa tgaagtttta
     1441 aatcaatcta aagtatatat gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg
     1501 aggcacctat ctcagcgatc tgtctatttc gttcatccat agttgcctga ctccccgtcg
     1561 tgtagataac tacgatacgg gagggcttac catctggccc cagtgctgca atgataccgc
     1621 gagaaccacg ctcaccggct ccagatttat cagcaataaa ccagccagcc ggaagggccg
     1681 agcgcagaag tggtcctgca actttatccg cctccatcca gtctattaat tgttgccggg
     1741 aagctagagt aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc attgctacag
     1801 gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt cagctccggt tcccaacgat
     1861 caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc
     1921 cgatcgttgt cagaagtaag ttggccgcag tgttatcact catggttatg gcagcactgc
     1981 ataattctct tactgtcatg ccatccgtaa gatgcttttc tgtgactggt gagtactcaa
     2041 ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac
     2101 gggataatac cgcgccacat agcagaactt taaaagtgct catcattgga aaacgttctt
     2161 cggggcgaaa actctcaagg atcttaccgc tgttgagatc cagttcgatg taacccactc
     2221 gtgcacccaa ctgatcttca gcatctttta ctttcaccag cgtttctggg tgagcaaaaa
     2281 caggaaggca aaatgccgca aaaaagggaa taagggcgac acggaaatgt tgaatactca
     2341 tactcttcct ttttcaatat tattgaagca tttatcaggg ttattgtctc atgagcggat
     2401 acatatttga atgtatttag aaaaataaac aaataggggt tccgcgcaca tttccccgaa
     2461 aagtgccacc taaattgtaa gcgttaatat tttgttaaaa ttcgcgttaa atttttgtta
     2521 aatcagctca ttttttaacc aataggccga aatcggcaaa atcccttata aa
//
