LOCUS       sequence-247655-        2703 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  MS2-modified guide RNA targeting the FT promoter with GreenGate D-E 
            flanking sequences.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 2703)
  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 2703)
  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: pMH183
FEATURES             Location/Qualifiers
     source          1..2703
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     270..292
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     284..301
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     285..301
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     312..331
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        312..330
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     misc_RNA        563..581
                     /label=MS2 stem loop
                     /note="stem loop that binds the bacteriophage MS2 coat 
                     protein"
     misc_RNA        633..651
                     /label=MS2 stem loop
                     /note="stem loop that binds the bacteriophage MS2 coat 
                     protein"
     primer_bind     complement(838..855)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(948..1536)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1028..1047)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(1707..2498,2499..2567))
                     /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     2330..2349
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(2568..2672)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 taagcgttaa tattttgtta aaattcgcgt taaatttttg ttaaatcagc tcatttttta
       61 accaataggc cgaaatcggc aaaatccctt ataaatcaaa agaatagacc gagatagggt
      121 tgagtggccg ctacagggcg ctcccattcg ccattcaggc tgcgcaactg ttgggaaggg
      181 cgtttcggtg cgggcctctt cgctattacg ccagctggcg aaagggggat gtgctgcaag
      241 gcgattaagt tgggtaacgc cagggttttc ccagtcacga cgttgtaaaa cgacggccag
      301 tgagcgcgac gtaatacgac tcactatagg gcgaattggc ggaaggccgt caaggccgca
      361 tgaagggcgc gccaagtgaa gcttggtctc atcaggtgga tccaaagaac aatagtattt
      421 cttatatagg cccatttaag ttgaaaacaa tcttcaaaag tcccacatcg cttagataag
      481 aaaacgaagc tgagtttata tacagctaga gtcgaagtag tgattggatt tgcattaact
      541 cgggtgtttt agagctaggc caacatgagg atcacccatg tctgcagggc ctagcaagtt
      601 aaaataaggc tagtccgtta tcaacttggc caacatgagg atcacccatg tctgcagggc
      661 caagtggcac cgagtcggtg cttttggtac cctgctgaga ccgaattctc attaattaac
      721 aacctgggcc tcatgggcct tccgctcact gcccgctttc cagtcgggaa acctgtcgtg
      781 ccagctgcat taacatggtc atagctgttt ccttgcgtat tgggcgctct ccgcttcctc
      841 gctcactgac tcgctgcgct cggtcgttcg ggtaaagcct ggggtgccta atgagcaaaa
      901 ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc
      961 cgcccccctg acgagcatca caaaaatcga cgctcaagtc agaggtggcg aaacccgaca
     1021 ggactataaa gataccaggc gtttccccct ggaagctccc tcgtgcgctc tcctgttccg
     1081 accctgccgc ttaccggata cctgtccgcc tttctccctt cgggaagcgt ggcgctttct
     1141 catagctcac gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt
     1201 gtgcacgaac cccccgttca gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag
     1261 tccaacccgg taagacacga cttatcgcca ctggcagcag ccactggtaa caggattagc
     1321 agagcgaggt atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa ctacggctac
     1381 actagaagaa cagtatttgg tatctgcgct ctgctgaagc cagttacctt cggaaaaaga
     1441 gttggtagct cttgatccgg caaacaaacc accgctggta gcggtggttt ttttgtttgc
     1501 aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag atcctttgat cttttctacg
     1561 gggtctgacg ctcagtggaa cgaaaactca cgttaaggga ttttggtcat gagattatca
     1621 aaaaggatct tcacctagat ccttttaaat taaaaatgaa gttttaaatc aatctaaagt
     1681 atatatgagt aaacttggtc tgacagttac caatgcttaa tcagtgaggc acctatctca
     1741 gcgatctgtc tatttcgttc atccatagtt gcctgactcc ccgtcgtgta gataactacg
     1801 atacgggagg gcttaccatc tggccccagt gctgcaatga taccgcgaga accacgctca
     1861 ccggctccag atttatcagc aataaaccag ccagccggaa gggccgagcg cagaagtggt
     1921 cctgcaactt tatccgcctc catccagtct attaattgtt gccgggaagc tagagtaagt
     1981 agttcgccag ttaatagttt gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca
     2041 cgctcgtcgt ttggtatggc ttcattcagc tccggttccc aacgatcaag gcgagttaca
     2101 tgatccccca tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat cgttgtcaga
     2161 agtaagttgg ccgcagtgtt atcactcatg gttatggcag cactgcataa ttctcttact
     2221 gtcatgccat ccgtaagatg cttttctgtg actggtgagt actcaaccaa gtcattctga
     2281 gaatagtgta tgcggcgacc gagttgctct tgcccggcgt caatacggga taataccgcg
     2341 ccacatagca gaactttaaa agtgctcatc attggaaaac gttcttcggg gcgaaaactc
     2401 tcaaggatct taccgctgtt gagatccagt tcgatgtaac ccactcgtgc acccaactga
     2461 tcttcagcat cttttacttt caccagcgtt tctgggtgag caaaaacagg aaggcaaaat
     2521 gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa tactcatact cttccttttt
     2581 caatattatt gaagcattta tcagggttat tgtctcatga gcggatacat atttgaatgt
     2641 atttagaaaa ataaacaaat aggggttccg cgcacatttc cccgaaaagt gccacctaaa
     2701 ttg
//
