LOCUS       sequence-239400-        2703 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  MS2-modified guide RNA targeting the FT promoter with GreenGate E-F 
            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: pMH184
FEATURES             Location/Qualifiers
     source          1..2703
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     105..127
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     119..136
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     120..136
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     147..166
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        147..165
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     misc_RNA        398..416
                     /label=MS2 stem loop
                     /note="stem loop that binds the bacteriophage MS2 coat 
                     protein"
     misc_RNA        468..486
                     /label=MS2 stem loop
                     /note="stem loop that binds the bacteriophage MS2 coat 
                     protein"
     primer_bind     complement(673..690)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(783..1371)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(863..882)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(1542..2333,2334..2402))
                     /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     2165..2184
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(2403..2507)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 aactgttggg aagggcgttt cggtgcgggc ctcttcgcta ttacgccagc tggcgaaagg
       61 gggatgtgct gcaaggcgat taagttgggt aacgccaggg ttttcccagt cacgacgttg
      121 taaaacgacg gccagtgagc gcgacgtaat acgactcact atagggcgaa ttggcggaag
      181 gccgtcaagg ccgcatgaag ggcgcgccaa gtgaagcttg gtctcactgc aaggatccaa
      241 agaacaatag tatttcttat ataggcccat ttaagttgaa aacaatcttc aaaagtccca
      301 catcgcttag ataagaaaac gaagctgagt ttatatacag ctagagtcga agtagtgatt
      361 gtccagtgta ttagtgtggt gttttagagc taggccaaca tgaggatcac ccatgtctgc
      421 agggcctagc aagttaaaat aaggctagtc cgttatcaac ttggccaaca tgaggatcac
      481 ccatgtctgc agggccaagt ggcaccgagt cggtgctttt ggtaccacta tgagaccgaa
      541 ttctcattaa ttaacaacct gggcctcatg ggccttccgc tcactgcccg ctttccagtc
      601 gggaaacctg tcgtgccagc tgcattaaca tggtcatagc tgtttccttg cgtattgggc
      661 gctctccgct tcctcgctca ctgactcgct gcgctcggtc gttcgggtaa agcctggggt
      721 gcctaatgag caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg
      781 tttttccata ggctccgccc ccctgacgag catcacaaaa atcgacgctc aagtcagagg
      841 tggcgaaacc cgacaggact ataaagatac caggcgtttc cccctggaag ctccctcgtg
      901 cgctctcctg ttccgaccct gccgcttacc ggatacctgt ccgcctttct cccttcggga
      961 agcgtggcgc tttctcatag ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc
     1021 tccaagctgg gctgtgtgca cgaacccccc gttcagcccg accgctgcgc cttatccggt
     1081 aactatcgtc ttgagtccaa cccggtaaga cacgacttat cgccactggc agcagccact
     1141 ggtaacagga ttagcagagc gaggtatgta ggcggtgcta cagagttctt gaagtggtgg
     1201 cctaactacg gctacactag aagaacagta tttggtatct gcgctctgct gaagccagtt
     1261 accttcggaa aaagagttgg tagctcttga tccggcaaac aaaccaccgc tggtagcggt
     1321 ggtttttttg tttgcaagca gcagattacg cgcagaaaaa aaggatctca agaagatcct
     1381 ttgatctttt ctacggggtc tgacgctcag tggaacgaaa actcacgtta agggattttg
     1441 gtcatgagat tatcaaaaag gatcttcacc tagatccttt taaattaaaa atgaagtttt
     1501 aaatcaatct aaagtatata tgagtaaact tggtctgaca gttaccaatg cttaatcagt
     1561 gaggcaccta tctcagcgat ctgtctattt cgttcatcca tagttgcctg actccccgtc
     1621 gtgtagataa ctacgatacg ggagggctta ccatctggcc ccagtgctgc aatgataccg
     1681 cgagaaccac gctcaccggc tccagattta tcagcaataa accagccagc cggaagggcc
     1741 gagcgcagaa gtggtcctgc aactttatcc gcctccatcc agtctattaa ttgttgccgg
     1801 gaagctagag taagtagttc gccagttaat agtttgcgca acgttgttgc cattgctaca
     1861 ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat tcagctccgg ttcccaacga
     1921 tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag cggttagctc cttcggtcct
     1981 ccgatcgttg tcagaagtaa gttggccgca gtgttatcac tcatggttat ggcagcactg
     2041 cataattctc ttactgtcat gccatccgta agatgctttt ctgtgactgg tgagtactca
     2101 accaagtcat tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata
     2161 cgggataata ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg aaaacgttct
     2221 tcggggcgaa aactctcaag gatcttaccg ctgttgagat ccagttcgat gtaacccact
     2281 cgtgcaccca actgatcttc agcatctttt actttcacca gcgtttctgg gtgagcaaaa
     2341 acaggaaggc aaaatgccgc aaaaaaggga ataagggcga cacggaaatg ttgaatactc
     2401 atactcttcc tttttcaata ttattgaagc atttatcagg gttattgtct catgagcgga
     2461 tacatatttg aatgtattta gaaaaataaa caaatagggg ttccgcgcac atttccccga
     2521 aaagtgccac ctaaattgta agcgttaata ttttgttaaa attcgcgtta aatttttgtt
     2581 aaatcagctc attttttaac caataggccg aaatcggcaa aatcccttat aaatcaaaag
     2641 aatagaccga gatagggttg agtggccgct acagggcgct cccattcgcc attcaggctg
     2701 cgc
//
