LOCUS       sequence-234542-        2800 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  MS2-NLS (no stop codon) with GreenGate C-D flanking sequences.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 2800)
  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 2800)
  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: pMH186
FEATURES             Location/Qualifiers
     source          1..2800
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     278..300
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     292..309
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     293..309
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     320..339
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        320..338
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             343..354
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     CDS             396..782
                     /codon_start=1
                     /product="bacteriophage MS2 coat protein"
                     /label=MS2-N55K
                     /note="binds with high affinity to a specific stem-loop 
                     structure in the viral RNA (Lim et al., 1994)"
                     /translation="ASNFTQFVLVDNGGTGDVTVAPSNFANGVAEWISSNSRSQAYKVT
                     CSVRQSSAQKRKYTIKVEVPKVATQTVGGVELPVAAWRSYLNMELTIPIFATNSDCELI
                     VKAMQGLLKDGNPIPSAIAANSGIY"
     CDS             837..857
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     primer_bind     complement(1006..1023)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1116..1704)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1196..1215)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(1878..2672)
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     promoter        complement(2673..2777)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 ctaaattgta agcgttaata ttttgttaaa attcgcgtta aatttttgtt aaatcagctc
       61 attttttaac caataggccg aaatcggcaa aatcccttat aaatcaaaag aatagaccga
      121 gatagggttg agtggccgct acagggcgct cccattcgcc attcaggctg cgcaactgtt
      181 gggaagggcg tttcggtgcg ggcctcttcg ctattacgcc agctggcgaa agggggatgt
      241 gctgcaaggc gattaagttg ggtaacgcca gggttttccc agtcacgacg ttgtaaaacg
      301 acggccagtg agcgcgacgt aatacgactc actatagggc gaattgaagg aaggccgtca
      361 aggccgcata gtgaagcttg gtctcaggct ccatggcttc taattttact caatttgttt
      421 tggttgataa tggaggaact ggagatgtta ctgttgctcc ttctaatttt gctaatggag
      481 ttgctgaatg gatttcttct aattcaagat ctcaggctta taaggttact tgttctgtta
      541 gacaatcttc tgctcaaaag agaaagtata ctattaaggt tgaagttcct aaggttgcta
      601 ctcaaactgt tggaggagtt gaattgcctg ttgctgcttg gagatcttat ttgaatatgg
      661 aattgactat tcctattttt gctactaatt ctgattgtga attgattgtt aaggctatgc
      721 aaggattgtt gaaggatgga aatcctattc cttctgctat tgctgctaat tctggaattt
      781 attctgctgg aggaggagga tctggaggag gaggatctgg aggaggagga tctggaccta
      841 agaagaagag aaaggttgct gctgctggat cttcagtgag accgaattct cctgggcctc
      901 atgggccttc ctttcactgc ccgctttcca gtcgggaaac ctgtcgtgcc agctgcatta
      961 acatggtcat agctgtttcc ttgcgtattg ggcgctctcc gcttcctcgc tcactgactc
     1021 gctgcgctcg gtcgttcggg taaagcctgg ggtgcctaat gagcaaaagg ccagcaaaag
     1081 gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg cccccctgac
     1141 gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg actataaaga
     1201 taccaggcgt ttccccctgg aagctccctc gtgcgctctc ctgttccgac cctgccgctt
     1261 accggatacc tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca tagctcacgc
     1321 tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc
     1381 cccgttcagc ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc caacccggta
     1441 agacacgact tatcgccact ggcagcagcc actggtaaca ggattagcag agcgaggtat
     1501 gtaggcggtg ctacagagtt cttgaagtgg tggcctaact acggctacac tagaagaaca
     1561 gtatttggta tctgcgctct gctgaagcca gttaccttcg gaaaaagagt tggtagctct
     1621 tgatccggca aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt
     1681 acgcgcagaa aaaaaggatc tcaagaagat cctttgatct tttctacggg gtctgacgct
     1741 cagtggaacg aaaactcacg ttaagggatt ttggtcatga gattatcaaa aaggatcttc
     1801 acctagatcc ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa
     1861 acttggtctg acagttatta gaaaaattca tccagcagac gataaaacgc aatacgctgg
     1921 ctatccggtg ccgcaatgcc atacagcacc agaaaacgat ccgcccattc gccgcccagt
     1981 tcttccgcaa tatcacgggt ggccagcgca atatcctgat aacgatccgc cacgcccaga
     2041 cggccgcaat caataaagcc gctaaaacgg ccattttcca ccataatgtt cggcaggcac
     2101 gcatcaccat gggtcaccac cagatcttcg ccatccggca tgctcgcttt cagacgcgca
     2161 aacagctctg ccggtgccag gccctgatgt tcttcatcca gatcatcctg atccaccagg
     2221 cccgcttcca tacgggtacg cgcacgttca atacgatgtt tcgcctgatg atcaaacgga
     2281 caggtcgccg ggtccagggt atgcagacga cgcatggcat ccgccataat gctcactttt
     2341 tctgccggcg ccagatggct agacagcaga tcctgacccg gcacttcgcc cagcagcagc
     2401 caatcacggc ccgcttcggt caccacatcc agcaccgccg cacacggaac accggtggtg
     2461 gccagccagc tcagacgcgc cgcttcatcc tgcagctcgt tcagcgcacc gctcagatcg
     2521 gttttcacaa acagcaccgg acgaccctgc gcgctcagac gaaacaccgc cgcatcagag
     2581 cagccaatgg tctgctgcgc ccaatcatag ccaaacagac gttccaccca cgctgccggg
     2641 ctacccgcat gcaggccatc ctgttcaatc atactcttcc tttttcaata ttattgaagc
     2701 atttatcagg gttattgtct catgagcgga tacatatttg aatgtattta gaaaaataaa
     2761 caaatagggg ttccgcgcac atttccccga aaagtgccac
//
