LOCUS       sequence-250396-        3705 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of plant nuclear envelope LINC protein fused to 
            fluorescent protein and affinity tags for cell biology.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3705)
  AUTHORS   Gumber HK, McKenna JF, Tolmie AF, Jalovec AM, Kartick AC, Graumann 
            K, Bass HW
  TITLE     MLKS2 is an ARM domain and F-actin-associated KASH protein that 
            functions in stomatal complex development and meiotic chromosome 
            segregation.
  JOURNAL   Nucleus. 2019 Dec;10(1):144-166. doi: 10.1080/19491034.2019.1629795.
  PUBMED    31221013
REFERENCE   2  (bases 1 to 3705)
  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"; doi: 
            "10.1080/19491034.2019"; journalName: "Nucleus"; date: "2019-12"; 
            volume: "10"; issue: "1"; pages: "144-166"
COMMENT     Sequence Label: MLKS2delARMec
FEATURES             Location/Qualifiers
     source          1..3705
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     92..109
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(268..295)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(387..473)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     522..544
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     536..553
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     537..553
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    569..668
                     /gene="mutant version of attL"
                     /label=attL1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     CDS             join(671..673,674..676,677..1387)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             1397..1420
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             1424..1450
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     protein_bind    complement(1830..1929)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(1946..1965)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1947..1965)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1970..1986)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1970..1986)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2088..2107)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             2099..2908
                     /codon_start=1
                     /gene="aph(3')-Ia"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYGKP
                     DAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGKTA
                     FQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDASD
                     FDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGIAD
                     RYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     complement(2166..2185)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      3055..3643
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3544..3563
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 ctttcctgcg ttatcccctg attctgtgga taaccgtatt accgcctttg agtgagctga
       61 taccgctcgc cgcagccgaa cgaccgagcg cagcgagtca gtgagcgagg aagcggaaga
      121 gcgcccaata cgcaaaccgc ctctccccgc gcgttggccg attcattaat gcagctggca
      181 cgacaggttt cccgactgga aagcgggcag tgagcgcaac gcaattaata cgcgtaccgc
      241 tagccaggaa gagtttgtag aaacgcaaaa aggccatccg tcaggatggc cttctgctta
      301 gtttgatgcc tggcagttta tggcgggcgt cctgcccgcc accctccggg ccgttgcttc
      361 acaacgttca aatccgctcc cggcggattt gtcctactca ggagagcgtt caccgacaaa
      421 caacagataa aacgaaaggc ccagtcttcc gactgagcct ttcgttttat ttgatgcctg
      481 gcagttccct actctcgcgt taacgctagc atggatgttt tcccagtcac gacgttgtaa
      541 aacgacggcc agtcttaagc tcgggcccca aataatgatt ttattttgac tgatagtgac
      601 ctgttcgttg caacaaattg atgagcaatg cttttttata atgccaactt tgtacaaaaa
      661 agcaggcttc atggtctcca agggcgagga gctgttcacc ggggtggtcc ccatcctggt
      721 ggagctggat ggcgacgtta acgggcacaa gttctcggtg tctggcgagg gggagggcga
      781 cgctacctac gggaagctca cgctgaagtt catctgcacg accggcaagc tgccggtgcc
      841 ctggccaaca ctcgtcacaa ctctgactta cggcgtccag tgcttctcgc gctaccctga
      901 tcacatgaag cagcatgact tcttcaagtc ggctatgccc gagggctacg tgcaggagcg
      961 gacgatcttc ttcaaggacg atggcaacta caagacacgc gcggaggtca agttcgaggg
     1021 ggacactctc gttaacagga tcgagctgaa gggcattgac ttcaaggagg atggcaatat
     1081 cctcgggcac aagctggagt acaactacaa ttcccataac gtgtacatca tggccgacaa
     1141 gcagaagaac ggcattaagg tcaatttcaa gatccggcac aacattgagg atggcagcgt
     1201 tcagctcgcg gaccattacc agcagaacac accgatcggg gatggccctg tcctcctgcc
     1261 cgacaatcac tacctctcca ctcagagcgc cctgtcgaag gacccgaacg agaagaggga
     1321 ccatatggtg ctcctggagt tcgtcacagc cgcgggcatc actctcggga tggatgagct
     1381 gtacaagggg gcgggcgact acaaggacga tgacgataag gcttacccct acgacgtgcc
     1441 ggattacgcc ggggctggca tggggaaggg cctcgtcatt tcttcagagc agatgcagtc
     1501 gctgcattct gacaataagg gcgatgagat gaaggacgag aacgagacca tctacatgca
     1561 gaatagcaag ggcaggacac agactctctc gaacgctgac gcctcagtcc tgtccacaac
     1621 tgaggccgag aacacaagca tcaagacttc gcctgagatt gagctgaatg gggacgatgt
     1681 ttgcatcacc tccagcattg gcaagacgcg gaagtaccgc gcgaagttca ccctcttcct
     1741 gtccgtcatc gttattttcc tcgctatcat tacgatgctg atccgcattg agaacgacga
     1801 tgacagcgcg tacctggtgc cgacctgaga cccagctttc ttgtacaaag ttggcattat
     1861 aagaaagcat tgcttatcaa tttgttgcaa cgaacaggtc actatcagtc aaaataaaat
     1921 cattatttgc catccagctg atatccccta tagtgagtcg tattacatgg tcatagctgt
     1981 ttcctggcag ctctggcccg tgtctcaaaa tctctgatgt tacattgcac aagataaaaa
     2041 tatatcatca tgaacaataa aactgtctgc ttacataaac agtaatacaa ggggtgttat
     2101 gagccatatt caacgggaaa cgtcgaggcc gcgattaaat tccaacatgg atgctgattt
     2161 atatgggtat aaatgggctc gcgataatgt cgggcaatca ggtgcgacaa tctatcgctt
     2221 gtatgggaag cccgatgcgc cagagttgtt tctgaaacat ggcaaaggta gcgttgccaa
     2281 tgatgttaca gatgagatgg tcagactaaa ctggctgacg gaatttatgc ctcttccgac
     2341 catcaagcat tttatccgta ctcctgatga tgcatggtta ctcaccactg cgatccccgg
     2401 aaaaacagca ttccaggtat tagaagaata tcctgattca ggtgaaaata ttgttgatgc
     2461 gctggcagtg ttcctgcgcc ggttgcattc gattcctgtt tgtaattgtc cttttaacag
     2521 cgatcgcgta tttcgtctcg ctcaggcgca atcacgaatg aataacggtt tggttgatgc
     2581 gagtgatttt gatgacgagc gtaatggctg gcctgttgaa caagtctgga aagaaatgca
     2641 taaacttttg ccattctcac cggattcagt cgtcactcat ggtgatttct cacttgataa
     2701 ccttattttt gacgagggga aattaatagg ttgtattgat gttggacgag tcggaatcgc
     2761 agaccgatac caggatcttg ccatcctatg gaactgcctc ggtgagtttt ctccttcatt
     2821 acagaaacgg ctttttcaaa aatatggtat tgataatcct gatatgaata aattgcagtt
     2881 tcatttgatg ctcgatgagt ttttctaatc agaattggtt aattggttgt aacactggca
     2941 gagcattacg ctgacttgac gggacggcgc aagctcatga ccaaaatccc ttaacgtgag
     3001 ttacgcgtcg ttccactgag cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga
     3061 tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt
     3121 ggtttgtttg ccggatcaag agctaccaac tctttttccg aaggtaactg gcttcagcag
     3181 agcgcagata ccaaatactg ttcttctagt gtagccgtag ttaggccacc acttcaagaa
     3241 ctctgtagca ccgcctacat acctcgctct gctaatcctg ttaccagtgg ctgctgccag
     3301 tggcgataag tcgtgtctta ccgggttgga ctcaagacga tagttaccgg ataaggcgca
     3361 gcggtcgggc tgaacggggg gttcgtgcac acagcccagc ttggagcgaa cgacctacac
     3421 cgaactgaga tacctacagc gtgagctatg agaaagcgcc acgcttcccg aagggagaaa
     3481 ggcggacagg tatccggtaa gcggcagggt cggaacagga gagcgcacga gggagcttcc
     3541 agggggaaac gcctggtatc tttatagtcc tgtcgggttt cgccacctct gacttgagcg
     3601 tcgatttttg tgatgctcgt caggggggcg gagcctatgg aaaaacgcca gcaacgcggc
     3661 ctttttacgg ttcctggcct tttgctggcc ttttgctcac atgtt
//
