LOCUS       sequence-255656-        5217 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 5217)
  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 5217)
  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: MLKS2delKASHec
FEATURES             Location/Qualifiers
     source          1..5217
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     1..17
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    33..132
                     /gene="mutant version of attL"
                     /label=attL1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     CDS             join(135..137,138..140,141..851)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             861..884
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             888..914
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     protein_bind    complement(2806..2905)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(2922..2941)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2923..2941)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2946..2962)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2946..2962)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3064..3083)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             3075..3884
                     /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(3142..3161)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      4031..4619
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4520..4539
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4773..4790
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(4949..4976)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(5068..5154)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     join(5203..5217,1..8)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     join(5217,1..17)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
ORIGIN
        1 gtaaaacgac ggccagtctt aagctcgggc cccaaataat gattttattt tgactgatag
       61 tgacctgttc gttgcaacaa attgatgagc aatgcttttt tataatgcca actttgtaca
      121 aaaaagcagg cttcatggtc tccaaggggg aggagctgtt caccggggtg gtccccatcc
      181 tggtggagct ggatggcgac gttaacgggc acaagttctc ggtgtctggc gagggggagg
      241 gcgacgctac ctacgggaag ctcacgctga agttcatctg cacgaccggc aagctgccgg
      301 tgccctggcc aacactcgtc acaactctga cttacggcgt ccagtgcttc tcgcgctacc
      361 ctgatcacat gaagcagcat gacttcttca agtcggctat gcccgagggc tacgtgcagg
      421 agcggacgat cttcttcaag gacgatggca actacaagac acgcgcggag gtcaagttcg
      481 agggggacac tctcgttaac aggatcgagc tgaagggcat tgacttcaag gaggatggca
      541 atatcctcgg gcacaagctg gagtacaact acaattccca taacgtgtac atcatggccg
      601 acaagcagaa gaacggcatt aaggtcaatt tcaagatccg gcacaacatt gaggatggca
      661 gcgttcagct cgcggaccat taccagcaga acacaccgat cggggatggc cctgtcctcc
      721 tgcccgacaa tcactacctc tccactcaga gcgccctgtc gaaggacccg aacgagaaga
      781 gggaccatat ggtgctcctg gagttcgtca cagccgcggg catcactctc gggatggatg
      841 agctgtacaa gggggcgggc gactacaagg acgatgacga taaggcttac ccctacgacg
      901 tgccggatta cgccggggct ggcatggggc gctcactctc cccgctcctg aggcaggagc
      961 tggacaacct ggataaggac gccgattctc gcagggctgc catgaaggct ctcaagtcat
     1021 acgcccgcca cctggatagc aagtcgatcc cccatttcct ggccgaggtc tctgacacga
     1081 ccgccgctgg cgcgggggcc ggcagcggga tgccaaccgg cggggagttc acgatctcgc
     1141 tctacgaggt cctggccagg gttcacgggc ggaacatcgt gccccagatc ggcaatatta
     1201 tggcgaccat tatgtgcacg ctctccagct cgggcgggtc attcccactg caccaggcct
     1261 gctccaaggt tgtgccagct atcgctaggc atggcattga tccgtcggcc ccagacggcg
     1321 agaaggccgg gatcattgct agcctctgcc gcccactgtg ctcggctctc atgggcggga
     1381 gccaggacgg cggcgctgcc agcggcgctg ccctctgcct gaaggccctg gttgagtctt
     1441 caaactggag gttcgcgtcg ggcgagctgg tgaatgaggt ctgcctgaag gtggccggcg
     1501 ctatgcacga ctcggccaca cggtcgaacg ctcatatggg gctggccatg gcgctcgtga
     1561 agcacaatgg cctcatcgct gaggcctacg cgcgctctat cgtcaggtcc ggcctccaga
     1621 ttctggacgg ggataccgct gactccagct cgcagaagag gctgtccgcc atccagatga
     1681 ttaacttctt catgaagttc gtggacccga ggtgcctgtc ttcagagctg ggccgggtta
     1741 tcgatgtgat ggagcgctgc cagaatgacc ggatgccctt cgtgcggggc gcggctttcg
     1801 aggcctctca gtcagcgcgc aacattgccg cgcagaaggg ctccaggcac gaggttggga
     1861 cctctcctat ggtgggctca aacttccata agaggaggga gaagtcccct tgccggagcc
     1921 tctggggcgc caagaggtcc cccgctgcct ctacgatggc cgctgctagc cctgtgcagt
     1981 tccgctctcc ggagtcacag gtcgttgatt ccagcatcat gaatggctcc acactccacg
     2041 actccccagt gagcgtcggc cagtcgtctt gcaacttcga ccagagcagg cggactaacc
     2101 gcaggctgtg gtcgaatgac gcggttgatg tgtctctgaa ggacggcctc ttcatccagc
     2161 tgtgctccaa cagcaattcg tacgaggacg atctgggcga ggtttgcgat agcgaggtga
     2221 ccgacgctaa cttcgagtgc accaatacgt tcgccgggtt cgtctctccc tcacacaatg
     2281 gctccatcag cagggataag acgccttcac cgcgggcctc ggacaggccc atcaacattg
     2341 acgatgtgaa gatctactcc acgccaagga agctcctgcg gctcctgcag tcatcctacg
     2401 actcggattc tgcctcaaac atcgggcaga gcacggccaa gctcaggggc ctctcctccc
     2461 ccgaccagga gcacaagggc ctcgtcattt cttcagagca gatgcagtcg ctgcattctg
     2521 acaataaggg cgatgagatg aaggacgaga acgagaccat ctacatgcag aatagcaagg
     2581 gcaggacaca gactctctcg aacgctgacg cctcagtcct gtccacaact gaggccgaga
     2641 acacaagcat caagacttcg cctgagattg agctgaatgg ggacgatgtt tgcatcacct
     2701 ccagcattgg caagacgcgg aagtaccgcg cgaagttcac cctcttcctg tccgtcatcg
     2761 ttattttcct cgctatcatt acgatgctga tcgctgccgc atgagaccca gctttcttgt
     2821 acaaagttgg cattataaga aagcattgct tatcaatttg ttgcaacgaa caggtcacta
     2881 tcagtcaaaa taaaatcatt atttgccatc cagctgatat cccctatagt gagtcgtatt
     2941 acatggtcat agctgtttcc tggcagctct ggcccgtgtc tcaaaatctc tgatgttaca
     3001 ttgcacaaga taaaaatata tcatcatgaa caataaaact gtctgcttac ataaacagta
     3061 atacaagggg tgttatgagc catattcaac gggaaacgtc gaggccgcga ttaaattcca
     3121 acatggatgc tgatttatat gggtataaat gggctcgcga taatgtcggg caatcaggtg
     3181 cgacaatcta tcgcttgtat gggaagcccg atgcgccaga gttgtttctg aaacatggca
     3241 aaggtagcgt tgccaatgat gttacagatg agatggtcag actaaactgg ctgacggaat
     3301 ttatgcctct tccgaccatc aagcatttta tccgtactcc tgatgatgca tggttactca
     3361 ccactgcgat ccccggaaaa acagcattcc aggtattaga agaatatcct gattcaggtg
     3421 aaaatattgt tgatgcgctg gcagtgttcc tgcgccggtt gcattcgatt cctgtttgta
     3481 attgtccttt taacagcgat cgcgtatttc gtctcgctca ggcgcaatca cgaatgaata
     3541 acggtttggt tgatgcgagt gattttgatg acgagcgtaa tggctggcct gttgaacaag
     3601 tctggaaaga aatgcataaa cttttgccat tctcaccgga ttcagtcgtc actcatggtg
     3661 atttctcact tgataacctt atttttgacg aggggaaatt aataggttgt attgatgttg
     3721 gacgagtcgg aatcgcagac cgataccagg atcttgccat cctatggaac tgcctcggtg
     3781 agttttctcc ttcattacag aaacggcttt ttcaaaaata tggtattgat aatcctgata
     3841 tgaataaatt gcagtttcat ttgatgctcg atgagttttt ctaatcagaa ttggttaatt
     3901 ggttgtaaca ctggcagagc attacgctga cttgacggga cggcgcaagc tcatgaccaa
     3961 aatcccttaa cgtgagttac gcgtcgttcc actgagcgtc agaccccgta gaaaagatca
     4021 aaggatcttc ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac
     4081 caccgctacc agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg
     4141 taactggctt cagcagagcg cagataccaa atactgttct tctagtgtag ccgtagttag
     4201 gccaccactt caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac
     4261 cagtggctgc tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt
     4321 taccggataa ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg
     4381 agcgaacgac ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc
     4441 ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc
     4501 gcacgaggga gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc
     4561 acctctgact tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa
     4621 acgccagcaa cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgt
     4681 tctttcctgc gttatcccct gattctgtgg ataaccgtat taccgccttt gagtgagctg
     4741 ataccgctcg ccgcagccga acgaccgagc gcagcgagtc agtgagcgag gaagcggaag
     4801 agcgcccaat acgcaaaccg cctctccccg cgcgttggcc gattcattaa tgcagctggc
     4861 acgacaggtt tcccgactgg aaagcgggca gtgagcgcaa cgcaattaat acgcgtaccg
     4921 ctagccagga agagtttgta gaaacgcaaa aaggccatcc gtcaggatgg ccttctgctt
     4981 agtttgatgc ctggcagttt atggcgggcg tcctgcccgc caccctccgg gccgttgctt
     5041 cacaacgttc aaatccgctc ccggcggatt tgtcctactc aggagagcgt tcaccgacaa
     5101 acaacagata aaacgaaagg cccagtcttc cgactgagcc tttcgtttta tttgatgcct
     5161 ggcagttccc tactctcgcg ttaacgctag catggatgtt ttcccagtca cgacgtt
//
