LOCUS       sequence-255539-        4093 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of plant LINC fluorescent fusion protein for cell 
            biology.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4093)
  AUTHORS   Gumber HK, McKenna JF, Estrada AL, Tolmie AF, Graumann K, Bass HW
  TITLE     Identification and characterization of genes encoding the nuclear 
            envelope LINC complex in the monocot species Zea mays.
  JOURNAL   J Cell Sci. 2019 Feb 11;132(3). pii: jcs.221390. doi: 
            10.1242/jcs.221390.
  PUBMED    30659121
REFERENCE   2  (bases 1 to 4093)
  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: "J Cell 
            Sci."; date: "2019-02-11"; volume: "132(3). pii: jcs.221390. doi"; 
            pages: " 10.1242/jcs.221390"
COMMENT     Sequence Label: MLKG1delKASHec
FEATURES             Location/Qualifiers
     source          1..4093
                     /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(1682..1781)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(1798..1817)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1799..1817)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1822..1838)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1822..1838)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1940..1959)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             1951..2760
                     /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(2018..2037)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      2907..3495
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3396..3415
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3649..3666
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(3825..3852)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(3944..4030)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     join(4079..4093,1..8)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     join(4093,1..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 caccggcgtg gtcccaatcc
      181 tcgtcgagct ggacggggat gttaacggcc acaagttctc cgtgagcggc gagggggagg
      241 gcgacgccac ctacgggaag ctcacgctga agttcatttg cacgaccggc aagctgccgg
      301 tgccctggcc aaccctggtc acaactctga cgtacggcgt ccagtgcttc tcccgctacc
      361 cggaccacat gaagcagcat gatttcttca agagcgccat gccggagggc tacgttcagg
      421 agcgcaccat cttcttcaag gacgatggca actacaagac aagggcggag gttaagttcg
      481 agggcgacac tctcgtgaac cggatcgagc tgaagggcat tgacttcaag gaggatggga
      541 atattctcgg ccacaagctg gagtacaact acaattcgca taacgtctac atcatggctg
      601 acaagcagaa gaacggcatt aaggttaatt tcaagatccg ccacaatatt gaggacggct
      661 ccgtgcagct cgcggatcat taccagcaga acacgccgat cggggacggc cctgtcctcc
      721 tgccggataa tcactacctc tcgacgcagt ctgcgctgtc aaaggacccg aacgagaaga
      781 gggatcatat ggttctcctg gagttcgtga cagccgcggg gatcactctc ggcatggacg
      841 agctgtacaa gggggcgggc gactacaagg acgatgacga taaggcttac ccctacgacg
      901 tgccagatta cgctggggcc ggcggatcca tggacatgcc gccggcggcg cagggacgga
      961 gggcgatcgc cgtggtggac cggaagcttg acgagctctg cgcctgcctc gacgacgcga
     1021 tgtcgtccag gcagcggagc ccgtacgcgg aggagcggct ccgcgcggag atcaagtcga
     1081 gggccgactt ccttcggtcg ctcatggccg ccgaggtgga ctgccacggc gggacgcgcc
     1141 cggagcacct ggctgaggcc gaggcgcgct tcgccgtcct agaggccacg ttcgatcggt
     1201 gggcgcggcg cgcagtcgct gcacctgaac cagtcgtcga ggagaaggag gaggaggccg
     1261 actccttggg gtcctcccgg tccacgtgct cctgcaccga gtcgtgccag gaggcgacgg
     1321 agggggacgc catgcgcgac gccgtggaaa cacgcggcaa ggaggggacg tcggatggca
     1381 tcgccgagga caacgacgcc gaatgcgaga cggtggtaat ccgcagggag gaggcggcgg
     1441 cggttcctgt tgccaagaag cgggatgagg ctatgggaac acgcgaggag gtggcgctgg
     1501 acgccattgc caagaagaag ggcgataccg atcgcgatgc cgcggcggag acggggagga
     1561 ggaccgtaga ccagcggcgc tggtggaggc ggagtgctgc ctggtgcggt gcagccggcg
     1621 tcgtcgcgtt ggtcgcggtc gggttggcgg tggagttggt cgcggtgtag cctgcaggca
     1681 gacccagctt tcttgtacaa agttggcatt ataagaaagc attgcttatc aatttgttgc
     1741 aacgaacagg tcactatcag tcaaaataaa atcattattt gccatccagc tgatatcccc
     1801 tatagtgagt cgtattacat ggtcatagct gtttcctggc agctctggcc cgtgtctcaa
     1861 aatctctgat gttacattgc acaagataaa aatatatcat catgaacaat aaaactgtct
     1921 gcttacataa acagtaatac aaggggtgtt atgagccata ttcaacggga aacgtcgagg
     1981 ccgcgattaa attccaacat ggatgctgat ttatatgggt ataaatgggc tcgcgataat
     2041 gtcgggcaat caggtgcgac aatctatcgc ttgtatggga agcccgatgc gccagagttg
     2101 tttctgaaac atggcaaagg tagcgttgcc aatgatgtta cagatgagat ggtcagacta
     2161 aactggctga cggaatttat gcctcttccg accatcaagc attttatccg tactcctgat
     2221 gatgcatggt tactcaccac tgcgatcccc ggaaaaacag cattccaggt attagaagaa
     2281 tatcctgatt caggtgaaaa tattgttgat gcgctggcag tgttcctgcg ccggttgcat
     2341 tcgattcctg tttgtaattg tccttttaac agcgatcgcg tatttcgtct cgctcaggcg
     2401 caatcacgaa tgaataacgg tttggttgat gcgagtgatt ttgatgacga gcgtaatggc
     2461 tggcctgttg aacaagtctg gaaagaaatg cataaacttt tgccattctc accggattca
     2521 gtcgtcactc atggtgattt ctcacttgat aaccttattt ttgacgaggg gaaattaata
     2581 ggttgtattg atgttggacg agtcggaatc gcagaccgat accaggatct tgccatccta
     2641 tggaactgcc tcggtgagtt ttctccttca ttacagaaac ggctttttca aaaatatggt
     2701 attgataatc ctgatatgaa taaattgcag tttcatttga tgctcgatga gtttttctaa
     2761 tcagaattgg ttaattggtt gtaacactgg cagagcatta cgctgacttg acgggacggc
     2821 gcaagctcat gaccaaaatc ccttaacgtg agttacgcgt cgttccactg agcgtcagac
     2881 cccgtagaaa agatcaaagg atcttcttga gatccttttt ttctgcgcgt aatctgctgc
     2941 ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt tgccggatca agagctacca
     3001 actctttttc cgaaggtaac tggcttcagc agagcgcaga taccaaatac tgttcttcta
     3061 gtgtagccgt agttaggcca ccacttcaag aactctgtag caccgcctac atacctcgct
     3121 ctgctaatcc tgttaccagt ggctgctgcc agtggcgata agtcgtgtct taccgggttg
     3181 gactcaagac gatagttacc ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc
     3241 acacagccca gcttggagcg aacgacctac accgaactga gatacctaca gcgtgagcta
     3301 tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca ggtatccggt aagcggcagg
     3361 gtcggaacag gagagcgcac gagggagctt ccagggggaa acgcctggta tctttatagt
     3421 cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg
     3481 cggagcctat ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc cttttgctgg
     3541 ccttttgctc acatgttctt tcctgcgtta tcccctgatt ctgtggataa ccgtattacc
     3601 gcctttgagt gagctgatac cgctcgccgc agccgaacga ccgagcgcag cgagtcagtg
     3661 agcgaggaag cggaagagcg cccaatacgc aaaccgcctc tccccgcgcg ttggccgatt
     3721 cattaatgca gctggcacga caggtttccc gactggaaag cgggcagtga gcgcaacgca
     3781 attaatacgc gtaccgctag ccaggaagag tttgtagaaa cgcaaaaagg ccatccgtca
     3841 ggatggcctt ctgcttagtt tgatgcctgg cagtttatgg cgggcgtcct gcccgccacc
     3901 ctccgggccg ttgcttcaca acgttcaaat ccgctcccgg cggatttgtc ctactcagga
     3961 gagcgttcac cgacaaacaa cagataaaac gaaaggccca gtcttccgac tgagcctttc
     4021 gttttatttg atgcctggca gttccctact ctcgcgttaa cgctagcatg gatgttttcc
     4081 cagtcacgac gtt
//
