LOCUS       sequence-250382-        4197 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 4197)
  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 4197)
  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: MLKP2del4ec
FEATURES             Location/Qualifiers
     source          1..4197
                     /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(2322..2421)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(2438..2457)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2439..2457)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2462..2478)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2462..2478)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2580..2599)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             2591..3400
                     /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(2658..2677)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      3547..4135
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4036..4055
                     /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 agggggagga gctgttcacc ggcgtggtcc cgatcctggt
      721 ggagctggac ggggatgtca acggccacaa gttctcagtg tccggcgagg gggagggcga
      781 cgcgacatac gggaagctca ctctgaagtt catttgcacg accggcaagc tcccggttcc
      841 ctggccaacc ctggtgacaa ctctcacgta cggcgtccag tgcttctccc gctacccgga
      901 ccacatgaag cagcatgatt tcttcaagag cgccatgcca gagggctacg tgcaggagcg
      961 caccatcttc ttcaaggacg atggcaacta caagaccagg gcggaggtga agttcgaggg
     1021 ggacacgctg gtcaaccgga tcgagctgaa gggcattgac ttcaaggagg atgggaatat
     1081 tctgggccac aagctggagt acaactacaa ttcgcataac gtttacatca tggctgacaa
     1141 gcagaagaac ggcattaagg tcaatttcaa gatccgccac aatattgagg acggctccgt
     1201 ccagctggcc gatcattacc agcagaacac accgatcggg gacggccctg tcctcctgcc
     1261 cgataatcac tacctgtcta ctcagtcagc cctctccaag gacccgaacg agaagaggga
     1321 tcatatggtt ctcctggagt tcgtgacggc ggccgggatc accctgggca tggacgagct
     1381 gtacaagggg gccggcgatt acaaggacga tgacgataag gcgtacccct acgacgtccc
     1441 agattacgcg ggggctggca tgctcatgca ctacgacgtg gataggtcgg aggacaggca
     1501 gtccgaggag gtccagtctg cgtaccgcag ggacagcgat atcggcgagg agatgatcat
     1561 ttccagcgag gagcagaacg acgagaatga gtggtcgtgg gtgaaccagg acagggatgg
     1621 tggcccactg gctgagtctg tttcgtctct ccacatgacc cagcaggtgc tggagaatga
     1681 gatccagaag ctctcggagc tggggaagga gctggaggcc gaggagagca cgtcgggcaa
     1741 caaggaccag gatgtcattg ttctgccata cgctgaggtg gacatgctca agctgaatga
     1801 gaagatggag cacctggagc agaaggtgaa ggaggtctct aacacaatca gggagaagga
     1861 cctccggctg tcaaagctcc agatcctgat tggcacagcg gacagggcca ctgtggagga
     1921 ggctgctgcg agcattgatc agctggagac cgagttcgag caccatctgc aggagatcct
     1981 ggaggctgag attcagtacc tcgtcatgat gaaggcccgc cagaattggc aggttagggc
     2041 ggaggaccag atggctctgg aggagcacag ggcctcagct gcgggcgata acacacggat
     2101 gctcctgaag ctccgcgaga ctgagagcaa gatcgttatg ctcaaggagc aggtggacaa
     2161 gctggaggtt agggagaagg atctctaccg gaccacggag gtgctgaaga tgcagtcacg
     2221 caccttcaag gtctccctct tcggcctggt tcagctcatc atgatgtgcc tcagcctgaa
     2281 gatgttcttc gctcaggtgt cggccccttt cgacgagtga gacccagctt tcttgtacaa
     2341 agttggcatt ataagaaagc attgcttatc aatttgttgc aacgaacagg tcactatcag
     2401 tcaaaataaa atcattattt gccatccagc tgatatcccc tatagtgagt cgtattacat
     2461 ggtcatagct gtttcctggc agctctggcc cgtgtctcaa aatctctgat gttacattgc
     2521 acaagataaa aatatatcat catgaacaat aaaactgtct gcttacataa acagtaatac
     2581 aaggggtgtt atgagccata ttcaacggga aacgtcgagg ccgcgattaa attccaacat
     2641 ggatgctgat ttatatgggt ataaatgggc tcgcgataat gtcgggcaat caggtgcgac
     2701 aatctatcgc ttgtatggga agcccgatgc gccagagttg tttctgaaac atggcaaagg
     2761 tagcgttgcc aatgatgtta cagatgagat ggtcagacta aactggctga cggaatttat
     2821 gcctcttccg accatcaagc attttatccg tactcctgat gatgcatggt tactcaccac
     2881 tgcgatcccc ggaaaaacag cattccaggt attagaagaa tatcctgatt caggtgaaaa
     2941 tattgttgat gcgctggcag tgttcctgcg ccggttgcat tcgattcctg tttgtaattg
     3001 tccttttaac agcgatcgcg tatttcgtct cgctcaggcg caatcacgaa tgaataacgg
     3061 tttggttgat gcgagtgatt ttgatgacga gcgtaatggc tggcctgttg aacaagtctg
     3121 gaaagaaatg cataaacttt tgccattctc accggattca gtcgtcactc atggtgattt
     3181 ctcacttgat aaccttattt ttgacgaggg gaaattaata ggttgtattg atgttggacg
     3241 agtcggaatc gcagaccgat accaggatct tgccatccta tggaactgcc tcggtgagtt
     3301 ttctccttca ttacagaaac ggctttttca aaaatatggt attgataatc ctgatatgaa
     3361 taaattgcag tttcatttga tgctcgatga gtttttctaa tcagaattgg ttaattggtt
     3421 gtaacactgg cagagcatta cgctgacttg acgggacggc gcaagctcat gaccaaaatc
     3481 ccttaacgtg agttacgcgt cgttccactg agcgtcagac cccgtagaaa agatcaaagg
     3541 atcttcttga gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc
     3601 gctaccagcg gtggtttgtt tgccggatca agagctacca actctttttc cgaaggtaac
     3661 tggcttcagc agagcgcaga taccaaatac tgttcttcta gtgtagccgt agttaggcca
     3721 ccacttcaag aactctgtag caccgcctac atacctcgct ctgctaatcc tgttaccagt
     3781 ggctgctgcc agtggcgata agtcgtgtct taccgggttg gactcaagac gatagttacc
     3841 ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc acacagccca gcttggagcg
     3901 aacgacctac accgaactga gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc
     3961 cgaagggaga aaggcggaca ggtatccggt aagcggcagg gtcggaacag gagagcgcac
     4021 gagggagctt ccagggggaa acgcctggta tctttatagt cctgtcgggt ttcgccacct
     4081 ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc
     4141 cagcaacgcg gcctttttac ggttcctggc cttttgctgg ccttttgctc acatgtt
//
