LOCUS       sequence-250383-        4251 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 4251)
  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 4251)
  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: MLKP1del4ec
FEATURES             Location/Qualifiers
     source          1..4251
                     /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(2376..2475)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(2492..2511)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2493..2511)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2516..2532)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2516..2532)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2634..2653)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             2645..3454
                     /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(2712..2731)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      3601..4189
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4090..4109
                     /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 ggggatgtta acggccacaa gttctcagtg tccggcgagg gggagggcga
      781 cgccacctac gggaagctca cgctgaagtt catttgcacg accggcaagc tgccggtgcc
      841 ctggccaacc ctggtcacaa ctctgacgta cggcgtccag tgcttctccc gctacccgga
      901 ccacatgaag cagcatgatt tcttcaagtc cgctatgcca gagggctacg tgcaggagcg
      961 cacgatcttc ttcaaggacg atggcaacta caagaccagg gccgaggtca agttcgaggg
     1021 ggacacgctc gttaaccgga tcgagctgaa gggcattgac ttcaaggagg atgggaatat
     1081 cctcggccac aagctggagt acaactacaa ttcgcataac gtctacatca tggcggacaa
     1141 gcagaagaac gggattaagg ttaatttcaa gatccggcac aatattgagg acggcagcgt
     1201 gcagctcgct gatcattacc agcagaacac accgatcggg gacggccctg tcctcctgcc
     1261 cgataatcac tacctctcta ctcagtcagc gctgtccaag gacccgaacg agaagcgcga
     1321 tcatatggtc ctcctggagt tcgttacagc cgcggggatc actctcggca tggacgagct
     1381 gtacaagggg gctggcgatt acaaggacga tgacgataag gcctacccct acgacgtgcc
     1441 agattacgcc ggggctggca tgagggcggg cgacgcgttc agcaactacc gctgcaggac
     1501 catgtcgatg aattacgacg gcgataggag cgaggatcgg cagtcggagg aggtgcagtc
     1561 tgcctacaag cgggacagcg atgtcggcga ggagatgatc atttccagcg aggagcagaa
     1621 cgacgagaat gagtggagct gggtcaacca ggaccggaat ggcgatccct tcgcggagtc
     1681 ggtttcgtct ctccacacca cgcagcagat cctggagaat gagattcgca agctgtcgga
     1741 gctggggaag gagctggagg cggaggagag cacctcgggc aacaaggacc aggatgttat
     1801 cctcctgcca tactctgagg ccgacgtgct ggagctgtca gagaagatgg agcacctgga
     1861 gcagaaggtc aaggaggcgt cgaacaccat tcgcgagaag gacctcatgc tgtctaagct
     1921 gcagatcctc attggcacgg cggatcgggc tgccctggag gaggagctgg aggaggccgc
     1981 tgcttccatc gaccagctgg agacggagct ggagcgccac ctccaggaga agctggaggc
     2041 ggagattcag tgcctcctga tgctcaaggc ttcccagaac tggcaggttc gcgcggagga
     2101 cagggtggcc ctggaggagc atagggctct gtctgccggc gataagacac ggatgctcct
     2161 gaagctgcgc gagactgaga gcaagatcgt catgctcaag gagcaggtgg acaagctgga
     2221 ggtcagggag aaggagctga accggacaac tgaggtgctg aagatgcagt cacggacatt
     2281 ccgcgcgtcc ctgttcggcc tcgttcagct cgtgatgctc tgcctgtccc tcaaggtttt
     2341 cttcgcgcgc gtgagcgctc ctttcgacga gtgagaccca gctttcttgt acaaagttgg
     2401 cattataaga aagcattgct tatcaatttg ttgcaacgaa caggtcacta tcagtcaaaa
     2461 taaaatcatt atttgccatc cagctgatat cccctatagt gagtcgtatt acatggtcat
     2521 agctgtttcc tggcagctct ggcccgtgtc tcaaaatctc tgatgttaca ttgcacaaga
     2581 taaaaatata tcatcatgaa caataaaact gtctgcttac ataaacagta atacaagggg
     2641 tgttatgagc catattcaac gggaaacgtc gaggccgcga ttaaattcca acatggatgc
     2701 tgatttatat gggtataaat gggctcgcga taatgtcggg caatcaggtg cgacaatcta
     2761 tcgcttgtat gggaagcccg atgcgccaga gttgtttctg aaacatggca aaggtagcgt
     2821 tgccaatgat gttacagatg agatggtcag actaaactgg ctgacggaat ttatgcctct
     2881 tccgaccatc aagcatttta tccgtactcc tgatgatgca tggttactca ccactgcgat
     2941 ccccggaaaa acagcattcc aggtattaga agaatatcct gattcaggtg aaaatattgt
     3001 tgatgcgctg gcagtgttcc tgcgccggtt gcattcgatt cctgtttgta attgtccttt
     3061 taacagcgat cgcgtatttc gtctcgctca ggcgcaatca cgaatgaata acggtttggt
     3121 tgatgcgagt gattttgatg acgagcgtaa tggctggcct gttgaacaag tctggaaaga
     3181 aatgcataaa cttttgccat tctcaccgga ttcagtcgtc actcatggtg atttctcact
     3241 tgataacctt atttttgacg aggggaaatt aataggttgt attgatgttg gacgagtcgg
     3301 aatcgcagac cgataccagg atcttgccat cctatggaac tgcctcggtg agttttctcc
     3361 ttcattacag aaacggcttt ttcaaaaata tggtattgat aatcctgata tgaataaatt
     3421 gcagtttcat ttgatgctcg atgagttttt ctaatcagaa ttggttaatt ggttgtaaca
     3481 ctggcagagc attacgctga cttgacggga cggcgcaagc tcatgaccaa aatcccttaa
     3541 cgtgagttac gcgtcgttcc actgagcgtc agaccccgta gaaaagatca aaggatcttc
     3601 ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc
     3661 agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt
     3721 cagcagagcg cagataccaa atactgttct tctagtgtag ccgtagttag gccaccactt
     3781 caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc
     3841 tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa
     3901 ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac
     3961 ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg
     4021 gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga
     4081 gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact
     4141 tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa
     4201 cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgt t
//
