LOCUS       sequence-255504-        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     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(1786..1885)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(1902..1921)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1903..1921)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1926..1942)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1926..1942)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2044..2063)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             2055..2864
                     /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(2122..2141)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      3011..3599
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3500..3519
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3753..3770
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(3929..3956)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(4048..4134)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     join(4183..4197,1..8)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     join(4197,1..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 cttaatggtc tccaaggggg aggagctgtt caccggcgtg gtcccgatcc
      181 tggtggagct ggacggggat gtcaacggcc acaagttctc agtgtccggc gagggggagg
      241 gcgacgcgac atacgggaag ctcactctga agttcatttg cacgaccggc aagctcccgg
      301 ttccctggcc aaccctggtg acaactctca cgtacggcgt ccagtgcttc tcccgctacc
      361 cggaccacat gaagcagcat gatttcttca agagcgccat gccagagggc tacgtgcagg
      421 agcgcaccat cttcttcaag gacgatggca actacaagac cagggcggag gtgaagttcg
      481 agggggacac gctggtcaac cggatcgagc tgaagggcat tgacttcaag gaggatggga
      541 atattctggg ccacaagctg gagtacaact acaattcgca taacgtttac atcatggctg
      601 acaagcagaa gaacggcatt aaggtcaatt tcaagatccg ccacaatatt gaggacggct
      661 ccgtccagct ggccgatcat taccagcaga acacaccgat cggggacggc cctgtcctcc
      721 tgcccgataa tcactacctg tctactcagt cagccctctc caaggacccg aacgagaaga
      781 gggatcatat ggttctcctg gagttcgtga cggcggccgg gatcaccctg ggcatggacg
      841 agctgtacaa gggggccggc gattacaagg acgatgacga taaggcgtac ccctacgacg
      901 tcccagatta cgcgggggct ggcatgctca tgcactacga cgtggatagg tcggaggaca
      961 ggcagtccga ggaggtccag tctgcgtacc gcagggacag cgatatcggc gaggagatga
     1021 tcatttccag cgaggagcag aacgacgaga atgagtggtc gtgggtgaac caggacaggg
     1081 atggtggccc actggctgag tctgtttcgt ctctccacat gacccagcag gtgctggaga
     1141 atgagatcca gaagctctcg gagctgggga aggagctgga ggccgaggag agcacgtcgg
     1201 gcaacaagga ccaggatgtc attgttctgc catacgctga ggtggacatg ctcaagctga
     1261 atgagaagat ggagcacctg gagcagaagg tgaaggaggt ctctaacaca atcagggaga
     1321 aggacctccg gctgtcaaag ctccagatcc tgattggcac agcggacagg gccactgtgg
     1381 aggaggctgc tgcgagcatt gatcagctgg agaccgagtt cgagcaccat ctgcaggaga
     1441 tcctggaggc tgagattcag tacctcgtca tgatgaaggc ccgccagaat tggcaggtta
     1501 gggcggagga ccagatggct ctggaggagc acagggcctc agctgcgggc gataacacac
     1561 ggatgctcct gaagctccgc gagactgaga gcaagatcgt tatgctcaag gagcaggtgg
     1621 acaagctgga ggttagggag aaggatctct accggaccac ggaggtgctg aagatgcagt
     1681 cacgcacctt caaggtctcc ctcttcggcc tggttcagct catcatgatg tgcctcagcc
     1741 tgaagatgtt cttcgctcag gtgtcggccc ctttcgacga gtgaaaccca gctttcttgt
     1801 acaaagttgg cattataaga aagcattgct tatcaatttg ttgcaacgaa caggtcacta
     1861 tcagtcaaaa taaaatcatt atttgccatc cagctgatat cccctatagt gagtcgtatt
     1921 acatggtcat agctgtttcc tggcagctct ggcccgtgtc tcaaaatctc tgatgttaca
     1981 ttgcacaaga taaaaatata tcatcatgaa caataaaact gtctgcttac ataaacagta
     2041 atacaagggg tgttatgagc catattcaac gggaaacgtc gaggccgcga ttaaattcca
     2101 acatggatgc tgatttatat gggtataaat gggctcgcga taatgtcggg caatcaggtg
     2161 cgacaatcta tcgcttgtat gggaagcccg atgcgccaga gttgtttctg aaacatggca
     2221 aaggtagcgt tgccaatgat gttacagatg agatggtcag actaaactgg ctgacggaat
     2281 ttatgcctct tccgaccatc aagcatttta tccgtactcc tgatgatgca tggttactca
     2341 ccactgcgat ccccggaaaa acagcattcc aggtattaga agaatatcct gattcaggtg
     2401 aaaatattgt tgatgcgctg gcagtgttcc tgcgccggtt gcattcgatt cctgtttgta
     2461 attgtccttt taacagcgat cgcgtatttc gtctcgctca ggcgcaatca cgaatgaata
     2521 acggtttggt tgatgcgagt gattttgatg acgagcgtaa tggctggcct gttgaacaag
     2581 tctggaaaga aatgcataaa cttttgccat tctcaccgga ttcagtcgtc actcatggtg
     2641 atttctcact tgataacctt atttttgacg aggggaaatt aataggttgt attgatgttg
     2701 gacgagtcgg aatcgcagac cgataccagg atcttgccat cctatggaac tgcctcggtg
     2761 agttttctcc ttcattacag aaacggcttt ttcaaaaata tggtattgat aatcctgata
     2821 tgaataaatt gcagtttcat ttgatgctcg atgagttttt ctaatcagaa ttggttaatt
     2881 ggttgtaaca ctggcagagc attacgctga cttgacggga cggcgcaagc tcatgaccaa
     2941 aatcccttaa cgtgagttac gcgtcgttcc actgagcgtc agaccccgta gaaaagatca
     3001 aaggatcttc ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac
     3061 caccgctacc agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg
     3121 taactggctt cagcagagcg cagataccaa atactgttct tctagtgtag ccgtagttag
     3181 gccaccactt caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac
     3241 cagtggctgc tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt
     3301 taccggataa ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg
     3361 agcgaacgac ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc
     3421 ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc
     3481 gcacgaggga gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc
     3541 acctctgact tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa
     3601 acgccagcaa cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgt
     3661 tctttcctgc gttatcccct gattctgtgg ataaccgtat taccgccttt gagtgagctg
     3721 ataccgctcg ccgcagccga acgaccgagc gcagcgagtc agtgagcgag gaagcggaag
     3781 agcgcccaat acgcaaaccg cctctccccg cgcgttggcc gattcattaa tgcagctggc
     3841 acgacaggtt tcccgactgg aaagcgggca gtgagcgcaa cgcaattaat acgcgtaccg
     3901 ctagccagga agagtttgta gaaacgcaaa aaggccatcc gtcaggatgg ccttctgctt
     3961 agtttgatgc ctggcagttt atggcgggcg tcctgcccgc caccctccgg gccgttgctt
     4021 cacaacgttc aaatccgctc ccggcggatt tgtcctactc aggagagcgt tcaccgacaa
     4081 acaacagata aaacgaaagg cccagtcttc cgactgagcc tttcgtttta tttgatgcct
     4141 ggcagttccc tactctcgcg ttaacgctag catggatgtt ttcccagtca cgacgtt
//
