LOCUS       sequence-255537-        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     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(1840..1939)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(1956..1975)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1957..1975)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1980..1996)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1980..1996)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2098..2117)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             2109..2918
                     /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(2176..2195)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      3065..3653
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3554..3573
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3807..3824
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(3983..4010)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(4102..4188)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     join(4237..4251,1..8)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     join(4251,1..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 gttaacggcc acaagttctc agtgtccggc gagggggagg
      241 gcgacgccac ctacgggaag ctcacgctga agttcatttg cacgaccggc aagctgccgg
      301 tgccctggcc aaccctggtc acaactctga cgtacggcgt ccagtgcttc tcccgctacc
      361 cggaccacat gaagcagcat gatttcttca agtccgctat gccagagggc tacgtgcagg
      421 agcgcacgat cttcttcaag gacgatggca actacaagac cagggccgag gtcaagttcg
      481 agggggacac gctcgttaac cggatcgagc tgaagggcat tgacttcaag gaggatggga
      541 atatcctcgg ccacaagctg gagtacaact acaattcgca taacgtctac atcatggcgg
      601 acaagcagaa gaacgggatt aaggttaatt tcaagatccg gcacaatatt gaggacggca
      661 gcgtgcagct cgctgatcat taccagcaga acacaccgat cggggacggc cctgtcctcc
      721 tgcccgataa tcactacctc tctactcagt cagcgctgtc caaggacccg aacgagaagc
      781 gcgatcatat ggtcctcctg gagttcgtta cagccgcggg gatcactctc ggcatggacg
      841 agctgtacaa gggggctggc gattacaagg acgatgacga taaggcctac ccctacgacg
      901 tgccagatta cgccggggct ggcatgaggg cgggcgacgc gttcagcaac taccgctgca
      961 ggaccatgtc gatgaattac gacggcgata ggagcgagga tcggcagtcg gaggaggtgc
     1021 agtctgccta caagcgggac agcgatgtcg gcgaggagat gatcatttcc agcgaggagc
     1081 agaacgacga gaatgagtgg agctgggtca accaggaccg gaatggcgat cccttcgcgg
     1141 agtcggtttc gtctctccac accacgcagc agatcctgga gaatgagatt cgcaagctgt
     1201 cggagctggg gaaggagctg gaggcggagg agagcacctc gggcaacaag gaccaggatg
     1261 ttatcctcct gccatactct gaggccgacg tgctggagct gtcagagaag atggagcacc
     1321 tggagcagaa ggtcaaggag gcgtcgaaca ccattcgcga gaaggacctc atgctgtcta
     1381 agctgcagat cctcattggc acggcggatc gggctgccct ggaggaggag ctggaggagg
     1441 ccgctgcttc catcgaccag ctggagacgg agctggagcg ccacctccag gagaagctgg
     1501 aggcggagat tcagtgcctc ctgatgctca aggcttccca gaactggcag gttcgcgcgg
     1561 aggacagggt ggccctggag gagcataggg ctctgtctgc cggcgataag acacggatgc
     1621 tcctgaagct gcgcgagact gagagcaaga tcgtcatgct caaggagcag gtggacaagc
     1681 tggaggtcag ggagaaggag ctgaaccgga caactgaggt gctgaagatg cagtcacgga
     1741 cattccgcgc gtccctgttc ggcctcgttc agctcgtgat gctctgcctg tccctcaagg
     1801 ttttcttcgc gcgcgtgagc gctcctttcg acgagtgaaa cccagctttc ttgtacaaag
     1861 ttggcattat aagaaagcat tgcttatcaa tttgttgcaa cgaacaggtc actatcagtc
     1921 aaaataaaat cattatttgc catccagctg atatccccta tagtgagtcg tattacatgg
     1981 tcatagctgt ttcctggcag ctctggcccg tgtctcaaaa tctctgatgt tacattgcac
     2041 aagataaaaa tatatcatca tgaacaataa aactgtctgc ttacataaac agtaatacaa
     2101 ggggtgttat gagccatatt caacgggaaa cgtcgaggcc gcgattaaat tccaacatgg
     2161 atgctgattt atatgggtat aaatgggctc gcgataatgt cgggcaatca ggtgcgacaa
     2221 tctatcgctt gtatgggaag cccgatgcgc cagagttgtt tctgaaacat ggcaaaggta
     2281 gcgttgccaa tgatgttaca gatgagatgg tcagactaaa ctggctgacg gaatttatgc
     2341 ctcttccgac catcaagcat tttatccgta ctcctgatga tgcatggtta ctcaccactg
     2401 cgatccccgg aaaaacagca ttccaggtat tagaagaata tcctgattca ggtgaaaata
     2461 ttgttgatgc gctggcagtg ttcctgcgcc ggttgcattc gattcctgtt tgtaattgtc
     2521 cttttaacag cgatcgcgta tttcgtctcg ctcaggcgca atcacgaatg aataacggtt
     2581 tggttgatgc gagtgatttt gatgacgagc gtaatggctg gcctgttgaa caagtctgga
     2641 aagaaatgca taaacttttg ccattctcac cggattcagt cgtcactcat ggtgatttct
     2701 cacttgataa ccttattttt gacgagggga aattaatagg ttgtattgat gttggacgag
     2761 tcggaatcgc agaccgatac caggatcttg ccatcctatg gaactgcctc ggtgagtttt
     2821 ctccttcatt acagaaacgg ctttttcaaa aatatggtat tgataatcct gatatgaata
     2881 aattgcagtt tcatttgatg ctcgatgagt ttttctaatc agaattggtt aattggttgt
     2941 aacactggca gagcattacg ctgacttgac gggacggcgc aagctcatga ccaaaatccc
     3001 ttaacgtgag ttacgcgtcg ttccactgag cgtcagaccc cgtagaaaag atcaaaggat
     3061 cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc
     3121 taccagcggt ggtttgtttg ccggatcaag agctaccaac tctttttccg aaggtaactg
     3181 gcttcagcag agcgcagata ccaaatactg ttcttctagt gtagccgtag ttaggccacc
     3241 acttcaagaa ctctgtagca ccgcctacat acctcgctct gctaatcctg ttaccagtgg
     3301 ctgctgccag tggcgataag tcgtgtctta ccgggttgga ctcaagacga tagttaccgg
     3361 ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac acagcccagc ttggagcgaa
     3421 cgacctacac cgaactgaga tacctacagc gtgagctatg agaaagcgcc acgcttcccg
     3481 aagggagaaa ggcggacagg tatccggtaa gcggcagggt cggaacagga gagcgcacga
     3541 gggagcttcc agggggaaac gcctggtatc tttatagtcc tgtcgggttt cgccacctct
     3601 gacttgagcg tcgatttttg tgatgctcgt caggggggcg gagcctatgg aaaaacgcca
     3661 gcaacgcggc ctttttacgg ttcctggcct tttgctggcc ttttgctcac atgttctttc
     3721 ctgcgttatc ccctgattct gtggataacc gtattaccgc ctttgagtga gctgataccg
     3781 ctcgccgcag ccgaacgacc gagcgcagcg agtcagtgag cgaggaagcg gaagagcgcc
     3841 caatacgcaa accgcctctc cccgcgcgtt ggccgattca ttaatgcagc tggcacgaca
     3901 ggtttcccga ctggaaagcg ggcagtgagc gcaacgcaat taatacgcgt accgctagcc
     3961 aggaagagtt tgtagaaacg caaaaaggcc atccgtcagg atggccttct gcttagtttg
     4021 atgcctggca gtttatggcg ggcgtcctgc ccgccaccct ccgggccgtt gcttcacaac
     4081 gttcaaatcc gctcccggcg gatttgtcct actcaggaga gcgttcaccg acaaacaaca
     4141 gataaaacga aaggcccagt cttccgactg agcctttcgt tttatttgat gcctggcagt
     4201 tccctactct cgcgttaacg ctagcatgga tgttttccca gtcacgacgt t
//
