LOCUS       sequence-255905-        4129 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 4129)
  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 4129)
  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: ECPK1D
FEATURES             Location/Qualifiers
     source          1..4129
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    16..115
                     /gene="mutant version of attL"
                     /label=attL1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     CDS             join(118..120,121..123,124..834)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             844..867
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             871..897
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     protein_bind    complement(1701..1800)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(1817..1836)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1818..1836)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1841..1857)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1841..1857)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1959..1978)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             1970..2779
                     /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(2037..2056)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      2926..3514
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3415..3434
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3668..3685
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(3844..3871)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(3963..4049)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     4098..4120
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     4112..4129
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     4113..4129
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
ORIGIN
        1 cttaagctcg ggccccaaat aatgatttta ttttgactga tagtgacctg ttcgttgcaa
       61 caaattgatg agcaatgctt ttttataatg ccaactttgt acaaaaaagc aggcttcatg
      121 gtctccaagg gggaggagct gttcaccggc gtggtcccaa tcctcgtcga gctggacggg
      181 gatgttaacg gccacaagtt ctccgtgagc ggcgaggggg agggcgacgc cacctacggg
      241 aagctcacgc tgaagttcat ttgcacgacc ggcaagctgc cggtgccctg gccaaccctg
      301 gtcacaactc tgacgtacgg cgtccagtgc ttctcccgct acccggacca catgaagcag
      361 catgatttct tcaagagcgc catgccggag ggctacgttc aggagcgcac catcttcttc
      421 aaggacgatg gcaactacaa gacaagggcg gaggttaagt tcgagggcga cactctcgtg
      481 aaccggatcg agctgaaggg cattgacttc aaggaggatg ggaatattct cggccacaag
      541 ctggagtaca actacaattc gcataacgtc tacatcatgg ctgacaagca gaagaacggc
      601 attaaggtta atttcaagat ccgccacaat attgaggacg gctccgtgca gctcgcggat
      661 cattaccagc agaacacgcc gatcggggac ggccctgtcc tcctgccgga taatcactac
      721 ctctcgacgc agtctgcgct gtcaaaggac ccgaacgaga agagggatca tatggttctc
      781 ctggagttcg tgacagccgc ggggatcact ctcggcatgg acgagctgta caagggggcg
      841 ggcgactaca aggacgatga cgataaggct tacccctacg acgtgccaga ttacgctggg
      901 gccggcggat ccatggacat gccgccggcg gcgcagggac ggagggcgat cgccgtggtg
      961 gaccggaagc ttgacgagct ctgcgcctgc ctcgacgacg cgatgtcgtc caggcagcgg
     1021 agcccgtacg cggaggagcg gctccgcgcg gagatcaagt cgagggccga cttccttcgg
     1081 tcgctcatgg ccgccgaggt ggactgccac ggcgggacgc gcccggagca cctggctgag
     1141 gccgaggcgc gcttcgccgt cctagaggcc acgttcgatc ggtgggcgcg gcgcgcagtc
     1201 gctgcacctg aaccagtcgt cgaggagaag gaggaggagg ccgactcctt ggggtcctcc
     1261 cggtccacgt gctcctgcac cgagtcgtgc caggaggcga cggaggggga cgccatgcgc
     1321 gacgccgtgg aaacacgcgg caaggagggg acgtcggatg gcatcgccga ggacaacgac
     1381 gccgaatgcg agacggtggt aatccgcagg gaggaggcgg cggcggttcc tgttgccaag
     1441 aagcgggatg aggctatggg aacacgcgag gaggtggcgc tggacgccat tgccaagaag
     1501 aagggcgata ccgatcgcga tgccgcggcg gagacgggga ggaggaccgt agaccagcgg
     1561 cgctggtgga ggcggagtgc tgcctggtgc ggtgcagccg gcgtcgtcgc gttggtcgcg
     1621 gtcgggttgg cggtggagtt ggtcgcggtg gctgaccacg atgtgaatgc ttacgttgtt
     1681 ccgacgtagc ctgcaggcag acccagcttt cttgtacaaa gttggcatta taagaaagca
     1741 ttgcttatca atttgttgca acgaacaggt cactatcagt caaaataaaa tcattatttg
     1801 ccatccagct gatatcccct atagtgagtc gtattacatg gtcatagctg tttcctggca
     1861 gctctggccc gtgtctcaaa atctctgatg ttacattgca caagataaaa atatatcatc
     1921 atgaacaata aaactgtctg cttacataaa cagtaataca aggggtgtta tgagccatat
     1981 tcaacgggaa acgtcgaggc cgcgattaaa ttccaacatg gatgctgatt tatatgggta
     2041 taaatgggct cgcgataatg tcgggcaatc aggtgcgaca atctatcgct tgtatgggaa
     2101 gcccgatgcg ccagagttgt ttctgaaaca tggcaaaggt agcgttgcca atgatgttac
     2161 agatgagatg gtcagactaa actggctgac ggaatttatg cctcttccga ccatcaagca
     2221 ttttatccgt actcctgatg atgcatggtt actcaccact gcgatccccg gaaaaacagc
     2281 attccaggta ttagaagaat atcctgattc aggtgaaaat attgttgatg cgctggcagt
     2341 gttcctgcgc cggttgcatt cgattcctgt ttgtaattgt ccttttaaca gcgatcgcgt
     2401 atttcgtctc gctcaggcgc aatcacgaat gaataacggt ttggttgatg cgagtgattt
     2461 tgatgacgag cgtaatggct ggcctgttga acaagtctgg aaagaaatgc ataaactttt
     2521 gccattctca ccggattcag tcgtcactca tggtgatttc tcacttgata accttatttt
     2581 tgacgagggg aaattaatag gttgtattga tgttggacga gtcggaatcg cagaccgata
     2641 ccaggatctt gccatcctat ggaactgcct cggtgagttt tctccttcat tacagaaacg
     2701 gctttttcaa aaatatggta ttgataatcc tgatatgaat aaattgcagt ttcatttgat
     2761 gctcgatgag tttttctaat cagaattggt taattggttg taacactggc agagcattac
     2821 gctgacttga cgggacggcg caagctcatg accaaaatcc cttaacgtga gttacgcgtc
     2881 gttccactga gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag atcctttttt
     2941 tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt
     3001 gccggatcaa gagctaccaa ctctttttcc gaaggtaact ggcttcagca gagcgcagat
     3061 accaaatact gttcttctag tgtagccgta gttaggccac cacttcaaga actctgtagc
     3121 accgcctaca tacctcgctc tgctaatcct gttaccagtg gctgctgcca gtggcgataa
     3181 gtcgtgtctt accgggttgg actcaagacg atagttaccg gataaggcgc agcggtcggg
     3241 ctgaacgggg ggttcgtgca cacagcccag cttggagcga acgacctaca ccgaactgag
     3301 atacctacag cgtgagctat gagaaagcgc cacgcttccc gaagggagaa aggcggacag
     3361 gtatccggta agcggcaggg tcggaacagg agagcgcacg agggagcttc cagggggaaa
     3421 cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt
     3481 gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg
     3541 gttcctggcc ttttgctggc cttttgctca catgttcttt cctgcgttat cccctgattc
     3601 tgtggataac cgtattaccg cctttgagtg agctgatacc gctcgccgca gccgaacgac
     3661 cgagcgcagc gagtcagtga gcgaggaagc ggaagagcgc ccaatacgca aaccgcctct
     3721 ccccgcgcgt tggccgattc attaatgcag ctggcacgac aggtttcccg actggaaagc
     3781 gggcagtgag cgcaacgcaa ttaatacgcg taccgctagc caggaagagt ttgtagaaac
     3841 gcaaaaaggc catccgtcag gatggccttc tgcttagttt gatgcctggc agtttatggc
     3901 gggcgtcctg cccgccaccc tccgggccgt tgcttcacaa cgttcaaatc cgctcccggc
     3961 ggatttgtcc tactcaggag agcgttcacc gacaaacaac agataaaacg aaaggcccag
     4021 tcttccgact gagcctttcg ttttatttga tgcctggcag ttccctactc tcgcgttaac
     4081 gctagcatgg atgttttccc agtcacgacg ttgtaaaacg acggccagt
//
