LOCUS       sequence-255143-        4209 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 4209)
  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 4209)
  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: pHWBF07EC
FEATURES             Location/Qualifiers
     source          1..4209
                     /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(1798..1897)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(1914..1933)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1915..1933)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1938..1954)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1938..1954)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2056..2075)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             2067..2876
                     /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(2134..2153)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      3023..3611
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3512..3531
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3765..3782
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(3941..3968)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(4060..4146)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     join(4195..4209,1..8)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     join(4209,1..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 cttcatggtc 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 ggttgtgccg acgtgagacc
     1801 cagctttctt gtacaaagtt ggcattataa gaaagcattg cttatcaatt tgttgcaacg
     1861 aacaggtcac tatcagtcaa aataaaatca ttatttgcca tccagctgat atcccctata
     1921 gtgagtcgta ttacatggtc atagctgttt cctggcagct ctggcccgtg tctcaaaatc
     1981 tctgatgtta cattgcacaa gataaaaata tatcatcatg aacaataaaa ctgtctgctt
     2041 acataaacag taatacaagg ggtgttatga gccatattca acgggaaacg tcgaggccgc
     2101 gattaaattc caacatggat gctgatttat atgggtataa atgggctcgc gataatgtcg
     2161 ggcaatcagg tgcgacaatc tatcgcttgt atgggaagcc cgatgcgcca gagttgtttc
     2221 tgaaacatgg caaaggtagc gttgccaatg atgttacaga tgagatggtc agactaaact
     2281 ggctgacgga atttatgcct cttccgacca tcaagcattt tatccgtact cctgatgatg
     2341 catggttact caccactgcg atccccggaa aaacagcatt ccaggtatta gaagaatatc
     2401 ctgattcagg tgaaaatatt gttgatgcgc tggcagtgtt cctgcgccgg ttgcattcga
     2461 ttcctgtttg taattgtcct tttaacagcg atcgcgtatt tcgtctcgct caggcgcaat
     2521 cacgaatgaa taacggtttg gttgatgcga gtgattttga tgacgagcgt aatggctggc
     2581 ctgttgaaca agtctggaaa gaaatgcata aacttttgcc attctcaccg gattcagtcg
     2641 tcactcatgg tgatttctca cttgataacc ttatttttga cgaggggaaa ttaataggtt
     2701 gtattgatgt tggacgagtc ggaatcgcag accgatacca ggatcttgcc atcctatgga
     2761 actgcctcgg tgagttttct ccttcattac agaaacggct ttttcaaaaa tatggtattg
     2821 ataatcctga tatgaataaa ttgcagtttc atttgatgct cgatgagttt ttctaatcag
     2881 aattggttaa ttggttgtaa cactggcaga gcattacgct gacttgacgg gacggcgcaa
     2941 gctcatgacc aaaatccctt aacgtgagtt acgcgtcgtt ccactgagcg tcagaccccg
     3001 tagaaaagat caaaggatct tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc
     3061 aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc
     3121 tttttccgaa ggtaactggc ttcagcagag cgcagatacc aaatactgtt cttctagtgt
     3181 agccgtagtt aggccaccac ttcaagaact ctgtagcacc gcctacatac ctcgctctgc
     3241 taatcctgtt accagtggct gctgccagtg gcgataagtc gtgtcttacc gggttggact
     3301 caagacgata gttaccggat aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac
     3361 agcccagctt ggagcgaacg acctacaccg aactgagata cctacagcgt gagctatgag
     3421 aaagcgccac gcttcccgaa gggagaaagg cggacaggta tccggtaagc ggcagggtcg
     3481 gaacaggaga gcgcacgagg gagcttccag ggggaaacgc ctggtatctt tatagtcctg
     3541 tcgggtttcg ccacctctga cttgagcgtc gatttttgtg atgctcgtca ggggggcgga
     3601 gcctatggaa aaacgccagc aacgcggcct ttttacggtt cctggccttt tgctggcctt
     3661 ttgctcacat gttctttcct gcgttatccc ctgattctgt ggataaccgt attaccgcct
     3721 ttgagtgagc tgataccgct cgccgcagcc gaacgaccga gcgcagcgag tcagtgagcg
     3781 aggaagcgga agagcgccca atacgcaaac cgcctctccc cgcgcgttgg ccgattcatt
     3841 aatgcagctg gcacgacagg tttcccgact ggaaagcggg cagtgagcgc aacgcaatta
     3901 atacgcgtac cgctagccag gaagagtttg tagaaacgca aaaaggccat ccgtcaggat
     3961 ggccttctgc ttagtttgat gcctggcagt ttatggcggg cgtcctgccc gccaccctcc
     4021 gggccgttgc ttcacaacgt tcaaatccgc tcccggcgga tttgtcctac tcaggagagc
     4081 gttcaccgac aaacaacaga taaaacgaaa ggcccagtct tccgactgag cctttcgttt
     4141 tatttgatgc ctggcagttc cctactctcg cgttaacgct agcatggatg ttttcccagt
     4201 cacgacgtt
//
