LOCUS       sequence-250368-        4263 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 4263)
  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 4263)
  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: pHWBF09EC
FEATURES             Location/Qualifiers
     source          1..4263
                     /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(2388..2487)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(2504..2523)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2505..2523)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2528..2544)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2528..2544)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2646..2665)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             2657..3466
                     /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(2724..2743)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      3613..4201
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4102..4121
                     /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 ggccgtgccg acttgagacc cagctttctt
     2401 gtacaaagtt ggcattataa gaaagcattg cttatcaatt tgttgcaacg aacaggtcac
     2461 tatcagtcaa aataaaatca ttatttgcca tccagctgat atcccctata gtgagtcgta
     2521 ttacatggtc atagctgttt cctggcagct ctggcccgtg tctcaaaatc tctgatgtta
     2581 cattgcacaa gataaaaata tatcatcatg aacaataaaa ctgtctgctt acataaacag
     2641 taatacaagg ggtgttatga gccatattca acgggaaacg tcgaggccgc gattaaattc
     2701 caacatggat gctgatttat atgggtataa atgggctcgc gataatgtcg ggcaatcagg
     2761 tgcgacaatc tatcgcttgt atgggaagcc cgatgcgcca gagttgtttc tgaaacatgg
     2821 caaaggtagc gttgccaatg atgttacaga tgagatggtc agactaaact ggctgacgga
     2881 atttatgcct cttccgacca tcaagcattt tatccgtact cctgatgatg catggttact
     2941 caccactgcg atccccggaa aaacagcatt ccaggtatta gaagaatatc ctgattcagg
     3001 tgaaaatatt gttgatgcgc tggcagtgtt cctgcgccgg ttgcattcga ttcctgtttg
     3061 taattgtcct tttaacagcg atcgcgtatt tcgtctcgct caggcgcaat cacgaatgaa
     3121 taacggtttg gttgatgcga gtgattttga tgacgagcgt aatggctggc ctgttgaaca
     3181 agtctggaaa gaaatgcata aacttttgcc attctcaccg gattcagtcg tcactcatgg
     3241 tgatttctca cttgataacc ttatttttga cgaggggaaa ttaataggtt gtattgatgt
     3301 tggacgagtc ggaatcgcag accgatacca ggatcttgcc atcctatgga actgcctcgg
     3361 tgagttttct ccttcattac agaaacggct ttttcaaaaa tatggtattg ataatcctga
     3421 tatgaataaa ttgcagtttc atttgatgct cgatgagttt ttctaatcag aattggttaa
     3481 ttggttgtaa cactggcaga gcattacgct gacttgacgg gacggcgcaa gctcatgacc
     3541 aaaatccctt aacgtgagtt acgcgtcgtt ccactgagcg tcagaccccg tagaaaagat
     3601 caaaggatct tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa
     3661 accaccgcta ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc tttttccgaa
     3721 ggtaactggc ttcagcagag cgcagatacc aaatactgtt cttctagtgt agccgtagtt
     3781 aggccaccac ttcaagaact ctgtagcacc gcctacatac ctcgctctgc taatcctgtt
     3841 accagtggct gctgccagtg gcgataagtc gtgtcttacc gggttggact caagacgata
     3901 gttaccggat aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac agcccagctt
     3961 ggagcgaacg acctacaccg aactgagata cctacagcgt gagctatgag aaagcgccac
     4021 gcttcccgaa gggagaaagg cggacaggta tccggtaagc ggcagggtcg gaacaggaga
     4081 gcgcacgagg gagcttccag ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg
     4141 ccacctctga cttgagcgtc gatttttgtg atgctcgtca ggggggcgga gcctatggaa
     4201 aaacgccagc aacgcggcct ttttacggtt cctggccttt tgctggcctt ttgctcacat
     4261 gtt
//
