LOCUS       sequence-255145-        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     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(1852..1951)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(1968..1987)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1969..1987)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1992..2008)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1992..2008)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2110..2129)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             2121..2930
                     /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(2188..2207)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      3077..3665
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3566..3585
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3819..3836
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(3995..4022)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(4114..4200)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     join(4249..4263,1..8)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     join(4263,1..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 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 acgaggccgt gccgacttga gacccagctt
     1861 tcttgtacaa agttggcatt ataagaaagc attgcttatc aatttgttgc aacgaacagg
     1921 tcactatcag tcaaaataaa atcattattt gccatccagc tgatatcccc tatagtgagt
     1981 cgtattacat ggtcatagct gtttcctggc agctctggcc cgtgtctcaa aatctctgat
     2041 gttacattgc acaagataaa aatatatcat catgaacaat aaaactgtct gcttacataa
     2101 acagtaatac aaggggtgtt atgagccata ttcaacggga aacgtcgagg ccgcgattaa
     2161 attccaacat ggatgctgat ttatatgggt ataaatgggc tcgcgataat gtcgggcaat
     2221 caggtgcgac aatctatcgc ttgtatggga agcccgatgc gccagagttg tttctgaaac
     2281 atggcaaagg tagcgttgcc aatgatgtta cagatgagat ggtcagacta aactggctga
     2341 cggaatttat gcctcttccg accatcaagc attttatccg tactcctgat gatgcatggt
     2401 tactcaccac tgcgatcccc ggaaaaacag cattccaggt attagaagaa tatcctgatt
     2461 caggtgaaaa tattgttgat gcgctggcag tgttcctgcg ccggttgcat tcgattcctg
     2521 tttgtaattg tccttttaac agcgatcgcg tatttcgtct cgctcaggcg caatcacgaa
     2581 tgaataacgg tttggttgat gcgagtgatt ttgatgacga gcgtaatggc tggcctgttg
     2641 aacaagtctg gaaagaaatg cataaacttt tgccattctc accggattca gtcgtcactc
     2701 atggtgattt ctcacttgat aaccttattt ttgacgaggg gaaattaata ggttgtattg
     2761 atgttggacg agtcggaatc gcagaccgat accaggatct tgccatccta tggaactgcc
     2821 tcggtgagtt ttctccttca ttacagaaac ggctttttca aaaatatggt attgataatc
     2881 ctgatatgaa taaattgcag tttcatttga tgctcgatga gtttttctaa tcagaattgg
     2941 ttaattggtt gtaacactgg cagagcatta cgctgacttg acgggacggc gcaagctcat
     3001 gaccaaaatc ccttaacgtg agttacgcgt cgttccactg agcgtcagac cccgtagaaa
     3061 agatcaaagg atcttcttga gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa
     3121 aaaaaccacc gctaccagcg gtggtttgtt tgccggatca agagctacca actctttttc
     3181 cgaaggtaac tggcttcagc agagcgcaga taccaaatac tgttcttcta gtgtagccgt
     3241 agttaggcca ccacttcaag aactctgtag caccgcctac atacctcgct ctgctaatcc
     3301 tgttaccagt ggctgctgcc agtggcgata agtcgtgtct taccgggttg gactcaagac
     3361 gatagttacc ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc acacagccca
     3421 gcttggagcg aacgacctac accgaactga gatacctaca gcgtgagcta tgagaaagcg
     3481 ccacgcttcc cgaagggaga aaggcggaca ggtatccggt aagcggcagg gtcggaacag
     3541 gagagcgcac gagggagctt ccagggggaa acgcctggta tctttatagt cctgtcgggt
     3601 ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg cggagcctat
     3661 ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc cttttgctgg ccttttgctc
     3721 acatgttctt tcctgcgtta tcccctgatt ctgtggataa ccgtattacc gcctttgagt
     3781 gagctgatac cgctcgccgc agccgaacga ccgagcgcag cgagtcagtg agcgaggaag
     3841 cggaagagcg cccaatacgc aaaccgcctc tccccgcgcg ttggccgatt cattaatgca
     3901 gctggcacga caggtttccc gactggaaag cgggcagtga gcgcaacgca attaatacgc
     3961 gtaccgctag ccaggaagag tttgtagaaa cgcaaaaagg ccatccgtca ggatggcctt
     4021 ctgcttagtt tgatgcctgg cagtttatgg cgggcgtcct gcccgccacc ctccgggccg
     4081 ttgcttcaca acgttcaaat ccgctcccgg cggatttgtc ctactcagga gagcgttcac
     4141 cgacaaacaa cagataaaac gaaaggccca gtcttccgac tgagcctttc gttttatttg
     4201 atgcctggca gttccctact ctcgcgttaa cgctagcatg gatgttttcc cagtcacgac
     4261 gtt
//
