LOCUS       sequence-255538-        4248 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 4248)
  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 4248)
  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: MLKP1delKASHec
FEATURES             Location/Qualifiers
     source          1..4248
                     /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(1837..1936)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(1953..1972)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1954..1972)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1977..1993)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1977..1993)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2095..2114)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             2106..2915
                     /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(2173..2192)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      3062..3650
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3551..3570
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3804..3821
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(3980..4007)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(4099..4185)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     join(4234..4248,1..8)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     join(4248,1..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 gctgctgccg catgagaccc agctttcttg tacaaagttg
     1861 gcattataag aaagcattgc ttatcaattt gttgcaacga acaggtcact atcagtcaaa
     1921 ataaaatcat tatttgccat ccagctgata tcccctatag tgagtcgtat tacatggtca
     1981 tagctgtttc ctggcagctc tggcccgtgt ctcaaaatct ctgatgttac attgcacaag
     2041 ataaaaatat atcatcatga acaataaaac tgtctgctta cataaacagt aatacaaggg
     2101 gtgttatgag ccatattcaa cgggaaacgt cgaggccgcg attaaattcc aacatggatg
     2161 ctgatttata tgggtataaa tgggctcgcg ataatgtcgg gcaatcaggt gcgacaatct
     2221 atcgcttgta tgggaagccc gatgcgccag agttgtttct gaaacatggc aaaggtagcg
     2281 ttgccaatga tgttacagat gagatggtca gactaaactg gctgacggaa tttatgcctc
     2341 ttccgaccat caagcatttt atccgtactc ctgatgatgc atggttactc accactgcga
     2401 tccccggaaa aacagcattc caggtattag aagaatatcc tgattcaggt gaaaatattg
     2461 ttgatgcgct ggcagtgttc ctgcgccggt tgcattcgat tcctgtttgt aattgtcctt
     2521 ttaacagcga tcgcgtattt cgtctcgctc aggcgcaatc acgaatgaat aacggtttgg
     2581 ttgatgcgag tgattttgat gacgagcgta atggctggcc tgttgaacaa gtctggaaag
     2641 aaatgcataa acttttgcca ttctcaccgg attcagtcgt cactcatggt gatttctcac
     2701 ttgataacct tatttttgac gaggggaaat taataggttg tattgatgtt ggacgagtcg
     2761 gaatcgcaga ccgataccag gatcttgcca tcctatggaa ctgcctcggt gagttttctc
     2821 cttcattaca gaaacggctt tttcaaaaat atggtattga taatcctgat atgaataaat
     2881 tgcagtttca tttgatgctc gatgagtttt tctaatcaga attggttaat tggttgtaac
     2941 actggcagag cattacgctg acttgacggg acggcgcaag ctcatgacca aaatccctta
     3001 acgtgagtta cgcgtcgttc cactgagcgt cagaccccgt agaaaagatc aaaggatctt
     3061 cttgagatcc tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac
     3121 cagcggtggt ttgtttgccg gatcaagagc taccaactct ttttccgaag gtaactggct
     3181 tcagcagagc gcagatacca aatactgttc ttctagtgta gccgtagtta ggccaccact
     3241 tcaagaactc tgtagcaccg cctacatacc tcgctctgct aatcctgtta ccagtggctg
     3301 ctgccagtgg cgataagtcg tgtcttaccg ggttggactc aagacgatag ttaccggata
     3361 aggcgcagcg gtcgggctga acggggggtt cgtgcacaca gcccagcttg gagcgaacga
     3421 cctacaccga actgagatac ctacagcgtg agctatgaga aagcgccacg cttcccgaag
     3481 ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg
     3541 agcttccagg gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc cacctctgac
     3601 ttgagcgtcg atttttgtga tgctcgtcag gggggcggag cctatggaaa aacgccagca
     3661 acgcggcctt tttacggttc ctggcctttt gctggccttt tgctcacatg ttctttcctg
     3721 cgttatcccc tgattctgtg gataaccgta ttaccgcctt tgagtgagct gataccgctc
     3781 gccgcagccg aacgaccgag cgcagcgagt cagtgagcga ggaagcggaa gagcgcccaa
     3841 tacgcaaacc gcctctcccc gcgcgttggc cgattcatta atgcagctgg cacgacaggt
     3901 ttcccgactg gaaagcgggc agtgagcgca acgcaattaa tacgcgtacc gctagccagg
     3961 aagagtttgt agaaacgcaa aaaggccatc cgtcaggatg gccttctgct tagtttgatg
     4021 cctggcagtt tatggcgggc gtcctgcccg ccaccctccg ggccgttgct tcacaacgtt
     4081 caaatccgct cccggcggat ttgtcctact caggagagcg ttcaccgaca aacaacagat
     4141 aaaacgaaag gcccagtctt ccgactgagc ctttcgtttt atttgatgcc tggcagttcc
     4201 ctactctcgc gttaacgcta gcatggatgt tttcccagtc acgacgtt
//
