LOCUS       sequence-250385-        4093 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 4093)
  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 4093)
  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: MLKG1delKASHec
FEATURES             Location/Qualifiers
     source          1..4093
                     /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(2218..2317)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(2334..2353)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2335..2353)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2358..2374)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2358..2374)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2476..2495)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             2487..3296
                     /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(2554..2573)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      3443..4031
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3932..3951
                     /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 caatcctcgt
      721 cgagctggac ggggatgtta acggccacaa gttctccgtg agcggcgagg gggagggcga
      781 cgccacctac gggaagctca cgctgaagtt catttgcacg accggcaagc tgccggtgcc
      841 ctggccaacc ctggtcacaa ctctgacgta cggcgtccag tgcttctccc gctacccgga
      901 ccacatgaag cagcatgatt tcttcaagag cgccatgccg gagggctacg ttcaggagcg
      961 caccatcttc ttcaaggacg atggcaacta caagacaagg gcggaggtta agttcgaggg
     1021 cgacactctc gtgaaccgga tcgagctgaa gggcattgac ttcaaggagg atgggaatat
     1081 tctcggccac aagctggagt acaactacaa ttcgcataac gtctacatca tggctgacaa
     1141 gcagaagaac ggcattaagg ttaatttcaa gatccgccac aatattgagg acggctccgt
     1201 gcagctcgcg gatcattacc agcagaacac gccgatcggg gacggccctg tcctcctgcc
     1261 ggataatcac tacctctcga cgcagtctgc gctgtcaaag gacccgaacg agaagaggga
     1321 tcatatggtt ctcctggagt tcgtgacagc cgcggggatc actctcggca tggacgagct
     1381 gtacaagggg gcgggcgact acaaggacga tgacgataag gcttacccct acgacgtgcc
     1441 agattacgct ggggccggcg gatccatgga catgccgccg gcggcgcagg gacggagggc
     1501 gatcgccgtg gtggaccgga agcttgacga gctctgcgcc tgcctcgacg acgcgatgtc
     1561 gtccaggcag cggagcccgt acgcggagga gcggctccgc gcggagatca agtcgagggc
     1621 cgacttcctt cggtcgctca tggccgccga ggtggactgc cacggcggga cgcgcccgga
     1681 gcacctggct gaggccgagg cgcgcttcgc cgtcctagag gccacgttcg atcggtgggc
     1741 gcggcgcgca gtcgctgcac ctgaaccagt cgtcgaggag aaggaggagg aggccgactc
     1801 cttggggtcc tcccggtcca cgtgctcctg caccgagtcg tgccaggagg cgacggaggg
     1861 ggacgccatg cgcgacgccg tggaaacacg cggcaaggag gggacgtcgg atggcatcgc
     1921 cgaggacaac gacgccgaat gcgagacggt ggtaatccgc agggaggagg cggcggcggt
     1981 tcctgttgcc aagaagcggg atgaggctat gggaacacgc gaggaggtgg cgctggacgc
     2041 cattgccaag aagaagggcg ataccgatcg cgatgccgcg gcggagacgg ggaggaggac
     2101 cgtagaccag cggcgctggt ggaggcggag tgctgcctgg tgcggtgcag ccggcgtcgt
     2161 cgcgttggtc gcggtcgggt tggcggtgga gttggtcgcg gtgtagcctg caggcagacc
     2221 cagctttctt gtacaaagtt ggcattataa gaaagcattg cttatcaatt tgttgcaacg
     2281 aacaggtcac tatcagtcaa aataaaatca ttatttgcca tccagctgat atcccctata
     2341 gtgagtcgta ttacatggtc atagctgttt cctggcagct ctggcccgtg tctcaaaatc
     2401 tctgatgtta cattgcacaa gataaaaata tatcatcatg aacaataaaa ctgtctgctt
     2461 acataaacag taatacaagg ggtgttatga gccatattca acgggaaacg tcgaggccgc
     2521 gattaaattc caacatggat gctgatttat atgggtataa atgggctcgc gataatgtcg
     2581 ggcaatcagg tgcgacaatc tatcgcttgt atgggaagcc cgatgcgcca gagttgtttc
     2641 tgaaacatgg caaaggtagc gttgccaatg atgttacaga tgagatggtc agactaaact
     2701 ggctgacgga atttatgcct cttccgacca tcaagcattt tatccgtact cctgatgatg
     2761 catggttact caccactgcg atccccggaa aaacagcatt ccaggtatta gaagaatatc
     2821 ctgattcagg tgaaaatatt gttgatgcgc tggcagtgtt cctgcgccgg ttgcattcga
     2881 ttcctgtttg taattgtcct tttaacagcg atcgcgtatt tcgtctcgct caggcgcaat
     2941 cacgaatgaa taacggtttg gttgatgcga gtgattttga tgacgagcgt aatggctggc
     3001 ctgttgaaca agtctggaaa gaaatgcata aacttttgcc attctcaccg gattcagtcg
     3061 tcactcatgg tgatttctca cttgataacc ttatttttga cgaggggaaa ttaataggtt
     3121 gtattgatgt tggacgagtc ggaatcgcag accgatacca ggatcttgcc atcctatgga
     3181 actgcctcgg tgagttttct ccttcattac agaaacggct ttttcaaaaa tatggtattg
     3241 ataatcctga tatgaataaa ttgcagtttc atttgatgct cgatgagttt ttctaatcag
     3301 aattggttaa ttggttgtaa cactggcaga gcattacgct gacttgacgg gacggcgcaa
     3361 gctcatgacc aaaatccctt aacgtgagtt acgcgtcgtt ccactgagcg tcagaccccg
     3421 tagaaaagat caaaggatct tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc
     3481 aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc
     3541 tttttccgaa ggtaactggc ttcagcagag cgcagatacc aaatactgtt cttctagtgt
     3601 agccgtagtt aggccaccac ttcaagaact ctgtagcacc gcctacatac ctcgctctgc
     3661 taatcctgtt accagtggct gctgccagtg gcgataagtc gtgtcttacc gggttggact
     3721 caagacgata gttaccggat aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac
     3781 agcccagctt ggagcgaacg acctacaccg aactgagata cctacagcgt gagctatgag
     3841 aaagcgccac gcttcccgaa gggagaaagg cggacaggta tccggtaagc ggcagggtcg
     3901 gaacaggaga gcgcacgagg gagcttccag ggggaaacgc ctggtatctt tatagtcctg
     3961 tcgggtttcg ccacctctga cttgagcgtc gatttttgtg atgctcgtca ggggggcgga
     4021 gcctatggaa aaacgccagc aacgcggcct ttttacggtt cctggccttt tgctggcctt
     4081 ttgctcacat gtt
//
