LOCUS       sequence-250369-        4129 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 4129)
  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 4129)
  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: ECPK1D
FEATURES             Location/Qualifiers
     source          1..4129
                     /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(2254..2353)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(2370..2389)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2371..2389)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2394..2410)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2394..2410)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2512..2531)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             2523..3332
                     /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(2590..2609)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      3479..4067
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3968..3987
                     /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 gtggctgacc acgatgtgaa
     2221 tgcttacgtt gttccgacgt agcctgcagg cagacccagc tttcttgtac aaagttggca
     2281 ttataagaaa gcattgctta tcaatttgtt gcaacgaaca ggtcactatc agtcaaaata
     2341 aaatcattat ttgccatcca gctgatatcc cctatagtga gtcgtattac atggtcatag
     2401 ctgtttcctg gcagctctgg cccgtgtctc aaaatctctg atgttacatt gcacaagata
     2461 aaaatatatc atcatgaaca ataaaactgt ctgcttacat aaacagtaat acaaggggtg
     2521 ttatgagcca tattcaacgg gaaacgtcga ggccgcgatt aaattccaac atggatgctg
     2581 atttatatgg gtataaatgg gctcgcgata atgtcgggca atcaggtgcg acaatctatc
     2641 gcttgtatgg gaagcccgat gcgccagagt tgtttctgaa acatggcaaa ggtagcgttg
     2701 ccaatgatgt tacagatgag atggtcagac taaactggct gacggaattt atgcctcttc
     2761 cgaccatcaa gcattttatc cgtactcctg atgatgcatg gttactcacc actgcgatcc
     2821 ccggaaaaac agcattccag gtattagaag aatatcctga ttcaggtgaa aatattgttg
     2881 atgcgctggc agtgttcctg cgccggttgc attcgattcc tgtttgtaat tgtcctttta
     2941 acagcgatcg cgtatttcgt ctcgctcagg cgcaatcacg aatgaataac ggtttggttg
     3001 atgcgagtga ttttgatgac gagcgtaatg gctggcctgt tgaacaagtc tggaaagaaa
     3061 tgcataaact tttgccattc tcaccggatt cagtcgtcac tcatggtgat ttctcacttg
     3121 ataaccttat ttttgacgag gggaaattaa taggttgtat tgatgttgga cgagtcggaa
     3181 tcgcagaccg ataccaggat cttgccatcc tatggaactg cctcggtgag ttttctcctt
     3241 cattacagaa acggcttttt caaaaatatg gtattgataa tcctgatatg aataaattgc
     3301 agtttcattt gatgctcgat gagtttttct aatcagaatt ggttaattgg ttgtaacact
     3361 ggcagagcat tacgctgact tgacgggacg gcgcaagctc atgaccaaaa tcccttaacg
     3421 tgagttacgc gtcgttccac tgagcgtcag accccgtaga aaagatcaaa ggatcttctt
     3481 gagatccttt ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag
     3541 cggtggtttg tttgccggat caagagctac caactctttt tccgaaggta actggcttca
     3601 gcagagcgca gataccaaat actgttcttc tagtgtagcc gtagttaggc caccacttca
     3661 agaactctgt agcaccgcct acatacctcg ctctgctaat cctgttacca gtggctgctg
     3721 ccagtggcga taagtcgtgt cttaccgggt tggactcaag acgatagtta ccggataagg
     3781 cgcagcggtc gggctgaacg gggggttcgt gcacacagcc cagcttggag cgaacgacct
     3841 acaccgaact gagataccta cagcgtgagc tatgagaaag cgccacgctt cccgaaggga
     3901 gaaaggcgga caggtatccg gtaagcggca gggtcggaac aggagagcgc acgagggagc
     3961 ttccaggggg aaacgcctgg tatctttata gtcctgtcgg gtttcgccac ctctgacttg
     4021 agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac gccagcaacg
     4081 cggccttttt acggttcctg gccttttgct ggccttttgc tcacatgtt
//
