LOCUS       sequence-258110-        6823 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 6823)
  AUTHORS   McKenna JF, Gumber HK, Turpin ZM, Jalovec AM, Kartick AC, Graumann 
            K, Bass HW
  TITLE     Maize (Zea mays L.) Nucleoskeletal Proteins Regulate Nuclear 
            Envelope Remodeling and Function in Stomatal Complex Development and
            Pollen Viability.
  JOURNAL   Front Plant Sci. 2021 Feb 17;12:645218. doi: 
            10.3389/fpls.2021.645218. eCollection 2021.
  PUBMED    33679862
REFERENCE   2  (bases 1 to 6823)
  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: "Front Plant
            Sci."; date: "2021-02-17"; volume: "12:645218. doi"; pages: " 
            10.3389/fpls.2021.645218. eCollection 2021"
COMMENT     Sequence Label: NCH1ec
FEATURES             Location/Qualifiers
     source          1..6823
                     /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(4412..4511)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(4528..4547)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(4529..4547)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(4552..4568)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4552..4568)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4670..4689)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             4681..5490
                     /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(4748..4767)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      5637..6225
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6126..6145
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6379..6396
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(6555..6582)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(6674..6760)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     join(6809..6823,1..8)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     join(6823,1..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 gtcccaatcc
      181 tcgtcgagct ggacggggat gttaacggcc acaagttctc cgtgagcggc gagggggagg
      241 gcgacgccac ctacgggaag ctcacgctga agttcatttg cacgaccggc aagctgccgg
      301 tgccctggcc aaccctggtc acaactctga cgtacggcgt ccagtgcttc tcccgctacc
      361 cggaccacat gaagcagcat gatttcttca agagcgccat gccggagggc tacgttcagg
      421 agcgcaccat cttcttcaag gacgatggca actacaagac aagggcggag gttaagttcg
      481 agggcgacac tctcgtgaac cggatcgagc tgaagggcat tgacttcaag gaggatggga
      541 atattctcgg ccacaagctg gagtacaact acaattcgca taacgtctac atcatggctg
      601 acaagcagaa gaacggcatt aaggttaatt tcaagatccg ccacaatatt gaggacggct
      661 ccgtgcagct cgcggatcat taccagcaga acacgccgat cggggacggc cctgtcctcc
      721 tgccggataa tcactacctc tcgacgcagt ctgcgctgtc aaaggacccg aacgagaaga
      781 gggatcatat ggttctcctg gagttcgtga cagccgcggg gatcactctc ggcatggacg
      841 agctgtacaa gggggcgggc gactacaagg acgatgacga taaggcttac ccctacgacg
      901 tgccagatta cgctggggcc ggcggatcca tgtttactcc gcagggcaaa ggatggaccg
      961 gctggtcgac gccaactccg gccaaccagc ggagcggtgg tggggctcct gctgcgtcgg
     1021 cgccccttgg aaagggcaaa ggaagggtcg ccgagctcga gcaggagttg catgaatatc
     1081 agtacaatat ggggcttctt ttgattgaaa agaaagaatg ggcagcaaaa tttgaagaaa
     1141 tcagtgaagt gctgacacag aaagaggaga tactgaagag ggagcaggcg gcacatctga
     1201 atgctatatc agagtatgaa aggcgggagg agaacatgcg gaaagcacta ggtgttgaga
     1261 agcaatgtgt cgctgatctt gaaaaagcct tgcgtgatat acgagcagag atagctgagg
     1321 ttaaatttac atctgagaaa aagataacag atgctcagtc tcttgaggct agtttggagg
     1381 aaaaatcctt ggagatcgag ggaaaacttc atgctgcaga tgccaaactt gcagaggcaa
     1441 accgcaagaa gtctcaagct gaccgtgatt tagaagaggc ggaagctcgt caacgtagac
     1501 tggagaagga gaaactgtat tttgagacag agcggaaggc ccgagaaaag caacttaagg
     1561 agcaggagga atcactgcag gaatgggaga aaaagctgaa ggaaagtcag aatagactta
     1621 atgagttgca gagatctata aatgagcgag aggagagagc aaataagaat gatcaacttt
     1681 tcaagataaa acaggacgaa ctggaggagg cacgaaggac agtggaggcc gctaaagtta
     1741 ctctgaaagt caaagaggat gatattaaca aaagactaaa tgaattacat ttacaggaaa
     1801 aggatgctga ctcaaagcgc agtgcattgg aagagcaggg gaaaaaattg gatgagagag
     1861 aggcaaaggt cactaataga gaaaaggagg ggctccagaa gcttcttgag gatcatcagg
     1921 tggagcttga atccaaaaga cgggattttg aattagaatt agagagggaa aggaaatctt
     1981 ttgatcaaaa tatgacacag aagcaagcgg atttattgaa aagggagaaa gatgttaaat
     2041 ctttggaggc taaactttct aaaagtgagc aggcattgaa tgataagaag aaatcaatgg
     2101 aaaacttgca gaatgatctt gatgccaaat ctaaggctct gaagagttgg gatgaatcat
     2161 tgaaaaatga cgagaaaagg ttactgaaag aaaagcagca aatggatcat gaaagagaac
     2221 aattggagac atacaaattg gagcttgaaa agatcaaatc agcattagaa gctgagaagg
     2281 aaaaaatatc agaagagcag aataatctga aactcacagc gcaagagagg caggagcata
     2341 gcttgttgat agcaaaactt aagaaggaga ttgaggaata cagaatgcga agcaactccc
     2401 tatctgagga aatggaggat ttaaggaagc aacggcagaa atttgaagaa gagtgggaac
     2461 aactagacga aaagagagcc ctccttgtag aggaggacaa aaggttgaac attgagagga
     2521 tgaatttaga acggtggcgt gacaacgaag aaaaaaggtt aaatgatatg aaactcaaaa
     2581 tggatgaaga atacaagcag cagctggaga gtcttgagcg caaggaaaag gctttgagtg
     2641 atgatatgaa gcacaagcaa atggaaaatg atgaatttct gaagggagaa cgtgctgatg
     2701 ttcaacgtaa attgcaactc aaacggcatg aattggagat ggagatggag caaaaacaag
     2761 ccacgaagga gaaggaactg gaagaaaagg aaaatgagtt aaataagaaa atagattttg
     2821 tggaaaataa actccgacat gcaattgagt tgaatgagtc caagattgag aagttattat
     2881 tggagaaaag ggaggtgcag atggaaagag aattactgct ggaggaaaga aagaaaacgg
     2941 agactgacaa ggcagatatt agaagggata ttgagagtct gcactcattg agcaagagtt
     3001 tgaaggaacg tcgagaggca tataacaggg acaggagcag actcatcgaa ctatttgaaa
     3061 agtacaaggc gtgcaaaaac tgtgggattt ctatttttga gggattggat tctctgctgc
     3121 ttaaagacag tgctgaaatt gagcatccaa gtttggctgt tgagggagat gatcatgcct
     3181 tgaccaccga tacatcaggt ccagatacag gaacccttgt taattcaggt ggtcgttttt
     3241 ctttgttaca gaagtgttca aggcttttca agttctcccc tagaaaaaag ggtgagcagt
     3301 catcagaaca accatcagag aggaatatat cttttggtgc aagactcgag gaagcaacac
     3361 agagcgatgg agattatgtg ccaactcctg tctatgagat agcccacgat tctttcaatg
     3421 cagaggatga gctcccttct gacggtgaaa ccagggagaa tgaagagtct gaaaggcatg
     3481 acatagctga taatgcccag atggagtctt ctgttggtgt agcagataac agtatagata
     3541 ttctcggtac taagtcattt gatggtgcta acgacatggc agttgaagca actattgtct
     3601 ctgttgatca aaatggcgag gactctgtag ttcctgctga agcaggtgta gagcctgaaa
     3661 catcagagca aggtcggcgc cagcaaaacc agagaggcag aagaaaaggt ggtatgaaaa
     3721 gaacacggtc tgttaatgct gtagttgaag atgctaagat gatacttggc gaggtttttg
     3781 aggagaagac tgatgatcag ggagatacag tcaaggtggg tgctacacga aagcggcggt
     3841 ttgctggagc tacaataagt gaacaagatg aagaagggag cgaagcacat tcagaaagtg
     3901 tttcacttgg agggcagcgc cgtaagaggc ggcaaactgc tggggcagtg acagatgccc
     3961 caggagaaag acgttacaat ttgagacaca gtagagctgc aaatgctggc gctgcaacag
     4021 cccaagcaga caaaaaaaaa tcttcaaaag caagaaataa acatacagta gaagcttctg
     4081 ctgatgacac cgaaggaaca tcaaaggttg ttgaggaacc tgcccctgag agcaagaggg
     4141 tatctgaatc tgcagactac gatgcatccc agttgcatga gttttcccag gttgaagttg
     4201 gtgatgccca tgctgtagac ggtgagggtg ctggtgagga agatggtgat attttggatg
     4261 gccaagatgc actgcctgat gtgccaatga caccttctgg aagcgagttt ggagcggaac
     4321 aggaagatga ggacgatgat gattcagaga ggcgaaacca atcaatcagc aagaagctat
     4381 ggtcattctt cactacatga cctgcaggca gacccagctt tcttgtacaa agttggcatt
     4441 ataagaaagc attgcttatc aatttgttgc aacgaacagg tcactatcag tcaaaataaa
     4501 atcattattt gccatccagc tgatatcccc tatagtgagt cgtattacat ggtcatagct
     4561 gtttcctggc agctctggcc cgtgtctcaa aatctctgat gttacattgc acaagataaa
     4621 aatatatcat catgaacaat aaaactgtct gcttacataa acagtaatac aaggggtgtt
     4681 atgagccata ttcaacggga aacgtcgagg ccgcgattaa attccaacat ggatgctgat
     4741 ttatatgggt ataaatgggc tcgcgataat gtcgggcaat caggtgcgac aatctatcgc
     4801 ttgtatggga agcccgatgc gccagagttg tttctgaaac atggcaaagg tagcgttgcc
     4861 aatgatgtta cagatgagat ggtcagacta aactggctga cggaatttat gcctcttccg
     4921 accatcaagc attttatccg tactcctgat gatgcatggt tactcaccac tgcgatcccc
     4981 ggaaaaacag cattccaggt attagaagaa tatcctgatt caggtgaaaa tattgttgat
     5041 gcgctggcag tgttcctgcg ccggttgcat tcgattcctg tttgtaattg tccttttaac
     5101 agcgatcgcg tatttcgtct cgctcaggcg caatcacgaa tgaataacgg tttggttgat
     5161 gcgagtgatt ttgatgacga gcgtaatggc tggcctgttg aacaagtctg gaaagaaatg
     5221 cataaacttt tgccattctc accggattca gtcgtcactc atggtgattt ctcacttgat
     5281 aaccttattt ttgacgaggg gaaattaata ggttgtattg atgttggacg agtcggaatc
     5341 gcagaccgat accaggatct tgccatccta tggaactgcc tcggtgagtt ttctccttca
     5401 ttacagaaac ggctttttca aaaatatggt attgataatc ctgatatgaa taaattgcag
     5461 tttcatttga tgctcgatga gtttttctaa tcagaattgg ttaattggtt gtaacactgg
     5521 cagagcatta cgctgacttg acgggacggc gcaagctcat gaccaaaatc ccttaacgtg
     5581 agttacgcgt cgttccactg agcgtcagac cccgtagaaa agatcaaagg atcttcttga
     5641 gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg
     5701 gtggtttgtt tgccggatca agagctacca actctttttc cgaaggtaac tggcttcagc
     5761 agagcgcaga taccaaatac tgttcttcta gtgtagccgt agttaggcca ccacttcaag
     5821 aactctgtag caccgcctac atacctcgct ctgctaatcc tgttaccagt ggctgctgcc
     5881 agtggcgata agtcgtgtct taccgggttg gactcaagac gatagttacc ggataaggcg
     5941 cagcggtcgg gctgaacggg gggttcgtgc acacagccca gcttggagcg aacgacctac
     6001 accgaactga gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga
     6061 aaggcggaca ggtatccggt aagcggcagg gtcggaacag gagagcgcac gagggagctt
     6121 ccagggggaa acgcctggta tctttatagt cctgtcgggt ttcgccacct ctgacttgag
     6181 cgtcgatttt tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg
     6241 gcctttttac ggttcctggc cttttgctgg ccttttgctc acatgttctt tcctgcgtta
     6301 tcccctgatt ctgtggataa ccgtattacc gcctttgagt gagctgatac cgctcgccgc
     6361 agccgaacga ccgagcgcag cgagtcagtg agcgaggaag cggaagagcg cccaatacgc
     6421 aaaccgcctc tccccgcgcg ttggccgatt cattaatgca gctggcacga caggtttccc
     6481 gactggaaag cgggcagtga gcgcaacgca attaatacgc gtaccgctag ccaggaagag
     6541 tttgtagaaa cgcaaaaagg ccatccgtca ggatggcctt ctgcttagtt tgatgcctgg
     6601 cagtttatgg cgggcgtcct gcccgccacc ctccgggccg ttgcttcaca acgttcaaat
     6661 ccgctcccgg cggatttgtc ctactcagga gagcgttcac cgacaaacaa cagataaaac
     6721 gaaaggccca gtcttccgac tgagcctttc gttttatttg atgcctggca gttccctact
     6781 ctcgcgttaa cgctagcatg gatgttttcc cagtcacgac gtt
//
