LOCUS       sequence-251055-        6261 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 6261)
  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 6261)
  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: NCH2ec
FEATURES             Location/Qualifiers
     source          1..6261
                     /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"
     RBS             3893..3901
                     /label=Shine-Dalgarno sequence
                     /note="full consensus sequence for ribosome-binding sites 
                     upstream of start codons in E. coli; complementary to a 
                     region in the 3' end of the 16S rRNA (Chen et al., 1994)"
     protein_bind    complement(4386..4485)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(4502..4521)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(4503..4521)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(4526..4542)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4526..4542)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4644..4663)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             4655..5464
                     /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(4722..4741)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      5611..6199
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6100..6119
                     /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 gatccatggc gagcccgcgc tcggccggcg gtgacgaggc
     1501 gatctggatg aagctgcgcg aggccgggtt cgacgaggac gctgtcaggc gccgcgacaa
     1561 ggcggccctc atcggctaca tctcgcggct cgagtccgag atctatgaat atcagcacaa
     1621 tcttgggctt attttattgg agcggaagga gttgacgtcc aaatacgaac aactcaaagc
     1681 ttcttttgaa gccactgaga tcatactcaa gcgagagcga gcttcgcagc agtctgcttt
     1741 atctgaaaca agaaagaggg aagaaaacct aaaaaaaaat ctagccattc agaaggagtg
     1801 catatcaaat cttgaaaagg cacttcatga tatgcgcggg gaaacagctg aaataaaagt
     1861 ttcatatgaa gcaaaactgg ctgaatccct tcaaatgatg gagacggcac agaaaaagtt
     1921 tgatgaagca gaagagaaac ttctcactgc aaaatcatta gaagcagact gcattcgaac
     1981 tcgcaacacc tctctgagaa gactgcaaga cattgaagat cgcgaagatc agctaagaag
     2041 ataccaaaca tcttttgaac ttgagaatgc atcgaaagag aaagagatta acctcctgag
     2101 gaaatcgttg gatgatacca aaaaaatttt gcatgaaaag gagcaatgtt tactgaaaga
     2161 acaagtactt cttaatcaga gggatgacag catccttgag agacttgcgt acataactag
     2221 ctcagagaaa aggttggagg aggaaaagtt aaatctcgaa gatgaaagaa aggttctttt
     2281 ggaagagaaa aataagctgg atctgaacat gcaggcaatt atatccaggg aggaagctat
     2341 aattcagaag gaatccatac ttgacaaacg tgaaagtgaa ctattgattc tccaagagac
     2401 cattgcaagc aaagaaaggg ctgaaataga aaggctccgt caagaggaag aaattgcctt
     2461 ggtgaggagg aggcaggaat ttgacactga catggaaatt aagctcactt cttttgagga
     2521 agagattgat gcaaggaaag ccttgctcga tcaaagggaa accaccatca atgagaagga
     2581 ggatgcagtg gcgcaaaggg aacagaatct caaccttagg tttgctgagt tagcgaacaa
     2641 ggaagaatct ttggtgaaga aatcagatga gttaagggag gaagagaaaa ggctctcctc
     2701 tgagagagag acactgcaca ttgatctgca aaaagaaaag gaggaaattc ataacatgaa
     2761 attggattta gagaaggaaa aatctttctt tgaggaggag aagcgtgaag cgattcaagc
     2821 tcaggagaat ctagcaataa cccaaaatga gcgagaagat ttacaaagtt tgcaggtgaa
     2881 acttaaagat gaaattgata gtctgagagc ccaaaaggtt gacctaatgg ctgatgcaga
     2941 aaggctatta tcagaaaaag aaaggtttga gattgaatgg gagttgatcg atgagaagaa
     3001 ggaggaacta caaaaggagg cagccaggat tgctgaagaa cgaagagtaa tagatgagca
     3061 tctcaagagc gaatttgata ttatcaaaca agaaaaggaa gatctccgtg ttcagttaaa
     3121 aattagtaca gaatcccttg ctcatgaaca tgctgaattc atgaataaga tgcagcaaga
     3181 gcatgctagt tggcttagca ggatacaatt agaaagagaa gatctgaaga aagatattga
     3241 tatccagaga accgagttac tgaattctgc aaaggctagg cagatggaaa tagattcata
     3301 tttgagggaa aaagaagagg agtttgaaca gaaaaaatcc aaggaacttg aatacatcaa
     3361 ttctgaaaag gaaactatca gctcaaaact agaacatgtc aggcttgaat tgcaaaaact
     3421 tgaggaggag aggaaaaggt ctatgctgga gcgagagaga agagaggaag agctgtctga
     3481 aacaaagaag actatagacg cgcttaatga gcaacgcgag aagttacaag agcaaagaaa
     3541 actgcttcat tcagaccgaa aatcaattac acagcaaatg cagctgctta atgagttaga
     3601 agagctgaag attgaatctg agaacaggca actgtctttg agacagtgtg ggaaatcaaa
     3661 gcatgctggt gtggaaaacc tggaagacaa cggtgttcac ctgtctcctg atgaggatca
     3721 aaatgctagt ccaaaacaaa ctactgtaaa gaaattggaa gtgtcccctt cagtctccac
     3781 accaatttct tgggttaaaa aatgtgctca ggtgatattc aaacgctctc cagagaagag
     3841 tgctgatccc aacaatgata tacctccaaa gttgggaaat gtcaatgact gtacctcctt
     3901 agctactgcc tattcagatg gattgtttgc ttgtcacctg gaaaatggtg ctgagaaagt
     3961 accacaagct atcgacggtc tgaaagttgg gaaaaagagg cttaataatg cactctctca
     4021 tggtgactcc gaaatttcac agccaaaacg caagcagcaa aagagcgcta ctcagatttt
     4081 gagagtgatt ggaggagaaa ttgattccaa ctgcccatca gttctagaag aaaaatgttc
     4141 aaagaatgaa catgatgcag tcccagttgg tttgtgtggg aagggaccac aaaatcccca
     4201 agcagagcat tcagaggata tttcagtgtc tgcatctgaa ccgtcgaacg gacctggtgt
     4261 tgtggactct gaggacaagg atggagcaga tgaggagtct gacgatgaag gcgaggacga
     4321 ggaggaagag aagacgtcat cggcgaagaa gctgtggcgt ttcctcatca catgactgca
     4381 ggcagaccca gctttcttgt acaaagttgg cattataaga aagcattgct tatcaatttg
     4441 ttgcaacgaa caggtcacta tcagtcaaaa taaaatcatt atttgccatc cagctgatat
     4501 cccctatagt gagtcgtatt acatggtcat agctgtttcc tggcagctct ggcccgtgtc
     4561 tcaaaatctc tgatgttaca ttgcacaaga taaaaatata tcatcatgaa caataaaact
     4621 gtctgcttac ataaacagta atacaagggg tgttatgagc catattcaac gggaaacgtc
     4681 gaggccgcga ttaaattcca acatggatgc tgatttatat gggtataaat gggctcgcga
     4741 taatgtcggg caatcaggtg cgacaatcta tcgcttgtat gggaagcccg atgcgccaga
     4801 gttgtttctg aaacatggca aaggtagcgt tgccaatgat gttacagatg agatggtcag
     4861 actaaactgg ctgacggaat ttatgcctct tccgaccatc aagcatttta tccgtactcc
     4921 tgatgatgca tggttactca ccactgcgat ccccggaaaa acagcattcc aggtattaga
     4981 agaatatcct gattcaggtg aaaatattgt tgatgcgctg gcagtgttcc tgcgccggtt
     5041 gcattcgatt cctgtttgta attgtccttt taacagcgat cgcgtatttc gtctcgctca
     5101 ggcgcaatca cgaatgaata acggtttggt tgatgcgagt gattttgatg acgagcgtaa
     5161 tggctggcct gttgaacaag tctggaaaga aatgcataaa cttttgccat tctcaccgga
     5221 ttcagtcgtc actcatggtg atttctcact tgataacctt atttttgacg aggggaaatt
     5281 aataggttgt attgatgttg gacgagtcgg aatcgcagac cgataccagg atcttgccat
     5341 cctatggaac tgcctcggtg agttttctcc ttcattacag aaacggcttt ttcaaaaata
     5401 tggtattgat aatcctgata tgaataaatt gcagtttcat ttgatgctcg atgagttttt
     5461 ctaatcagaa ttggttaatt ggttgtaaca ctggcagagc attacgctga cttgacggga
     5521 cggcgcaagc tcatgaccaa aatcccttaa cgtgagttac gcgtcgttcc actgagcgtc
     5581 agaccccgta gaaaagatca aaggatcttc ttgagatcct ttttttctgc gcgtaatctg
     5641 ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt tgtttgccgg atcaagagct
     5701 accaactctt tttccgaagg taactggctt cagcagagcg cagataccaa atactgttct
     5761 tctagtgtag ccgtagttag gccaccactt caagaactct gtagcaccgc ctacatacct
     5821 cgctctgcta atcctgttac cagtggctgc tgccagtggc gataagtcgt gtcttaccgg
     5881 gttggactca agacgatagt taccggataa ggcgcagcgg tcgggctgaa cggggggttc
     5941 gtgcacacag cccagcttgg agcgaacgac ctacaccgaa ctgagatacc tacagcgtga
     6001 gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg
     6061 cagggtcgga acaggagagc gcacgaggga gcttccaggg ggaaacgcct ggtatcttta
     6121 tagtcctgtc gggtttcgcc acctctgact tgagcgtcga tttttgtgat gctcgtcagg
     6181 ggggcggagc ctatggaaaa acgccagcaa cgcggccttt ttacggttcc tggccttttg
     6241 ctggcctttt gctcacatgt t
//
