LOCUS       sequence-258111-        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     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"
     RBS             3357..3365
                     /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(3850..3949)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(3966..3985)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(3967..3985)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(3990..4006)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3990..4006)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4108..4127)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             4119..4928
                     /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(4186..4205)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      5075..5663
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5564..5583
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5817..5834
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(5993..6020)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(6112..6198)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     join(6247..6261,1..8)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     join(6261,1..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 tggcgagccc gcgctcggcc ggcggtgacg
      961 aggcgatctg gatgaagctg cgcgaggccg ggttcgacga ggacgctgtc aggcgccgcg
     1021 acaaggcggc cctcatcggc tacatctcgc ggctcgagtc cgagatctat gaatatcagc
     1081 acaatcttgg gcttatttta ttggagcgga aggagttgac gtccaaatac gaacaactca
     1141 aagcttcttt tgaagccact gagatcatac tcaagcgaga gcgagcttcg cagcagtctg
     1201 ctttatctga aacaagaaag agggaagaaa acctaaaaaa aaatctagcc attcagaagg
     1261 agtgcatatc aaatcttgaa aaggcacttc atgatatgcg cggggaaaca gctgaaataa
     1321 aagtttcata tgaagcaaaa ctggctgaat cccttcaaat gatggagacg gcacagaaaa
     1381 agtttgatga agcagaagag aaacttctca ctgcaaaatc attagaagca gactgcattc
     1441 gaactcgcaa cacctctctg agaagactgc aagacattga agatcgcgaa gatcagctaa
     1501 gaagatacca aacatctttt gaacttgaga atgcatcgaa agagaaagag attaacctcc
     1561 tgaggaaatc gttggatgat accaaaaaaa ttttgcatga aaaggagcaa tgtttactga
     1621 aagaacaagt acttcttaat cagagggatg acagcatcct tgagagactt gcgtacataa
     1681 ctagctcaga gaaaaggttg gaggaggaaa agttaaatct cgaagatgaa agaaaggttc
     1741 ttttggaaga gaaaaataag ctggatctga acatgcaggc aattatatcc agggaggaag
     1801 ctataattca gaaggaatcc atacttgaca aacgtgaaag tgaactattg attctccaag
     1861 agaccattgc aagcaaagaa agggctgaaa tagaaaggct ccgtcaagag gaagaaattg
     1921 ccttggtgag gaggaggcag gaatttgaca ctgacatgga aattaagctc acttcttttg
     1981 aggaagagat tgatgcaagg aaagccttgc tcgatcaaag ggaaaccacc atcaatgaga
     2041 aggaggatgc agtggcgcaa agggaacaga atctcaacct taggtttgct gagttagcga
     2101 acaaggaaga atctttggtg aagaaatcag atgagttaag ggaggaagag aaaaggctct
     2161 cctctgagag agagacactg cacattgatc tgcaaaaaga aaaggaggaa attcataaca
     2221 tgaaattgga tttagagaag gaaaaatctt tctttgagga ggagaagcgt gaagcgattc
     2281 aagctcagga gaatctagca ataacccaaa atgagcgaga agatttacaa agtttgcagg
     2341 tgaaacttaa agatgaaatt gatagtctga gagcccaaaa ggttgaccta atggctgatg
     2401 cagaaaggct attatcagaa aaagaaaggt ttgagattga atgggagttg atcgatgaga
     2461 agaaggagga actacaaaag gaggcagcca ggattgctga agaacgaaga gtaatagatg
     2521 agcatctcaa gagcgaattt gatattatca aacaagaaaa ggaagatctc cgtgttcagt
     2581 taaaaattag tacagaatcc cttgctcatg aacatgctga attcatgaat aagatgcagc
     2641 aagagcatgc tagttggctt agcaggatac aattagaaag agaagatctg aagaaagata
     2701 ttgatatcca gagaaccgag ttactgaatt ctgcaaaggc taggcagatg gaaatagatt
     2761 catatttgag ggaaaaagaa gaggagtttg aacagaaaaa atccaaggaa cttgaataca
     2821 tcaattctga aaaggaaact atcagctcaa aactagaaca tgtcaggctt gaattgcaaa
     2881 aacttgagga ggagaggaaa aggtctatgc tggagcgaga gagaagagag gaagagctgt
     2941 ctgaaacaaa gaagactata gacgcgctta atgagcaacg cgagaagtta caagagcaaa
     3001 gaaaactgct tcattcagac cgaaaatcaa ttacacagca aatgcagctg cttaatgagt
     3061 tagaagagct gaagattgaa tctgagaaca ggcaactgtc tttgagacag tgtgggaaat
     3121 caaagcatgc tggtgtggaa aacctggaag acaacggtgt tcacctgtct cctgatgagg
     3181 atcaaaatgc tagtccaaaa caaactactg taaagaaatt ggaagtgtcc ccttcagtct
     3241 ccacaccaat ttcttgggtt aaaaaatgtg ctcaggtgat attcaaacgc tctccagaga
     3301 agagtgctga tcccaacaat gatatacctc caaagttggg aaatgtcaat gactgtacct
     3361 ccttagctac tgcctattca gatggattgt ttgcttgtca cctggaaaat ggtgctgaga
     3421 aagtaccaca agctatcgac ggtctgaaag ttgggaaaaa gaggcttaat aatgcactct
     3481 ctcatggtga ctccgaaatt tcacagccaa aacgcaagca gcaaaagagc gctactcaga
     3541 ttttgagagt gattggagga gaaattgatt ccaactgccc atcagttcta gaagaaaaat
     3601 gttcaaagaa tgaacatgat gcagtcccag ttggtttgtg tgggaaggga ccacaaaatc
     3661 cccaagcaga gcattcagag gatatttcag tgtctgcatc tgaaccgtcg aacggacctg
     3721 gtgttgtgga ctctgaggac aaggatggag cagatgagga gtctgacgat gaaggcgagg
     3781 acgaggagga agagaagacg tcatcggcga agaagctgtg gcgtttcctc atcacatgac
     3841 tgcaggcaga cccagctttc ttgtacaaag ttggcattat aagaaagcat tgcttatcaa
     3901 tttgttgcaa cgaacaggtc actatcagtc aaaataaaat cattatttgc catccagctg
     3961 atatccccta tagtgagtcg tattacatgg tcatagctgt ttcctggcag ctctggcccg
     4021 tgtctcaaaa tctctgatgt tacattgcac aagataaaaa tatatcatca tgaacaataa
     4081 aactgtctgc ttacataaac agtaatacaa ggggtgttat gagccatatt caacgggaaa
     4141 cgtcgaggcc gcgattaaat tccaacatgg atgctgattt atatgggtat aaatgggctc
     4201 gcgataatgt cgggcaatca ggtgcgacaa tctatcgctt gtatgggaag cccgatgcgc
     4261 cagagttgtt tctgaaacat ggcaaaggta gcgttgccaa tgatgttaca gatgagatgg
     4321 tcagactaaa ctggctgacg gaatttatgc ctcttccgac catcaagcat tttatccgta
     4381 ctcctgatga tgcatggtta ctcaccactg cgatccccgg aaaaacagca ttccaggtat
     4441 tagaagaata tcctgattca ggtgaaaata ttgttgatgc gctggcagtg ttcctgcgcc
     4501 ggttgcattc gattcctgtt tgtaattgtc cttttaacag cgatcgcgta tttcgtctcg
     4561 ctcaggcgca atcacgaatg aataacggtt tggttgatgc gagtgatttt gatgacgagc
     4621 gtaatggctg gcctgttgaa caagtctgga aagaaatgca taaacttttg ccattctcac
     4681 cggattcagt cgtcactcat ggtgatttct cacttgataa ccttattttt gacgagggga
     4741 aattaatagg ttgtattgat gttggacgag tcggaatcgc agaccgatac caggatcttg
     4801 ccatcctatg gaactgcctc ggtgagtttt ctccttcatt acagaaacgg ctttttcaaa
     4861 aatatggtat tgataatcct gatatgaata aattgcagtt tcatttgatg ctcgatgagt
     4921 ttttctaatc agaattggtt aattggttgt aacactggca gagcattacg ctgacttgac
     4981 gggacggcgc aagctcatga ccaaaatccc ttaacgtgag ttacgcgtcg ttccactgag
     5041 cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa
     5101 tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg ccggatcaag
     5161 agctaccaac tctttttccg aaggtaactg gcttcagcag agcgcagata ccaaatactg
     5221 ttcttctagt gtagccgtag ttaggccacc acttcaagaa ctctgtagca ccgcctacat
     5281 acctcgctct gctaatcctg ttaccagtgg ctgctgccag tggcgataag tcgtgtctta
     5341 ccgggttgga ctcaagacga tagttaccgg ataaggcgca gcggtcgggc tgaacggggg
     5401 gttcgtgcac acagcccagc ttggagcgaa cgacctacac cgaactgaga tacctacagc
     5461 gtgagctatg agaaagcgcc acgcttcccg aagggagaaa ggcggacagg tatccggtaa
     5521 gcggcagggt cggaacagga gagcgcacga gggagcttcc agggggaaac gcctggtatc
     5581 tttatagtcc tgtcgggttt cgccacctct gacttgagcg tcgatttttg tgatgctcgt
     5641 caggggggcg gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct
     5701 tttgctggcc ttttgctcac atgttctttc ctgcgttatc ccctgattct gtggataacc
     5761 gtattaccgc ctttgagtga gctgataccg ctcgccgcag ccgaacgacc gagcgcagcg
     5821 agtcagtgag cgaggaagcg gaagagcgcc caatacgcaa accgcctctc cccgcgcgtt
     5881 ggccgattca ttaatgcagc tggcacgaca ggtttcccga ctggaaagcg ggcagtgagc
     5941 gcaacgcaat taatacgcgt accgctagcc aggaagagtt tgtagaaacg caaaaaggcc
     6001 atccgtcagg atggccttct gcttagtttg atgcctggca gtttatggcg ggcgtcctgc
     6061 ccgccaccct ccgggccgtt gcttcacaac gttcaaatcc gctcccggcg gatttgtcct
     6121 actcaggaga gcgttcaccg acaaacaaca gataaaacga aaggcccagt cttccgactg
     6181 agcctttcgt tttatttgat gcctggcagt tccctactct cgcgttaacg ctagcatgga
     6241 tgttttccca gtcacgacgt t
//
