LOCUS       sequence-258112-        4396 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 4396)
  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 4396)
  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: MNEAP1ec
FEATURES             Location/Qualifiers
     source          1..4396
                     /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(1985..2084)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(2101..2120)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2102..2120)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2125..2141)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2125..2141)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2243..2262)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             2254..3063
                     /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(2321..2340)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      3210..3798
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3699..3718
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3952..3969
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(4128..4155)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(4247..4333)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     join(4382..4396,1..8)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     join(4396,1..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 tgtctgtctc ggagaaggtg gtgctatcat
      961 catcagtttc ttcgtcgtca gatttagatc cgttgctgaa gggtcttacg gagaagaagt
     1021 tgagctttag gaggaacgtg gtgtcattgg cagccgagct caaggatgtg aggagcaagc
     1081 ttgcttccca ggagcagctg tttgtaaggg aatctcaaac tagaaagctt gtggatacaa
     1141 aagcaaggag catggaagag gaagtaagca agttgcagaa atgcttacag gacaaggatg
     1201 agcaactgcg ttcatccaca agtagcacgg aacagtacct ccatgagttg gatgatctta
     1261 gaacacaact gtcatttact caagctacag cagaagcaag tgctgcatca gctaagtcag
     1321 cccagatgca gtgcttatca ttgctgaaag agctgaatga gaaggacatc tcgctcaaag
     1381 agcacgggct tcgagtgaac aagctaggtg agcagcttga tcttctccag agggaactac
     1441 aagcaaggga gctctcgcaa atgcaactta aagatgaagt tataagaatc gagaccgaca
     1501 tcatggatgc agttgccaaa gctgggtcta ggtctgacaa agataatgag cttttaaaga
     1561 tcttatctga tgtttcacca aggaatgtcc agaacctcaa caaccttttg aatgctaagg
     1621 atacagaaat tgcgaggttg agagaggaaa tcagaatatt gtctgctcac tggacgaaca
     1681 aaaccaagga acttgagtca catttagaaa agcaccggag aactgatcag gagctgaaga
     1741 agagggttct aaagcttgaa ttttgccttc aagaatccca gtctcagatg cgaaaattaa
     1801 agaggatggg cgagagaaga gacaaggctc tcaaggagct catggaccag gtggccacga
     1861 agcagccaaa cagcatgtgc catggcaaca aggagaattt ctgggagggc cagggcttca
     1921 agttcatcgt gtctatgtcg atgctggcac tagtgatcct cgcaaagcga tgacctgcag
     1981 gcagacccag ctttcttgta caaagttggc attataagaa agcattgctt atcaatttgt
     2041 tgcaacgaac aggtcactat cagtcaaaat aaaatcatta tttgccatcc agctgatatc
     2101 ccctatagtg agtcgtatta catggtcata gctgtttcct ggcagctctg gcccgtgtct
     2161 caaaatctct gatgttacat tgcacaagat aaaaatatat catcatgaac aataaaactg
     2221 tctgcttaca taaacagtaa tacaaggggt gttatgagcc atattcaacg ggaaacgtcg
     2281 aggccgcgat taaattccaa catggatgct gatttatatg ggtataaatg ggctcgcgat
     2341 aatgtcgggc aatcaggtgc gacaatctat cgcttgtatg ggaagcccga tgcgccagag
     2401 ttgtttctga aacatggcaa aggtagcgtt gccaatgatg ttacagatga gatggtcaga
     2461 ctaaactggc tgacggaatt tatgcctctt ccgaccatca agcattttat ccgtactcct
     2521 gatgatgcat ggttactcac cactgcgatc cccggaaaaa cagcattcca ggtattagaa
     2581 gaatatcctg attcaggtga aaatattgtt gatgcgctgg cagtgttcct gcgccggttg
     2641 cattcgattc ctgtttgtaa ttgtcctttt aacagcgatc gcgtatttcg tctcgctcag
     2701 gcgcaatcac gaatgaataa cggtttggtt gatgcgagtg attttgatga cgagcgtaat
     2761 ggctggcctg ttgaacaagt ctggaaagaa atgcataaac ttttgccatt ctcaccggat
     2821 tcagtcgtca ctcatggtga tttctcactt gataacctta tttttgacga ggggaaatta
     2881 ataggttgta ttgatgttgg acgagtcgga atcgcagacc gataccagga tcttgccatc
     2941 ctatggaact gcctcggtga gttttctcct tcattacaga aacggctttt tcaaaaatat
     3001 ggtattgata atcctgatat gaataaattg cagtttcatt tgatgctcga tgagtttttc
     3061 taatcagaat tggttaattg gttgtaacac tggcagagca ttacgctgac ttgacgggac
     3121 ggcgcaagct catgaccaaa atcccttaac gtgagttacg cgtcgttcca ctgagcgtca
     3181 gaccccgtag aaaagatcaa aggatcttct tgagatcctt tttttctgcg cgtaatctgc
     3241 tgcttgcaaa caaaaaaacc accgctacca gcggtggttt gtttgccgga tcaagagcta
     3301 ccaactcttt ttccgaaggt aactggcttc agcagagcgc agataccaaa tactgttctt
     3361 ctagtgtagc cgtagttagg ccaccacttc aagaactctg tagcaccgcc tacatacctc
     3421 gctctgctaa tcctgttacc agtggctgct gccagtggcg ataagtcgtg tcttaccggg
     3481 ttggactcaa gacgatagtt accggataag gcgcagcggt cgggctgaac ggggggttcg
     3541 tgcacacagc ccagcttgga gcgaacgacc tacaccgaac tgagatacct acagcgtgag
     3601 ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc
     3661 agggtcggaa caggagagcg cacgagggag cttccagggg gaaacgcctg gtatctttat
     3721 agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg
     3781 gggcggagcc tatggaaaaa cgccagcaac gcggcctttt tacggttcct ggccttttgc
     3841 tggccttttg ctcacatgtt ctttcctgcg ttatcccctg attctgtgga taaccgtatt
     3901 accgcctttg agtgagctga taccgctcgc cgcagccgaa cgaccgagcg cagcgagtca
     3961 gtgagcgagg aagcggaaga gcgcccaata cgcaaaccgc ctctccccgc gcgttggccg
     4021 attcattaat gcagctggca cgacaggttt cccgactgga aagcgggcag tgagcgcaac
     4081 gcaattaata cgcgtaccgc tagccaggaa gagtttgtag aaacgcaaaa aggccatccg
     4141 tcaggatggc cttctgctta gtttgatgcc tggcagttta tggcgggcgt cctgcccgcc
     4201 accctccggg ccgttgcttc acaacgttca aatccgctcc cggcggattt gtcctactca
     4261 ggagagcgtt caccgacaaa caacagataa aacgaaaggc ccagtcttcc gactgagcct
     4321 ttcgttttat ttgatgcctg gcagttccct actctcgcgt taacgctagc atggatgttt
     4381 tcccagtcac gacgtt
//
