LOCUS       sequence-251057-        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     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(2521..2620)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(2637..2656)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2638..2656)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2661..2677)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2661..2677)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2779..2798)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             2790..3599
                     /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(2857..2876)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      3746..4334
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4235..4254
                     /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 gatccatgtc tgtctcggag aaggtggtgc tatcatcatc
     1501 agtttcttcg tcgtcagatt tagatccgtt gctgaagggt cttacggaga agaagttgag
     1561 ctttaggagg aacgtggtgt cattggcagc cgagctcaag gatgtgagga gcaagcttgc
     1621 ttcccaggag cagctgtttg taagggaatc tcaaactaga aagcttgtgg atacaaaagc
     1681 aaggagcatg gaagaggaag taagcaagtt gcagaaatgc ttacaggaca aggatgagca
     1741 actgcgttca tccacaagta gcacggaaca gtacctccat gagttggatg atcttagaac
     1801 acaactgtca tttactcaag ctacagcaga agcaagtgct gcatcagcta agtcagccca
     1861 gatgcagtgc ttatcattgc tgaaagagct gaatgagaag gacatctcgc tcaaagagca
     1921 cgggcttcga gtgaacaagc taggtgagca gcttgatctt ctccagaggg aactacaagc
     1981 aagggagctc tcgcaaatgc aacttaaaga tgaagttata agaatcgaga ccgacatcat
     2041 ggatgcagtt gccaaagctg ggtctaggtc tgacaaagat aatgagcttt taaagatctt
     2101 atctgatgtt tcaccaagga atgtccagaa cctcaacaac cttttgaatg ctaaggatac
     2161 agaaattgcg aggttgagag aggaaatcag aatattgtct gctcactgga cgaacaaaac
     2221 caaggaactt gagtcacatt tagaaaagca ccggagaact gatcaggagc tgaagaagag
     2281 ggttctaaag cttgaatttt gccttcaaga atcccagtct cagatgcgaa aattaaagag
     2341 gatgggcgag agaagagaca aggctctcaa ggagctcatg gaccaggtgg ccacgaagca
     2401 gccaaacagc atgtgccatg gcaacaagga gaatttctgg gagggccagg gcttcaagtt
     2461 catcgtgtct atgtcgatgc tggcactagt gatcctcgca aagcgatgac ctgcaggcag
     2521 acccagcttt cttgtacaaa gttggcatta taagaaagca ttgcttatca atttgttgca
     2581 acgaacaggt cactatcagt caaaataaaa tcattatttg ccatccagct gatatcccct
     2641 atagtgagtc gtattacatg gtcatagctg tttcctggca gctctggccc gtgtctcaaa
     2701 atctctgatg ttacattgca caagataaaa atatatcatc atgaacaata aaactgtctg
     2761 cttacataaa cagtaataca aggggtgtta tgagccatat tcaacgggaa acgtcgaggc
     2821 cgcgattaaa ttccaacatg gatgctgatt tatatgggta taaatgggct cgcgataatg
     2881 tcgggcaatc aggtgcgaca atctatcgct tgtatgggaa gcccgatgcg ccagagttgt
     2941 ttctgaaaca tggcaaaggt agcgttgcca atgatgttac agatgagatg gtcagactaa
     3001 actggctgac ggaatttatg cctcttccga ccatcaagca ttttatccgt actcctgatg
     3061 atgcatggtt actcaccact gcgatccccg gaaaaacagc attccaggta ttagaagaat
     3121 atcctgattc aggtgaaaat attgttgatg cgctggcagt gttcctgcgc cggttgcatt
     3181 cgattcctgt ttgtaattgt ccttttaaca gcgatcgcgt atttcgtctc gctcaggcgc
     3241 aatcacgaat gaataacggt ttggttgatg cgagtgattt tgatgacgag cgtaatggct
     3301 ggcctgttga acaagtctgg aaagaaatgc ataaactttt gccattctca ccggattcag
     3361 tcgtcactca tggtgatttc tcacttgata accttatttt tgacgagggg aaattaatag
     3421 gttgtattga tgttggacga gtcggaatcg cagaccgata ccaggatctt gccatcctat
     3481 ggaactgcct cggtgagttt tctccttcat tacagaaacg gctttttcaa aaatatggta
     3541 ttgataatcc tgatatgaat aaattgcagt ttcatttgat gctcgatgag tttttctaat
     3601 cagaattggt taattggttg taacactggc agagcattac gctgacttga cgggacggcg
     3661 caagctcatg accaaaatcc cttaacgtga gttacgcgtc gttccactga gcgtcagacc
     3721 ccgtagaaaa gatcaaagga tcttcttgag atcctttttt tctgcgcgta atctgctgct
     3781 tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa gagctaccaa
     3841 ctctttttcc gaaggtaact ggcttcagca gagcgcagat accaaatact gttcttctag
     3901 tgtagccgta gttaggccac cacttcaaga actctgtagc accgcctaca tacctcgctc
     3961 tgctaatcct gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt accgggttgg
     4021 actcaagacg atagttaccg gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca
     4081 cacagcccag cttggagcga acgacctaca ccgaactgag atacctacag cgtgagctat
     4141 gagaaagcgc cacgcttccc gaagggagaa aggcggacag gtatccggta agcggcaggg
     4201 tcggaacagg agagcgcacg agggagcttc cagggggaaa cgcctggtat ctttatagtc
     4261 ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt gtgatgctcg tcaggggggc
     4321 ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc ttttgctggc
     4381 cttttgctca catgtt
//
