LOCUS       sequence-251053-        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     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(4948..5047)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(5064..5083)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(5065..5083)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(5088..5104)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5088..5104)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5206..5225)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             5217..6026
                     /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(5284..5303)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      6173..6761
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6662..6681
                     /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 gatccatgtt tactccgcag ggcaaaggat ggaccggctg
     1501 gtcgacgcca actccggcca accagcggag cggtggtggg gctcctgctg cgtcggcgcc
     1561 ccttggaaag ggcaaaggaa gggtcgccga gctcgagcag gagttgcatg aatatcagta
     1621 caatatgggg cttcttttga ttgaaaagaa agaatgggca gcaaaatttg aagaaatcag
     1681 tgaagtgctg acacagaaag aggagatact gaagagggag caggcggcac atctgaatgc
     1741 tatatcagag tatgaaaggc gggaggagaa catgcggaaa gcactaggtg ttgagaagca
     1801 atgtgtcgct gatcttgaaa aagccttgcg tgatatacga gcagagatag ctgaggttaa
     1861 atttacatct gagaaaaaga taacagatgc tcagtctctt gaggctagtt tggaggaaaa
     1921 atccttggag atcgagggaa aacttcatgc tgcagatgcc aaacttgcag aggcaaaccg
     1981 caagaagtct caagctgacc gtgatttaga agaggcggaa gctcgtcaac gtagactgga
     2041 gaaggagaaa ctgtattttg agacagagcg gaaggcccga gaaaagcaac ttaaggagca
     2101 ggaggaatca ctgcaggaat gggagaaaaa gctgaaggaa agtcagaata gacttaatga
     2161 gttgcagaga tctataaatg agcgagagga gagagcaaat aagaatgatc aacttttcaa
     2221 gataaaacag gacgaactgg aggaggcacg aaggacagtg gaggccgcta aagttactct
     2281 gaaagtcaaa gaggatgata ttaacaaaag actaaatgaa ttacatttac aggaaaagga
     2341 tgctgactca aagcgcagtg cattggaaga gcaggggaaa aaattggatg agagagaggc
     2401 aaaggtcact aatagagaaa aggaggggct ccagaagctt cttgaggatc atcaggtgga
     2461 gcttgaatcc aaaagacggg attttgaatt agaattagag agggaaagga aatcttttga
     2521 tcaaaatatg acacagaagc aagcggattt attgaaaagg gagaaagatg ttaaatcttt
     2581 ggaggctaaa ctttctaaaa gtgagcaggc attgaatgat aagaagaaat caatggaaaa
     2641 cttgcagaat gatcttgatg ccaaatctaa ggctctgaag agttgggatg aatcattgaa
     2701 aaatgacgag aaaaggttac tgaaagaaaa gcagcaaatg gatcatgaaa gagaacaatt
     2761 ggagacatac aaattggagc ttgaaaagat caaatcagca ttagaagctg agaaggaaaa
     2821 aatatcagaa gagcagaata atctgaaact cacagcgcaa gagaggcagg agcatagctt
     2881 gttgatagca aaacttaaga aggagattga ggaatacaga atgcgaagca actccctatc
     2941 tgaggaaatg gaggatttaa ggaagcaacg gcagaaattt gaagaagagt gggaacaact
     3001 agacgaaaag agagccctcc ttgtagagga ggacaaaagg ttgaacattg agaggatgaa
     3061 tttagaacgg tggcgtgaca acgaagaaaa aaggttaaat gatatgaaac tcaaaatgga
     3121 tgaagaatac aagcagcagc tggagagtct tgagcgcaag gaaaaggctt tgagtgatga
     3181 tatgaagcac aagcaaatgg aaaatgatga atttctgaag ggagaacgtg ctgatgttca
     3241 acgtaaattg caactcaaac ggcatgaatt ggagatggag atggagcaaa aacaagccac
     3301 gaaggagaag gaactggaag aaaaggaaaa tgagttaaat aagaaaatag attttgtgga
     3361 aaataaactc cgacatgcaa ttgagttgaa tgagtccaag attgagaagt tattattgga
     3421 gaaaagggag gtgcagatgg aaagagaatt actgctggag gaaagaaaga aaacggagac
     3481 tgacaaggca gatattagaa gggatattga gagtctgcac tcattgagca agagtttgaa
     3541 ggaacgtcga gaggcatata acagggacag gagcagactc atcgaactat ttgaaaagta
     3601 caaggcgtgc aaaaactgtg ggatttctat ttttgaggga ttggattctc tgctgcttaa
     3661 agacagtgct gaaattgagc atccaagttt ggctgttgag ggagatgatc atgccttgac
     3721 caccgataca tcaggtccag atacaggaac ccttgttaat tcaggtggtc gtttttcttt
     3781 gttacagaag tgttcaaggc ttttcaagtt ctcccctaga aaaaagggtg agcagtcatc
     3841 agaacaacca tcagagagga atatatcttt tggtgcaaga ctcgaggaag caacacagag
     3901 cgatggagat tatgtgccaa ctcctgtcta tgagatagcc cacgattctt tcaatgcaga
     3961 ggatgagctc ccttctgacg gtgaaaccag ggagaatgaa gagtctgaaa ggcatgacat
     4021 agctgataat gcccagatgg agtcttctgt tggtgtagca gataacagta tagatattct
     4081 cggtactaag tcatttgatg gtgctaacga catggcagtt gaagcaacta ttgtctctgt
     4141 tgatcaaaat ggcgaggact ctgtagttcc tgctgaagca ggtgtagagc ctgaaacatc
     4201 agagcaaggt cggcgccagc aaaaccagag aggcagaaga aaaggtggta tgaaaagaac
     4261 acggtctgtt aatgctgtag ttgaagatgc taagatgata cttggcgagg tttttgagga
     4321 gaagactgat gatcagggag atacagtcaa ggtgggtgct acacgaaagc ggcggtttgc
     4381 tggagctaca ataagtgaac aagatgaaga agggagcgaa gcacattcag aaagtgtttc
     4441 acttggaggg cagcgccgta agaggcggca aactgctggg gcagtgacag atgccccagg
     4501 agaaagacgt tacaatttga gacacagtag agctgcaaat gctggcgctg caacagccca
     4561 agcagacaaa aaaaaatctt caaaagcaag aaataaacat acagtagaag cttctgctga
     4621 tgacaccgaa ggaacatcaa aggttgttga ggaacctgcc cctgagagca agagggtatc
     4681 tgaatctgca gactacgatg catcccagtt gcatgagttt tcccaggttg aagttggtga
     4741 tgcccatgct gtagacggtg agggtgctgg tgaggaagat ggtgatattt tggatggcca
     4801 agatgcactg cctgatgtgc caatgacacc ttctggaagc gagtttggag cggaacagga
     4861 agatgaggac gatgatgatt cagagaggcg aaaccaatca atcagcaaga agctatggtc
     4921 attcttcact acatgacctg caggcagacc cagctttctt gtacaaagtt ggcattataa
     4981 gaaagcattg cttatcaatt tgttgcaacg aacaggtcac tatcagtcaa aataaaatca
     5041 ttatttgcca tccagctgat atcccctata gtgagtcgta ttacatggtc atagctgttt
     5101 cctggcagct ctggcccgtg tctcaaaatc tctgatgtta cattgcacaa gataaaaata
     5161 tatcatcatg aacaataaaa ctgtctgctt acataaacag taatacaagg ggtgttatga
     5221 gccatattca acgggaaacg tcgaggccgc gattaaattc caacatggat gctgatttat
     5281 atgggtataa atgggctcgc gataatgtcg ggcaatcagg tgcgacaatc tatcgcttgt
     5341 atgggaagcc cgatgcgcca gagttgtttc tgaaacatgg caaaggtagc gttgccaatg
     5401 atgttacaga tgagatggtc agactaaact ggctgacgga atttatgcct cttccgacca
     5461 tcaagcattt tatccgtact cctgatgatg catggttact caccactgcg atccccggaa
     5521 aaacagcatt ccaggtatta gaagaatatc ctgattcagg tgaaaatatt gttgatgcgc
     5581 tggcagtgtt cctgcgccgg ttgcattcga ttcctgtttg taattgtcct tttaacagcg
     5641 atcgcgtatt tcgtctcgct caggcgcaat cacgaatgaa taacggtttg gttgatgcga
     5701 gtgattttga tgacgagcgt aatggctggc ctgttgaaca agtctggaaa gaaatgcata
     5761 aacttttgcc attctcaccg gattcagtcg tcactcatgg tgatttctca cttgataacc
     5821 ttatttttga cgaggggaaa ttaataggtt gtattgatgt tggacgagtc ggaatcgcag
     5881 accgatacca ggatcttgcc atcctatgga actgcctcgg tgagttttct ccttcattac
     5941 agaaacggct ttttcaaaaa tatggtattg ataatcctga tatgaataaa ttgcagtttc
     6001 atttgatgct cgatgagttt ttctaatcag aattggttaa ttggttgtaa cactggcaga
     6061 gcattacgct gacttgacgg gacggcgcaa gctcatgacc aaaatccctt aacgtgagtt
     6121 acgcgtcgtt ccactgagcg tcagaccccg tagaaaagat caaaggatct tcttgagatc
     6181 ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg
     6241 tttgtttgcc ggatcaagag ctaccaactc tttttccgaa ggtaactggc ttcagcagag
     6301 cgcagatacc aaatactgtt cttctagtgt agccgtagtt aggccaccac ttcaagaact
     6361 ctgtagcacc gcctacatac ctcgctctgc taatcctgtt accagtggct gctgccagtg
     6421 gcgataagtc gtgtcttacc gggttggact caagacgata gttaccggat aaggcgcagc
     6481 ggtcgggctg aacggggggt tcgtgcacac agcccagctt ggagcgaacg acctacaccg
     6541 aactgagata cctacagcgt gagctatgag aaagcgccac gcttcccgaa gggagaaagg
     6601 cggacaggta tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg gagcttccag
     6661 ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg ccacctctga cttgagcgtc
     6721 gatttttgtg atgctcgtca ggggggcgga gcctatggaa aaacgccagc aacgcggcct
     6781 ttttacggtt cctggccttt tgctggcctt ttgctcacat gtt
//
