LOCUS       sequence-250366-        4647 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 4647)
  AUTHORS   Gumber HK, McKenna JF, Estrada AL, Tolmie AF, Graumann K, Bass HW
  TITLE     Identification and characterization of genes encoding the nuclear 
            envelope LINC complex in the monocot species Zea mays.
  JOURNAL   J Cell Sci. 2019 Feb 11;132(3). pii: jcs.221390. doi: 
            10.1242/jcs.221390.
  PUBMED    30659121
REFERENCE   2  (bases 1 to 4647)
  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: "J Cell 
            Sci."; date: "2019-02-11"; volume: "132(3). pii: jcs.221390. doi"; 
            pages: " 10.1242/jcs.221390"
COMMENT     Sequence Label: pHWBF06EC
FEATURES             Location/Qualifiers
     source          1..4647
                     /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             1388..1411
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             1415..1441
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     protein_bind    complement(2772..2871)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(2888..2907)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2889..2907)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2912..2928)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2912..2928)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3030..3049)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             3041..3850
                     /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(3108..3127)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      3997..4585
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4486..4505
                     /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 atggtgtcca agggcgagga ggacaacatg gccatcatta aggagttcat
      721 gcgcttcaag gtccacatgg agggctccgt taatgggcat gagttcgaga tcgaggggga
      781 gggcgagggg aggccctacg agggcacaca gactgccaag ctgaaggtga cgaagggcgg
      841 gcctctcccc ttcgcctggg acatcctgtc gcctcagttc atgtacggct ctaaggccta
      901 cgtgaagcac cccgctgaca ttccagatta cctcaagctg agcttcccgg agggcttcaa
      961 gtgggagcgg gttatgaact tcgaggacgg cggggtggtc acggtgaccc aggactccag
     1021 cctccaggat ggcgagttca tctacaaggt caagctgcgc ggcacaaatt tcccttccga
     1081 cgggccggtt atgcagaaga agactatggg ctgggaggcc tcgtctgagc ggatgtaccc
     1141 ggaggatggc gcgctcaagg gggagattaa gcagcgcctc aagctgaagg acggtggcca
     1201 ctacgatgct gaggtgaaga ccacgtacaa ggccaagaag ccggtccagc tcccaggcgc
     1261 ctacaacgtt aatatcaagc tggacattac atcgcacaac gaggattaca ctatcgtgga
     1321 gcagtacgag agggccgagg gcaggcattc tacgggcggg atggacgagc tgtacaaggg
     1381 cgccggggat tacaaggacg atgacgataa ggcgtaccct tacgacgtcc ccgattacgc
     1441 gggcgccggg atggcttctc cctcattcgc ggccgctgct acctctccgg cgtcatcccc
     1501 actcacgctg gacgctatcc cactggcctc aaggcccgcc ccaggcgccg ctgcggccca
     1561 gcggaagcgc tcggtgttcc tcctgaacca ccggcctcat ccggtctcgc ccaccccgcc
     1621 cctgcagcca accgctgcgg cctccgctca gcacgccagg aggaagaaga cctctcatcc
     1681 acctaggccc aggtggcaga ccgtgctctc catcgctgcc aagaatgccg ctctcctggc
     1741 ggccctcctg tacctcggcg acctggcctg gaggtggagc cacccgcccc cacctagccc
     1801 gcccccagac cgggctgccc tggagggcta cgcggctcgc gtcgatgagg ttgaggccag
     1861 cctcacacgg actttccgca tgatccaggt ccagctggag gcggttgaca ggaagattga
     1921 tggcgaagtg ggggcggcgc ggggcgacct cctggcgctc ctggaggata agaggctggc
     1981 tctggagagg cagctcacgc gcctggacgc tagggccggc gagctggggg atgcgctcgc
     2041 tggcctgaag aggatggagt tcctccggaa ggacgagttc gagaagttct gggatgaggt
     2101 gaagggctca ctgggctcct cctccgagtc ggaggtggac ctcgatcagg tccgggccct
     2161 ggcgcgcgag atcgtcatgc gcgagattga gaagcacgct gcggacggca tcgggcgggt
     2221 ggattacgct gtggcttccg gcgggggcag ggttgtgcgg catagcgagg cctacgttcc
     2281 caagaggggc ttcatggtgt ggatgtccgg cgtggacgtt gggcccaagc cagagaagat
     2341 gctgcagcct agcttcggcg agcccgggca gtgcttcgct ctccagggct caaacgggtt
     2401 cgtggaggtc aagctgaagt ccggcatcat tccagaggcg gtgaccctgg agcacgtttc
     2461 taaggacgtg gcctacgatc gctcaacggc gcctaagggc tgcagggtct acgggtggta
     2521 cgaggagacg cccggcgaga cccagtcggg gcatgccgct aagatggcgc tcgctgagtt
     2581 cgcctacgac ctggagaaga acaatgttca gacattcgac gtgactgcgc cggatgttgg
     2641 cgtgatcaac atggtgcgcc tcgacttcac atcgaatcac ggctcatccc agctcacttg
     2701 catctaccgc ctgcgggtgc acggccatga gcctgtcagc cccggcagct cggctgggtc
     2761 gcaggcctga gacccagctt tcttgtacaa agttggcatt ataagaaagc attgcttatc
     2821 aatttgttgc aacgaacagg tcactatcag tcaaaataaa atcattattt gccatccagc
     2881 tgatatcccc tatagtgagt cgtattacat ggtcatagct gtttcctggc agctctggcc
     2941 cgtgtctcaa aatctctgat gttacattgc acaagataaa aatatatcat catgaacaat
     3001 aaaactgtct gcttacataa acagtaatac aaggggtgtt atgagccata ttcaacggga
     3061 aacgtcgagg ccgcgattaa attccaacat ggatgctgat ttatatgggt ataaatgggc
     3121 tcgcgataat gtcgggcaat caggtgcgac aatctatcgc ttgtatggga agcccgatgc
     3181 gccagagttg tttctgaaac atggcaaagg tagcgttgcc aatgatgtta cagatgagat
     3241 ggtcagacta aactggctga cggaatttat gcctcttccg accatcaagc attttatccg
     3301 tactcctgat gatgcatggt tactcaccac tgcgatcccc ggaaaaacag cattccaggt
     3361 attagaagaa tatcctgatt caggtgaaaa tattgttgat gcgctggcag tgttcctgcg
     3421 ccggttgcat tcgattcctg tttgtaattg tccttttaac agcgatcgcg tatttcgtct
     3481 cgctcaggcg caatcacgaa tgaataacgg tttggttgat gcgagtgatt ttgatgacga
     3541 gcgtaatggc tggcctgttg aacaagtctg gaaagaaatg cataaacttt tgccattctc
     3601 accggattca gtcgtcactc atggtgattt ctcacttgat aaccttattt ttgacgaggg
     3661 gaaattaata ggttgtattg atgttggacg agtcggaatc gcagaccgat accaggatct
     3721 tgccatccta tggaactgcc tcggtgagtt ttctccttca ttacagaaac ggctttttca
     3781 aaaatatggt attgataatc ctgatatgaa taaattgcag tttcatttga tgctcgatga
     3841 gtttttctaa tcagaattgg ttaattggtt gtaacactgg cagagcatta cgctgacttg
     3901 acgggacggc gcaagctcat gaccaaaatc ccttaacgtg agttacgcgt cgttccactg
     3961 agcgtcagac cccgtagaaa agatcaaagg atcttcttga gatccttttt ttctgcgcgt
     4021 aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt tgccggatca
     4081 agagctacca actctttttc cgaaggtaac tggcttcagc agagcgcaga taccaaatac
     4141 tgttcttcta gtgtagccgt agttaggcca ccacttcaag aactctgtag caccgcctac
     4201 atacctcgct ctgctaatcc tgttaccagt ggctgctgcc agtggcgata agtcgtgtct
     4261 taccgggttg gactcaagac gatagttacc ggataaggcg cagcggtcgg gctgaacggg
     4321 gggttcgtgc acacagccca gcttggagcg aacgacctac accgaactga gatacctaca
     4381 gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca ggtatccggt
     4441 aagcggcagg gtcggaacag gagagcgcac gagggagctt ccagggggaa acgcctggta
     4501 tctttatagt cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc
     4561 gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc
     4621 cttttgctgg ccttttgctc acatgtt
//
