LOCUS       sequence-250391-        5250 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of plant nuclear envelope LINC protein fused to 
            fluorescent protein and affinity tags for cell biology.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5250)
  AUTHORS   Gumber HK, McKenna JF, Tolmie AF, Jalovec AM, Kartick AC, Graumann 
            K, Bass HW
  TITLE     MLKS2 is an ARM domain and F-actin-associated KASH protein that 
            functions in stomatal complex development and meiotic chromosome 
            segregation.
  JOURNAL   Nucleus. 2019 Dec;10(1):144-166. doi: 10.1080/19491034.2019.1629795.
  PUBMED    31221013
REFERENCE   2  (bases 1 to 5250)
  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"; doi: 
            "10.1080/19491034.2019"; journalName: "Nucleus"; date: "2019-12"; 
            volume: "10"; issue: "1"; pages: "144-166"
COMMENT     Sequence Label: MLKS2ec
FEATURES             Location/Qualifiers
     source          1..5250
                     /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(3375..3474)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(3491..3510)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(3492..3510)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(3515..3531)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3515..3531)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3633..3652)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             3644..4453
                     /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(3711..3730)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      4600..5188
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5089..5108
                     /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 agggcgagga gctgttcacc ggggtggtcc ccatcctggt
      721 ggagctggat ggcgacgtta acgggcacaa gttctcggtg tctggcgagg gggagggcga
      781 cgctacctac gggaagctca cgctgaagtt catctgcacg accggcaagc tgccggtgcc
      841 ctggccaaca ctcgtcacaa ctctgactta cggcgtccag tgcttctcgc gctaccctga
      901 tcacatgaag cagcatgact tcttcaagtc ggctatgccc gagggctacg tgcaggagcg
      961 gacgatcttc ttcaaggacg atggcaacta caagacacgc gcggaggtca agttcgaggg
     1021 ggacactctc gttaacagga tcgagctgaa gggcattgac ttcaaggagg atggcaatat
     1081 cctcgggcac aagctggagt acaactacaa ttcccataac gtgtacatca tggccgacaa
     1141 gcagaagaac ggcattaagg tcaatttcaa gatccggcac aacattgagg atggcagcgt
     1201 tcagctcgcg gaccattacc agcagaacac accgatcggg gatggccctg tcctcctgcc
     1261 cgacaatcac tacctctcca ctcagagcgc cctgtcgaag gacccgaacg agaagaggga
     1321 ccatatggtg ctcctggagt tcgtcacagc cgcgggcatc actctcggga tggatgagct
     1381 gtacaagggg gcgggcgact acaaggacga tgacgataag gcttacccct acgacgtgcc
     1441 ggattacgcc ggggctggca tggggcgctc actctccccg ctcctgaggc aggagctgga
     1501 caacctggat aaggacgccg attctcgcag ggctgccatg aaggctctca agtcatacgc
     1561 ccgccacctg gatagcaagt cgatccccca tttcctggcc gaggtctctg acacgaccgc
     1621 cgctggcgcg ggggccggca gcgggatgcc aaccggcggg gagttcacga tctcgctcta
     1681 cgaggtcctg gccagggttc acgggcggaa catcgtgccc cagatcggca atattatggc
     1741 gaccattatg tgcacgctct ccagctcggg cgggtcattc ccactgcacc aggcctgctc
     1801 caaggttgtg ccagctatcg ctaggcatgg cattgatccg tcggccccag acggcgagaa
     1861 ggccgggatc attgctagcc tctgccgccc actgtgctcg gctctcatgg gcgggagcca
     1921 ggacggcggc gctgccagcg gcgctgccct ctgcctgaag gccctggttg agtcttcaaa
     1981 ctggaggttc gcgtcgggcg agctggtgaa tgaggtctgc ctgaaggtgg ccggcgctat
     2041 gcacgactcg gccacacggt cgaacgctca tatggggctg gccatggcgc tcgtgaagca
     2101 caatggcctc atcgctgagg cctacgcgcg ctctatcgtc aggtccggcc tccagattct
     2161 ggacggggat accgctgact ccagctcgca gaagaggctg tccgccatcc agatgattaa
     2221 cttcttcatg aagttcgtgg acccgaggtg cctgtcttca gagctgggcc gggttatcga
     2281 tgtgatggag cgctgccaga atgaccggat gcccttcgtg cggggcgcgg ctttcgaggc
     2341 ctctcagtca gcgcgcaaca ttgccgcgca gaagggctcc aggcacgagg ttgggacctc
     2401 tcctatggtg ggctcaaact tccataagag gagggagaag tccccttgcc ggagcctctg
     2461 gggcgccaag aggtcccccg ctgcctctac gatggccgct gctagccctg tgcagttccg
     2521 ctctccggag tcacaggtcg ttgattccag catcatgaat ggctccacac tccacgactc
     2581 cccagtgagc gtcggccagt cgtcttgcaa cttcgaccag agcaggcgga ctaaccgcag
     2641 gctgtggtcg aatgacgcgg ttgatgtgtc tctgaaggac ggcctcttca tccagctgtg
     2701 ctccaacagc aattcgtacg aggacgatct gggcgaggtt tgcgatagcg aggtgaccga
     2761 cgctaacttc gagtgcacca atacgttcgc cgggttcgtc tctccctcac acaatggctc
     2821 catcagcagg gataagacgc cttcaccgcg ggcctcggac aggcccatca acattgacga
     2881 tgtgaagatc tactccacgc caaggaagct cctgcggctc ctgcagtcat cctacgactc
     2941 ggattctgcc tcaaacatcg ggcagagcac ggccaagctc aggggcctct cctcccccga
     3001 ccaggagcac aagggcctcg tcatttcttc agagcagatg cagtcgctgc attctgacaa
     3061 taagggcgat gagatgaagg acgagaacga gaccatctac atgcagaata gcaagggcag
     3121 gacacagact ctctcgaacg ctgacgcctc agtcctgtcc acaactgagg ccgagaacac
     3181 aagcatcaag acttcgcctg agattgagct gaatggggac gatgtttgca tcacctccag
     3241 cattggcaag acgcggaagt accgcgcgaa gttcaccctc ttcctgtccg tcatcgttat
     3301 tttcctcgct atcattacga tgctgatccg cattgagaac gacgatgaca gcgcgtacct
     3361 ggtgccgacc tgagacccag ctttcttgta caaagttggc attataagaa agcattgctt
     3421 atcaatttgt tgcaacgaac aggtcactat cagtcaaaat aaaatcatta tttgccatcc
     3481 agctgatatc ccctatagtg agtcgtatta catggtcata gctgtttcct ggcagctctg
     3541 gcccgtgtct caaaatctct gatgttacat tgcacaagat aaaaatatat catcatgaac
     3601 aataaaactg tctgcttaca taaacagtaa tacaaggggt gttatgagcc atattcaacg
     3661 ggaaacgtcg aggccgcgat taaattccaa catggatgct gatttatatg ggtataaatg
     3721 ggctcgcgat aatgtcgggc aatcaggtgc gacaatctat cgcttgtatg ggaagcccga
     3781 tgcgccagag ttgtttctga aacatggcaa aggtagcgtt gccaatgatg ttacagatga
     3841 gatggtcaga ctaaactggc tgacggaatt tatgcctctt ccgaccatca agcattttat
     3901 ccgtactcct gatgatgcat ggttactcac cactgcgatc cccggaaaaa cagcattcca
     3961 ggtattagaa gaatatcctg attcaggtga aaatattgtt gatgcgctgg cagtgttcct
     4021 gcgccggttg cattcgattc ctgtttgtaa ttgtcctttt aacagcgatc gcgtatttcg
     4081 tctcgctcag gcgcaatcac gaatgaataa cggtttggtt gatgcgagtg attttgatga
     4141 cgagcgtaat ggctggcctg ttgaacaagt ctggaaagaa atgcataaac ttttgccatt
     4201 ctcaccggat tcagtcgtca ctcatggtga tttctcactt gataacctta tttttgacga
     4261 ggggaaatta ataggttgta ttgatgttgg acgagtcgga atcgcagacc gataccagga
     4321 tcttgccatc ctatggaact gcctcggtga gttttctcct tcattacaga aacggctttt
     4381 tcaaaaatat ggtattgata atcctgatat gaataaattg cagtttcatt tgatgctcga
     4441 tgagtttttc taatcagaat tggttaattg gttgtaacac tggcagagca ttacgctgac
     4501 ttgacgggac ggcgcaagct catgaccaaa atcccttaac gtgagttacg cgtcgttcca
     4561 ctgagcgtca gaccccgtag aaaagatcaa aggatcttct tgagatcctt tttttctgcg
     4621 cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca gcggtggttt gtttgccgga
     4681 tcaagagcta ccaactcttt ttccgaaggt aactggcttc agcagagcgc agataccaaa
     4741 tactgttctt ctagtgtagc cgtagttagg ccaccacttc aagaactctg tagcaccgcc
     4801 tacatacctc gctctgctaa tcctgttacc agtggctgct gccagtggcg ataagtcgtg
     4861 tcttaccggg ttggactcaa gacgatagtt accggataag gcgcagcggt cgggctgaac
     4921 ggggggttcg tgcacacagc ccagcttgga gcgaacgacc tacaccgaac tgagatacct
     4981 acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc
     5041 ggtaagcggc agggtcggaa caggagagcg cacgagggag cttccagggg gaaacgcctg
     5101 gtatctttat agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg
     5161 ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac gcggcctttt tacggttcct
     5221 ggccttttgc tggccttttg ctcacatgtt
//
