LOCUS       sequence-255563-        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     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(2839..2938)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(2955..2974)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2956..2974)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2979..2995)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2979..2995)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3097..3116)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             3108..3917
                     /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(3175..3194)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      4064..4652
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4553..4572
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4806..4823
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(4982..5009)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(5101..5187)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     join(5236..5250,1..8)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     join(5250,1..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 caccggggtg gtccccatcc
      181 tggtggagct ggatggcgac gttaacgggc acaagttctc ggtgtctggc gagggggagg
      241 gcgacgctac ctacgggaag ctcacgctga agttcatctg cacgaccggc aagctgccgg
      301 tgccctggcc aacactcgtc acaactctga cttacggcgt ccagtgcttc tcgcgctacc
      361 ctgatcacat gaagcagcat gacttcttca agtcggctat gcccgagggc tacgtgcagg
      421 agcggacgat cttcttcaag gacgatggca actacaagac acgcgcggag gtcaagttcg
      481 agggggacac tctcgttaac aggatcgagc tgaagggcat tgacttcaag gaggatggca
      541 atatcctcgg gcacaagctg gagtacaact acaattccca taacgtgtac atcatggccg
      601 acaagcagaa gaacggcatt aaggtcaatt tcaagatccg gcacaacatt gaggatggca
      661 gcgttcagct cgcggaccat taccagcaga acacaccgat cggggatggc cctgtcctcc
      721 tgcccgacaa tcactacctc tccactcaga gcgccctgtc gaaggacccg aacgagaaga
      781 gggaccatat ggtgctcctg gagttcgtca cagccgcggg catcactctc gggatggatg
      841 agctgtacaa gggggcgggc gactacaagg acgatgacga taaggcttac ccctacgacg
      901 tgccggatta cgccggggct ggcatggggc gctcactctc cccgctcctg aggcaggagc
      961 tggacaacct ggataaggac gccgattctc gcagggctgc catgaaggct ctcaagtcat
     1021 acgcccgcca cctggatagc aagtcgatcc cccatttcct ggccgaggtc tctgacacga
     1081 ccgccgctgg cgcgggggcc ggcagcggga tgccaaccgg cggggagttc acgatctcgc
     1141 tctacgaggt cctggccagg gttcacgggc ggaacatcgt gccccagatc ggcaatatta
     1201 tggcgaccat tatgtgcacg ctctccagct cgggcgggtc attcccactg caccaggcct
     1261 gctccaaggt tgtgccagct atcgctaggc atggcattga tccgtcggcc ccagacggcg
     1321 agaaggccgg gatcattgct agcctctgcc gcccactgtg ctcggctctc atgggcggga
     1381 gccaggacgg cggcgctgcc agcggcgctg ccctctgcct gaaggccctg gttgagtctt
     1441 caaactggag gttcgcgtcg ggcgagctgg tgaatgaggt ctgcctgaag gtggccggcg
     1501 ctatgcacga ctcggccaca cggtcgaacg ctcatatggg gctggccatg gcgctcgtga
     1561 agcacaatgg cctcatcgct gaggcctacg cgcgctctat cgtcaggtcc ggcctccaga
     1621 ttctggacgg ggataccgct gactccagct cgcagaagag gctgtccgcc atccagatga
     1681 ttaacttctt catgaagttc gtggacccga ggtgcctgtc ttcagagctg ggccgggtta
     1741 tcgatgtgat ggagcgctgc cagaatgacc ggatgccctt cgtgcggggc gcggctttcg
     1801 aggcctctca gtcagcgcgc aacattgccg cgcagaaggg ctccaggcac gaggttggga
     1861 cctctcctat ggtgggctca aacttccata agaggaggga gaagtcccct tgccggagcc
     1921 tctggggcgc caagaggtcc cccgctgcct ctacgatggc cgctgctagc cctgtgcagt
     1981 tccgctctcc ggagtcacag gtcgttgatt ccagcatcat gaatggctcc acactccacg
     2041 actccccagt gagcgtcggc cagtcgtctt gcaacttcga ccagagcagg cggactaacc
     2101 gcaggctgtg gtcgaatgac gcggttgatg tgtctctgaa ggacggcctc ttcatccagc
     2161 tgtgctccaa cagcaattcg tacgaggacg atctgggcga ggtttgcgat agcgaggtga
     2221 ccgacgctaa cttcgagtgc accaatacgt tcgccgggtt cgtctctccc tcacacaatg
     2281 gctccatcag cagggataag acgccttcac cgcgggcctc ggacaggccc atcaacattg
     2341 acgatgtgaa gatctactcc acgccaagga agctcctgcg gctcctgcag tcatcctacg
     2401 actcggattc tgcctcaaac atcgggcaga gcacggccaa gctcaggggc ctctcctccc
     2461 ccgaccagga gcacaagggc ctcgtcattt cttcagagca gatgcagtcg ctgcattctg
     2521 acaataaggg cgatgagatg aaggacgaga acgagaccat ctacatgcag aatagcaagg
     2581 gcaggacaca gactctctcg aacgctgacg cctcagtcct gtccacaact gaggccgaga
     2641 acacaagcat caagacttcg cctgagattg agctgaatgg ggacgatgtt tgcatcacct
     2701 ccagcattgg caagacgcgg aagtaccgcg cgaagttcac cctcttcctg tccgtcatcg
     2761 ttattttcct cgctatcatt acgatgctga tccgcattga gaacgacgat gacagcgcgt
     2821 acctggtgcc gacctgagac ccagctttct tgtacaaagt tggcattata agaaagcatt
     2881 gcttatcaat ttgttgcaac gaacaggtca ctatcagtca aaataaaatc attatttgcc
     2941 atccagctga tatcccctat agtgagtcgt attacatggt catagctgtt tcctggcagc
     3001 tctggcccgt gtctcaaaat ctctgatgtt acattgcaca agataaaaat atatcatcat
     3061 gaacaataaa actgtctgct tacataaaca gtaatacaag gggtgttatg agccatattc
     3121 aacgggaaac gtcgaggccg cgattaaatt ccaacatgga tgctgattta tatgggtata
     3181 aatgggctcg cgataatgtc gggcaatcag gtgcgacaat ctatcgcttg tatgggaagc
     3241 ccgatgcgcc agagttgttt ctgaaacatg gcaaaggtag cgttgccaat gatgttacag
     3301 atgagatggt cagactaaac tggctgacgg aatttatgcc tcttccgacc atcaagcatt
     3361 ttatccgtac tcctgatgat gcatggttac tcaccactgc gatccccgga aaaacagcat
     3421 tccaggtatt agaagaatat cctgattcag gtgaaaatat tgttgatgcg ctggcagtgt
     3481 tcctgcgccg gttgcattcg attcctgttt gtaattgtcc ttttaacagc gatcgcgtat
     3541 ttcgtctcgc tcaggcgcaa tcacgaatga ataacggttt ggttgatgcg agtgattttg
     3601 atgacgagcg taatggctgg cctgttgaac aagtctggaa agaaatgcat aaacttttgc
     3661 cattctcacc ggattcagtc gtcactcatg gtgatttctc acttgataac cttatttttg
     3721 acgaggggaa attaataggt tgtattgatg ttggacgagt cggaatcgca gaccgatacc
     3781 aggatcttgc catcctatgg aactgcctcg gtgagttttc tccttcatta cagaaacggc
     3841 tttttcaaaa atatggtatt gataatcctg atatgaataa attgcagttt catttgatgc
     3901 tcgatgagtt tttctaatca gaattggtta attggttgta acactggcag agcattacgc
     3961 tgacttgacg ggacggcgca agctcatgac caaaatccct taacgtgagt tacgcgtcgt
     4021 tccactgagc gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat cctttttttc
     4081 tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct accagcggtg gtttgtttgc
     4141 cggatcaaga gctaccaact ctttttccga aggtaactgg cttcagcaga gcgcagatac
     4201 caaatactgt tcttctagtg tagccgtagt taggccacca cttcaagaac tctgtagcac
     4261 cgcctacata cctcgctctg ctaatcctgt taccagtggc tgctgccagt ggcgataagt
     4321 cgtgtcttac cgggttggac tcaagacgat agttaccgga taaggcgcag cggtcgggct
     4381 gaacgggggg ttcgtgcaca cagcccagct tggagcgaac gacctacacc gaactgagat
     4441 acctacagcg tgagctatga gaaagcgcca cgcttcccga agggagaaag gcggacaggt
     4501 atccggtaag cggcagggtc ggaacaggag agcgcacgag ggagcttcca gggggaaacg
     4561 cctggtatct ttatagtcct gtcgggtttc gccacctctg acttgagcgt cgatttttgt
     4621 gatgctcgtc aggggggcgg agcctatgga aaaacgccag caacgcggcc tttttacggt
     4681 tcctggcctt ttgctggcct tttgctcaca tgttctttcc tgcgttatcc cctgattctg
     4741 tggataaccg tattaccgcc tttgagtgag ctgataccgc tcgccgcagc cgaacgaccg
     4801 agcgcagcga gtcagtgagc gaggaagcgg aagagcgccc aatacgcaaa ccgcctctcc
     4861 ccgcgcgttg gccgattcat taatgcagct ggcacgacag gtttcccgac tggaaagcgg
     4921 gcagtgagcg caacgcaatt aatacgcgta ccgctagcca ggaagagttt gtagaaacgc
     4981 aaaaaggcca tccgtcagga tggccttctg cttagtttga tgcctggcag tttatggcgg
     5041 gcgtcctgcc cgccaccctc cgggccgttg cttcacaacg ttcaaatccg ctcccggcgg
     5101 atttgtccta ctcaggagag cgttcaccga caaacaacag ataaaacgaa aggcccagtc
     5161 ttccgactga gcctttcgtt ttatttgatg cctggcagtt ccctactctc gcgttaacgc
     5221 tagcatggat gttttcccag tcacgacgtt
//
