LOCUS       sequence-272647-        6239 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of C. elegans KNL1.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6239)
  AUTHORS   Kern DM, Kim T, Rigney M, Hattersley N, Desai A, Cheeseman IM
  TITLE     The outer kinetochore protein KNL-1 contains a defined 
            oligomerization domain in nematodes.
  JOURNAL   Mol Biol Cell. 2015 Jan 15;26(2):229-37. doi: 
            10.1091/mbc.E14-06-1125. Epub 2014 Nov 19.
  PUBMED    25411336
REFERENCE   2  (bases 1 to 6239)
  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: 
            "10.1091/mbc.E14-06-1125"; journalName: "Mol Biol Cell"; date: 
            "2015-01-15- 15"; volume: "26"; issue: "2"; pages: "229-37"
COMMENT     Sequence Label: pDK134
FEATURES             Location/Qualifiers
     source          1..6239
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     51..70
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     154..173
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        154..172
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     RBS             246..268
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             288..305
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             309..341
                     /codon_start=1
                     /product="leader peptide from bacteriophage T7 gene 10"
                     /label=T7 tag (gene 10 leader)
                     /note="promotes efficient translation in E. coli"
                     /translation="MASMTGGQQMG"
     CDS             join(345..353,354..368)
                     /codon_start=1
                     /product="Xpress(TM) epitope tag, including an enterokinase
                     recognition and cleavage site"
                     /label=Xpress(TM) tag
                     /translation="DLYDDDDK"
     CDS             369..392
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     primer_bind     complement(1922..1940)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      1936..1983
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     promoter        complement(join(2349..2354,2355..2371,2372..2377))
                     /label=tet promoter
                     /note="E. coli promoter for tetracycline efflux protein 
                     gene"
     primer_bind     complement(2404..2422)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        2490..2594
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2595..2663,2664..3455)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(2813..2832)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3626..4214
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4115..4134
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4368..4385
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    4400..4539
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(4465..4484)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4625..4647
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(4641..4832)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
ORIGIN
        1 ccgaaacaag cgctcatgag cccgaagtgg cgagcccgat cttccccatc ggtgatgtcg
       61 gcgatatagg cgccagcaac cgcacctgtg gcgccggtga tgccggccac gatgcgtccg
      121 gcgtagagga tcgagatctc gatcccgcga aattaatacg actcactata gggagaccac
      181 aacggtttcc ctctagtgcc ggctccggag agctccaatt ggaattcgct agctctagaa
      241 ataattttgt ttaactttaa gaaggagata tacgtatgcg gggttctcat catcatcatc
      301 atcatggtat ggctagcatg actggtggac agcaaatggg tcgggatctg tacgacgatg
      361 acgataagct ggaagttctg ttccaggggc cccatatgtc gatggagcct cgtaagaagc
      421 ggaactcgat tctcaaggtg cggcaagccg tcgaaaccat cgaggaaacc gtcatgaaca
      481 gtgggcctag ttccacaaca actaatcgac gagtcagctt tcataacgtg aagcatgtca
      541 agcagtatga cagggaccat ggtaaaattc ttgacgccac accagttaag gagaagatta
      601 ctgacactat tggatcagat ggtattttga caccacgtgg cggaaacatg gatatttccg
      661 aatctccggc ctgcacgtcc tcatttcaag tgttcggcgg tggtaatctc gataaaacta
      721 tggatatgtc tctcgaaaca actatcaacg agaacaacga aacggcgaga ttgtttgaaa
      781 ccacaagaga tccaacacta ttatacgaaa agatcgtcga aaccacaaca aaagttaccg
      841 agcgaattgt tagtatgcca ctggatgata ccttagcaat gttcaataca acgaatcaag
      901 aagataagga tatgtcagtt gatcgttcag ttcttttcac gattcccaaa gttccgaagc
      961 ataacgctac aatgaataga actataccga tggacctcga tgaatcaaaa gcagcgggcg
     1021 gccagtgcga tgaaacgatg aatgtgttca atttcacaaa cttggaagcc gctgaaatgg
     1081 atacgagtaa attagatgaa aataatacca tgaatgctat ccggattccg attaattcaa
     1141 acgtcatgcc tgtagacatg gacatcactg aacatcacac tttaattgaa gaaaagaaaa
     1201 atgatacatt cgggccaagt caactgatgg acatttcggc gccacaagtt caagttaatg
     1261 atactttggc cattttcaac agtccgagag acatctgtaa taagggtttg ggtgttcctc
     1321 agaatctaat aaatatcgcc tcgaacgtcg tacctgtgga catggacatc actgatcagg
     1381 ccgtattaaa cgcggagaag aaaaatgatc aattcgagac aagtcagctt atggacattt
     1441 ctattccgaa agttctagta aatgacacta tggcgatgtt caacagcccg aaacacgtca
     1501 gtaagagcag catggatctc gagaaaacga ttgaagccgc tgacaaatca acgaaatacc
     1561 cgagtatcgc agatgaggtg gaagatttag acatggatat ggatatcact gaacaacaac
     1621 catgtgaggc tggtaatcag cagaacgacg gcttgcaact tcaaaaggag gatttaatgg
     1681 acatttcggt gattcgagat tcacctgcag taaacgacac catggctgtg ttccagagtc
     1741 ctgccagagt aaagatcgga gcgaacaact cgatcattga ttcgcagaaa tctatcgtgt
     1801 tcggtgacga aatgagcatt gacgagacac aataaggatc cagatctaag cttggtacca
     1861 cgcgtgcgcg ctgatccggc tgctaacaaa gcccgaaagg aagctgagtt ggctgctgcc
     1921 accgctgagc aataactagc ataacccctt ggggcctcta aacgggtctt gaggggtttt
     1981 ttgctgaaag gaggaactat atccggatat ccacaggacg ggtgtggtcg ccatgatcgc
     2041 gtagtcgata gtggctccaa gtagcgaagc gagcaggact gggcggcggc caaagcggtc
     2101 ggacagtgct ccgagaacgg gtgcgcatag aaattgcatc aacgcatata gcgctagcag
     2161 cacgccatag tgactggcga tgctgtcgga atggacgata tcccgcaaga ggcccggcag
     2221 taccggcata accaagccta tgcctacagc atccagggtg acggtgccga ggatgacgat
     2281 gagcgcattg ttagatttca tacacggtgc ctgactgcgt tagcaattta actgtgataa
     2341 actaccgcat taaagcttat cgatgataag ctgtcaaaca tgagaattct tgaagacgaa
     2401 agggcctcgt gatacgccta tttttatagg ttaatgtcat gataataatg gtttcttaga
     2461 cgtcaggtgg cacttttcgg ggaaatgtgc gcggaacccc tatttgttta tttttctaaa
     2521 tacattcaaa tatgtatccg ctcatgagac aataaccctg ataaatgctt caataatatt
     2581 gaaaaaggaa gagtatgagt attcaacatt tccgtgtcgc ccttattccc ttttttgcgg
     2641 cattttgcct tcctgttttt gctcacccag aaacgctggt gaaagtaaaa gatgctgaag
     2701 atcagttggg tgcacgagtg ggttacatcg aactggatct caacagcggt aagatccttg
     2761 agagttttcg ccccgaagaa cgttttccaa tgatgagcac ttttaaagtt ctgctatgtg
     2821 gcgcggtatt atcccgtgtt gacgccgggc aagagcaact cggtcgccgc atacactatt
     2881 ctcagaatga cttggttgag tactcaccag tcacagaaaa gcatcttacg gatggcatga
     2941 cagtaagaga attatgcagt gctgccataa ccatgagtga taacactgcg gccaacttac
     3001 ttctgacaac gatcggagga ccgaaggagc taaccgcttt tttgcacaac atgggggatc
     3061 atgtaactcg ccttgatcgt tgggaaccgg agctgaatga agccatacca aacgacgagc
     3121 gtgacaccac gatgcctgca gcaatggcaa caacgttgcg caaactatta actggcgaac
     3181 tacttactct agcttcccgg caacaattaa tagactggat ggaggcggat aaagttgcag
     3241 gaccacttct gcgctcggcc cttccggctg gctggtttat tgctgataaa tctggagccg
     3301 gtgagcgtgg gtctcgcggt atcattgcag cactggggcc agatggtaag ccctcccgta
     3361 tcgtagttat ctacacgacg gggagtcagg caactatgga tgaacgaaat agacagatcg
     3421 ctgagatagg tgcctcactg attaagcatt ggtaactgtc agaccaagtt tactcatata
     3481 tactttagat tgatttaaaa cttcattttt aatttaaaag gatctaggtg aagatccttt
     3541 ttgataatct catgaccaaa atcccttaac gtgagttttc gttccactga gcgtcagacc
     3601 ccgtagaaaa gatcaaagga tcttcttgag atcctttttt tctgcgcgta atctgctgct
     3661 tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa gagctaccaa
     3721 ctctttttcc gaaggtaact ggcttcagca gagcgcagat accaaatact gtccttctag
     3781 tgtagccgta gttaggccac cacttcaaga actctgtagc accgcctaca tacctcgctc
     3841 tgctaatcct gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt accgggttgg
     3901 actcaagacg atagttaccg gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca
     3961 cacagcccag cttggagcga acgacctaca ccgaactgag atacctacag cgtgagctat
     4021 gagaaagcgc cacgcttccc gaagggagaa aggcggacag gtatccggta agcggcaggg
     4081 tcggaacagg agagcgcacg agggagcttc cagggggaaa cgcctggtat ctttatagtc
     4141 ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt gtgatgctcg tcaggggggc
     4201 ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc ttttgctggc
     4261 cttttgctca catgttcttt cctgcgttat cccctgattc tgtggataac cgtattaccg
     4321 cctttgagtg agctgatacc gctcgccgca gccgaacgac cgagcgcagc gagtcagtga
     4381 gcgaggaagc ggaagagcgc ctgatgcggt attttctcct tacgcatctg tgcggtattt
     4441 cacaccgcaa tggtgcactc tcagtacaat ctgctctgat gccgcatagt taagccagta
     4501 tacactccgc tatcgctacg tgactgggtc atggctgcgc cccgacaccc gccaacaccc
     4561 gctgacgcgc cctgacgggc ttgtctgctc ccggcatccg cttacagaca agctgtgacc
     4621 gtctccggga gctgcatgtg tcagaggttt tcaccgtcat caccgaaacg cgcgaggcag
     4681 ctgcggtaaa gctcatcagc gtggtcgtga agcgattcac agatgtctgc ctgttcatcc
     4741 gcgtccagct cgttgagttt ctccagaagc gttaatgtct ggcttctgat aaagcgggcc
     4801 atgttaaggg cggttttttc ctgtttggtc actgatgcct ccgtgtaagg gggatttctg
     4861 ttcatggggg taatgatacc gatgaaacga gagaggatgc tcacgatacg ggttactgat
     4921 gatgaacatg cccggttact ggaacgttgt gagggtaaac aactggcggt atggatgcgg
     4981 cgggaccaga gaaaaatcac tcagggtcaa tgccagcgct tcgttaatac agatgtaggt
     5041 gttccacagg gtagccagca gcatcctgcg atgcagatcc ggaacataat ggtgcagggc
     5101 gctgacttcc gcgtttccag actttacgaa acacggaaac cgaagaccat tcatgttgtt
     5161 gctcaggtcg cagacgtttt gcagcagcag tcgcttcacg ttcgctcgcg tatcggtgat
     5221 tcattctgct aaccagtaag gcaaccccgc cagcctagcc gggtcctcaa cgacaggagc
     5281 acgatcatgc gcacccgtgg ccaggaccca acgctgcccg agatgcgccg cgtgcggctg
     5341 ctggagatgg cggacgcgat ggatatgttc tgccaagggt tggtttgcgc attcacagtt
     5401 ctccgcaaga attgattggc tccaattctt ggagtggtga atccgttagc gaggtgccgc
     5461 cggcttccat tcaggtcgag gtggcccggc tccatgcacc gcgacgcaac gcggggaggc
     5521 agacaaggta tagggcggcg cctacaatcc atgccaaccc gttccatgtg ctcgccgagg
     5581 cggcataaat cgccgtgacg atcagcggtc caatgatcga agttaggctg gtaagagccg
     5641 cgagcgatcc ttgaagctgt ccctgatggt cgtcatctac ctgcctggac agcatggcct
     5701 gcaacgcggg catcccgatg ccgccggaag cgagaagaat cataatgggg aaggccatcc
     5761 agcctcgcgt cgcgaacgcc agcaagacgt agcccagcgc gtcggccgcc atgccggcga
     5821 taatggcctg cttctcgccg aaacgtttgg tggcgggacc agtgacgaag gcttgagcga
     5881 gggcgtgcaa gattccgaat accgcaagcg acaggccgat catcgtcgcg ctccagcgaa
     5941 agcggtcctc gccgaaaatg acccagagcg ctgccggcac ctgtcctacg agttgcatga
     6001 taaagaagac agtcataagt gcggcgacga tagtcatgcc ccgcgcccac cggaaggagc
     6061 tgactgggtt gaaggctctc aagggcatcg gtcgacgctc tcccttatgc gactcctgca
     6121 ttaggaagca gcccagtagt aggttgaggc cgttgagcac cgccgccgca aggaatggtg
     6181 catgcaagga gatggcgccc aacagtcccc cggccacggg gcctgccacc atacccacg
//
