LOCUS       sequence-272650-        7739 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Bacterial expression of C. elegans KNL1.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7739)
  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 7739)
  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: pDK540
FEATURES             Location/Qualifiers
     source          1..7739
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     184..203
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     287..306
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        287..305
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     RBS             379..401
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             3448..3465
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(3555..3573)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      3569..3616
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     promoter        complement(join(3982..3987,3988..4004,4005..4010))
                     /label=tet promoter
                     /note="E. coli promoter for tetracycline efflux protein 
                     gene"
     primer_bind     complement(4037..4055)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4123..4227
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4228..4296,4297..5088)
                     /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(4446..4465)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5259..5847
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5748..5767
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6001..6018
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    6033..6172
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(6098..6117)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     6258..6280
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(6274..6465)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
ORIGIN
        1 atgcgactcc tgcattagga agcagcccag tagtaggttg aggccgttga gcaccgccgc
       61 cgcaaggaat ggtgcatgca aggagatggc gcccaacagt cccccggcca cggggcctgc
      121 caccataccc acgccgaaac aagcgctcat gagcccgaag tggcgagccc gatcttcccc
      181 atcggtgatg tcggcgatat aggcgccagc aaccgcacct gtggcgccgg tgatgccggc
      241 cacgatgcgt ccggcgtaga ggatcgagat ctcgatcccg cgaaattaat acgactcact
      301 atagggagac cacaacggtt tccctctagt gccggctccg gagagctcca attggaattc
      361 gctagctcta gaaataattt tgtttaactt taagaaggag atatacatat gtcgatggag
      421 cctcgtaaga agcggaactc gattctcaag gtgcggcaag ccgtcgaaac catcgaggaa
      481 accgtcatga acagtgggcc tagttccaca acaactaatc gacgagtcag ctttcataac
      541 gtgaagcatg tcaagcagta tgacagggac catggtaaaa ttcttgacgc cacaccagtt
      601 aaggagaaga ttactgacac tattggatca gatggtattt tgacaccacg tggcggaaac
      661 atggatattt ccgaatctcc ggcctgcacg tcctcatttc aagtgttcgg cggtggtaat
      721 ctcgataaaa ctatggatat gtctctcgaa acaactatca acgagaacaa cgaaacggcg
      781 agattgtttg aaaccacaag agatccaaca ctattagcag aaaagatcgt cgaaaccaca
      841 acaaaagtta ccgagcgaat tgttagtatg ccactggatg ataccttagc aatgttcaat
      901 acaacgaatc aagaagataa ggatatgtca gttgatcgtt cagttctttt cacgattccc
      961 aaagttccga agcataacgc tacaatgaat agaactatac cgatggacct cgatgaatca
     1021 aaagcagcgg gcggccagtg cgatgaaacg atgaatgtgt tcaatttcac aaacttggaa
     1081 gccgctgaaa tggatacgag taaattagat gaaaataata ccatgaatgc tatccggatt
     1141 ccgattaatt caaacgtcat gcctgtagac atggacatca ctgaacatca cactttaatt
     1201 gaagaaaaga aaaatgatac attcgggcca agtcaactga tggacatttc ggcgccacaa
     1261 gttcaagtta atgatacttt ggccattttc aacagtccga gagacatctg taataagggt
     1321 ttgggtgttc ctcagaatct aataaatatc gcctcgaacg tcgtacctgt ggacatggac
     1381 atcactgatc aggccgtatt aaacgcggag aagaaaaatg atcaattcga gacaagtcag
     1441 cttatggaca tttctattcc gaaagttcta gtaaatgaca ctatggcgat gttcaacagc
     1501 ccgaaacacg tcagtaagag cagcatggat ctcgagaaaa cgattgaagc cgctgacaaa
     1561 tcaacgaaat acccgagtat cgcagatgag gtggaagatt tagacatgga tatggatatc
     1621 actgaacaac aaccatgtga ggctggtaat cagcagaacg acggcttgca acttcaaaag
     1681 gaggatttaa tggacatttc ggtgattcga gattcacctg cagtaaacga caccatggct
     1741 gtgttccaga gtcctgccag agtaaagatc ggagcgaaca actcgatcat tgattcgcag
     1801 aaatctatcg tgttcggtga cgaaatgagc attgacgaga cacaaaatga tggaaccttg
     1861 acgttgccaa agtcgaatgt agaagtgact acaactaatg atgtctacac gtctctcgag
     1921 cggcaagagg aaaatgcttc agaaaacgta tccatgataa acgaatcttc tgttcattcg
     1981 gaaatcgaca aaaagtcgtt tatgctcatc gaagaagaaa gggcttttat gcactcctcc
     2041 atgattgatg tagcacaaaa gttggaagac gatggttcgt cgaagacgcc agtcatcctt
     2101 gcttcacagt cagcttctct tgccactaaa gaaccatcag cccttcacaa ctcgagtgca
     2161 actctcaaca attcgatgga attggacaac aatactcttc ttaaaactat gcaaattaca
     2221 acgtgtgaag acattagcat ggtccatgag tctattgctg ttgaactgaa cagtaacaaa
     2281 gagcaggagc aattcggaga tgagactttg cagaaaaatg atacctcgaa tactggcgcg
     2341 aatttcacat tccaaggcca taatgaaaca tcgcaaatca tgaacaatgt cgactcggaa
     2401 gcagtgaaca cgtccaagat ttcaacatat tcggctttca atttgagcat caaccagtct
     2461 atctctaaac gacgtcgatc tcttctgaat tctgctcgtg aatctcctcg tcgtgttgcg
     2521 ttggagaatt ctataatgtc gatgaatggg caaacagtgg aagctctgac agaatatcga
     2581 cagaataaaa ctatgcagac gagtcaagat tcgatgccga gtatgagttt gaacgattcg
     2641 ggaagagata ttctcgcgat gaatacatca gtccgctctc ctcatctgaa ttcttcaaaa
     2701 actgctgccc caggaacacc atcattgatg tcacaaaatg tacaacttcc acccccatct
     2761 cctcaattcg aaatgccaga cttcgatcca gctgtggtca acgttgtata tttaacatct
     2821 gaagatccgt ccactgaaca acatccagaa gctctcaaat ttcagcgtat tgttgaaaac
     2881 gagaaaatga aagtacaaca cgagattgat tctctgaatt caaccaatca actttctgct
     2941 gagaaaattg atatgttgaa gactaaggag ctcttgaagt ttagtcatga tgagcgagaa
     3001 gcgattatga ttgcaagaaa agacgcggaa atcaagtttt tggagcttcg tctgaaattt
     3061 gcactcgaaa aaaaaattga aagtgaccag gaaattgctg aactagaaca aggaaattcg
     3121 aaaatggctg agcagctaag aggtctcgat aagatggctg tcgttcaaaa agaactagaa
     3181 aagctgagaa gtcttcctcc atcacgcgaa gagagcggga aaatccgaaa ggagtggatg
     3241 gagatgaagc aatgggaatt cgaccagaaa atgaaagcac tccgaaatgt acgctcaaac
     3301 atgattgcac ttcgttcaga gaaaaatgct ctcgaaatga aagtcgcgga agaacacgag
     3361 aagtttgccc agaggaacga tttgaagaaa agtcgaatgc tggtgttctc taaggctgtt
     3421 aagaaaattg tgaacttcgg cagcagccat catcatcatc atcactaggg atccagatct
     3481 aagcttggta ccacgcgtgc gcgctgatcc ggctgctaac aaagcccgaa aggaagctga
     3541 gttggctgct gccaccgctg agcaataact agcataaccc cttggggcct ctaaacgggt
     3601 cttgaggggt tttttgctga aaggaggaac tatatccgga tatccacagg acgggtgtgg
     3661 tcgccatgat cgcgtagtcg atagtggctc caagtagcga agcgagcagg actgggcggc
     3721 ggccaaagcg gtcggacagt gctccgagaa cgggtgcgca tagaaattgc atcaacgcat
     3781 atagcgctag cagcacgcca tagtgactgg cgatgctgtc ggaatggacg atatcccgca
     3841 agaggcccgg cagtaccggc ataaccaagc ctatgcctac agcatccagg gtgacggtgc
     3901 cgaggatgac gatgagcgca ttgttagatt tcatacacgg tgcctgactg cgttagcaat
     3961 ttaactgtga taaactaccg cattaaagct tatcgatgat aagctgtcaa acatgagaat
     4021 tcttgaagac gaaagggcct cgtgatacgc ctatttttat aggttaatgt catgataata
     4081 atggtttctt agacgtcagg tggcactttt cggggaaatg tgcgcggaac ccctatttgt
     4141 ttatttttct aaatacattc aaatatgtat ccgctcatga gacaataacc ctgataaatg
     4201 cttcaataat attgaaaaag gaagagtatg agtattcaac atttccgtgt cgcccttatt
     4261 cccttttttg cggcattttg ccttcctgtt tttgctcacc cagaaacgct ggtgaaagta
     4321 aaagatgctg aagatcagtt gggtgcacga gtgggttaca tcgaactgga tctcaacagc
     4381 ggtaagatcc ttgagagttt tcgccccgaa gaacgttttc caatgatgag cacttttaaa
     4441 gttctgctat gtggcgcggt attatcccgt gttgacgccg ggcaagagca actcggtcgc
     4501 cgcatacact attctcagaa tgacttggtt gagtactcac cagtcacaga aaagcatctt
     4561 acggatggca tgacagtaag agaattatgc agtgctgcca taaccatgag tgataacact
     4621 gcggccaact tacttctgac aacgatcgga ggaccgaagg agctaaccgc ttttttgcac
     4681 aacatggggg atcatgtaac tcgccttgat cgttgggaac cggagctgaa tgaagccata
     4741 ccaaacgacg agcgtgacac cacgatgcct gcagcaatgg caacaacgtt gcgcaaacta
     4801 ttaactggcg aactacttac tctagcttcc cggcaacaat taatagactg gatggaggcg
     4861 gataaagttg caggaccact tctgcgctcg gcccttccgg ctggctggtt tattgctgat
     4921 aaatctggag ccggtgagcg tgggtctcgc ggtatcattg cagcactggg gccagatggt
     4981 aagccctccc gtatcgtagt tatctacacg acggggagtc aggcaactat ggatgaacga
     5041 aatagacaga tcgctgagat aggtgcctca ctgattaagc attggtaact gtcagaccaa
     5101 gtttactcat atatacttta gattgattta aaacttcatt tttaatttaa aaggatctag
     5161 gtgaagatcc tttttgataa tctcatgacc aaaatccctt aacgtgagtt ttcgttccac
     5221 tgagcgtcag accccgtaga aaagatcaaa ggatcttctt gagatccttt ttttctgcgc
     5281 gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg tttgccggat
     5341 caagagctac caactctttt tccgaaggta actggcttca gcagagcgca gataccaaat
     5401 actgtccttc tagtgtagcc gtagttaggc caccacttca agaactctgt agcaccgcct
     5461 acatacctcg ctctgctaat cctgttacca gtggctgctg ccagtggcga taagtcgtgt
     5521 cttaccgggt tggactcaag acgatagtta ccggataagg cgcagcggtc gggctgaacg
     5581 gggggttcgt gcacacagcc cagcttggag cgaacgacct acaccgaact gagataccta
     5641 cagcgtgagc tatgagaaag cgccacgctt cccgaaggga gaaaggcgga caggtatccg
     5701 gtaagcggca gggtcggaac aggagagcgc acgagggagc ttccaggggg aaacgcctgg
     5761 tatctttata gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc
     5821 tcgtcagggg ggcggagcct atggaaaaac gccagcaacg cggccttttt acggttcctg
     5881 gccttttgct ggccttttgc tcacatgttc tttcctgcgt tatcccctga ttctgtggat
     5941 aaccgtatta ccgcctttga gtgagctgat accgctcgcc gcagccgaac gaccgagcgc
     6001 agcgagtcag tgagcgagga agcggaagag cgcctgatgc ggtattttct ccttacgcat
     6061 ctgtgcggta tttcacaccg caatggtgca ctctcagtac aatctgctct gatgccgcat
     6121 agttaagcca gtatacactc cgctatcgct acgtgactgg gtcatggctg cgccccgaca
     6181 cccgccaaca cccgctgacg cgccctgacg ggcttgtctg ctcccggcat ccgcttacag
     6241 acaagctgtg accgtctccg ggagctgcat gtgtcagagg ttttcaccgt catcaccgaa
     6301 acgcgcgagg cagctgcggt aaagctcatc agcgtggtcg tgaagcgatt cacagatgtc
     6361 tgcctgttca tccgcgtcca gctcgttgag tttctccaga agcgttaatg tctggcttct
     6421 gataaagcgg gccatgttaa gggcggtttt ttcctgtttg gtcactgatg cctccgtgta
     6481 agggggattt ctgttcatgg gggtaatgat accgatgaaa cgagagagga tgctcacgat
     6541 acgggttact gatgatgaac atgcccggtt actggaacgt tgtgagggta aacaactggc
     6601 ggtatggatg cggcgggacc agagaaaaat cactcagggt caatgccagc gcttcgttaa
     6661 tacagatgta ggtgttccac agggtagcca gcagcatcct gcgatgcaga tccggaacat
     6721 aatggtgcag ggcgctgact tccgcgtttc cagactttac gaaacacgga aaccgaagac
     6781 cattcatgtt gttgctcagg tcgcagacgt tttgcagcag cagtcgcttc acgttcgctc
     6841 gcgtatcggt gattcattct gctaaccagt aaggcaaccc cgccagccta gccgggtcct
     6901 caacgacagg agcacgatca tgcgcacccg tggccaggac ccaacgctgc ccgagatgcg
     6961 ccgcgtgcgg ctgctggaga tggcggacgc gatggatatg ttctgccaag ggttggtttg
     7021 cgcattcaca gttctccgca agaattgatt ggctccaatt cttggagtgg tgaatccgtt
     7081 agcgaggtgc cgccggcttc cattcaggtc gaggtggccc ggctccatgc accgcgacgc
     7141 aacgcgggga ggcagacaag gtatagggcg gcgcctacaa tccatgccaa cccgttccat
     7201 gtgctcgccg aggcggcata aatcgccgtg acgatcagcg gtccaatgat cgaagttagg
     7261 ctggtaagag ccgcgagcga tccttgaagc tgtccctgat ggtcgtcatc tacctgcctg
     7321 gacagcatgg cctgcaacgc gggcatcccg atgccgccgg aagcgagaag aatcataatg
     7381 gggaaggcca tccagcctcg cgtcgcgaac gccagcaaga cgtagcccag cgcgtcggcc
     7441 gccatgccgg cgataatggc ctgcttctcg ccgaaacgtt tggtggcggg accagtgacg
     7501 aaggcttgag cgagggcgtg caagattccg aataccgcaa gcgacaggcc gatcatcgtc
     7561 gcgctccagc gaaagcggtc ctcgccgaaa atgacccaga gcgctgccgg cacctgtcct
     7621 acgagttgca tgataaagaa gacagtcata agtgcggcga cgatagtcat gccccgcgcc
     7681 caccggaagg agctgactgg gttgaaggct ctcaagggca tcggtcgacg ctctccctt
//
