LOCUS       sequence-272303-        5432 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  bacterial expression of human CENP-N 1-240.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5432)
  AUTHORS   McKinley KL, Sekulic N, Guo LY, Tsinman T, Black BE, Cheeseman IM
  TITLE     The CENP-L-N Complex Forms a Critical Node in an Integrated Meshwork
            of Interactions at the Centromere-Kinetochore Interface.
  JOURNAL   Mol Cell. 2015 Dec 17;60(6):886-98. doi: 
            10.1016/j.molcel.2015.10.027. Epub 2015 Nov 19.
  PUBMED    26698661
REFERENCE   2  (bases 1 to 5432)
  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: 
            "10.1016/j.molcel.2015.10.027"; journalName: "Mol Cell"; date: 
            "2015-12-17- 17"; volume: "60"; issue: "6"; pages: "886-98"
COMMENT     Sequence Label: pDK637
FEATURES             Location/Qualifiers
     source          1..5432
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     58..77
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     161..180
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        161..179
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     RBS             253..275
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             1012..1032
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=7xHis
                     /translation="HHHHHHH"
     primer_bind     complement(1122..1140)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      1136..1183
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     promoter        complement(join(1549..1554,1555..1571,1572..1577))
                     /label=tet promoter
                     /note="E. coli promoter for tetracycline efflux protein 
                     gene"
     primer_bind     complement(1604..1622)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        1690..1794
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1795..1863,1864..2655)
                     /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(2013..2032)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2826..3414
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3315..3334
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3568..3585
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    3600..3739
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(3665..3684)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3825..3847
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(3841..4032)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
ORIGIN
        1 acccacgccg aaacaagcgc tcatgagccc gaagtggcga gcccgatctt ccccatcggt
       61 gatgtcggcg atataggcgc cagcaaccgc acctgtggcg ccggtgatgc cggccacgat
      121 gcgtccggcg tagaggatcg agatctcgat cccgcgaaat taatacgact cactataggg
      181 agaccacaac ggtttccctc tagtgccggc tccggagagc tccaattgga attcgctagc
      241 tctagaaata attttgttta actttaagaa ggagatatac atatggatga gactgttgct
      301 gagttcatca agaggaccat cttgaaaatc cccatgaatg aactgacaac aatcctgaag
      361 gcctgggatt ttttgtctga aaatcaactg cagactgtaa atttccgaca gagaaaggaa
      421 tctgtagttc agcacttgat ccatctgtgt gaggaaaagc gtgcaagtat cagtgatgct
      481 gccctgttag acatcattta tatgcaattt catcagcacc agaaagtttg ggatgttttt
      541 cagatgagta aaggaccagg tgaagatgtt gacctttttg atatgaaaca atttaaaaat
      601 tcgttcaaga aaattcttca gagagcatta aaaaatgtga cagtcagctt cagagaaact
      661 gaggagaatg cagtctggat tcgaattgcc tggggaacac agtacacaaa gccaaaccag
      721 tacaaaccta cctacgtggt gtactactcc cagactccgt acgccttcac gtcctcctcc
      781 atgctgaggc gcaatacacc gcttctgggt caggcgctga caattgctag caaacaccat
      841 cagattgtga aaatggacct gagaagtcgg tatctggact ctcttaaggc tattgttttt
      901 aaacagtata atcagacctt tgaaactcac aactctacga cacctctaca ggaaagaagc
      961 cttggactag atataaatat ggattcaagg atcattcatg aaggcagcag ccatcatcat
     1021 catcatcacc actgaggatc cagatctaag cttggtacca cgcgtgcgcg ctgatccggc
     1081 tgctaacaaa gcccgaaagg aagctgagtt ggctgctgcc accgctgagc aataactagc
     1141 ataacccctt ggggcctcta aacgggtctt gaggggtttt ttgctgaaag gaggaactat
     1201 atccggatat ccacaggacg ggtgtggtcg ccatgatcgc gtagtcgata gtggctccaa
     1261 gtagcgaagc gagcaggact gggcggcggc caaagcggtc ggacagtgct ccgagaacgg
     1321 gtgcgcatag aaattgcatc aacgcatata gcgctagcag cacgccatag tgactggcga
     1381 tgctgtcgga atggacgata tcccgcaaga ggcccggcag taccggcata accaagccta
     1441 tgcctacagc atccagggtg acggtgccga ggatgacgat gagcgcattg ttagatttca
     1501 tacacggtgc ctgactgcgt tagcaattta actgtgataa actaccgcat taaagcttat
     1561 cgatgataag ctgtcaaaca tgagaattct tgaagacgaa agggcctcgt gatacgccta
     1621 tttttatagg ttaatgtcat gataataatg gtttcttaga cgtcaggtgg cacttttcgg
     1681 ggaaatgtgc gcggaacccc tatttgttta tttttctaaa tacattcaaa tatgtatccg
     1741 ctcatgagac aataaccctg ataaatgctt caataatatt gaaaaaggaa gagtatgagt
     1801 attcaacatt tccgtgtcgc ccttattccc ttttttgcgg cattttgcct tcctgttttt
     1861 gctcacccag aaacgctggt gaaagtaaaa gatgctgaag atcagttggg tgcacgagtg
     1921 ggttacatcg aactggatct caacagcggt aagatccttg agagttttcg ccccgaagaa
     1981 cgttttccaa tgatgagcac ttttaaagtt ctgctatgtg gcgcggtatt atcccgtgtt
     2041 gacgccgggc aagagcaact cggtcgccgc atacactatt ctcagaatga cttggttgag
     2101 tactcaccag tcacagaaaa gcatcttacg gatggcatga cagtaagaga attatgcagt
     2161 gctgccataa ccatgagtga taacactgcg gccaacttac ttctgacaac gatcggagga
     2221 ccgaaggagc taaccgcttt tttgcacaac atgggggatc atgtaactcg ccttgatcgt
     2281 tgggaaccgg agctgaatga agccatacca aacgacgagc gtgacaccac gatgcctgca
     2341 gcaatggcaa caacgttgcg caaactatta actggcgaac tacttactct agcttcccgg
     2401 caacaattaa tagactggat ggaggcggat aaagttgcag gaccacttct gcgctcggcc
     2461 cttccggctg gctggtttat tgctgataaa tctggagccg gtgagcgtgg gtctcgcggt
     2521 atcattgcag cactggggcc agatggtaag ccctcccgta tcgtagttat ctacacgacg
     2581 gggagtcagg caactatgga tgaacgaaat agacagatcg ctgagatagg tgcctcactg
     2641 attaagcatt ggtaactgtc agaccaagtt tactcatata tactttagat tgatttaaaa
     2701 cttcattttt aatttaaaag gatctaggtg aagatccttt ttgataatct catgaccaaa
     2761 atcccttaac gtgagttttc gttccactga gcgtcagacc ccgtagaaaa gatcaaagga
     2821 tcttcttgag atcctttttt tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg
     2881 ctaccagcgg tggtttgttt gccggatcaa gagctaccaa ctctttttcc gaaggtaact
     2941 ggcttcagca gagcgcagat accaaatact gtccttctag tgtagccgta gttaggccac
     3001 cacttcaaga actctgtagc accgcctaca tacctcgctc tgctaatcct gttaccagtg
     3061 gctgctgcca gtggcgataa gtcgtgtctt accgggttgg actcaagacg atagttaccg
     3121 gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca cacagcccag cttggagcga
     3181 acgacctaca ccgaactgag atacctacag cgtgagctat gagaaagcgc cacgcttccc
     3241 gaagggagaa aggcggacag gtatccggta agcggcaggg tcggaacagg agagcgcacg
     3301 agggagcttc cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc
     3361 tgacttgagc gtcgattttt gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc
     3421 agcaacgcgg cctttttacg gttcctggcc ttttgctggc cttttgctca catgttcttt
     3481 cctgcgttat cccctgattc tgtggataac cgtattaccg cctttgagtg agctgatacc
     3541 gctcgccgca gccgaacgac cgagcgcagc gagtcagtga gcgaggaagc ggaagagcgc
     3601 ctgatgcggt attttctcct tacgcatctg tgcggtattt cacaccgcaa tggtgcactc
     3661 tcagtacaat ctgctctgat gccgcatagt taagccagta tacactccgc tatcgctacg
     3721 tgactgggtc atggctgcgc cccgacaccc gccaacaccc gctgacgcgc cctgacgggc
     3781 ttgtctgctc ccggcatccg cttacagaca agctgtgacc gtctccggga gctgcatgtg
     3841 tcagaggttt tcaccgtcat caccgaaacg cgcgaggcag ctgcggtaaa gctcatcagc
     3901 gtggtcgtga agcgattcac agatgtctgc ctgttcatcc gcgtccagct cgttgagttt
     3961 ctccagaagc gttaatgtct ggcttctgat aaagcgggcc atgttaaggg cggttttttc
     4021 ctgtttggtc actgatgcct ccgtgtaagg gggatttctg ttcatggggg taatgatacc
     4081 gatgaaacga gagaggatgc tcacgatacg ggttactgat gatgaacatg cccggttact
     4141 ggaacgttgt gagggtaaac aactggcggt atggatgcgg cgggaccaga gaaaaatcac
     4201 tcagggtcaa tgccagcgct tcgttaatac agatgtaggt gttccacagg gtagccagca
     4261 gcatcctgcg atgcagatcc ggaacataat ggtgcagggc gctgacttcc gcgtttccag
     4321 actttacgaa acacggaaac cgaagaccat tcatgttgtt gctcaggtcg cagacgtttt
     4381 gcagcagcag tcgcttcacg ttcgctcgcg tatcggtgat tcattctgct aaccagtaag
     4441 gcaaccccgc cagcctagcc gggtcctcaa cgacaggagc acgatcatgc gcacccgtgg
     4501 ccaggaccca acgctgcccg agatgcgccg cgtgcggctg ctggagatgg cggacgcgat
     4561 ggatatgttc tgccaagggt tggtttgcgc attcacagtt ctccgcaaga attgattggc
     4621 tccaattctt ggagtggtga atccgttagc gaggtgccgc cggcttccat tcaggtcgag
     4681 gtggcccggc tccatgcacc gcgacgcaac gcggggaggc agacaaggta tagggcggcg
     4741 cctacaatcc atgccaaccc gttccatgtg ctcgccgagg cggcataaat cgccgtgacg
     4801 atcagcggtc caatgatcga agttaggctg gtaagagccg cgagcgatcc ttgaagctgt
     4861 ccctgatggt cgtcatctac ctgcctggac agcatggcct gcaacgcggg catcccgatg
     4921 ccgccggaag cgagaagaat cataatgggg aaggccatcc agcctcgcgt cgcgaacgcc
     4981 agcaagacgt agcccagcgc gtcggccgcc atgccggcga taatggcctg cttctcgccg
     5041 aaacgtttgg tggcgggacc agtgacgaag gcttgagcga gggcgtgcaa gattccgaat
     5101 accgcaagcg acaggccgat catcgtcgcg ctccagcgaa agcggtcctc gccgaaaatg
     5161 acccagagcg ctgccggcac ctgtcctacg agttgcatga taaagaagac agtcataagt
     5221 gcggcgacga tagtcatgcc ccgcgcccac cggaaggagc tgactgggtt gaaggctctc
     5281 aagggcatcg gtcgacgctc tcccttatgc gactcctgca ttaggaagca gcccagtagt
     5341 aggttgaggc cgttgagcac cgccgccgca aggaatggtg catgcaagga gatggcgccc
     5401 aacagtcccc cggccacggg gcctgccacc at
//
