LOCUS       sequence-271966-        6174 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Baculovirus expression of CENP-T and CENP-W.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6174)
  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 6174)
  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: pKM410
FEATURES             Location/Qualifiers
     source          1..6174
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        complement(join(64..69,70..86,87..92))
                     /gene="intI1 (promoter lies within the coding sequence)"
                     /label=Pc promoter
                     /note="class 1 integron promoter"
     protein_bind    complement(163..196)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     promoter        273..364
                     /gene="polh from Autographa californica"
                     /label=polyhedrin promoter
                     /note="promoter for the baculovirus polyhedrin gene"
     CDS             407..1060
                     /codon_start=1
                     /product="glutathione S-transferase from Schistosoma 
                     japonicum"
                     /label=GST
                     /translation="MSPILGYWKIKGLVQPTRLLLEYLEEKYEEHLYERDEGDKWRNKK
                     FELGLEFPNLPYYIDGDVKLTQSMAIIRYIADKHNMLGGCPKERAEISMLEGAVLDIRY
                     GVSRIAYSKDFETLKVDFLSKLPEMLKMFEDRLCHKTYLNGDHVTHPDFMLYDALDVVL
                     YMDPMCLDAFPKLVCFKKRIEAIPQIDKYLKSSKYIAWPLQGWQATFGGGDHPPK"
     primer_bind     1018..1040
                     /label=pGEX 5'
                     /note="pGEX vectors, Glutathione-S-transferase, forward 
                     primer"
     CDS             1067..1090
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     polyA_signal    2986..3120
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     promoter        3292..3383
                     /gene="polh from Autographa californica"
                     /label=polyhedrin promoter
                     /note="promoter for the baculovirus polyhedrin gene"
     polyA_signal    3893..4027
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     mobile_element  4148..4313
                     /mobile_element_type="transposon:Tn7"
                     /label=Tn7L
                     /note="mini-Tn7 element (left end of the Tn7 transposon)"
     rep_origin      4503..5091
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4992..5011
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     mobile_element  5226..5449
                     /mobile_element_type="transposon:Tn7"
                     /label=Tn7R
                     /note="mini-Tn7 element (right end of the Tn7 transposon)"
     CDS             complement(5516..6049)
                     /codon_start=1
                     /gene="aacC1"
                     /product="gentamycin acetyltransferase"
                     /label=GmR
                     /note="confers resistance to gentamycin"
                     /translation="MLRSSNDVTQQGSRPKTKLGGSSMGIIRTCRLGPDQVKSMRAALD
                     LFGREFGDVATYSQHQPDSDYLGNLLRSKTFIALAAFDQEAVVGALAAYVLPKFEQPRS
                     EIYIYDLAVSGEHRRQGIATALINLLKHEANALGAYVIYVQADYGDDPAVALYTKLGIR
                     EEVMHFDIDPSTAT"
ORIGIN
        1 ccaccgctgc gttcggtcaa ggttctggac cagttgcgtg agcgcatacg ctacttgcat
       61 tacagtttac gaaccgaaca ggcttatgtc aactgggttc gtgccttcat ccgtttccac
      121 ggtgtgcgtc acccggcaac cttgggcagc agcgaagtcg ccataacttc gtatagcata
      181 cattatacga agttatctgt aactataacg gtcctaaggt agcgagttta aacactagta
      241 tcgattcgcg acctactccg gaatattaat agatcatgga gataattaaa atgataacca
      301 tctcgcaaat aaataagtat tttactgttt tcgtaacagt tttgtaataa aaaaacctat
      361 aaatattccg gattattcat accgtcccac catcgggcgc ggatctatgt cccctatact
      421 aggttattgg aaaattaagg gccttgtgca acccactcga cttcttttgg aatatcttga
      481 agaaaaatat gaagagcatt tgtatgagcg cgatgaaggt gataaatggc gaaacaaaaa
      541 gtttgaattg ggtttggagt ttcccaatct tccttattat attgatggtg atgttaaatt
      601 aacacagtct atggccatca tacgttatat agctgacaag cacaacatgt tgggtggttg
      661 tccaaaagag cgtgcagaga tttcaatgct tgaaggagcg gttttggata ttagatacgg
      721 tgtttcgaga attgcatata gtaaagactt tgaaactctc aaagttgatt ttcttagcaa
      781 gctacctgaa atgctgaaaa tgttcgaaga tcgtttatgt cataaaacat atttaaatgg
      841 tgatcatgta acccatcctg acttcatgtt gtatgacgct cttgatgttg ttttatacat
      901 ggacccaatg tgcctggatg cgttcccaaa attagtttgt tttaaaaaac gtattgaagc
      961 tatcccacaa attgataagt acttgaaatc cagcaagtat atagcatggc ctttgcaggg
     1021 ctggcaagcc acgtttggtg gtggcgacca tcctccaaaa tcggatctgg aagttctgtt
     1081 ccaggggccc ctgggatcca tggcagacca caaccctgac agcgattcta cacctcgtac
     1141 actgctgcgt cgtgtgctgg atactgctga tcctcgtact cctcgtcgtc ctcgtagcgc
     1201 tcgtgctggt gctcgccgtg ccctgctgga aacagcctct cctcgtaaac tgagtggcca
     1261 gacccgtact attgcccgtg ggcgttcaca tggagcccgc tcggtcggtc gtagtgctca
     1321 cattcaagcc tcgggacacc tggaagaaca aacccctcgt accctgctga aaaacatcct
     1381 gctgactgcc cctgaaagct ctattctgat gccggaaagc gtcgttaaac cggttcctgc
     1441 tcctcaagcc gttcaacctt ctcgtcaaga gtcctcctgt ggatctctgg agctgcaact
     1501 gcctgaactg gagcctccta caacactggc accggggctg ctggcgcctg gtcgtcgtaa
     1561 acaacgtctg cgtctgagcg tgtttcaaca aggtgttgat cagggcctga gtctgtctca
     1621 agaacctcaa ggcaacgctg atgcttctag tctgacccgt tctctgaacc tgacctttgc
     1681 tactccactg caacctcaaa gcgtgcaacg tcctggactg gctcgccgcc ctcctgctcg
     1741 ccgtgccgtt gatgtgggtg cttttctgcg tgacctgcgt gatacaagcc tggcacctcc
     1801 gaatattgtg ctggaggata cacagccgtt ttcacagcct atggttggta gccctaacgt
     1861 ttatcactca ctgccgtgta cccctcacac tggtgccgaa gatgctgaac aagctgctgg
     1921 tcgtaaaaca caaagctctg gaccgggact gcaaaaaaac tctccgggga aacctgccca
     1981 attcctggcg ggtgaagctg aagaagtcaa cgcctttgct ctgggatttc tgtctaccag
     2041 ctcgggtgtt agcggagaag atgaggtgga acctctgcat gatggtgtgg aagaagccga
     2101 gaagaaaatg gaagaggaag gggtgtctgt ctctgaaatg gaagcgactg gtgctcaagg
     2161 accaagtcgt gtcgaagagg cagaaggcca taccgaggtt accgaagccg aaggaagcca
     2221 agggacagct gaagctgatg gacctggggc ctcaagtggg gatgaagatg ctagtggccg
     2281 tgccgcctca cctgaatctg cttctagtac accggaaagc ctgcaagcac gtcgtcatca
     2341 ccaatttctg gaacctgctc cggctcctgg tgctgctgtt ctgtctagtg aacctgctga
     2401 acctctgctg gttcgtcatc cgcctcgtcc tcgtacaact ggaccacgtc ctcgccaaga
     2461 ccctcataaa gcaggactga gccattatgt taaactgttc tcgttctatg ccaaaatgcc
     2521 gatggagcgt aaagctctgg aaatggtcga aaaatgcctg gacaaatatt tccaacatct
     2581 gtgtgacgat ctggaagtgt ttgccgctca tgctgggcgt aaaacagtga aacctgagga
     2641 tctggagctg ctgatgcgtc gtcaaggact ggttacggat caagtgtcgc tgcatgtcct
     2701 ggtggaacgt catctgccgc tggaatatcg ccaactgctg attccttgtg cttatagcgg
     2761 gaactccgtc tttcctgccc agtaagaatt caaaggccta cgtcgacgag ctcacttgtc
     2821 gcggccgctt tcgaatctag agcctgcagt ctcgacaagc ttgtcgagaa gtactagagg
     2881 atcataatca gccataccac atttgtagag gttttacttg ctttaaaaaa cctcccacac
     2941 ctccccctga acctgaaaca taaaatgaat gcaattgttg ttgttaactt gtttattgca
     3001 gcttataatg gttacaaata aagcaatagc atcacaaatt tcacaaataa agcatttttt
     3061 tcactgcatt ctagttgtgg tttgtccaaa ctcatcaatg tatcttatca tgtctggatc
     3121 tgatcactgc ttgagcctag aagatccggc tgctaacaaa gcccgaaagg aagctgagtt
     3181 ggctgctgcc accgctgagc aataactatc ataaccccta gggtataccc atctaaggta
     3241 gcgagtttaa acactagtat cgattcgcga cctactccgg aatattaata gatcatggag
     3301 ataattaaaa tgataaccat ctcgcaaata aataagtatt ttactgtttt cgtaacagtt
     3361 ttgtaataaa aaaacctata aatattccgg attattcata ccgtcccacc atcgggcgcg
     3421 gatccatggc actgtctacg attgtgagtc agcgtaaaca aatcaaacgt aaagcccctc
     3481 gtggtttcct gaaacgtgtg tttaaacgta aaaaacctca gctgcgcctg gaaaaatcag
     3541 gtgatctgct ggttcatctg aattgcctgt tattcgtcca ccgtctggca gaagaatccc
     3601 gtactaatgc ctgcgcttcg aaatgtcgtg tcatcaacaa agaacatgtg ctggctgccg
     3661 ctaaagtcat tctgaaaaaa tcccgtggct gagaattcaa aggcctacgt cgacgagctc
     3721 acttgtcgcg gccgctttcg aatctagagc ctgcagtctc gacaagcttg tcgagaagta
     3781 ctagaggatc ataatcagcc ataccacatt tgtagaggtt ttacttgctt taaaaaacct
     3841 cccacacctc cccctgaacc tgaaacataa aatgaatgca attgttgttg ttaacttgtt
     3901 tattgcagct tataatggtt acaaataaag caatagcatc acaaatttca caaataaagc
     3961 atttttttca ctgcattcta gttgtggttt gtccaaactc atcaatgtat cttatcatgt
     4021 ctggatctga tcactgcttg agcctagaag atccggctgc taacaaagcc cgaaaggaag
     4081 ctgagttggc tgctgccacc gctgagcaat aactatcata acccctaggg tatacccatc
     4141 taattggaac cagataagtg aaatctagtt ccaaactatt ttgtcatttt taattttcgt
     4201 attagcttac gacgctacac ccagttccca tctattttgt cactcttccc taaataatcc
     4261 ttaaaaactc catttccacc cctcccagtt cccaactatt ttgtccgccc acaaccggtt
     4321 gacttgggtc aactgtcaga ccaagtttac tcatatatac tttagattga tttaaaactt
     4381 catttttaat ttaaaaggat ctaggtgaag atcctttttg ataatctcat gaccaaaatc
     4441 ccttaacgtg agttttcgtt ccactgagcg tcagaccccg tagaaaagat caaaggatct
     4501 tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta
     4561 ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc tttttccgaa ggtaactggc
     4621 ttcagcagag cgcagatacc aaatactgtt cttctagtgt agccgtagtt aggccaccac
     4681 ttcaagaact ctgtagcacc gcctacatac ctcgctctgc taatcctgtt accagtggct
     4741 gctgccagtg gcgataagtc gtgtcttacc gggttggact caagacgata gttaccggat
     4801 aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac agcccagctt ggagcgaacg
     4861 acctacaccg aactgagata cctacagcgt gagctatgag aaagcgccac gcttcccgaa
     4921 gggagaaagg cggacaggta tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg
     4981 gagcttccag ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg ccacctctga
     5041 cttgagcgtc gatttttgtg atgctcgtca ggggggcgga gcctatggaa aaacgccagc
     5101 aacgcggcct ttttacggtt cctggccttt tgctggcctt ttgctcacat gttctttcct
     5161 gcgttatccc ctgattgact tgggtcgctc ttcctgtgga tgcgcagatg ccctgcgtaa
     5221 gcgggtgtgg gcggacaata aagtcttaaa ctgaacaaaa tagatctaaa ctatgacaat
     5281 aaagtcttaa actagacaga atagttgtaa actgaaatca gtccagttat gctgtgaaaa
     5341 agcatactgg acttttgtta tggctaaagc aaactcttca ttttctgaag tgcaaattgc
     5401 ccgtcgtatt aaagaggggc gtggccaagg gcatgtaaag actatattcg cggcgttgtg
     5461 acaatttacc gaacaactcc gcggccggga agccgatctc ggcttgaacg aattgttagg
     5521 tggcggtact tgggtcgata tcaaagtgca tcacttcttc ccgtatgccc aactttgtat
     5581 agagagccac tgcgggatcg tcaccgtaat ctgcttgcac gtagatcaca taagcaccaa
     5641 gcgcgttggc ctcatgcttg aggagattga tgagcgcggt ggcaatgccc tgcctccggt
     5701 gctcgccgga gactgcgaga tcatagatat agatctcact acgcggctgc tcaaacttgg
     5761 gcagaacgta agccgcgaga gcgccaacaa ccgcttcttg gtcgaaggca gcaagcgcga
     5821 tgaatgtctt actacggagc aagttcccga ggtaatcgga gtccggctga tgttgggagt
     5881 aggtggctac gtctccgaac tcacgaccga aaagatcaag agcagcccgc atggatttga
     5941 cttggtcagg gccgagccta catgtgcgaa tgatgcccat acttgagcca cctaactttg
     6001 ttttagggcg actgccctgc tgcgtaacat cgttgctgct gcgtaacatc gttgctgctc
     6061 cataacatca aacatcgacc cacggcgtaa cgcgcttgct gcttggatgc ccgaggcata
     6121 gactgtacaa aaaaacagtc ataacaagcc atgaaaaccg ccactgcgcc gtta
//
