LOCUS       sequence-272753-        8991 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Baculovirus expresion of Astrin, LC8, and SKAP.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8991)
  AUTHORS   Kern DM, Monda JK, Su KC, Wilson-Kubalek EM, Cheeseman IM
  TITLE     Astrin-SKAP complex reconstitution reveals its kinetochore 
            interaction with microtubule-bound Ndc80.
  JOURNAL   Elife. 2017 Aug 25;6. pii: 26866. doi: 10.7554/eLife.26866.
  PUBMED    28841134
REFERENCE   2  (bases 1 to 8991)
  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"; journalName: "Elife."; 
            date: "2017-08-25"; volume: "6. pii: 26866. doi"; pages: " 
            10.7554/eLife.26866"
COMMENT     Sequence Label: pDK521
FEATURES             Location/Qualifiers
     source          1..8991
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        complement(join(37..42,43..59,60..65))
                     /gene="intI1 (promoter lies within the coding sequence)"
                     /label=Pc promoter
                     /note="class 1 integron promoter"
     protein_bind    complement(136..169)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     promoter        246..337
                     /gene="polh from Autographa californica"
                     /label=polyhedrin promoter
                     /note="promoter for the baculovirus polyhedrin gene"
     CDS             complement(1492..1503)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     CDS             2525..3238
                     /codon_start=1
                     /product="GFP variant that folds robustly even when fused 
                     to poorly folded proteins (Pedelacq et al., 2006)"
                     /label=superfolder GFP
                     /translation="MRKGEELFTGVVPILVELDGDVNGHKFSVRGEGEGDATNGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFARYPDHMKQHDFFKSAMPEGYVQERTISFKDDG
                     TYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNFNSHNVYITADKQKNGIKA
                     NFKIRHNVEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSVLSKDPNEKRDHMVLLE
                     FVTAAGITHGMDELYK"
     CDS             3248..3268
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=7xHis
                     /translation="HHHHHHH"
     polyA_signal    3401..3535
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     promoter        3707..3798
                     /gene="polh from Autographa californica"
                     /label=polyhedrin promoter
                     /note="promoter for the baculovirus polyhedrin gene"
     polyA_signal    4779..4913
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     promoter        5085..5176
                     /gene="polh from Autographa californica"
                     /label=polyhedrin promoter
                     /note="promoter for the baculovirus polyhedrin gene"
     regulatory      5234..5243
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     polyA_signal    5752..5886
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     promoter        6058..6149
                     /gene="polh from Autographa californica"
                     /label=polyhedrin promoter
                     /note="promoter for the baculovirus polyhedrin gene"
     polyA_signal    6683..6817
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     mobile_element  6938..7103
                     /mobile_element_type="transposon:Tn7"
                     /label=Tn7L
                     /note="mini-Tn7 element (left end of the Tn7 transposon)"
     rep_origin      7293..7881
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7782..7801
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     mobile_element  8016..8239
                     /mobile_element_type="transposon:Tn7"
                     /label=Tn7R
                     /note="mini-Tn7 element (right end of the Tn7 transposon)"
     CDS             complement(8306..8839)
                     /codon_start=1
                     /gene="aacC1"
                     /product="gentamycin acetyltransferase"
                     /label=GmR
                     /note="confers resistance to gentamycin"
                     /translation="MLRSSNDVTQQGSRPKTKLGGSSMGIIRTCRLGPDQVKSMRAALD
                     LFGREFGDVATYSQHQPDSDYLGNLLRSKTFIALAAFDQEAVVGALAAYVLPKFEQPRS
                     EIYIYDLAVSGEHRRQGIATALINLLKHEANALGAYVIYVQADYGDDPAVALYTKLGIR
                     EEVMHFDIDPSTAT"
ORIGIN
        1 gaccagttgc gtgagcgcat acgctacttg cattacagtt tacgaaccga acaggcttat
       61 gtcaactggg ttcgtgcctt catccgtttc cacggtgtgc gtcacccggc aaccttgggc
      121 agcagcgaag tcgccataac ttcgtatagc atacattata cgaagttatc tgtaactata
      181 acggtcctaa ggtagcgagt ttaaacacta gtatcgattc gcgacctact ccggaatatt
      241 aatagatcat ggagataatt aaaatgataa ccatctcgca aataaataag tattttactg
      301 ttttcgtaac agttttgtaa taaaaaaacc tataaatatt ccggattatt cataccgtcc
      361 caccatcggg cgcggatcca actcctaaaa aaccgccacc atgtggcgag tgaaaaaact
      421 gagcctcagc ctgtcgcctt cgccccagac gggaaaacca tctatgagaa ctcctctccg
      481 tgaacttacc ctgcagcccg gtgccctcac caactctgga aaaagatccc ccgcttgctc
      541 ctcgctgacc ccatcactgt gcaagctggg gctgcaggaa ggcagcaaca actcatctcc
      601 agtggatttt gtaaataaca agaggacaga cttatcttca gaacatttca gtcattcctc
      661 aaagtggcta gaaacttgtc agcatgaatc agatgagcag cctctagatc caattcccca
      721 aattagctct actcctaaaa cgtctgagga agcagtagac ccactgggca attatatggt
      781 taaaaccatc gtccttgtac catctccact ggggcagcaa caagacatga tatttgaggc
      841 ccgtttagat accatggcag agacaaacag catatcttta aatggacctt tgagaacaga
      901 cgatctggtg agagaggagg tggcaccctg catgggagac aggttttcag aagttgctgc
      961 tgtatctgag aaacctatct ttcaggaatc tccgtcccat ctcttagagg agtctccacc
     1021 aaatccctgt tctgaacaac tacattgctc caaggaaagc ctgagcagta gaactgaggc
     1081 tgtgcgtgag gacttagtac cttctgaaag taacgccttc ttgccttcct ctgttctctg
     1141 gctttcccct tcaactgcct tggcagcaga tttccgtgtc aatcatgtgg acccagagga
     1201 ggaaattgta gagcatggag ctatggagga aagagaaatg aggtttccca cacatcctaa
     1261 ggagtctgaa acagaagatc aagcacttgt ctcaagtgtg gaagatattc tgtccacatg
     1321 cctgacacca aatctagtag aaatggaatc ccaagaagct ccaggcccag cagtagaaga
     1381 tgttggtagg attcttggct ctgatacaga gtcttggatg tccccgctgg cctggctgga
     1441 aaaaggtgta aatacctccg tcatgctgga aaatctccgc caaagcttat cccttccctc
     1501 gatgcttcgg gatgctgcaa ttggcactac ccctttctct acttgctcgg tggggacttg
     1561 gtttactcct tcagcaccac aggaaaagag tacaaacaca tcccagacag gcctggttgg
     1621 caccaagcac agtacttctg agacagagca gctcctgtgt ggccggcctc cagatctgac
     1681 tgccttgtct cgacatgact tggaagataa cctgctgagc tctcttgtca ttctggaggt
     1741 tctctcccgc cagcttcggg actggaagag ccagctggct gtccctcacc cagaaaccca
     1801 ggacagtagc acacagactg acacatctca cagtgggata actaataaac ttcagcatct
     1861 taaggagagc catgagatgg gacaggccct acagcaggcc agaaatgtca tgcaatcatg
     1921 ggtgcttatc tctaaagagc tgatatcctt gcttcaccta tccctgttgc atttagaaga
     1981 agataagact actgtgagtc aggagtctcg gcgtgcagaa acattggtct gttgctgttt
     2041 tgatttgctg aagaaattga gggcaaagct ccagagcctc aaagcagaaa gggaggaggc
     2101 aaggcacaga gaggaaatgg ctctcagagg caaggatgcg gcagagatag tgttggaggc
     2161 tttctgtgca cacgccagcc agcgcatcag ccagctggaa caggacctag catccatgcg
     2221 ggaattcaga ggccttctga aggatgccca gacccaactg gtagggcttc atgccaagca
     2281 agaagagctg gttcagcaga cagtgagtct tacttctacc ttgcaacaag actggaggag
     2341 tatgcaactg gattatacaa catggacagc tttgctgagt cggtcccgac aactcacaga
     2401 gaaactcaca gtcaagagcc agcaagccct gcaggaacgt gatgtggcaa ttgaggaaaa
     2461 gcaggaggtt tctagggtgt ctagaggtgg atctggagct gcagctggat ctggttctgg
     2521 ttctatgcgt aaaggcgaag agctgttcac tggtgtcgtc cctattctgg tggaactgga
     2581 tggtgatgtc aacggtcata agttttccgt gcgtggcgag ggtgaaggtg acgcaactaa
     2641 tggtaaactg acgctgaagt tcatctgtac tactggtaaa ctgccggtac cttggccgac
     2701 tctggtaacg acgctgactt atggtgttca gtgctttgct cgttatccgg accatatgaa
     2761 gcagcatgac ttcttcaagt ccgccatgcc ggaaggctat gtgcaggaac gcacgatttc
     2821 ctttaaggat gacggcacgt acaaaacgcg tgcggaagtg aaatttgaag gcgataccct
     2881 ggtaaaccgc attgagctga aaggcattga ctttaaagaa gacggcaata tcctgggcca
     2941 taagctggaa tacaatttta acagccacaa tgtttacatc accgccgata aacaaaaaaa
     3001 tggcattaaa gcgaatttta aaattcgcca caacgtggag gatggcagcg tgcagctggc
     3061 tgatcactac cagcaaaaca ctccaatcgg tgatggtcct gttctgctgc cagacaatca
     3121 ctatctgagc acgcaaagcg ttctgtctaa agatccgaac gagaaacgcg atcatatggt
     3181 tctgctggag ttcgtaaccg cagcgggcat cacgcatggt atggatgaac tgtacaaagg
     3241 cagcagccat catcatcatc atcaccactg aaagcttgtc gagaagtact agaggatcat
     3301 aatcagccat accacatttg tagaggtttt acttgcttta aaaaacctcc cacacctccc
     3361 cctgaacctg aaacataaaa tgaatgcaat tgttgttgtt aacttgttta ttgcagctta
     3421 taatggttac aaataaagca atagcatcac aaatttcaca aataaagcat ttttttcact
     3481 gcattctagt tgtggtttgt ccaaactcat caatgtatct tatcatgtct ggatctgatc
     3541 actgcttgag cctagaagat ccggctgcta acaaagcccg aaaggaagct gagttggctg
     3601 ctgccaccgc tgagcaataa ctatcataac ccctagggta tacccatcta aggtagcgag
     3661 tttaaacact agtatcgatt cgcgacctac tccggaatat taatagatca tggagataat
     3721 taaaatgata accatctcgc aaataaataa gtattttact gttttcgtaa cagttttgta
     3781 ataaaaaaac ctataaatat tccggattat tcataccgtc ccaccatcgg gcgcggatcc
     3841 aactcctaaa aaaccgccac catgaccagt gtggttaaga cagtgtatag cctgcagccc
     3901 ccctctgcgc tgagcggcgg ccagccggca gacacacaaa ctcgggccac ttctaagagt
     3961 ctcttacctg ttaggtccaa agaagtcgat gtttccaaac agcttcattc aggaggtcca
     4021 gagaatgatg ttacaaaaat caccaaactg agacgagaga atgggcaaat gaaagctact
     4081 gacactgcca ccagaaggaa tgtcagaaaa ggctacaaac cactgagtaa gcaaaaatca
     4141 gaggaagagc tcaaggacaa gaaccagctg ttagaagccg tcaacaagca gttgcaccag
     4201 aagttgactg aaactcaggg agagctgaag gacctgaccc agaaggtaga gctgctggag
     4261 aagtttcggg acaactgttt ggcaattttg gagagcaagg gccttgatcc agctttaggc
     4321 agtgagaccc tggcatcacg acaagaatcc actactgatc acatggactc tatgttgctg
     4381 ttagaaactt tgcaagagga gctgaagctt tttaacgaaa cagccaaaaa gcagatggag
     4441 gagttacagg ccttaaaggt aaagctggag atgaaagagg aaagagtccg attcctagaa
     4501 cagcaaacct tatgtaacaa tcaagtaaat gatttaacaa cagcccttaa ggaaatggag
     4561 cagctattag aaatgtaaga attcaaaggc ctacgtcgac gagctcactt gtcgcggccg
     4621 ctttcgaatc tagagcctgc agtctcgaca agcttgtcga gaagtactag aggatcataa
     4681 tcagccatac cacatttgta gaggttttac ttgctttaaa aaacctccca cacctccccc
     4741 tgaacctgaa acataaaatg aatgcaattg ttgttgttaa cttgtttatt gcagcttata
     4801 atggttacaa ataaagcaat agcatcacaa atttcacaaa taaagcattt ttttcactgc
     4861 attctagttg tggtttgtcc aaactcatca atgtatctta tcatgtctgg atctgatcac
     4921 tgcttgagcc tagaagatcc ggctgctaac aaagcccgaa aggaagctga gttggctgct
     4981 gccaccgctg agcaataact atcataaccc ctagggtata cccatctaag gtagcgagtt
     5041 taaacactag tatcgattcg cgacctactc cggaatatta atagatcatg gagataatta
     5101 aaatgataac catctcgcaa ataaataagt attttactgt tttcgtaaca gttttgtaat
     5161 aaaaaaacct ataaatattc cggattattc ataccgtccc accatcgggc gcggatccaa
     5221 ctcctaaaaa accgccacca tggcccatta caaagccgcc gactcgaagc gtgagcagtt
     5281 ccggaggtac ttggagaagt cgggggtgct ggacacgctg actaaggtgt tggtagcctt
     5341 atatgaagaa ccagagaaac ctaacagtgc tttggatttt ttaaagcatc acttaggagc
     5401 tgctactcca gaaaatccag aaatagagct gcttcgccta gaactggccg aaatgaaaga
     5461 gaagtatgaa gctattgtag aagaaaataa aaaactgaaa gcaaagcttg ctcagtatga
     5521 accacctcag gaggagaagc gtgctgaata ggaattcaaa ggcctacgtc gacgagctca
     5581 cttgtcgcgg ccgctttcga atctagagcc tgcagtctcg acaagcttgt cgagaagtac
     5641 tagaggatca taatcagcca taccacattt gtagaggttt tacttgcttt aaaaaacctc
     5701 ccacacctcc ccctgaacct gaaacataaa atgaatgcaa ttgttgttgt taacttgttt
     5761 attgcagctt ataatggtta caaataaagc aatagcatca caaatttcac aaataaagca
     5821 tttttttcac tgcattctag ttgtggtttg tccaaactca tcaatgtatc ttatcatgtc
     5881 tggatctgat cactgcttga gcctagaaga tccggctgct aacaaagccc gaaaggaagc
     5941 tgagttggct gctgccaccg ctgagcaata actatcataa cccctagggt atacccatct
     6001 aaggtagcga gtttaaacac tagtatcgat tcgcgaccta ctccggaata ttaatagatc
     6061 atggagataa ttaaaatgat aaccatctcg caaataaata agtattttac tgttttcgta
     6121 acagttttgt aataaaaaaa cctataaata ttccggatta ttcataccgt cccaccatcg
     6181 ggcgcggatc caactcctaa aaaaccgcca ccatgtgcga ccgaaaggcc gtgatcaaaa
     6241 atgcggacat gtcggaagag atgcaacagg actcggtgga gtgcgctact caggcgctgg
     6301 agaaatacaa catagagaag gacattgcgg ctcatatcaa gaaggaattt gacaagaagt
     6361 acaatcccac ctggcattgc atcgtgggga ggaacttcgg tagttatgtg acacatgaaa
     6421 ccaaacactt catctacttc tacctgggcc aagtggccat tcttctgttc aaatctggtt
     6481 aagaattcaa aggcctacgt cgacgagctc acttgtcgcg gccgctttcg aatctagagc
     6541 ctgcagtctc gacaagcttg tcgagaagta ctagaggatc ataatcagcc ataccacatt
     6601 tgtagaggtt ttacttgctt taaaaaacct cccacacctc cccctgaacc tgaaacataa
     6661 aatgaatgca attgttgttg ttaacttgtt tattgcagct tataatggtt acaaataaag
     6721 caatagcatc acaaatttca caaataaagc atttttttca ctgcattcta gttgtggttt
     6781 gtccaaactc atcaatgtat cttatcatgt ctggatctga tcactgcttg agcctagaag
     6841 atccggctgc taacaaagcc cgaaaggaag ctgagttggc tgctgccacc gctgagcaat
     6901 aactatcata acccctaggg tatacccatc taattggaac cagataagtg aaatctagtt
     6961 ccaaactatt ttgtcatttt taattttcgt attagcttac gacgctacac ccagttccca
     7021 tctattttgt cactcttccc taaataatcc ttaaaaactc catttccacc cctcccagtt
     7081 cccaactatt ttgtccgccc acaaccggtt gacttgggtc aactgtcaga ccaagtttac
     7141 tcatatatac tttagattga tttaaaactt catttttaat ttaaaaggat ctaggtgaag
     7201 atcctttttg ataatctcat gaccaaaatc ccttaacgtg agttttcgtt ccactgagcg
     7261 tcagaccccg tagaaaagat caaaggatct tcttgagatc ctttttttct gcgcgtaatc
     7321 tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc ggatcaagag
     7381 ctaccaactc tttttccgaa ggtaactggc ttcagcagag cgcagatacc aaatactgtt
     7441 cttctagtgt agccgtagtt aggccaccac ttcaagaact ctgtagcacc gcctacatac
     7501 ctcgctctgc taatcctgtt accagtggct gctgccagtg gcgataagtc gtgtcttacc
     7561 gggttggact caagacgata gttaccggat aaggcgcagc ggtcgggctg aacggggggt
     7621 tcgtgcacac agcccagctt ggagcgaacg acctacaccg aactgagata cctacagcgt
     7681 gagctatgag aaagcgccac gcttcccgaa gggagaaagg cggacaggta tccggtaagc
     7741 ggcagggtcg gaacaggaga gcgcacgagg gagcttccag ggggaaacgc ctggtatctt
     7801 tatagtcctg tcgggtttcg ccacctctga cttgagcgtc gatttttgtg atgctcgtca
     7861 ggggggcgga gcctatggaa aaacgccagc aacgcggcct ttttacggtt cctggccttt
     7921 tgctggcctt ttgctcacat gttctttcct gcgttatccc ctgattgact tgggtcgctc
     7981 ttcctgtgga tgcgcagatg ccctgcgtaa gcgggtgtgg gcggacaata aagtcttaaa
     8041 ctgaacaaaa tagatctaaa ctatgacaat aaagtcttaa actagacaga atagttgtaa
     8101 actgaaatca gtccagttat gctgtgaaaa agcatactgg acttttgtta tggctaaagc
     8161 aaactcttca ttttctgaag tgcaaattgc ccgtcgtatt aaagaggggc gtggccaagg
     8221 gcatgtaaag actatattcg cggcgttgtg acaatttacc gaacaactcc gcggccggga
     8281 agccgatctc ggcttgaacg aattgttagg tggcggtact tgggtcgata tcaaagtgca
     8341 tcacttcttc ccgtatgccc aactttgtat agagagccac tgcgggatcg tcaccgtaat
     8401 ctgcttgcac gtagatcaca taagcaccaa gcgcgttggc ctcatgcttg aggagattga
     8461 tgagcgcggt ggcaatgccc tgcctccggt gctcgccgga gactgcgaga tcatagatat
     8521 agatctcact acgcggctgc tcaaacttgg gcagaacgta agccgcgaga gcgccaacaa
     8581 ccgcttcttg gtcgaaggca gcaagcgcga tgaatgtctt actacggagc aagttcccga
     8641 ggtaatcgga gtccggctga tgttgggagt aggtggctac gtctccgaac tcacgaccga
     8701 aaagatcaag agcagcccgc atggatttga cttggtcagg gccgagccta catgtgcgaa
     8761 tgatgcccat acttgagcca cctaactttg ttttagggcg actgccctgc tgcgtaacat
     8821 cgttgctgct gcgtaacatc gttgctgctc cataacatca aacatcgacc cacggcgtaa
     8881 cgcgcttgct gcttggatgc ccgaggcata gactgtacaa aaaaacagtc ataacaagcc
     8941 atgaaaaccg ccactgcgcc gttaccaccg ctgcgttcgg tcaaggttct g
//
