LOCUS       sequence-272291-        9207 bp    DNA     circular SYN 15-APR-2025
DEFINITION  Baculovirus expression of CENP-HKM and His-CENP-I.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9207)
  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 9207)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 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: pKM377
FEATURES             Location/Qualifiers
     source          1..9207
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        complement(join(47..52,53..69,70..75))
                     /gene="intI1 (promoter lies within the coding sequence)"
                     /label=Pc promoter
                     /note="class 1 integron promoter"
     protein_bind    complement(146..179)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     promoter        256..347
                     /gene="polh from Autographa californica"
                     /label=polyhedrin promoter
                     /note="promoter for the baculovirus polyhedrin gene"
     polyA_signal    1318..1452
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     promoter        1624..1715
                     /gene="polh from Autographa californica"
                     /label=polyhedrin promoter
                     /note="promoter for the baculovirus polyhedrin gene"
     polyA_signal    2752..2886
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     promoter        3058..3149
                     /gene="polh from Autographa californica"
                     /label=polyhedrin promoter
                     /note="promoter for the baculovirus polyhedrin gene"
     regulatory      3630..3639
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     polyA_signal    3956..4090
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     promoter        4262..4353
                     /gene="polh from Autographa californica"
                     /label=polyhedrin promoter
                     /note="promoter for the baculovirus polyhedrin gene"
     CDS             4399..4419
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=7xHis
                     /translation="HHHHHHH"
     polyA_signal    6909..7043
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     mobile_element  7164..7329
                     /mobile_element_type="transposon:Tn7"
                     /label=Tn7L
                     /note="mini-Tn7 element (left end of the Tn7 transposon)"
     rep_origin      7519..8107
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     8008..8027
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     mobile_element  8242..8465
                     /mobile_element_type="transposon:Tn7"
                     /label=Tn7R
                     /note="mini-Tn7 element (right end of the Tn7 transposon)"
     CDS             complement(8532..9065)
                     /codon_start=1
                     /gene="aacC1"
                     /product="gentamycin acetyltransferase"
                     /label=GmR
                     /note="confers resistance to gentamycin"
                     /translation="MLRSSNDVTQQGSRPKTKLGGSSMGIIRTCRLGPDQVKSMRAALD
                     LFGREFGDVATYSQHQPDSDYLGNLLRSKTFIALAAFDQEAVVGALAAYVLPKFEQPRS
                     EIYIYDLAVSGEHRRQGIATALINLLKHEANALGAYVIYVQADYGDDPAVALYTKLGIR
                     EEVMHFDIDPSTAT"
ORIGIN
        1 caaggttctg gaccagttgc gtgagcgcat acgctacttg cattacagtt tacgaaccga
       61 acaggcttat gtcaactggg ttcgtgcctt catccgtttc cacggtgtgc gtcacccggc
      121 aaccttgggc agcagcgaag tcgccataac ttcgtatagc atacattata cgaagttatc
      181 tgtaactata acggtcctaa ggtagcgagt ttaaacacta gtatcgattc gcgacctact
      241 ccggaatatt aatagatcat ggagataatt aaaatgataa ccatctcgca aataaataag
      301 tattttactg ttttcgtaac agttttgtaa taaaaaaacc tataaatatt ccggattatt
      361 cataccgtcc caccatcggg cgcggatcca tggaggaaca gccacaaatg caggatgccg
      421 atgaaccagc agactcagga ggtgaaggtc gcgcaggagg cccgccacaa gttgccggag
      481 cacaagcagc gtgtagtgag gatcgtatga ccctgctgct gcgcctgcgt gcacaaacaa
      541 aacagcaact gctggagtat aaaagcatgg tcgatgccag cgaagaaaaa accccagagc
      601 aaatcatgca agagaaacaa atcgaggcca aaatcgagga cctggaaaac gaaattgaag
      661 aagtgaaagt cgcctttgag atcaaaaaac tggccctgga tcgtatgcgt ctgtctacag
      721 ccctgaaaaa aaacctggag aaaatcagtc gtcagtcaag cgttctgatg gacaacatga
      781 aacacctgct ggaactgaac aaactgatca tgaaatctca gcaagagtcg tgggatctgg
      841 aggagaaact gctggatatt cgtaaaaaac gcctgcagct gaaacaagca agcgagtcta
      901 aactgctcga gatccagacc gaaaaaaaca aacagaaaat cgacctggac agcatggaaa
      961 attccgagcg tatcaaaatc atccgccaga atctgcagat ggaaatcaaa atcaccaccg
     1021 tcatccaaca cgtctttcag aacctgatcc tgggtagcaa agtgaattgg gccgaggacc
     1081 ctgccctgaa agagattgtt ctgcaactgg agaaaaatgt ggacatgatg tgagtcgacg
     1141 agctcacttg tcgcggccgc tttcgaatct agagcctgca gtctcgacaa gcttgtcgag
     1201 aagtactaga ggatcataat cagccatacc acatttgtag aggttttact tgctttaaaa
     1261 aacctcccac acctccccct gaacctgaaa cataaaatga atgcaattgt tgttgttaac
     1321 ttgtttattg cagcttataa tggttacaaa taaagcaata gcatcacaaa tttcacaaat
     1381 aaagcatttt tttcactgca ttctagttgt ggtttgtcca aactcatcaa tgtatcttat
     1441 catgtctgga tctgatcact gcttgagcct agaagatccg gctgctaaca aagcccgaaa
     1501 ggaagctgag ttggctgctg ccaccgctga gcaataacta tcataacccc tagggtatac
     1561 ccatctaagg tagcgagttt aaacactagt atcgattcgc gacctactcc ggaatattaa
     1621 tagatcatgg agataattaa aatgataacc atctcgcaaa taaataagta ttttactgtt
     1681 ttcgtaacag ttttgtaata aaaaaaccta taaatattcc ggattattca taccgtccca
     1741 ccatcgggcg cggatccatg aaccaggagg atctggaccc tgatagtact accgatgtcg
     1801 gtgatgtgac caataccgaa gaagagctga ttcgtgagtg tgaagaaatg tggaaagaca
     1861 tggaggaatg ccaaaacaaa ctgagcctga ttggaaccga aactctgacc gatagcaatg
     1921 cccaactgag tctgctgatt atgcaggtga aatgcctgac tgctgaactg agccaatggc
     1981 agaaaaaaac accggaaacc atcccgctga ctgaagatgt gctgattacg ctgggtaaag
     2041 aagaatttca gaaactgcgc caggacctgg aaatggtgct gtctaccaaa gagtccaaaa
     2101 acgaaaaact gaaagaggac ctggagcgtg aacaacgttg gctggacgaa cagcagcaaa
     2161 tcatggaatc cctgaacgtt ctgcactctg agctgaaaaa caaagtggaa acctttagcg
     2221 aaagccgtat ctttaacgaa ctgaaaacaa aaatgctgaa catcaaagag tataaagaga
     2281 aactgctgag tacactgggg gaatttctgg aagatcactt ccctctgcct gatcgtagtg
     2341 ttaaaaaaaa aaaaaaaaac atccaagaaa gctccgtgaa cctgatcact ctgcacgaga
     2401 tgctggaaat cctgattaat cgcctgtttg acgtgccaca cgacccttat gtgaaaatct
     2461 ccgacagctt ctggcctcct tatgttgaac tgctgctgcg taatggtatt gcactgcgtc
     2521 atccagagga ccctacccgt attcgtctgg aggcgttcca tcaataagtc gacgagctca
     2581 cttgtcgcgg ccgctttcga atctagagcc tgcagtctcg acaagcttgt cgagaagtac
     2641 tagaggatca taatcagcca taccacattt gtagaggttt tacttgcttt aaaaaacctc
     2701 ccacacctcc ccctgaacct gaaacataaa atgaatgcaa ttgttgttgt taacttgttt
     2761 attgcagctt ataatggtta caaataaagc aatagcatca caaatttcac aaataaagca
     2821 tttttttcac tgcattctag ttgtggtttg tccaaactca tcaatgtatc ttatcatgtc
     2881 tggatctgat cactgcttga gcctagaaga tccggctgct aacaaagccc gaaaggaagc
     2941 tgagttggct gctgccaccg ctgagcaata actatcataa cccctagggt atacccatct
     3001 aaggtagcga gtttaaacac tagtatcgat tcgcgaccta ctccggaata ttaatagatc
     3061 atggagataa ttaaaatgat aaccatctcg caaataaata agtattttac tgttttcgta
     3121 acagttttgt aataaaaaaa cctataaata ttccggatta ttcataccgt cccaccatcg
     3181 ggcgcggatc caactcctaa aaaaccgcca ccatgtcggt gttgaggccc ctggacaagc
     3241 tgcccggcct gaacacggcc acaatcttgc tggtgggcac ggaggatgct cttctgcagc
     3301 agctggcgga ctcgatgctc aaagaggact gcgcctccga gctgaaggtc cacttggcaa
     3361 agtccctccc tttgccctcc agtgtgaatc ggccccgaat tgacctgatc gtgtttgtgg
     3421 ttaatcttca cagcaaatac agtctccaga acacagagga gtccctgcgc catgtggatg
     3481 ccagcttctt cttggggaag gtgtgtttcc tcgccacagg tgctgggcgg gagagccact
     3541 gcagcattca ccggcacacc gtggtgaagc tggcccacac ctatcaaagc cccctgctct
     3601 actgtgacct ggaggtggaa ggctttaggg ccaccatggc gcagcgcctg gtgcgcgtgc
     3661 tgcagatctg tgctggccac gtgcccggtg tctcagctct gaacctgctg tccctgctga
     3721 gaagctctga gggcccctcc ctggaggacc tgtgagaatt caaaggccta cgtcgacgag
     3781 ctcacttgtc gcggccgctt tcgaatctag agcctgcagt ctcgacaagc ttgtcgagaa
     3841 gtactagagg atcataatca gccataccac atttgtagag gttttacttg ctttaaaaaa
     3901 cctcccacac ctccccctga acctgaaaca taaaatgaat gcaattgttg ttgttaactt
     3961 gtttattgca gcttataatg gttacaaata aagcaatagc atcacaaatt tcacaaataa
     4021 agcatttttt tcactgcatt ctagttgtgg tttgtccaaa ctcatcaatg tatcttatca
     4081 tgtctggatc tgatcactgc ttgagcctag aagatccggc tgctaacaaa gcccgaaagg
     4141 aagctgagtt ggctgctgcc accgctgagc aataactatc ataaccccta gggtataccc
     4201 atctaaggta gcgagtttaa acactagtat cgattcgcga cctactccgg aatattaata
     4261 gatcatggag ataattaaaa tgataaccat ctcgcaaata aataagtatt ttactgtttt
     4321 cgtaacagtt ttgtaataaa aaaacctata aatattccgg attattcata ccgtcccacc
     4381 atcgggcgcg gatccatgca tcatcatcat catcatcatg gcagcggcag catgagccca
     4441 cagaaacgtg taaaaaatgt ccaggcccag aaccgtacca gtcaaggcag tagcagcttc
     4501 caaacaaccc tgtcagcatg gaaagtgaaa caggacccga gcaacagcaa aaacatctct
     4561 aaacatggcc agaataaccc agttggcgac tatgagcacg ccgatgatca agcagaagag
     4621 gacgccctgc aaatggcggt aggctatttc gaaaaaggcc ctatcaaagc cagtcagaac
     4681 aaagacaaaa cgctggagaa acacctgaaa accgtggaga acgtggcatg gaaaaatggt
     4741 ctggcgagcg aagaaatcga catcctgctg aatattgccc tgagcggcaa atttggtaat
     4801 gccgtgaata cccgtatcct gaaatgtatg atcccggcaa ccgtgattag tgaagattcc
     4861 gtcgtgaaag cggtgtcatg gctgtgtgtt ggcaaatgtt ccggtagcac gaaagttctg
     4921 ttctatcgct ggctggttgc catgttcgat tttatcgacc gtaaagagca gatcaacctg
     4981 ctgtatggct tcttctttgc ctcactgcag gacgatgccc tgtgtcctta tgtgtgtcac
     5041 ctgctgtatc tgctgaccaa aaaagaaaac gtcaaacctt tccgtgtccg taaactgctg
     5101 gacctgcaag ccaaaatggg tatgcagcct cacctgcaag cactgctgtc tctgtataaa
     5161 ttctttgccc cggcactgat ttccgttagt ctgccggttc gtaaaaaaat ctatttcaaa
     5221 aacagcgaga acctgtggaa aacggctctg ctggccgtta aacagcgtaa tcgtggtcct
     5281 tctcctgaac cgctgaaact gatgctggga ccggctaacg ttcgtccgct gaaacgtaaa
     5341 tggaactccc tgtcggtgat tccagtactg aactcaagct cgtatactaa agagtgtggt
     5401 aaaaaagaga tgagcctgag cgattgtctg aatcgtagtg gctccttccc tctggagcag
     5461 ctgcaatctt tcccacaact gctgcaaaac atccattgtc tggaactgcc tagccaaatg
     5521 ggctctgtgc tgaacaattc gctgctgctg cactatatca actgtgtccg tgatgaacca
     5581 gttctgctgc gcttctatta ttggctgtct cagactctgc aggaagagtg tatctggtat
     5641 aaagtgaaca actatgagcg tggtaaagag ttcactaact ttctcgacac cattattcgt
     5701 gccgaatgct ttctgcaaga gggcttctat tcctgtgaag cgttcctgta taaaagcctg
     5761 ccactgtggg atgggctgtg ttgtcgtagc cagtttctgc aactggtgag ctggattcct
     5821 tttagcagct ttagcgaggt taaaccgctg ctgtttgacc atctggcaca gctgttcttc
     5881 accagcacca tctatttcaa atgctccgtg ctgcaatccc tgaaagagct gctgcaaaat
     5941 tggctgctgt ggctgtctat ggatattcac atgaaaccgg tgaccaattc tcctctggaa
     6001 accactctgg gtggttctat gaatagcgtg agcaaactga tccactatgt gggttggctg
     6061 agtacaaccg caatgcgtct ggagagcaac aacacctttc tgctgcattt catcctggac
     6121 ttctatgaga aagtgtgtga catctatatc aattataacc tgcctctggt tgtgctgttc
     6181 cctcctggca tcttctatag cgccctgctg tctctggata ctagcatcct gaatcaactg
     6241 tgcttcatta tgcaccgtta tcgtaaaaac ctgacggcgg cgaaaaaaaa cgagctggta
     6301 cagaaaacca aaagcgagtt caacttcagc tccaaaacct atcaagagtt taatcattat
     6361 ctgacctcca tggtcgggtg tctgtggaca tcaaaaccgt tcggtaaagg catctatatc
     6421 gacccggaga tcctggaaaa aacgggtgta gccgagtata aaaactccct gaacgtggtc
     6481 caccatccgt catttctgtc ttatgctgtg tccttcctgc tgcaggaatc tcctgaagag
     6541 cgtaccgtta atgtgagcag tattcgtggc aaaaaatggt cgtggtatct ggactatctg
     6601 ttctctcaag gtctgcaagg gctgaaactg tttatccgta gcagcgttca ccatagcagt
     6661 atccctcgtg ctgaagggat caactgtaat aaccagtatc tggagtgaga attcaaaggc
     6721 ctacgtcgac gagctcactt gtcgcggccg ctttcgaatc tagagcctgc agtctcgaca
     6781 agcttgtcga gaagtactag aggatcataa tcagccatac cacatttgta gaggttttac
     6841 ttgctttaaa aaacctccca cacctccccc tgaacctgaa acataaaatg aatgcaattg
     6901 ttgttgttaa cttgtttatt gcagcttata atggttacaa ataaagcaat agcatcacaa
     6961 atttcacaaa taaagcattt ttttcactgc attctagttg tggtttgtcc aaactcatca
     7021 atgtatctta tcatgtctgg atctgatcac tgcttgagcc tagaagatcc ggctgctaac
     7081 aaagcccgaa aggaagctga gttggctgct gccaccgctg agcaataact atcataaccc
     7141 ctagggtata cccatctaat tggaaccaga taagtgaaat ctagttccaa actattttgt
     7201 catttttaat tttcgtatta gcttacgacg ctacacccag ttcccatcta ttttgtcact
     7261 cttccctaaa taatccttaa aaactccatt tccacccctc ccagttccca actattttgt
     7321 ccgcccacaa ccggttgact tgggtcaact gtcagaccaa gtttactcat atatacttta
     7381 gattgattta aaacttcatt tttaatttaa aaggatctag gtgaagatcc tttttgataa
     7441 tctcatgacc aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag accccgtaga
     7501 aaagatcaaa ggatcttctt gagatccttt ttttctgcgc gtaatctgct gcttgcaaac
     7561 aaaaaaacca ccgctaccag cggtggtttg tttgccggat caagagctac caactctttt
     7621 tccgaaggta actggcttca gcagagcgca gataccaaat actgttcttc tagtgtagcc
     7681 gtagttaggc caccacttca agaactctgt agcaccgcct acatacctcg ctctgctaat
     7741 cctgttacca gtggctgctg ccagtggcga taagtcgtgt cttaccgggt tggactcaag
     7801 acgatagtta ccggataagg cgcagcggtc gggctgaacg gggggttcgt gcacacagcc
     7861 cagcttggag cgaacgacct acaccgaact gagataccta cagcgtgagc tatgagaaag
     7921 cgccacgctt cccgaaggga gaaaggcgga caggtatccg gtaagcggca gggtcggaac
     7981 aggagagcgc acgagggagc ttccaggggg aaacgcctgg tatctttata gtcctgtcgg
     8041 gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct
     8101 atggaaaaac gccagcaacg cggccttttt acggttcctg gccttttgct ggccttttgc
     8161 tcacatgttc tttcctgcgt tatcccctga ttgacttggg tcgctcttcc tgtggatgcg
     8221 cagatgccct gcgtaagcgg gtgtgggcgg acaataaagt cttaaactga acaaaataga
     8281 tctaaactat gacaataaag tcttaaacta gacagaatag ttgtaaactg aaatcagtcc
     8341 agttatgctg tgaaaaagca tactggactt ttgttatggc taaagcaaac tcttcatttt
     8401 ctgaagtgca aattgcccgt cgtattaaag aggggcgtgg ccaagggcat gtaaagacta
     8461 tattcgcggc gttgtgacaa tttaccgaac aactccgcgg ccgggaagcc gatctcggct
     8521 tgaacgaatt gttaggtggc ggtacttggg tcgatatcaa agtgcatcac ttcttcccgt
     8581 atgcccaact ttgtatagag agccactgcg ggatcgtcac cgtaatctgc ttgcacgtag
     8641 atcacataag caccaagcgc gttggcctca tgcttgagga gattgatgag cgcggtggca
     8701 atgccctgcc tccggtgctc gccggagact gcgagatcat agatatagat ctcactacgc
     8761 ggctgctcaa acttgggcag aacgtaagcc gcgagagcgc caacaaccgc ttcttggtcg
     8821 aaggcagcaa gcgcgatgaa tgtcttacta cggagcaagt tcccgaggta atcggagtcc
     8881 ggctgatgtt gggagtaggt ggctacgtct ccgaactcac gaccgaaaag atcaagagca
     8941 gcccgcatgg atttgacttg gtcagggccg agcctacatg tgcgaatgat gcccatactt
     9001 gagccaccta actttgtttt agggcgactg ccctgctgcg taacatcgtt gctgctgcgt
     9061 aacatcgttg ctgctccata acatcaaaca tcgacccacg gcgtaacgcg cttgctgctt
     9121 ggatgcccga ggcatagact gtacaaaaaa acagtcataa caagccatga aaaccgccac
     9181 tgcgccgtta ccaccgctgc gttcggt
//
