LOCUS       sequence-270314-        4131 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Bacterial expression of His-TEV-Ska1 and Ska2.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4131)
  AUTHORS   Monda JK, Whitney IP, Tarasovetc EV, Wilson-Kubalek E, Milligan RA, 
            Grishchuk EL, Cheeseman IM
  TITLE     Microtubule Tip Tracking by the Spindle and Kinetochore Protein Ska1
            Requires Diverse Tubulin-Interacting Surfaces.
  JOURNAL   Curr Biol. 2017 Dec 4;27(23):3666-3675.e6. doi: 
            10.1016/j.cub.2017.10.018. Epub 2017 Nov 16.
  PUBMED    29153323
REFERENCE   2  (bases 1 to 4131)
  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.cub.2017.10.018"; journalName: "Curr Biol"; date: 
            "2017-12-4- 4"; volume: "27"; issue: "23"; pages: "3666-3675.e6"
COMMENT     Sequence Label: pIW086
FEATURES             Location/Qualifiers
     source          1..4131
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     163..182
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        163..181
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     RBS             213..235
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     primer_bind     633..650
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     634..650
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     RBS             681..703
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             723..740
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             768..788
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     primer_bind     complement(1697..1715)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      1711..1758
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     promoter        1825..1916
                     /gene="bla"
                     /label=AmpR promoter
     terminator      2781..2820
                     /label=fd terminator
                     /note="central terminator from bacteriophage fd (Otsuka and
                     Kunisawa, 1982)"
     rep_origin      2851..3439
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3340..3359
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3593..3610
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    3625..3764
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(3690..3709)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3850..3872
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(3866..4057)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
ORIGIN
        1 acgggttact gatgatgaac atgcccggtt actggaacgt tgtgagggta aacaactggc
       61 ggtatggatg cggcgggacc agagaaaaat cactcagggt caatgccagc gcttcgttaa
      121 tacagatgta ggtgttggca ccatgcatcg atcccgcgaa attaatacga ctcactatag
      181 ggagaccaca acggtttccc tctagaaata attttgttta actttaagaa ggagatatac
      241 atatggaggc ggaggtcgat aagctggaac tgatgttcca gaaagctgag tctgatctgg
      301 attacattca atacaggctg gaatatgaaa tcaagactaa tcatcctgat tcagcaagtg
      361 agaaaaatcc agttacactc ttaaaggaat tgtcagtgat aaagtctcga tatcaaactt
      421 tgtatgcccg ctttaaacca gttgctgttg agcagaaaga gagtaagagc cgcatttgtg
      481 ctactgtgaa aaagactatg aatatgatac aaaaactaca gaagcaaaca gacctggagc
      541 tgtcaccact gactaaagaa gagaaaactg cggcagagca attcaaattt cacatgccag
      601 atttatgagg atccgggccc tgtacaagat cctgtaaaac gacggccagt gaattggaat
      661 tcgctagctc tagaaataat tttgtttaac tttaagaagg agatatacat atgggcagca
      721 gccatcatca tcatcatcac agcagcggat ctggtggtgg tggtggtgaa aacctgtact
      781 tccagggatc catggcctcg tcagatctgg aacaattatg ctctcatgtt aatgaaaaga
      841 ttggcaatat taagaaaacc ttatcattaa gaaactgtgg ccaggaacct accttgaaaa
      901 ctgtattaaa taaaatagga gatgagatca ttgtaataaa tgaacttcta aataaattgg
      961 aattggaaat tcagtatcaa gaacaaacca acaattcact caaggaactc tgtgaatctc
     1021 ttgaagaaga ttacaaagac atagaacatc ttaaagaaaa cgttccttcc catttgcctc
     1081 aagtaacagt aacccagagc tgtgttaagg gatcagatct tgatcctgaa gaaccaatca
     1141 aagttgaaga acctgaaccc gtaaagaagc ctcccaaaga gcaaagaagt attaaggaaa
     1201 tgccatttat aacttgtgat gagttcaatg gtgttccttc gtacatgaaa tcccgcttaa
     1261 cctataatca aattaatgat gttattaaag aaatcaacaa ggcagtaatt agtaaatata
     1321 aaatcctaca tcagccagcc gcctctatga attctgtgac cagaaatctc tatcacagat
     1381 ttattgatga agaaacgaag gataccaaag gtcgttattt tatagtggaa gctgacataa
     1441 aggagttcac aactttgaaa gctgacaaga agtttcacgt gttactgaat attttacgac
     1501 actgccggag gctatcagag gtccgagggg gaggacttac tcgttatgtt ataacctgaa
     1561 agcttcgcca gggttttccc agtcgagctc gatatcggta ccagcggata acaatttcac
     1621 atccggattc gcgaacgcgt ctcgagagat ccggctgcta acaaagcccg aaaggaagct
     1681 gagttggctg ctgccaccgc tgagcaataa ctagcataac cccttggggc ctctaaacgg
     1741 gtcttgaggg gttttttgct gaaaggagga actatatccg actagtagcc cgcctaatga
     1801 gcgggctttt ttttaattcc cctatttgtt tatttttcta aatacattca aatatgtatc
     1861 cgctcatgag acaataaccc tgataaatgc ttcaataata ttgaaaaagg aagagtatga
     1921 gtattcaaca tttccgtgtc gcccttattc ccttttttgc ggcattttgc cttcctgttt
     1981 ttgctcaccc agaaacgctc gtgaaagtaa aagacgcaga ggaccaattg ggggcacgag
     2041 tgggatacat agaactggac ttgaatagcg gtaaaatcct tgagagtttt cgccctgaag
     2101 agcgttttcc aatgatgagc actttcaaag ttctgctatg tggagcagta ttatcccgtg
     2161 tagatgcggg gcaagagcaa ctcggacgac gaatacacta ttcgcagaat gacttggttg
     2221 aatactcccc agtgacagaa aagcacctta cggacggaat gacggtaaga gaattatgta
     2281 gtgccgccat aacgatgagt gataacactg cggcgaactt acttctgaca accatcggtg
     2341 gaccgaagga attaaccgct tttttgcaca atatgggaga ccatgtaact cgccttgacc
     2401 gttgggaacc agaactgaat gaagccatac caaacgacga gcgagacacc acaatgcctg
     2461 cggcaatggc aacaacatta cgcaaactat taactggcga actacttact ctggcttcac
     2521 ggcaacaatt aatagactgg cttgaagcgg ataaagttgc aggaccacta ctgcgttcgg
     2581 cacttcctgc tggctggttt attgctgata aatctggggc aggagagcgt ggttcacggg
     2641 gtatcattgc cgcacttgga ccagatggta agccttcccg tatcgtagtt atctacacga
     2701 cgggtagtca ggcaactatg gacgaacgaa atagacagat tgctgaaata ggggcttcac
     2761 tgattaagca ttggtaaacc gatacaatta aaggctcctt ttggagcctt tttttttgga
     2821 cggaccggta gaaaagatca aaggatcttc ttgagatcct ttttttctgc gcgtaatctg
     2881 ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt tgtttgccgg atcaagagct
     2941 accaactctt tttccgaagg taactggctt cagcagagcg cagataccaa atactgtcct
     3001 tctagtgtag ccgtagttag gccaccactt caagaactct gtagcaccgc ctacatacct
     3061 cgctctgcta atcctgttac cagtggctgc tgccagtggc gataagtcgt gtcttaccgg
     3121 gttggactca agacgatagt taccggataa ggcgcagcgg tcgggctgaa cggggggttc
     3181 gtgcacacag cccagcttgg agcgaacgac ctacaccgaa ctgagatacc tacagcgtga
     3241 gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg
     3301 cagggtcgga acaggagagc gcacgaggga gcttccaggg ggaaacgcct ggtatcttta
     3361 tagtcctgtc gggtttcgcc acctctgact tgagcgtcga tttttgtgat gctcgtcagg
     3421 ggggcggagc ctatggaaaa acgccagcaa cgcggccttt ttacggttcc tggccttttg
     3481 ctggcctttt gctcacatgt tctttcctgc gttatcccct gattctgtgg ataaccgtat
     3541 taccgccttt gagtgagctg ataccgctcg ccgcagccga acgaccgagc gcagcgagtc
     3601 agtgagcgag gaagcggaag agcgcctgat gcggtatttt ctccttacgc atctgtgcgg
     3661 tatttcacac cgcaatggtg cactctcagt acaatctgct ctgatgccgc atagttaagc
     3721 cagtatacac tccgctatcg ctacgtgact gggtcatggc tgcgccccga cacccgccaa
     3781 cacccgctga cgcgccctga cgggcttgtc tgctcccggc atccgcttac agacaagctg
     3841 tgaccgtctc cgggagctgc atgtgtcaga ggttttcacc gtcatcaccg aaacgcgcga
     3901 ggcagctgcg gtaaagctca tcagcgtggt cgtgaagcga ttcacagatg tctgcctgtt
     3961 catccgcgtc cagctcgttg agtttctcca gaagcgttaa tgtctggctt ctgataaagc
     4021 gggccatgtt aagggcggtt ttttcctgtt tggtcactga tgcctccgtg taagggggat
     4081 ttctgttcat ggggtaatga taccgatgaa acgagagagg atgctcacga t
//
