LOCUS       sequence-271205-        5172 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Bacterial expression of C. elegans Ska1 microtubule binding domain 
            (MTBD).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5172)
  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 5172)
  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: pJS263
FEATURES             Location/Qualifiers
     source          1..5172
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     79..98
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     182..201
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        182..200
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     RBS             274..296
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             316..333
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             337..369
                     /codon_start=1
                     /product="leader peptide from bacteriophage T7 gene 10"
                     /label=T7 tag (gene 10 leader)
                     /note="promotes efficient translation in E. coli"
                     /translation="MASMTGGQQMG"
     CDS             join(373..381,382..396)
                     /codon_start=1
                     /product="Xpress(TM) epitope tag, including an enterokinase
                     recognition and cleavage site"
                     /label=Xpress(TM) tag
                     /translation="DLYDDDDK"
     CDS             397..420
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     primer_bind     complement(894..912)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      908..955
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     promoter        complement(join(1321..1326,1327..1343,1344..1349))
                     /label=tet promoter
                     /note="E. coli promoter for tetracycline efflux protein 
                     gene"
     primer_bind     complement(1376..1394)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        1462..1566
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1567..1635,1636..2427)
                     /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(1785..1804)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2587..3175
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3076..3095
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3329..3346
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    3361..3500
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(3426..3445)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3586..3608
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(3602..3793)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
ORIGIN
        1 ggccacgggg cctgccacca tacccacgcc gaaacaagcg ctcatgagcc cgaagtggcg
       61 agcccgatct tccccatcgg tgatgtcggc gatataggcg ccagcaaccg cacctgtggc
      121 gccggtgatg ccggccacga tgcgtccggc gtagaggatc gagatctcga tcccgcgaaa
      181 ttaatacgac tcactatagg gagaccacaa cggtttccct ctagtgccgg ctccggagag
      241 ctccaattgg aattcgctag ctctagaaat aattttgttt aactttaaga aggagatata
      301 cgtatgcggg gttctcatca tcatcatcat catggtatgg ctagcatgac tggtggacag
      361 caaatgggtc gggatctgta cgacgatgac gataagctgg aagttctgtt ccaagggccc
      421 catatgaacg acattcgtat tgtcccacag atcactgatg aggagtttaa aaccatcccg
      481 aaatatcaac tgggtcgcct gacactggag atgatgaacg agattgtgag caaaatggac
      541 gatttcctga tgaaaaaatc caaaatcctg gggaaaacca acaaacaact gacccgtagc
      601 gatcgtgaag ttctggacaa ctggcgtgaa ctggaaatga aagcccgtaa acgtctgcct
      661 acaacgctgt tctttatcga gacagacatc cgtcctatgc tgcaagaccg cctgcgtccg
      721 tccttcgcca aagcaattcc ttgcctgcgc cacatccgcc gtattcgtga agaacgttgt
      781 ggtccgctga ccttctatta tccgtgagga tccagatcta agcttggtac cacgcgtgcg
      841 cgctgatccg gctgctaaca aagcccgaaa ggaagctgag ttggctgctg ccaccgctga
      901 gcaataacta gcataacccc ttggggcctc taaacgggtc ttgaggggtt ttttgctgaa
      961 aggaggaact atatccggat atccacagga cgggtgtggt cgccatgatc gcgtagtcga
     1021 tagtggctcc aagtagcgaa gcgagcagga ctgggcggcg gccaaagcgg tcggacagtg
     1081 ctccgagaac gggtgcgcat agaaattgca tcaacgcata tagcgctagc agcacgccat
     1141 agtgactggc gatgctgtcg gaatggacga tatcccgcaa gaggcccggc agtaccggca
     1201 taaccaagcc tatgcctaca gcatccaggg tgacggtgcc gaggatgacg atgagcgcat
     1261 tgttagattt catacacggt gcctgactgc gttagcaatt taactgtgat aaactaccgc
     1321 attaaagctt atcgatgata agctgtcaaa catgagaatt cttgaagacg aaagggcctc
     1381 gtgatacgcc tatttttata ggttaatgtc atgataataa tggtttctta gacgtcaggt
     1441 ggcacttttc ggggaaatgt gcgcggaacc cctatttgtt tatttttcta aatacattca
     1501 aatatgtatc cgctcatgag acaataaccc tgataaatgc ttcaataata ttgaaaaagg
     1561 aagagtatga gtattcaaca tttccgtgtc gcccttattc ccttttttgc ggcattttgc
     1621 cttcctgttt ttgctcaccc agaaacgctg gtgaaagtaa aagatgctga agatcagttg
     1681 ggtgcacgag tgggttacat cgaactggat ctcaacagcg gtaagatcct tgagagtttt
     1741 cgccccgaag aacgttttcc aatgatgagc acttttaaag ttctgctatg tggcgcggta
     1801 ttatcccgtg ttgacgccgg gcaagagcaa ctcggtcgcc gcatacacta ttctcagaat
     1861 gacttggttg agtactcacc agtcacagaa aagcatctta cggatggcat gacagtaaga
     1921 gaattatgca gtgctgccat aaccatgagt gataacactg cggccaactt acttctgaca
     1981 acgatcggag gaccgaagga gctaaccgct tttttgcaca acatggggga tcatgtaact
     2041 cgccttgatc gttgggaacc ggagctgaat gaagccatac caaacgacga gcgtgacacc
     2101 acgatgcctg cagcaatggc aacaacgttg cgcaaactat taactggcga actacttact
     2161 ctagcttccc ggcaacaatt aatagactgg atggaggcgg ataaagttgc aggaccactt
     2221 ctgcgctcgg cccttccggc tggctggttt attgctgata aatctggagc cggtgagcgt
     2281 gggtctcgcg gtatcattgc agcactgggg ccagatggta agccctcccg tatcgtagtt
     2341 atctacacga cggggagtca ggcaactatg gatgaacgaa atagacagat cgctgagata
     2401 ggtgcctcac tgattaagca ttggtaactg tcagaccaag tttactcata tatactttag
     2461 attgatttgc ggccgcaaaa ggatctaggt gaagatcctt tttgataatc tcatgaccaa
     2521 aatcccttaa cgtgagtttt cgttccactg agcgtcagac cccgtagaaa agatcaaagg
     2581 atcttcttga gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc
     2641 gctaccagcg gtggtttgtt tgccggatca agagctacca actctttttc cgaaggtaac
     2701 tggcttcagc agagcgcaga taccaaatac tgtccttcta gtgtagccgt agttaggcca
     2761 ccacttcaag aactctgtag caccgcctac atacctcgct ctgctaatcc tgttaccagt
     2821 ggctgctgcc agtggcgata agtcgtgtct taccgggttg gactcaagac gatagttacc
     2881 ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc acacagccca gcttggagcg
     2941 aacgacctac accgaactga gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc
     3001 cgaagggaga aaggcggaca ggtatccggt aagcggcagg gtcggaacag gagagcgcac
     3061 gagggagctt ccagggggaa acgcctggta tctttatagt cctgtcgggt ttcgccacct
     3121 ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc
     3181 cagcaacgcg gcctttttac ggttcctggc cttttgctgg ccttttgctc acatgttctt
     3241 tcctgcgtta tcccctgatt ctgtggataa ccgtattacc gcctttgagt gagctgatac
     3301 cgctcgccgc agccgaacga ccgagcgcag cgagtcagtg agcgaggaag cggaagagcg
     3361 cctgatgcgg tattttctcc ttacgcatct gtgcggtatt tcacaccgca atggtgcact
     3421 ctcagtacaa tctgctctga tgccgcatag ttaagccagt atacactccg ctatcgctac
     3481 gtgactgggt catggctgcg ccccgacacc cgccaacacc cgctgacgcg ccctgacggg
     3541 cttgtctgct cccggcatcc gcttacagac aagctgtgac cgtctccggg agctgcatgt
     3601 gtcagaggtt ttcaccgtca tcaccgaaac gcgcgaggca gctgcggtaa agctcatcag
     3661 cgtggtcgtg aagcgattca cagatgtctg cctgttcatc cgcgtccagc tcgttgagtt
     3721 tctccagaag cgttaatgtc tggcttctga taaagcgggc catgttaagg gcggtttttt
     3781 cctgtttggt cactgatgcc tccgtgtaag ggggatttct gttcatgggg gtaatgatac
     3841 cgatgaaacg agagaggatg ctcacgatac gggttactga tgatgaacat gcccggttac
     3901 tggaacgttg tgagggtaaa caactggcgg tatggatgcg gcgggaccag agaaaaatca
     3961 ctcagggtca atgccagcgc ttcgttaata cagatgtagg tgttccacag ggtagccagc
     4021 agcatcctgc gatgcagatc cggaacataa tggtgcaggg cgctgacttc cgcgtttcca
     4081 gactttacga aacacggaaa ccgaagacca ttcatgttgt tgctcaggtc gcagacgttt
     4141 tgcagcagca gtcgcttcac gttcgctcgc gtatcggtga ttcattctgc taaccagtaa
     4201 ggcaaccccg ccagcctagc cgggtcctca acgacaggag cacgatcatg cgcacccgtg
     4261 gccaggaccc aacgctgccc gagatgcgcc gcgtgcggct gctggagatg gcggacgcga
     4321 tggatatgtt ctgccaaggg ttggtttgcg cattcacagt tctccgcaag aattgattgg
     4381 ctccaattct tggagtggtg aatccgttag cgaggtgccg ccggcttcca ttcaggtcga
     4441 ggtggcccgg ctccatgcac cgcgacgcaa cgcggggagg cagacaaggt atagggcggc
     4501 gcctacaatc catgccaacc cgttccatgt gctcgccgag gcggcataaa tcgccgtgac
     4561 gatcagcggt ccaatgatcg aagttaggct ggtaagagcc gcgagcgatc cttgaagctg
     4621 tccctgatgg tcgtcatcta cctgcctgga cagcatggcc tgcaacgcgg gcatcccgat
     4681 gccgccggaa gcgagaagaa tcataatggg gaaggccatc cagcctcgcg tcgcgaacgc
     4741 cagcaagacg tagcccagcg cgtcggccgc catgccggcg ataatggcct gcttctcgcc
     4801 gaaacgtttg gtggcgggac cagtgacgaa ggcttgagcg agggcgtgca agattccgaa
     4861 taccgcaagc gacaggccga tcatcgtcgc gctccagcga aagcggtcct cgccgaaaat
     4921 gacccagagc gctgccggca cctgtcctac gagttgcatg ataaagaaga cagtcataag
     4981 tgcggcgacg atagtcatgc cccgcgccca ccggaaggag ctgactgggt tgaaggctct
     5041 caagggcatc ggtcgacgct ctcccttatg cgactcctgc attaggaagc agcccagtag
     5101 taggttgagg ccgttgagca ccgccgccgc aaggaatggt gcatgcaagg agatggcgcc
     5161 caacagtccc cc
//
