LOCUS       sequence-271339-        6205 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Bacterial expression of human Ska3.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6205)
  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 6205)
  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: pCB004
FEATURES             Location/Qualifiers
     source          1..6205
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        join(81..86,87..102,103..109)
                     /label=tac promoter
                     /note="strong E. coli promoter; hybrid between the trp and 
                     lac UV5 promoters"
     protein_bind    117..133
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     CDS             156..809
                     /codon_start=1
                     /product="glutathione S-transferase from Schistosoma 
                     japonicum"
                     /label=GST
                     /translation="MSPILGYWKIKGLVQPTRLLLEYLEEKYEEHLYERDEGDKWRNKK
                     FELGLEFPNLPYYIDGDVKLTQSMAIIRYIADKHNMLGGCPKERAEISMLEGAVLDIRY
                     GVSRIAYSKDFETLKVDFLSKLPEMLKMFEDRLCHKTYLNGDHVTHPDFMLYDALDVVL
                     YMDPMCLDAFPKLVCFKKRIEAIPQIDKYLKSSKYIAWPLQGWQATFGGGDHPPK"
     primer_bind     767..789
                     /label=pGEX 5'
                     /note="pGEX vectors, Glutathione-S-transferase, forward 
                     primer"
     CDS             816..839
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     primer_bind     complement(2153..2175)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(2320..2338)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        2406..2510
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2511..2579,2580..3371)
                     /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(2729..2748)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3542..4130
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4031..4050
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4284..4301
                     /label=L4440
                     /note="L4440 vector, forward primer"
     promoter        4374..4451
                     /gene="lacI"
                     /label=lacI promoter
     CDS             4452..5534
                     /codon_start=1
                     /gene="lacI"
                     /product="lac repressor"
                     /label=lacI
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
                     /translation="MKPVTLYDVAEYAGVSYQTVSRVVNQASHVSAKTREKVEAAMAEL
                     NYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSRADQLGASVVVSMVERSGV
                     EACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSIIFSH
                     EDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREGDWSA
                     MSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTEDSSC
                     YIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR
                     ALADSLMQLARQVSRLESGQ"
     primer_bind     complement(4471..4490)
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        join(5583..5588,5589..5606,5607..5613)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5621..5637
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     primer_bind     5645..5661
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     5645..5661
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             5657..6184
                     /codon_start=1
                     /gene="lacZ fragment"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /label=lacZ-alpha
                     /translation="MTMITDSLAVVLQRRDWENPGVTQLNRLAAHPPFASWRNSEEART
                     DRPSQQLRSLNGEWRFAWFPAPEAVPESWLECDLPEADTVVVPSNWQMHGYDAPIYTNV
                     TYPITVNPPFVPTENPTGCYSLTFNVDESWLQEGQTRIIFDGVGISLSTARCTNASGVR
                     QPSEAVVWLCRS"
     primer_bind     complement(5677..5694)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(5677..5693)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5686..5708)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
ORIGIN
        1 ctcaaggcgc actcccgttc tggataatgt tttttgcgcc gacatcataa cggttctggc
       61 aaatattctg aaatgagctg ttgacaatta atcatcggct cgtataatgt gtggaattgt
      121 gagcggataa caatttcaca caggaaacag tattcatgtc ccctatacta ggttattgga
      181 aaattaaggg ccttgtgcaa cccactcgac ttcttttgga atatcttgaa gaaaaatatg
      241 aagagcattt gtatgagcgc gatgaaggtg ataaatggcg aaacaaaaag tttgaattgg
      301 gtttggagtt tcccaatctt ccttattata ttgatggtga tgttaaatta acacagtcta
      361 tggccatcat acgttatata gctgacaagc acaacatgtt gggtggttgt ccaaaagagc
      421 gtgcagagat ttcaatgctt gaaggagcgg ttttggatat tagatacggt gtttcgagaa
      481 ttgcatatag taaagacttt gaaactctca aagttgattt tcttagcaag ctacctgaaa
      541 tgctgaaaat gttcgaagat cgtttatgtc ataaaacata tttaaatggt gatcatgtaa
      601 cccatcctga cttcatgttg tatgacgctc ttgatgttgt tttatacatg gacccaatgt
      661 gcctggatgc gttcccaaaa ttagtttgtt ttaaaaaacg tattgaagct atcccacaaa
      721 ttgataagta cttgaaatcc agcaagtata tagcatggcc tttgcagggc tggcaagcca
      781 cgtttggtgg tggcgaccat cctccaaaat cggatctgga agttctgttc caggggcccc
      841 tgggatccat ggaccctatc cggagcttct gcgggaagct gcggtctctg gccagcacgc
      901 tggactgcga gacggcccgg ctgcagcgag cgctggacgg agaggaaagc gactttgaag
      961 attatccaat gagaatttta tatgaccttc attcagaagt tcagactcta aaggatgata
     1021 ttaatattct tcttgataaa gcaagattgg aaaatcaaga aggcattgat ttcataaagg
     1081 caacaaaagt actaatggaa aaaaattcaa tggatattat gaaaataaga gagtatttcc
     1141 agaagtatgg atatagtcca cgtgtcaaga aaaattcagt acacgagcaa gaagccatta
     1201 actctgaccc agagttgtct aattgtgaaa attttcagaa gactgatgtg aaagatgatc
     1261 tgtctgatcc tcctgttgca agcagttgta tttctgagaa gtctccacgt agtccacaac
     1321 tttcagattt tggacttgag cggtacatcg tatcccaagt tctaccaaac cctccacagg
     1381 cagtgaacaa ctataaggaa gagcccgtaa ttgtaacccc acctaccaaa caatcactag
     1441 taaaagtact aaaaactcca aaatgtgcac taaaaatgga tgattttgag tgtgtaactc
     1501 ctaaattaga acactttggt atctctgaat atactatgtg tttaaatgaa gattacacaa
     1561 tgggacttaa aaatgcgagg aataataaaa gtgaggaggc catagataca gaatccaggc
     1621 tcaatgataa tgtttttgcc actcccagcc ccatcatcca gcagttggaa aaaagtgatg
     1681 ccgaatatac caactctcct ttggtaccta cattctgtac tcctggtttg aaaattccat
     1741 ctacaaagaa cagcatagct ttggtatcca caaattaccc attatcaaaa acaaatagtt
     1801 catcaaatga tttggaagtt gaagatcgta cttcgttggt tttaaattca gacacatgct
     1861 ttgagaattt aacagatccc tcttcaccta cgatttcttc ttatgagaat ctgctcagaa
     1921 cacctacacc tccagaagta actaaaattc cagaagatat tctccagctt ttatcaaaat
     1981 acaactcaaa cctagctact ccaatagcaa ttaaagcagt gccacccagt aaaaggttcc
     2041 ttaaacatgg acagaacatc cgagatgtca gcaacaaaga aaactgactc gagcggccgc
     2101 atcgtgactg actgacgatc tgcctcgcgc gtttcggtga tgacggtgaa aacctctgac
     2161 acatgcagct cccggagacg gtcacagctt gtctgtaagc ggatgccggg agcagacaag
     2221 cccgtcaggg cgcgtcagcg ggtgttggcg ggtgtcgggg cgcagccatg acccagtcac
     2281 gtagcgatag cggagtgtat aattcttgaa gacgaaaggg cctcgtgata cgcctatttt
     2341 tataggttaa tgtcatgata ataatggttt cttagacgtc aggtggcact tttcggggaa
     2401 atgtgcgcgg aacccctatt tgtttatttt tctaaataca ttcaaatatg tatccgctca
     2461 tgagacaata accctgataa atgcttcaat aatattgaaa aaggaagagt atgagtattc
     2521 aacatttccg tgtcgccctt attccctttt ttgcggcatt ttgccttcct gtttttgctc
     2581 acccagaaac gctggtgaaa gtaaaagatg ctgaagatca gttgggtgca cgagtgggtt
     2641 acatcgaact ggatctcaac agcggtaaga tccttgagag ttttcgcccc gaagaacgtt
     2701 ttccaatgat gagcactttt aaagttctgc tatgtggcgc ggtattatcc cgtgttgacg
     2761 ccgggcaaga gcaactcggt cgccgcatac actattctca gaatgacttg gttgagtact
     2821 caccagtcac agaaaagcat cttacggatg gcatgacagt aagagaatta tgcagtgctg
     2881 ccataaccat gagtgataac actgcggcca acttacttct gacaacgatc ggaggaccga
     2941 aggagctaac cgcttttttg cacaacatgg gggatcatgt aactcgcctt gatcgttggg
     3001 aaccggagct gaatgaagcc ataccaaacg acgagcgtga caccacgatg cctgcagcaa
     3061 tggcaacaac gttgcgcaaa ctattaactg gcgaactact tactctagct tcccggcaac
     3121 aattaataga ctggatggag gcggataaag ttgcaggacc acttctgcgc tcggcccttc
     3181 cggctggctg gtttattgct gataaatctg gagccggtga gcgtgggtct cgcggtatca
     3241 ttgcagcact ggggccagat ggtaagccct cccgtatcgt agttatctac acgacgggga
     3301 gtcaggcaac tatggatgaa cgaaatagac agatcgctga gataggtgcc tcactgatta
     3361 agcattggta actgtcagac caagtttact catatatact ttagattgat ttaaaacttc
     3421 atttttaatt taaaaggatc taggtgaaga tcctttttga taatctcatg accaaaatcc
     3481 cttaacgtga gttttcgttc cactgagcgt cagaccccgt agaaaagatc aaaggatctt
     3541 cttgagatcc tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac
     3601 cagcggtggt ttgtttgccg gatcaagagc taccaactct ttttccgaag gtaactggct
     3661 tcagcagagc gcagatacca aatactgtcc ttctagtgta gccgtagtta ggccaccact
     3721 tcaagaactc tgtagcaccg cctacatacc tcgctctgct aatcctgtta ccagtggctg
     3781 ctgccagtgg cgataagtcg tgtcttaccg ggttggactc aagacgatag ttaccggata
     3841 aggcgcagcg gtcgggctga acggggggtt cgtgcacaca gcccagcttg gagcgaacga
     3901 cctacaccga actgagatac ctacagcgtg agctatgaga aagcgccacg cttcccgaag
     3961 ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg
     4021 agcttccagg gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc cacctctgac
     4081 ttgagcgtcg atttttgtga tgctcgtcag gggggcggag cctatggaaa aacgccagca
     4141 acgcggcctt tttacggttc ctggcctttt gctggccttt tgctcacatg ttctttcctg
     4201 cgttatcccc tgattctgtg gataaccgta ttaccgcctt tgagtgagct gataccgctc
     4261 gccgcagccg aacgaccgag cgcagcgagt cagtgagcga ggaagcggaa gagcgcctga
     4321 tgcggtattt tctccttacg catctgtgcg gtatttcaca ccgcataaat tccgacacca
     4381 tcgaatggcg caaaaccttt cgcggtatgg catgatagcg cccggaagag agtcaattca
     4441 gggtggtgaa tgtgaaacca gtaacgttat acgatgtcgc agagtatgcc ggtgtctctt
     4501 atcagaccgt ttcccgcgtg gtgaaccagg ccagccacgt ttctgcgaaa acgcgggaaa
     4561 aagtggaagc ggcgatggcg gagctgaatt acattcccaa ccgcgtggca caacaactgg
     4621 cgggcaaaca gtcgttgctg attggcgttg ccacctccag tctggccctg cacgcgccgt
     4681 cgcaaattgt cgcggcgatt aaatctcgcg ccgatcaact gggtgccagc gtggtggtgt
     4741 cgatggtaga acgaagcggc gtcgaagcct gtaaagcggc ggtgcacaat cttctcgcgc
     4801 aacgcgtcag tgggctgatc attaactatc cgctggatga ccaggatgcc attgctgtgg
     4861 aagctgcctg cactaatgtt ccggcgttat ttcttgatgt ctctgaccag acacccatca
     4921 acagtattat tttctcccat gaagacggta cgcgactggg cgtggagcat ctggtcgcat
     4981 tgggtcacca gcaaatcgcg ctgttagcgg gcccattaag ttctgtctcg gcgcgtctgc
     5041 gtctggctgg ctggcataaa tatctcactc gcaatcaaat tcagccgata gcggaacggg
     5101 aaggcgactg gagtgccatg tccggttttc aacaaaccat gcaaatgctg aatgagggca
     5161 tcgttcccac tgcgatgctg gttgccaacg atcagatggc gctgggcgca atgcgcgcca
     5221 ttaccgagtc cgggctgcgc gttggtgcgg atatctcggt agtgggatac gacgataccg
     5281 aagacagctc atgttatatc ccgccgttaa ccaccatcaa acaggatttt cgcctgctgg
     5341 ggcaaaccag cgtggaccgc ttgctgcaac tctctcaggg ccaggcggtg aagggcaatc
     5401 agctgttgcc cgtctcactg gtgaaaagaa aaaccacgct agcgcccaat acgcaaaccg
     5461 cctctccccg cgcgttggcc gattcattaa tgcagctggc acgacaggtt tcccgactgg
     5521 aaagcgggca gtgagcgcaa cgcaattaat gtaagttagc tcactcatta ggcaccccag
     5581 gctttacact ttatgcttcc ggctcgtatg ttgtgtggaa ttgtgagcgg ataacaattt
     5641 cacacaggaa acagctatga ccatgattac ggattcactg gccgtcgttt tacaacgtcg
     5701 tgactgggaa aaccctggcg ttacccaact taatcgcctt gcagcacatc cccctttcgc
     5761 cagctggcgt aatagcgaag aggcccgcac cgatcgccct tcccaacagt tgcgcagcct
     5821 gaatggcgaa tggcgctttg cctggtttcc ggcaccagaa gcggtgccgg aaagctggct
     5881 ggagtgcgat cttcctgagg ccgatactgt cgtcgtcccc tcaaactggc agatgcacgg
     5941 ttacgatgcg cccatctaca ccaacgtgac ctatcccatt acggtcaatc cgccgtttgt
     6001 tcccacggag aatccgacgg gttgttactc gctcacattt aatgttgatg aaagctggct
     6061 acaggaaggc cagacgcgaa ttatttttga tggcgttgga attagcttat cgactgcacg
     6121 gtgcaccaat gcttctggcg tcaggcagcc atcggaagct gtggtatggc tgtgcaggtc
     6181 gtaaatcact gcataattcg tgtcg
//
