LOCUS       sequence-122256-        7511 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  For bacterial expression of recombinant human TACC3 tagged with GST 
            and His..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7511)
  AUTHORS   Nixon FM, Gutierrez-Caballero C, Hood FE, Booth DG, Prior IA, Royle 
            SJ
  TITLE     The mesh is a network of microtubule connectors that stabilizes 
            individual kinetochore fibers of the mitotic spindle.
  JOURNAL   Elife. 2015 Jun 19;4. doi: 10.7554/eLife.07635.
  PUBMED    26090906
REFERENCE   2  (bases 1 to 7511)
  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: "2015-06-19"; volume: "4. doi"; pages: " 10.7554/eLife.07635"
COMMENT     Sequence Label: GST-TACC3-His6
FEATURES             Location/Qualifiers
     source          1..7511
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        join(183..188,189..204,205..211)
                     /label=tac promoter
                     /note="strong E. coli promoter; hybrid between the trp and 
                     lac UV5 promoters"
     protein_bind    219..235
                     /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             258..911
                     /codon_start=1
                     /product="glutathione S-transferase from Schistosoma 
                     japonicum"
                     /label=GST
                     /translation="MSPILGYWKIKGLVQPTRLLLEYLEEKYEEHLYERDEGDKWRNKK
                     FELGLEFPNLPYYIDGDVKLTQSMAIIRYIADKHNMLGGCPKERAEISMLEGAVLDIRY
                     GVSRIAYSKDFETLKVDFLSKLPEMLKMFEDRLCHKTYLNGDHVTHPDFMLYDALDVVL
                     YMDPMCLDAFPKLVCFKKRIEAIPQIDKYLKSSKYIAWPLQGWQATFGGGDHPPK"
     primer_bind     869..891
                     /label=pGEX 5'
                     /note="pGEX vectors, Glutathione-S-transferase, forward 
                     primer"
     CDS             918..941
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     CDS             3480..3497
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(3561..3583)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(3728..3746)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3814..3918
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3919..3987,3988..4779)
                     /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(4137..4156)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4950..5538
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5439..5458
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5692..5709
                     /label=L4440
                     /note="L4440 vector, forward primer"
     promoter        5782..5859
                     /gene="lacI (mutant)"
                     /label=lacIq promoter
                     /note="In the lacIq allele, a single base change in the 
                     promoter boosts expression of the lacI gene about 10-fold."
     CDS             5860..6942
                     /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
                     YIPPSTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR
                     ALADSLMQLARQVSRLESGQ"
     primer_bind     complement(5879..5898)
                     /label=LacI-R
                     /note="LacI, reverse primer"
     protein_bind    6955..6976
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(6991..6996,6997..7014,7015..7021)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7029..7045
                     /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     7053..7069
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     7053..7069
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             join(7065..7511,1..81)
                     /codon_start=1
                     /gene="lacZ fragment"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /label=lacZ-alpha
                     /translation="MTMITDSLAVVLQRRDWENPGVTQLNRLAAHPPFASWRNSEEART
                     DRPSQQLRSLNGEWRFAWFPAPEAVPESWLECDLPEADTVVVPSNWQMHGYDAPIYTNV
                     TYPITVNPPFVPTENPTGCYSLTFNVDESWLQEGQTRIIFDGVGITLSTARCTNASGVR
                     QPSEAVVWLCRS"
     primer_bind     complement(7085..7102)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(7085..7101)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(7094..7116)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
ORIGIN
        1 acgttatcga ctgcacggtg caccaatgct tctggcgtca ggcagccatc ggaagctgtg
       61 gtatggctgt gcaggtcgta aatcactgca taattcgtgt cgctcaaggc gcactcccgt
      121 tctggataat gttttttgcg ccgacatcat aacggttctg gcaaatattc tgaaatgagc
      181 tgttgacaat taatcatcgg ctcgtataat gtgtggaatt gtgagcggat aacaatttca
      241 cacaggaaac agtattcatg tcccctatac taggttattg gaaaattaag ggccttgtgc
      301 aacccactcg acttcttttg gaatatcttg aagaaaaata tgaagagcat ttgtatgagc
      361 gcgatgaagg tgataaatgg cgaaacaaaa agtttgaatt gggtttggag tttcccaatc
      421 ttccttatta tattgatggt gatgttaaat taacacagtc tatggccatc atacgttata
      481 tagctgacaa gcacaacatg ttgggtggtt gtccaaaaga gcgtgcagag atttcaatgc
      541 ttgaaggagc ggttttggat attagatacg gtgtttcgag aattgcatat agtaaagact
      601 ttgaaactct caaagttgat tttcttagca agctacctga aatgctgaaa atgttcgaag
      661 atcgtttatg tcataaaaca tatttaaatg gtgatcatgt aacccatcct gacttcatgt
      721 tgtatgacgc tcttgatgtt gttttataca tggacccaat gtgcctggat gcgttcccaa
      781 aattagtttg ttttaaaaaa cgtattgaag ctatcccaca aattgataag tacttgaaat
      841 ccagcaagta tatagcatgg cctttgcagg gctggcaagc cacgtttggt ggtggcgacc
      901 atcctccaaa atcggatctg gaagttctgt tccaggggcc cctgggatcc ccggaattcc
      961 cgggtatgag tctgcaggtc ttaaacgaca aaaatgtcag caatgaaaaa aatacagaaa
     1021 attgcgactt cctgttttcg ccaccagaag ttaccggaag atcgtctgtt cttcgtgtgt
     1081 cacagaaaga aaatgtgcca cccaagaacc tggccaaagc tatgaaggtg acttttcaga
     1141 cacctctgcg ggatccacag acgcacagga ttctaagtcc tagcatggcc agcaaacttg
     1201 aggctccttt cactcaggat gacacccttg gactggaaaa ctcacacccg gtctggacac
     1261 agaaagagaa ccaacagctc atcaaggaag tggatgccaa aactactcat ggaattctac
     1321 agaaaccagt ggaggctgac accgacctcc tgggggatgc aagcccagcc tttgggagtg
     1381 gcagctccag cgagtctggc ccaggtgccc tggctgacct ggactgctca agctcttccc
     1441 agagcccagg aagttctgag aaccaaatgg tgtctccagg aaaagtgtct ggcagccctg
     1501 agcaagccgt ggaggaaaac cttagttcct attccttaga cagaagagtg acacccgcct
     1561 ctgagaccct agaagaccct tgcaggacag agtcccagca caaagcggag actccgcacg
     1621 gagccgagga agaatgcaaa gcggagactc cgcacggagc cgaggaggaa tgccggcacg
     1681 gtggggtctg tgctcccgca gcagtggcca cttcgcctcc tggtgcaatc cctaaggaag
     1741 cctgcggagg agcacccctg cagggtctgc ctggcgaagc cctgggctgc cctgcgggtg
     1801 tgggcacccc cgtgccagca gatggcactc agacccttac ctgtgcacac acctctgctc
     1861 ctgagagcac agccccaacc aaccacctgg tggctggcag ggccatgacc ctgagtcctc
     1921 aggaagaagt ggctgcaggc caaatggcca gctcctcgag gagcggacct gtaaaactag
     1981 aatttgatgt atctgatggc gccaccagca aaagggcacc cccaccaagg agactgggag
     2041 agaggtccgg cctcaagcct cccttgagga aagcagcagt gaggcagcaa aaggccccgc
     2101 aggaggtgga ggaggacgac ggtaggagcg gagcaggaga ggaccccccc atgccagctt
     2161 ctcggggctc ttaccacctc gactgggaca aaatggatga cccaaacttc atcccgttcg
     2221 gaggtgacac caagtctggt tgcagtgagg cccagccccc agaaagccct gagaccaggc
     2281 tgggccagcc agcggctgaa cagttgcatg ctgggcctgc cacggaggag ccaggtccct
     2341 gtctgagcca gcagctgcat tcagcctcag cggaggacac gcctgtggtg cagttggcag
     2401 ccgagacccc aacagcagag agcaaggaga gagccttgaa ctctgccagc acctcgcttc
     2461 ccacaagctg tccaggcagt gagccagtgc ccacccatca gcaggggcag cctgccttgg
     2521 agctgaaaga ggagagcttc agagaccccg ctgaggttct aggcacgggc gcggaggtgg
     2581 attacctgga gcagtttgga acttcctcgt ttaaggagtc ggccttgagg aagcagtcct
     2641 tatacctcaa gttcgacccc ctcctgaggg acagtcctgg tagaccagtg cccgtggcca
     2701 ccgagaccag cagcatgcac ggtgcaaatg agactccctc aggacgtccg cgggaagcca
     2761 agcttgtgga gttcgatttc ttgggagcac tggacattcc tgtgccaggc ccacccccag
     2821 gtgttcccgc gcctgggggc ccacccctgt ccaccggacc tatagtggac ctgctccagt
     2881 acagccagaa ggacctggat gcagtggtaa aggcgacaca ggaggagaac cgggagctga
     2941 ggagcaggtg tgaggagctc cacgggaaga acctggaact ggggaagatc atggacaggt
     3001 tcgaagaggt tgtgtaccag gccatggagg aagttcagaa gcagaaggaa ctttccaaag
     3061 ctgaaatcca gaaagttcta aaagaaaaag accaacttac cacagatctg aactccatgg
     3121 agaagtcctt ctccgacctc ttcaagcgtt ttgagaaaca gaaagaggtg atcgagggct
     3181 accgcaagaa cgaagagtca ctgaagaagt gcgtggagga ttacctggca aggatcaccc
     3241 aggagggcca gaggtaccaa gccctgaagg cccacgcgga ggagaagctg cagctggcaa
     3301 acgaggagat cgcccaggtc cggagcaagg cccaggcgga agcgttggcc ctccaggcca
     3361 gcctgaggaa ggagcagatg cgcatccagt cgctggagaa gacagtggag cagaagacta
     3421 aagagaacga ggagctgacc aggatctgcg acgacctcat ctccaagatg gagaagatcc
     3481 atcaccatca ccatcactga gcggccgcat cgtgactgac tgacgatctg cctcgcgcgt
     3541 ttcggtgatg acggtgaaaa cctctgacac atgcagctcc cggagacggt cacagcttgt
     3601 ctgtaagcgg atgccgggag cagacaagcc cgtcagggcg cgtcagcggg tgttggcggg
     3661 tgtcggggcg cagccatgac ccagtcacgt agcgatagcg gagtgtataa ttcttgaaga
     3721 cgaaagggcc tcgtgatacg cctattttta taggttaatg tcatgataat aatggtttct
     3781 tagacgtcag gtggcacttt tcggggaaat gtgcgcggaa cccctatttg tttatttttc
     3841 taaatacatt caaatatgta tccgctcatg agacaataac cctgataaat gcttcaataa
     3901 tattgaaaaa ggaagagtat gagtattcaa catttccgtg tcgcccttat tccctttttt
     3961 gcggcatttt gccttcctgt ttttgctcac ccagaaacgc tggtgaaagt aaaagatgct
     4021 gaagatcagt tgggtgcacg agtgggttac atcgaactgg atctcaacag cggtaagatc
     4081 cttgagagtt ttcgccccga agaacgtttt ccaatgatga gcacttttaa agttctgcta
     4141 tgtggcgcgg tattatcccg tgttgacgcc gggcaagagc aactcggtcg ccgcatacac
     4201 tattctcaga atgacttggt tgagtactca ccagtcacag aaaagcatct tacggatggc
     4261 atgacagtaa gagaattatg cagtgctgcc ataaccatga gtgataacac tgcggccaac
     4321 ttacttctga caacgatcgg aggaccgaag gagctaaccg cttttttgca caacatgggg
     4381 gatcatgtaa ctcgccttga tcgttgggaa ccggagctga atgaagccat accaaacgac
     4441 gagcgtgaca ccacgatgcc tgcagcaatg gcaacaacgt tgcgcaaact attaactggc
     4501 gaactactta ctctagcttc ccggcaacaa ttaatagact ggatggaggc ggataaagtt
     4561 gcaggaccac ttctgcgctc ggcccttccg gctggctggt ttattgctga taaatctgga
     4621 gccggtgagc gtgggtctcg cggtatcatt gcagcactgg ggccagatgg taagccctcc
     4681 cgtatcgtag ttatctacac gacggggagt caggcaacta tggatgaacg aaatagacag
     4741 atcgctgaga taggtgcctc actgattaag cattggtaac tgtcagacca agtttactca
     4801 tatatacttt agattgattt aaaacttcat ttttaattta aaaggatcta ggtgaagatc
     4861 ctttttgata atctcatgac caaaatccct taacgtgagt tttcgttcca ctgagcgtca
     4921 gaccccgtag aaaagatcaa aggatcttct tgagatcctt tttttctgcg cgtaatctgc
     4981 tgcttgcaaa caaaaaaacc accgctacca gcggtggttt gtttgccgga tcaagagcta
     5041 ccaactcttt ttccgaaggt aactggcttc agcagagcgc agataccaaa tactgtcctt
     5101 ctagtgtagc cgtagttagg ccaccacttc aagaactctg tagcaccgcc tacatacctc
     5161 gctctgctaa tcctgttacc agtggctgct gccagtggcg ataagtcgtg tcttaccggg
     5221 ttggactcaa gacgatagtt accggataag gcgcagcggt cgggctgaac ggggggttcg
     5281 tgcacacagc ccagcttgga gcgaacgacc tacaccgaac tgagatacct acagcgtgag
     5341 ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc
     5401 agggtcggaa caggagagcg cacgagggag cttccagggg gaaacgcctg gtatctttat
     5461 agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg
     5521 gggcggagcc tatggaaaaa cgccagcaac gcggcctttt tacggttcct ggccttttgc
     5581 tggccttttg ctcacatgtt ctttcctgcg ttatcccctg attctgtgga taaccgtatt
     5641 accgcctttg agtgagctga taccgctcgc cgcagccgaa cgaccgagcg cagcgagtca
     5701 gtgagcgagg aagcggaaga gcgcctgatg cggtattttc tccttacgca tctgtgcggt
     5761 atttcacacc gcataaattc cgacaccatc gaatggtgca aaacctttcg cggtatggca
     5821 tgatagcgcc cggaagagag tcaattcagg gtggtgaatg tgaaaccagt aacgttatac
     5881 gatgtcgcag agtatgccgg tgtctcttat cagaccgttt cccgcgtggt gaaccaggcc
     5941 agccacgttt ctgcgaaaac gcgggaaaaa gtggaagcgg cgatggcgga gctgaattac
     6001 attcccaacc gcgtggcaca acaactggcg ggcaaacagt cgttgctgat tggcgttgcc
     6061 acctccagtc tggccctgca cgcgccgtcg caaattgtcg cggcgattaa atctcgcgcc
     6121 gatcaactgg gtgccagcgt ggtggtgtcg atggtagaac gaagcggcgt cgaagcctgt
     6181 aaagcggcgg tgcacaatct tctcgcgcaa cgcgtcagtg ggctgatcat taactatccg
     6241 ctggatgacc aggatgccat tgctgtggaa gctgcctgca ctaatgttcc ggcgttattt
     6301 cttgatgtct ctgaccagac acccatcaac agtattattt tctcccatga agacggtacg
     6361 cgactgggcg tggagcatct ggtcgcattg ggtcaccagc aaatcgcgct gttagcgggc
     6421 ccattaagtt ctgtctcggc gcgtctgcgt ctggctggct ggcataaata tctcactcgc
     6481 aatcaaattc agccgatagc ggaacgggaa ggcgactgga gtgccatgtc cggttttcaa
     6541 caaaccatgc aaatgctgaa tgagggcatc gttcccactg cgatgctggt tgccaacgat
     6601 cagatggcgc tgggcgcaat gcgcgccatt accgagtccg ggctgcgcgt tggtgcggat
     6661 atctcggtag tgggatacga cgataccgaa gacagctcat gttatatccc gccgtcaacc
     6721 accatcaaac aggattttcg cctgctgggg caaaccagcg tggaccgctt gctgcaactc
     6781 tctcagggcc aggcggtgaa gggcaatcag ctgttgcccg tctcactggt gaaaagaaaa
     6841 accaccctgg cgcccaatac gcaaaccgcc tctccccgcg cgttggccga ttcattaatg
     6901 cagctggcac gacaggtttc ccgactggaa agcgggcagt gagcgcaacg caattaatgt
     6961 gagttagctc actcattagg caccccaggc tttacacttt atgcttccgg ctcgtatgtt
     7021 gtgtggaatt gtgagcggat aacaatttca cacaggaaac agctatgacc atgattacgg
     7081 attcactggc cgtcgtttta caacgtcgtg actgggaaaa ccctggcgtt acccaactta
     7141 atcgccttgc agcacatccc cctttcgcca gctggcgtaa tagcgaagag gcccgcaccg
     7201 atcgcccttc ccaacagttg cgcagcctga atggcgaatg gcgctttgcc tggtttccgg
     7261 caccagaagc ggtgccggaa agctggctgg agtgcgatct tcctgaggcc gatactgtcg
     7321 tcgtcccctc aaactggcag atgcacggtt acgatgcgcc catctacacc aacgtaacct
     7381 atcccattac ggtcaatccg ccgtttgttc ccacggagaa tccgacgggt tgttactcgc
     7441 tcacatttaa tgttgatgaa agctggctac aggaaggcca gacgcgaatt atttttgatg
     7501 gcgttggaat t
//
