LOCUS       sequence-271799-        7227 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Bacterial expression of GST-HJURP.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7227)
  AUTHORS   McKinley KL, Cheeseman IM
  TITLE     Polo-like kinase 1 licenses CENP-A deposition at centromeres.
  JOURNAL   Cell. 2014 Jul 17;158(2):397-411. doi: 10.1016/j.cell.2014.06.016.
  PUBMED    25036634
REFERENCE   2  (bases 1 to 7227)
  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.cell.2014.06"; journalName: "Cell"; date: "2014-07-17- 
            17"; volume: "158"; issue: "2"; pages: "397-411"
COMMENT     Sequence Label: pKM061
FEATURES             Location/Qualifiers
     source          1..7227
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     457..476
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     710..727
                     /label=L4440
                     /note="L4440 vector, forward primer"
     promoter        800..877
                     /gene="lacI"
                     /label=lacI promoter
     CDS             878..1960
                     /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(897..916)
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        join(2009..2014,2015..2032,2033..2039)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2047..2063
                     /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     2071..2087
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     2071..2087
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             2083..2610
                     /codon_start=1
                     /gene="lacZ fragment"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /label=lacZ-alpha
                     /translation="MTMITDSLAVVLQRRDWENPGVTQLNRLAAHPPFASWRNSEEART
                     DRPSQQLRSLNGEWRFAWFPAPEAVPESWLECDLPEADTVVVPSNWQMHGYDAPIYTNV
                     TYPITVNPPFVPTENPTGCYSLTFNVDESWLQEGQTRIIFDGVGISLSTARCTNASGVR
                     QPSEAVVWLCRS"
     primer_bind     complement(2103..2120)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(2103..2119)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2112..2134)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     promoter        join(2712..2717,2718..2733,2734..2740)
                     /label=tac promoter
                     /note="strong E. coli promoter; hybrid between the trp and 
                     lac UV5 promoters"
     protein_bind    2748..2764
                     /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             2787..3440
                     /codon_start=1
                     /product="glutathione S-transferase from Schistosoma 
                     japonicum"
                     /label=GST
                     /translation="MSPILGYWKIKGLVQPTRLLLEYLEEKYEEHLYERDEGDKWRNKK
                     FELGLEFPNLPYYIDGDVKLTQSMAIIRYIADKHNMLGGCPKERAEISMLEGAVLDIRY
                     GVSRIAYSKDFETLKVDFLSKLPEMLKMFEDRLCHKTYLNGDHVTHPDFMLYDALDVVL
                     YMDPMCLDAFPKLVCFKKRIEAIPQIDKYLKSSKYIAWPLQGWQATFGGGDHPPK"
     primer_bind     3398..3420
                     /label=pGEX 5'
                     /note="pGEX vectors, Glutathione-S-transferase, forward 
                     primer"
     CDS             3447..3470
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     primer_bind     complement(5806..5828)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(5973..5991)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        6059..6163
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6164..6232,6233..7024)
                     /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(6382..6401)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      join(7195..7227,1..556)
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
ORIGIN
        1 cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc aagagctacc
       61 aactcttttt ccgaaggtaa ctggcttcag cagagcgcag ataccaaata ctgtccttct
      121 agtgtagccg tagttaggcc accacttcaa gaactctgta gcaccgccta catacctcgc
      181 tctgctaatc ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt
      241 ggactcaaga cgatagttac cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg
      301 cacacagccc agcttggagc gaacgaccta caccgaactg agatacctac agcgtgagct
      361 atgagaaagc gccacgcttc ccgaagggag aaaggcggac aggtatccgg taagcggcag
      421 ggtcggaaca ggagagcgca cgagggagct tccaggggga aacgcctggt atctttatag
      481 tcctgtcggg tttcgccacc tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg
      541 gcggagccta tggaaaaacg ccagcaacgc ggccttttta cggttcctgg ccttttgctg
      601 gccttttgct cacatgttct ttcctgcgtt atcccctgat tctgtggata accgtattac
      661 cgcctttgag tgagctgata ccgctcgccg cagccgaacg accgagcgca gcgagtcagt
      721 gagcgaggaa gcggaagagc gcctgatgcg gtattttctc cttacgcatc tgtgcggtat
      781 ttcacaccgc ataaattccg acaccatcga atggcgcaaa acctttcgcg gtatggcatg
      841 atagcgcccg gaagagagtc aattcagggt ggtgaatgtg aaaccagtaa cgttatacga
      901 tgtcgcagag tatgccggtg tctcttatca gaccgtttcc cgcgtggtga accaggccag
      961 ccacgtttct gcgaaaacgc gggaaaaagt ggaagcggcg atggcggagc tgaattacat
     1021 tcccaaccgc gtggcacaac aactggcggg caaacagtcg ttgctgattg gcgttgccac
     1081 ctccagtctg gccctgcacg cgccgtcgca aattgtcgcg gcgattaaat ctcgcgccga
     1141 tcaactgggt gccagcgtgg tggtgtcgat ggtagaacga agcggcgtcg aagcctgtaa
     1201 agcggcggtg cacaatcttc tcgcgcaacg cgtcagtggg ctgatcatta actatccgct
     1261 ggatgaccag gatgccattg ctgtggaagc tgcctgcact aatgttccgg cgttatttct
     1321 tgatgtctct gaccagacac ccatcaacag tattattttc tcccatgaag acggtacgcg
     1381 actgggcgtg gagcatctgg tcgcattggg tcaccagcaa atcgcgctgt tagcgggccc
     1441 attaagttct gtctcggcgc gtctgcgtct ggctggctgg cataaatatc tcactcgcaa
     1501 tcaaattcag ccgatagcgg aacgggaagg cgactggagt gccatgtccg gttttcaaca
     1561 aaccatgcaa atgctgaatg agggcatcgt tcccactgcg atgctggttg ccaacgatca
     1621 gatggcgctg ggcgcaatgc gcgccattac cgagtccggg ctgcgcgttg gtgcggatat
     1681 ctcggtagtg ggatacgacg ataccgaaga cagctcatgt tatatcccgc cgttaaccac
     1741 catcaaacag gattttcgcc tgctggggca aaccagcgtg gaccgcttgc tgcaactctc
     1801 tcagggccag gcggtgaagg gcaatcagct gttgcccgtc tcactggtga aaagaaaaac
     1861 cacgctagcg cccaatacgc aaaccgcctc tccccgcgcg ttggccgatt cattaatgca
     1921 gctggcacga caggtttccc gactggaaag cgggcagtga gcgcaacgca attaatgtaa
     1981 gttagctcac tcattaggca ccccaggctt tacactttat gcttccggct cgtatgttgt
     2041 gtggaattgt gagcggataa caatttcaca caggaaacag ctatgaccat gattacggat
     2101 tcactggccg tcgttttaca acgtcgtgac tgggaaaacc ctggcgttac ccaacttaat
     2161 cgccttgcag cacatccccc tttcgccagc tggcgtaata gcgaagaggc ccgcaccgat
     2221 cgcccttccc aacagttgcg cagcctgaat ggcgaatggc gctttgcctg gtttccggca
     2281 ccagaagcgg tgccggaaag ctggctggag tgcgatcttc ctgaggccga tactgtcgtc
     2341 gtcccctcaa actggcagat gcacggttac gatgcgccca tctacaccaa cgtgacctat
     2401 cccattacgg tcaatccgcc gtttgttccc acggagaatc cgacgggttg ttactcgctc
     2461 acatttaatg ttgatgaaag ctggctacag gaaggccaga cgcgaattat ttttgatggc
     2521 gttggaatta gcttatcgac tgcacggtgc accaatgctt ctggcgtcag gcagccatcg
     2581 gaagctgtgg tatggctgtg caggtcgtaa atcactgcat aattcgtgtc gctcaaggcg
     2641 cactcccgtt ctggataatg ttttttgcgc cgacatcata acggttctgg caaatattct
     2701 gaaatgagct gttgacaatt aatcatcggc tcgtataatg tgtggaattg tgagcggata
     2761 acaatttcac acaggaaaca gtattcatgt cccctatact aggttattgg aaaattaagg
     2821 gccttgtgca acccactcga cttcttttgg aatatcttga agaaaaatat gaagagcatt
     2881 tgtatgagcg cgatgaaggt gataaatggc gaaacaaaaa gtttgaattg ggtttggagt
     2941 ttcccaatct tccttattat attgatggtg atgttaaatt aacacagtct atggccatca
     3001 tacgttatat agctgacaag cacaacatgt tgggtggttg tccaaaagag cgtgcagaga
     3061 tttcaatgct tgaaggagcg gttttggata ttagatacgg tgtttcgaga attgcatata
     3121 gtaaagactt tgaaactctc aaagttgatt ttcttagcaa gctacctgaa atgctgaaaa
     3181 tgttcgaaga tcgtttatgt cataaaacat atttaaatgg tgatcatgta acccatcctg
     3241 acttcatgtt gtatgacgct cttgatgttg ttttatacat ggacccaatg tgcctggatg
     3301 cgttcccaaa attagtttgt tttaaaaaac gtattgaagc tatcccacaa attgataagt
     3361 acttgaaatc cagcaagtat atagcatggc ctttgcaggg ctggcaagcc acgtttggtg
     3421 gtggcgacca tcctccaaaa tcggatctgg aagttctgtt ccaggggccc ctgggatccc
     3481 cggaattcat gctgggtacg ctgcgcgcca tggagggcga ggacgtggaa gacgaccagc
     3541 tgctgcagaa gctcagggcc agtcgccgcc gcttccagag gcgcatgcag cggctgatag
     3601 agaagtacaa ccagcccttc gaggacaccc cggtggtgca aatggccacg ctgacctacg
     3661 agacgccaca gggattgaga atttggggtg gaagactaat aaaggaaaga aacaaaggag
     3721 agatccagga ctcctccatg aagcccgcgg acaggacaga tggctccgtg caagctgcag
     3781 cctggggtcc tgagcttccc tcgcaccgca cagtcctggg agccgattca aaaagcggtg
     3841 aggtcgatgc cacgtcagac caggaagagt cagttgcttg ggccttagca cctgcagtgc
     3901 ctcaaagccc tttgaaaaat gaattaagaa ggaaatactt gacccaagtg gatatactgc
     3961 tacaaggtgc agagtatttt gagtgtgcag gtaacagagc tggaagggat gtacgtgtga
     4021 ctccgctgcc ttcactggcc tcacctgccg tgcctgcccc cggatactgc agtcgtatct
     4081 ccggaaagag tcctggtgac ccagcgaaac cagcttcatc tcccagagaa tgggatcctt
     4141 tgcatccttc ctccacagac atggccttag tacctagaaa tgacagcctc tccctacaag
     4201 agaccagtag cagcagcttc ttaagcagcc agccctttga agatgatgac atttgcaatg
     4261 tgaccatcag tgacctgtac gcagggatgc tgcactccat gagccggctg ttgagcacaa
     4321 agccatcaag catcatctcc accaaaacgt tcatcatgca aaactggaac tgcaggagga
     4381 ggcacagata taagagcagg atgaacaaaa catattgcaa aggagccaga cgttctcaga
     4441 ggagctccaa ggagaacttc ataccctgct ctgagcctgt gaaagggaca ggggcattaa
     4501 gagattgcaa gaacgtatta gatgtttctt gccgtaagac aggtttaaaa ttggaaaaag
     4561 cttttcttga agtcaacaga ccccaaatcc ataagttaga tccaagttgg aaggagcgca
     4621 aagtgacacc ctcgaagtat tcttccttga tttacttcga ctccagtgca acatataatc
     4681 ttgatgagga aaatagattt aggacattaa aatggttaat ttctcctgta aaaatagttt
     4741 ccagaccaac aatacgacag ggccatggag agaaccgtca gagggagatt gaaatccgat
     4801 ttgatcagct tcatcgggaa tattgcctga gtcccaggaa ccagcctcgc cggatgtgcc
     4861 tcccggactc ctgggccatg aacatgtaca gagggggtcc tgcgagtcct ggtggccttc
     4921 agggcttaga aacccgcagg ctgagtttac cttccagcaa agcaaaagca aaaagtttaa
     4981 gtgaggcttt tgaaaaccta ggcaaaagat ctctggaagc aggtaggtgc ctgcccaaga
     5041 gcgattcatc ttcatcactt ccaaagacca accccacaca cagcgcaact cgcccgcagc
     5101 agacatctga ccttcacgtt cagggaaata gttctggaat atttagaaag tcagtgtcac
     5161 ccagcaaaac tctttcagtc ccagataaag aagtgccagg ccacggaagg aatcgttacg
     5221 atgaaattaa agaagaattt gacaagcttc atcaaaagta ttgcctcaaa tctcctgggc
     5281 agatgacagt gcctttatgt attggagtgt ctacagataa agcaagtatg gaagttcgat
     5341 atcaaacaga aggcttctta ggaaaattaa atccagaccc tcacttccag ggtttccaga
     5401 agttgccatc atcacccctg gggtgcagaa aaagtctact gggctcaact gcaattgagg
     5461 ctccttcatc tacatgtgtt gctcgtgcca tcacgaggga tggcacgagg gaccatcagt
     5521 tccctgcaaa aagacccagg ctatcagaac cccagggctc cggacgccag ggcaattccc
     5581 tgggtgcctc agatggggtg gacaacaccg tcagaccggg agaccagggc agctcttcac
     5641 agcccaactc agaagagaga ggagagaaca cgtcttacag gatggaagag aaaagtgatt
     5701 tcatgctaga aaaattggaa actaaaagtg tgtaggtcga ctcgagcggc cgcatcgtga
     5761 ctgactgacg atctgcctcg cgcgtttcgg tgatgacggt gaaaacctct gacacatgca
     5821 gctcccggag acggtcacag cttgtctgta agcggatgcc gggagcagac aagcccgtca
     5881 gggcgcgtca gcgggtgttg gcgggtgtcg gggcgcagcc atgacccagt cacgtagcga
     5941 tagcggagtg tataattctt gaagacgaaa gggcctcgtg atacgcctat ttttataggt
     6001 taatgtcatg ataataatgg tttcttagac gtcaggtggc acttttcggg gaaatgtgcg
     6061 cggaacccct atttgtttat ttttctaaat acattcaaat atgtatccgc tcatgagaca
     6121 ataaccctga taaatgcttc aataatattg aaaaaggaag agtatgagta ttcaacattt
     6181 ccgtgtcgcc cttattccct tttttgcggc attttgcctt cctgtttttg ctcacccaga
     6241 aacgctggtg aaagtaaaag atgctgaaga tcagttgggt gcacgagtgg gttacatcga
     6301 actggatctc aacagcggta agatccttga gagttttcgc cccgaagaac gttttccaat
     6361 gatgagcact tttaaagttc tgctatgtgg cgcggtatta tcccgtgttg acgccgggca
     6421 agagcaactc ggtcgccgca tacactattc tcagaatgac ttggttgagt actcaccagt
     6481 cacagaaaag catcttacgg atggcatgac agtaagagaa ttatgcagtg ctgccataac
     6541 catgagtgat aacactgcgg ccaacttact tctgacaacg atcggaggac cgaaggagct
     6601 aaccgctttt ttgcacaaca tgggggatca tgtaactcgc cttgatcgtt gggaaccgga
     6661 gctgaatgaa gccataccaa acgacgagcg tgacaccacg atgcctgcag caatggcaac
     6721 aacgttgcgc aaactattaa ctggcgaact acttactcta gcttcccggc aacaattaat
     6781 agactggatg gaggcggata aagttgcagg accacttctg cgctcggccc ttccggctgg
     6841 ctggtttatt gctgataaat ctggagccgg tgagcgtggg tctcgcggta tcattgcagc
     6901 actggggcca gatggtaagc cctcccgtat cgtagttatc tacacgacgg ggagtcaggc
     6961 aactatggat gaacgaaata gacagatcgc tgagataggt gcctcactga ttaagcattg
     7021 gtaactgtca gaccaagttt actcatatat actttagatt gatttaaaac ttcattttta
     7081 atttaaaagg atctaggtga agatcctttt tgataatctc atgaccaaaa tcccttaacg
     7141 tgagttttcg ttccactgag cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga
     7201 tccttttttt ctgcgcgtaa tctgctg
//
