LOCUS       sequence-337481-        6306 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  pGEX2T-TUBG1 bacterial expression plasmid for production of 
            recombinant GST fusion protein.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6306)
  AUTHORS   Corvaisier M, Zhou J, Malycheva D, Cornella N, Chioureas D, 
            Gustafsson NMS, Rossello CA, Ayora S, Li T, Ekstrom-Holka K, 
            Jirstrom K, Lindstrom L, Alvarado-Kristensson M
  TITLE     The gamma-tubulin meshwork assists in the recruitment of PCNA to 
            chromatin in mammalian cells.
  JOURNAL   Commun Biol. 2021 Jun 22;4(1):767. doi: 10.1038/s42003-021-02280-1.
  PUBMED    34158617
REFERENCE   2  (bases 1 to 6306)
  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: "Commun 
            Biol."; date: "2021-06-22"; volume: "4(1):767. doi"; pages: " 
            10.1038/s42003-021-02280-1"
COMMENT     Sequence Label: pGEX2T-TUBG1
FEATURES             Location/Qualifiers
     source          1..6306
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(39..1121)
                     /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     1083..1102
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(1122..1199)
                     /gene="lacI"
                     /label=lacI promoter
     primer_bind     complement(1272..1289)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1443..2031)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1523..1542)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2202..2993,2994..3062))
                     /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     2825..2844
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3063..3167)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     3235..3253
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     3398..3420
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(4841..4858)
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
     CDS             complement(4865..5518)
                     /codon_start=1
                     /product="glutathione S-transferase from Schistosoma 
                     japonicum"
                     /label=GST
                     /translation="MSPILGYWKIKGLVQPTRLLLEYLEEKYEEHLYERDEGDKWRNKK
                     FELGLEFPNLPYYIDGDVKLTQSMAIIRYIADKHNMLGGCPKERAEISMLEGAVLDIRY
                     GVSRIAYSKDFETLKVDFLSKLPEMLKMFEDRLCHKTYLNGDHVTHPDFMLYDALDVVL
                     YMDPMCLDAFPKLVCFKKRIEAIPQIDKYLKSSKYIAWPLQGWQATFGGGDHPPK"
     primer_bind     complement(4885..4907)
                     /label=pGEX 5'
                     /note="pGEX vectors, Glutathione-S-transferase, forward 
                     primer"
     protein_bind    5541..5557
                     /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)."
     promoter        complement(join(5565..5571,5572..5587,5588..5593))
                     /label=tac promoter
                     /note="strong E. coli promoter; hybrid between the trp and 
                     lac UV5 promoters"
     CDS             complement(5695..6222)
                     /codon_start=1
                     /gene="lacZ fragment"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /label=lacZ-alpha
                     /translation="MTMITDSLAVVLQRRDWENPGVTQLNRLAAHPPFASWRNSEEART
                     DRPSQQLRSLNGEWRFAWFPAPEAVPESWLECDLPEADTVVVPSNWQMHGYDAPIYTNV
                     TYPITVNPPFVPTENPTGCYSLTFNVDESWLQEGQTRIIFDGVGISLSTARCTNASGVR
                     QPSEAVVWLCRS"
     primer_bind     6171..6193
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     6185..6202
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     6186..6202
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6218..6234)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6218..6234)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     protein_bind    6242..6258
                     /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)."
     promoter        complement(join(6266..6272,6273..6290,6291..6296))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
ORIGIN
        1 cctaatgagt gagctaactt acattaattg cgttgcgctc actgcccgct ttccagtcgg
       61 gaaacctgtc gtgccagctg cattaatgaa tcggccaacg cgcggggaga ggcggtttgc
      121 gtattgggcg ccagggtggt ttttcttttc accagtgaga cgggcaacag ctgattgccc
      181 ttcaccgcct ggccctgaga gagttgcagc aagcggtcca cgctggtttg ccccagcagg
      241 cgaaaatcct gtttgatggt ggttaacggc gggatataac atgagctgtc ttcggtatcg
      301 tcgtatccca ctaccgagat atccgcacca acgcgcagcc cggactcggt aatggcgcgc
      361 attgcgccca gcgccatctg atcgttggca accagcatcg cagtgggaac gatgccctca
      421 ttcagcattt gcatggtttg ttgaaaaccg gacatggcac tccagtcgcc ttcccgttcc
      481 gctatcggct gaatttgatt gcgagtgaga tatttatgcc agccagccag acgcagacgc
      541 gccgagacag aacttaatgg gcccgctaac agcgcgattt gctggtgacc caatgcgacc
      601 agatgctcca cgcccagtcg cgtaccgtct tcatgggaga aaataatact gttgatgggt
      661 gtctggtcag agacatcaag aaataacgcc ggaacattag tgcaggcagc ttccacagca
      721 atggcatcct ggtcatccag cggatagtta atgatcagcc cactgacgcg ttgcgcgaga
      781 agattgtgca ccgccgcttt acaggcttcg acgccgcttc gttctaccat cgacaccacc
      841 acgctggcac ccagttgatc ggcgcgagat ttaatcgccg cgacaatttg cgacggcgcg
      901 tgcagggcca gactggaggt ggcaacgcca atcagcaacg actgtttgcc cgccagttgt
      961 tgtgccacgc ggttgggaat gtaattcagc tccgccatcg ccgcttccac tttttcccgc
     1021 gttttcgcag aaacgtggct ggcctggttc accacgcggg aaacggtctg ataagagaca
     1081 ccggcatact ctgcgacatc gtataacgtt actggtttca cattcaccac cctgaattga
     1141 ctctcttccg ggcgctatca tgccataccg cgaaaggttt tgcgccattc gatggtgtcg
     1201 gaatttatgc ggtgtgaaat accgcacaga tgcgtaagga gaaaataccg catcaggcgc
     1261 tcttccgctt cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta
     1321 tcagctcact caaaggcggt aatacggtta tccacagaat caggggataa cgcaggaaag
     1381 aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg
     1441 tttttccata ggctccgccc ccctgacgag catcacaaaa atcgacgctc aagtcagagg
     1501 tggcgaaacc cgacaggact ataaagatac caggcgtttc cccctggaag ctccctcgtg
     1561 cgctctcctg ttccgaccct gccgcttacc ggatacctgt ccgcctttct cccttcggga
     1621 agcgtggcgc tttctcatag ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc
     1681 tccaagctgg gctgtgtgca cgaacccccc gttcagcccg accgctgcgc cttatccggt
     1741 aactatcgtc ttgagtccaa cccggtaaga cacgacttat cgccactggc agcagccact
     1801 ggtaacagga ttagcagagc gaggtatgta ggcggtgcta cagagttctt gaagtggtgg
     1861 cctaactacg gctacactag aaggacagta tttggtatct gcgctctgct gaagccagtt
     1921 accttcggaa aaagagttgg tagctcttga tccggcaaac aaaccaccgc tggtagcggt
     1981 ggtttttttg tttgcaagca gcagattacg cgcagaaaaa aaggatctca agaagatcct
     2041 ttgatctttt ctacggggtc tgacgctcag tggaacgaaa actcacgtta agggattttg
     2101 gtcatgagat tatcaaaaag gatcttcacc tagatccttt taaattaaaa atgaagtttt
     2161 aaatcaatct aaagtatata tgagtaaact tggtctgaca gttaccaatg cttaatcagt
     2221 gaggcaccta tctcagcgat ctgtctattt cgttcatcca tagttgcctg actccccgtc
     2281 gtgtagataa ctacgatacg ggagggctta ccatctggcc ccagtgctgc aatgataccg
     2341 cgagacccac gctcaccggc tccagattta tcagcaataa accagccagc cggaagggcc
     2401 gagcgcagaa gtggtcctgc aactttatcc gcctccatcc agtctattaa ttgttgccgg
     2461 gaagctagag taagtagttc gccagttaat agtttgcgca acgttgttgc cattgctgca
     2521 ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat tcagctccgg ttcccaacga
     2581 tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag cggttagctc cttcggtcct
     2641 ccgatcgttg tcagaagtaa gttggccgca gtgttatcac tcatggttat ggcagcactg
     2701 cataattctc ttactgtcat gccatccgta agatgctttt ctgtgactgg tgagtactca
     2761 accaagtcat tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaaca
     2821 cgggataata ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg aaaacgttct
     2881 tcggggcgaa aactctcaag gatcttaccg ctgttgagat ccagttcgat gtaacccact
     2941 cgtgcaccca actgatcttc agcatctttt actttcacca gcgtttctgg gtgagcaaaa
     3001 acaggaaggc aaaatgccgc aaaaaaggga ataagggcga cacggaaatg ttgaatactc
     3061 atactcttcc tttttcaata ttattgaagc atttatcagg gttattgtct catgagcgga
     3121 tacatatttg aatgtattta gaaaaataaa caaatagggg ttccgcgcac atttccccga
     3181 aaagtgccac ctgacgtcta agaaaccatt attatcatga cattaaccta taaaaatagg
     3241 cgtatcacga ggccctttcg tcttcaagaa ttatacactc cgctatcgct acgtgactgg
     3301 gtcatggctg cgccccgaca cccgccaaca cccgctgacg cgccctgacg ggcttgtctg
     3361 ctcccggcat ccgcttacag acaagctgtg accgtctccg ggagctgcat gtgtcagagg
     3421 ttttcaccgt catcaccgaa acgcgcgagg cagatcgtca gtcagtcacg atgaattctc
     3481 actgctcctg ggtgccccag gagatgtagt ctggccgtgt ggccgcatgg tactcatcga
     3541 tgagctgctg cacaatctcc ctggatgtgt ccatctcatc aaagttgtcc ttgaacatgt
     3601 cctccttgcg gaactgctcc aggaaggcct cccgcttacg cagcttgtca tactggcgac
     3661 aggttctctc gaagagcgag gagatgctgg tgtggttggc catcatgagc ccgctgaccc
     3721 ggtgggccga gggcaggtag ggagacttcc tcgacagggc cacctggatg ctggcggggc
     3781 cccacgggat gaagttggcc aacttgcgtt cccggatcct ctgcaagctc ttgtggacct
     3841 gggtggggtc cacctctccc tggatgatgt tgaggatggc gatgtagcag tggttggtct
     3901 ggcggtctcg gcctgtggac accatcacgt tcttgggctg cagcagccgc ctcatgacat
     3961 ccaggaccgt ggtcttcctc acgctggcca ctgactggtc cgtagtgaga ggggtgtagc
     4021 cggtcatgag gaagtggagc cgtggggtgg gaatgagcga ggcgatgagg ccgatgaggt
     4081 cattgttcat gtagccaggg tagcgcaggg tggtggtgct ggctgacatg atggtagaca
     4141 ccagctggtt gatctgggag aaggatgggt tctggatgtg caggcggtct gtggcaatcc
     4201 ggttcagggc tgtgttgtcc agcaccacca cacagtctgc attctgcgtc agcctcttga
     4261 gtgtgaggag tgaattgtaa ggctggacca ccacatcgct catctcgtcc tggttgggaa
     4321 acactgagta tgtctgcacc agcttcttag gatacctgtc attcagccgt tctaagaggt
     4381 aggaacccag tccagagcct gtccccccag caatggagtg acacagcaca aagccctcta
     4441 gactgtcact accatctgcc tcccggtcta tgatgtcaaa aatgtcctca tggatctttt
     4501 ctccctggga gaatccgctg gcccagttgt tgccagctcc tcctccatgt tccgacaggt
     4561 agatgttctc tgggttgtag agcttggcat agggggagtt gaggatggag tggatcaccc
     4621 ggggttccaa gtccagcagc acggcccggg ggatgtagtg ctcatcgtct gcctggtaga
     4681 aaaagacgtc cttgtggtca gtgccctcgg tggcgaactc ctccacgatg ccctcggggc
     4741 tgataccatg ctcggcgcac agctgtttcc agaactcgaa cccaatctga ttgccgcact
     4801 ggcccaactg tagggtgatg atttccctcg gcatagcttg ggatccacgc ggaaccagat
     4861 ccgattttgg aggatggtcg ccaccaccaa acgtggcttg ccagccctgc aaaggccatg
     4921 ctatatactt gctggatttc aagtacttat caatttgtgg gatagcttca atacgttttt
     4981 taaaacaaac taattttggg aacgcatcca ggcacattgg gtccatgtat aaaacaacat
     5041 caagagcgtc atacaacatg aagtcaggat gggttacatg atcaccattt aaatatgttt
     5101 tatgacataa acgatcttcg aacattttca gcatttcagg tagcttgcta agaaaatcaa
     5161 ctttgagagt ttcaaagtct ttactatatg caattctcga aacaccgtat ctaatatcca
     5221 aaaccgctcc ttcaagcatt gaaatctctg cacgctcttt tggacaacca cccaacatgt
     5281 tgtgcttgtc agctatataa cgtatgatgg ccatagactg tgttaattta acatcaccat
     5341 caatataata aggaagattg ggaaactcca aacccaattc aaactttttg tttcgccatt
     5401 tatcaccttc atcgcgctca tacaaatgct cttcatattt ttcttcaaga tattccaaaa
     5461 gaagtcgagt gggttgcaca aggcccttaa ttttccaata acctagtata ggggacatga
     5521 atactgtttc ctgtgtgaaa ttgttatccg ctcacaattc cacacattat acgagccgat
     5581 gattaattgt caacagctca tttcagaata tttgccagaa ccgttatgat gtcggcgcaa
     5641 aaaacattat ccagaacggg agtgcgcctt gagcgacacg aattatgcag tgatttacga
     5701 cctgcacagc cataccacag cttccgatgg ctgcctgacg ccagaagcat tggtgcaccg
     5761 tgcagtcgat aagctaattc caacgccatc aaaaataatt cgcgtctggc cttcctgtag
     5821 ccagctttca tcaacattaa atgtgagcga gtaacaaccc gtcggattct ccgtgggaac
     5881 aaacggcgga ttgaccgtaa tgggataggt cacgttggtg tagatgggcg catcgtaacc
     5941 gtgcatctgc cagtttgagg ggacgacgac agtatcggcc tcaggaagat cgcactccag
     6001 ccagctttcc ggcaccgctt ctggtgccgg aaaccaggca aagcgccatt cgccattcag
     6061 gctgcgcaac tgttgggaag ggcgatcggt gcgggcctct tcgctattac gccagctggc
     6121 gaaaggggga tgtgctgcaa ggcgattaag ttgggtaacg ccagggtttt cccagtcacg
     6181 acgttgtaaa acgacggcca gtgaatccgt aatcatggtc atagctgttt cctgtgtgaa
     6241 attgttatcc gctcacaatt ccacacaaca tacgagccgg aagcataaag tgtaaagcct
     6301 ggggtg
//
