LOCUS       sequence-354807-        7021 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Express GST-DAXX-Delta-DE-His in bacteria.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7021)
  AUTHORS   Huang L, Agrawal T, Zhu G, Yu S, Tao L, Lin J, Marmorstein R, 
            Shorter J, Yang X
  TITLE     DAXX represents a new type of protein-folding enabler.
  JOURNAL   Nature. 2021 Aug 18. pii: 10.1038/s41586-021-03824-5. doi: 
            10.1038/s41586-021-03824-5.
  PUBMED    34408321
REFERENCE   2  (bases 1 to 7021)
  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: "Nature. 
            2021 Aug 18. pii: 10.1038/s41586-021-03824-5. doi: 
            10.1038/s41586-021-03824-5."
COMMENT     Sequence Label: pGEX-DAXX-Delta-D/E-His
FEATURES             Location/Qualifiers
     source          1..7021
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        join(125..130,131..148,149..155)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    163..179
                     /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     187..203
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     187..203
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             199..726
                     /codon_start=1
                     /gene="lacZ fragment"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /label=lacZ-alpha
                     /translation="MTMITDSLAVVLQRRDWENPGVTQLNRLAAHPPFASWRNSEEART
                     DRPSQQLRSLNGEWRFAWFPAPEAVPESWLECDLPEADTVVVPSNWQMHGYDAPIYTNV
                     TYPITVNPPFVPTENPTGCYSLTFNVDESWLQEGQTRIIFDGVGISLSTARCTNASGVR
                     QPSEAVVWLCRS"
     primer_bind     complement(219..236)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(219..235)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(228..250)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     promoter        join(828..833,834..849,850..856)
                     /label=tac promoter
                     /note="strong E. coli promoter; hybrid between the trp and 
                     lac UV5 promoters"
     protein_bind    864..880
                     /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             903..1556
                     /codon_start=1
                     /product="glutathione S-transferase from Schistosoma 
                     japonicum"
                     /label=GST
                     /translation="MSPILGYWKIKGLVQPTRLLLEYLEEKYEEHLYERDEGDKWRNKK
                     FELGLEFPNLPYYIDGDVKLTQSMAIIRYIADKHNMLGGCPKERAEISMLEGAVLDIRY
                     GVSRIAYSKDFETLKVDFLSKLPEMLKMFEDRLCHKTYLNGDHVTHPDFMLYDALDVVL
                     YMDPMCLDAFPKLVCFKKRIEAIPQIDKYLKSSKYIAWPLQGWQATFGGGDHPPK"
     primer_bind     1514..1536
                     /label=pGEX 5'
                     /note="pGEX vectors, Glutathione-S-transferase, forward 
                     primer"
     CDS             1563..1580
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
     CDS             1581..1601
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     CDS             3612..3629
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(3716..3738)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(3883..3901)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3969..4073
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4074..4142,4143..4934)
                     /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(4292..4311)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5105..5693
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5594..5613
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5847..5864
                     /label=L4440
                     /note="L4440 vector, forward primer"
     promoter        5937..6014
                     /gene="lacI"
                     /label=lacI promoter
     CDS             join(6015..7021,1..76)
                     /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(6034..6053)
                     /label=LacI-R
                     /note="LacI, reverse primer"
ORIGIN
        1 cgcctctccc cgcgcgttgg ccgattcatt aatgcagctg gcacgacagg tttcccgact
       61 ggaaagcggg cagtgagcgc aacgcaatta atgtaagtta gctcactcat taggcacccc
      121 aggctttaca ctttatgctt ccggctcgta tgttgtgtgg aattgtgagc ggataacaat
      181 ttcacacagg aaacagctat gaccatgatt acggattcac tggccgtcgt tttacaacgt
      241 cgtgactggg aaaaccctgg cgttacccaa cttaatcgcc ttgcagcaca tccccctttc
      301 gccagctggc gtaatagcga agaggcccgc accgatcgcc cttcccaaca gttgcgcagc
      361 ctgaatggcg aatggcgctt tgcctggttt ccggcaccag aagcggtgcc ggaaagctgg
      421 ctggagtgcg atcttcctga ggccgatact gtcgtcgtcc cctcaaactg gcagatgcac
      481 ggttacgatg cgcccatcta caccaacgtg acctatccca ttacggtcaa tccgccgttt
      541 gttcccacgg agaatccgac gggttgttac tcgctcacat ttaatgttga tgaaagctgg
      601 ctacaggaag gccagacgcg aattattttt gatggcgttg gaattagctt atcgactgca
      661 cggtgcacca atgcttctgg cgtcaggcag ccatcggaag ctgtggtatg gctgtgcagg
      721 tcgtaaatca ctgcataatt cgtgtcgctc aaggcgcact cccgttctgg ataatgtttt
      781 ttgcgccgac atcataacgg ttctggcaaa tattctgaaa tgagctgttg acaattaatc
      841 atcggctcgt ataatgtgtg gaattgtgag cggataacaa tttcacacag gaaacagtat
      901 tcatgtcccc tatactaggt tattggaaaa ttaagggcct tgtgcaaccc actcgacttc
      961 ttttggaata tcttgaagaa aaatatgaag agcatttgta tgagcgcgat gaaggtgata
     1021 aatggcgaaa caaaaagttt gaattgggtt tggagtttcc caatcttcct tattatattg
     1081 atggtgatgt taaattaaca cagtctatgg ccatcatacg ttatatagct gacaagcaca
     1141 acatgttggg tggttgtcca aaagagcgtg cagagatttc aatgcttgaa ggagcggttt
     1201 tggatattag atacggtgtt tcgagaattg catatagtaa agactttgaa actctcaaag
     1261 ttgattttct tagcaagcta cctgaaatgc tgaaaatgtt cgaagatcgt ttatgtcata
     1321 aaacatattt aaatggtgat catgtaaccc atcctgactt catgttgtat gacgctcttg
     1381 atgttgtttt atacatggac ccaatgtgcc tggatgcgtt cccaaaatta gtttgtttta
     1441 aaaaacgtat tgaagctatc ccacaaattg ataagtactt gaaatccagc aagtatatag
     1501 catggccttt gcagggctgg caagccacgt ttggtggtgg cgaccatcct ccaaaatcgg
     1561 atctggttcc gcgtggatct gagaacctgt acttccaggg atccgccacc gctaacagca
     1621 tcatcgtgct ggatgatgat gacgaagatg aagcagctgc tcagccaggg ccctcccacc
     1681 cactccccaa tgcggcctca cctggggcag aagcccctag ctcctctgag cctcatgggg
     1741 ccagaggaag cagtagttcg ggcggcaaga aatgctacaa gctggagaat gagaagctgt
     1801 tcgaagagtt ccttgaactt tgtaagatgc agacagcaga ccaccctgag gtggtcccat
     1861 tcctctataa ccggcagcaa cgtgcccact ctctgttttt ggcctcggcg gagttctgca
     1921 acatcctctc tagggtcctg tctcgggccc ggagccggcc agccaagctc tatgtctaca
     1981 tcaatgagct ctgcactgtt ctcaaggccc actcagccaa aaagaagctg aacttggccc
     2041 ctgccgccac cacctccaat gagccctctg ggaataaccc tcccacacac ctctccttgg
     2101 accccacaaa tgctgaaaac actgcctctc agtctccaag gacccgtggt tcccggcggc
     2161 agatccagcg tttggagcag ctgctggcgc tctatgtggc agagatccgg cggctgcagg
     2221 aaaaggagtt ggatctctca gaattggatg acccagactc cgcatacctg caggaggcac
     2281 ggttgaagcg taagctgatc cgcctctttg ggcgactatg tgagctgaaa gactgctctt
     2341 cactgaccgg ccgtgtcata gagcagcgca tcccctaccg tggcacccgc tacccagagg
     2401 ttaacaggcg cattgagcgg ctcatcaaca agccagggcc tgataccttc cctgactatg
     2461 gggatgtgct tcgggctgta gagaaggcag ctgcccgaca cagccttggc ctcccccgac
     2521 agcagctcca gctcatggct caggatgcct tccgagatgt gggcatcagg ttacaggagc
     2581 gacgtcacct cgatctcatc tacaactttg gctgccacct cacagatgac tataggccag
     2641 gcgttgaccc tgcactatca gatcctgtgt tggcccggcg ccttcgggaa aaccggagtt
     2701 tggccatgag tcggctggat gaggtcatct ccaaatacgc aatgttgcaa gacaaaagtg
     2761 aggagggcga gagaaaaaag agaagagctc ggctccaagg cacctcttcc cactctgcag
     2821 acacccccga agcctccttg gattctggtg agggccctag tggaatggca cccatgtcct
     2881 cactacagat ctccaatgaa aagaacctgg aacctggcaa acagatcagc agatcttcag
     2941 gggagcagca aaacaaagga cgcatagtgt caccatcgtt actgtcagaa gaacccctgg
     3001 ccccctccag catagatgct gaaagcaatg gagaacagcc tgaggagctg accctggagg
     3061 aagaaagccc tgtgtctcag ctctttgagc tagagattga agctttgccc ctggataccc
     3121 cttcctctgt ggagacggac atttcctctt ccaggaagca atcagaggag cccttcacca
     3181 ctgtcttaga gaatggagca ggcatggtct cttctacttc cttcaatgga ggcgtctctc
     3241 ctcacaactg gggagattct ggtcccccct gcaaaaaatc tcggaaggag aagaagcaaa
     3301 caggatcagg gccattagga aacagctatg tggaaaggca aaggtcagtg catgagaaga
     3361 atgggaaaaa gatatgtacc ctgcccagcc caccttcccc cttggcttcc ttggccccag
     3421 ttgctgattc ctccacgagg gtggactctc ccagccatgg cctggtgacc agctccctct
     3481 gcatcccttc tccagcccgg ctgtcccaaa ccccccattc acagcctcct cggcctggta
     3541 cttgcaagac aagtgtggcc acacaatgcg atccagaaga gatcatcgtg ctctcagact
     3601 ctgatgtcga ccaccaccac caccaccact agctcgacaa gcttgatatc gcggccgcaa
     3661 ttcatcgtga ctgactgacg atctgcctcg cgcgtttcgg tgatgacggt gaaaacctct
     3721 gacacatgca gctcccggag acggtcacag cttgtctgta agcggatgcc gggagcagac
     3781 aagcccgtca gggcgcgtca gcgggtgttg gcgggtgtcg gggcgcagcc atgacccagt
     3841 cacgtagcga tagcggagtg tataattctt gaagacgaaa gggcctcgtg atacgcctat
     3901 ttttataggt taatgtcatg ataataatgg tttcttagac gtcaggtggc acttttcggg
     3961 gaaatgtgcg cggaacccct atttgtttat ttttctaaat acattcaaat atgtatccgc
     4021 tcatgagaca ataaccctga taaatgcttc aataatattg aaaaaggaag agtatgagta
     4081 ttcaacattt ccgtgtcgcc cttattccct tttttgcggc attttgcctt cctgtttttg
     4141 ctcacccaga aacgctggtg aaagtaaaag atgctgaaga tcagttgggt gcacgagtgg
     4201 gttacatcga actggatctc aacagcggta agatccttga gagttttcgc cccgaagaac
     4261 gttttccaat gatgagcact tttaaagttc tgctatgtgg cgcggtatta tcccgtgttg
     4321 acgccgggca agagcaactc ggtcgccgca tacactattc tcagaatgac ttggttgagt
     4381 actcaccagt cacagaaaag catcttacgg atggcatgac agtaagagaa ttatgcagtg
     4441 ctgccataac catgagtgat aacactgcgg ccaacttact tctgacaacg atcggaggac
     4501 cgaaggagct aaccgctttt ttgcacaaca tgggggatca tgtaactcgc cttgatcgtt
     4561 gggaaccgga gctgaatgaa gccataccaa acgacgagcg tgacaccacg atgcctgcag
     4621 caatggcaac aacgttgcgc aaactattaa ctggcgaact acttactcta gcttcccggc
     4681 aacaattaat agactggatg gaggcggata aagttgcagg accacttctg cgctcggccc
     4741 ttccggctgg ctggtttatt gctgataaat ctggagccgg tgagcgtggg tctcgcggta
     4801 tcattgcagc actggggcca gatggtaagc cctcccgtat cgtagttatc tacacgacgg
     4861 ggagtcaggc aactatggat gaacgaaata gacagatcgc tgagataggt gcctcactga
     4921 ttaagcattg gtaactgtca gaccaagttt actcatatat actttagatt gatttaaaac
     4981 ttcattttta atttaaaagg atctaggtga agatcctttt tgataatctc atgaccaaaa
     5041 tcccttaacg tgagttttcg ttccactgag cgtcagaccc cgtagaaaag atcaaaggat
     5101 cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc
     5161 taccagcggt ggtttgtttg ccggatcaag agctaccaac tctttttccg aaggtaactg
     5221 gcttcagcag agcgcagata ccaaatactg tccttctagt gtagccgtag ttaggccacc
     5281 acttcaagaa ctctgtagca ccgcctacat acctcgctct gctaatcctg ttaccagtgg
     5341 ctgctgccag tggcgataag tcgtgtctta ccgggttgga ctcaagacga tagttaccgg
     5401 ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac acagcccagc ttggagcgaa
     5461 cgacctacac cgaactgaga tacctacagc gtgagctatg agaaagcgcc acgcttcccg
     5521 aagggagaaa ggcggacagg tatccggtaa gcggcagggt cggaacagga gagcgcacga
     5581 gggagcttcc agggggaaac gcctggtatc tttatagtcc tgtcgggttt cgccacctct
     5641 gacttgagcg tcgatttttg tgatgctcgt caggggggcg gagcctatgg aaaaacgcca
     5701 gcaacgcggc ctttttacgg ttcctggcct tttgctggcc ttttgctcac atgttctttc
     5761 ctgcgttatc ccctgattct gtggataacc gtattaccgc ctttgagtga gctgataccg
     5821 ctcgccgcag ccgaacgacc gagcgcagcg agtcagtgag cgaggaagcg gaagagcgcc
     5881 tgatgcggta ttttctcctt acgcatctgt gcggtatttc acaccgcata aattccgaca
     5941 ccatcgaatg gcgcaaaacc tttcgcggta tggcatgata gcgcccggaa gagagtcaat
     6001 tcagggtggt gaatgtgaaa ccagtaacgt tatacgatgt cgcagagtat gccggtgtct
     6061 cttatcagac cgtttcccgc gtggtgaacc aggccagcca cgtttctgcg aaaacgcggg
     6121 aaaaagtgga agcggcgatg gcggagctga attacattcc caaccgcgtg gcacaacaac
     6181 tggcgggcaa acagtcgttg ctgattggcg ttgccacctc cagtctggcc ctgcacgcgc
     6241 cgtcgcaaat tgtcgcggcg attaaatctc gcgccgatca actgggtgcc agcgtggtgg
     6301 tgtcgatggt agaacgaagc ggcgtcgaag cctgtaaagc ggcggtgcac aatcttctcg
     6361 cgcaacgcgt cagtgggctg atcattaact atccgctgga tgaccaggat gccattgctg
     6421 tggaagctgc ctgcactaat gttccggcgt tatttcttga tgtctctgac cagacaccca
     6481 tcaacagtat tattttctcc catgaagacg gtacgcgact gggcgtggag catctggtcg
     6541 cattgggtca ccagcaaatc gcgctgttag cgggcccatt aagttctgtc tcggcgcgtc
     6601 tgcgtctggc tggctggcat aaatatctca ctcgcaatca aattcagccg atagcggaac
     6661 gggaaggcga ctggagtgcc atgtccggtt ttcaacaaac catgcaaatg ctgaatgagg
     6721 gcatcgttcc cactgcgatg ctggttgcca acgatcagat ggcgctgggc gcaatgcgcg
     6781 ccattaccga gtccgggctg cgcgttggtg cggatatctc ggtagtggga tacgacgata
     6841 ccgaagacag ctcatgttat atcccgccgt taaccaccat caaacaggat tttcgcctgc
     6901 tggggcaaac cagcgtggac cgcttgctgc aactctctca gggccaggcg gtgaagggca
     6961 atcagctgtt gcccgtctca ctggtgaaaa gaaaaaccac cctggcgccc aatacgcaaa
     7021 c
//
