LOCUS       sequence-445580-        6307 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Bacterial expression of USP27X (C87A) with GST tag.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6307)
  AUTHORS   Koch I, Slovik M, Zhang Y, Liu B, Rennie M, Konz E, Cogne B, Daana 
            M, Davids L, Diets IJ, Gold NB, Holtz AM, Isidor B, Mor-Shaked H, 
            Neira Fresneda J, Niederhoffer KY, Nizon M, Pfundt R, Simon M, 
            Stegmann A, Guillen Sacoto MJ, Wevers M, Barakat TS, Yanovsky-Dagan 
            S, Atanassov BS, Toth R, Gao C, Bustos F, Harel T
  TITLE     USP27X variants underlying X-linked intellectual disability disrupt 
            protein function via distinct mechanisms.
  JOURNAL   Life Sci Alliance. 2024 Jan 5;7(3):e202302258. doi: 
            10.26508/lsa.202302258. Print 2024 Mar.
  PUBMED    38182161
REFERENCE   2  (bases 1 to 6307)
  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: "Life Sci 
            Alliance."; date: "2024-01-5"; volume: "7(3):e202302258. doi"; 
            pages: " 10.26508/lsa.202302258. Print 2024 Mar"
COMMENT     Sequence Label: pGEX6P1 GST-hUSP27X C87A
FEATURES             Location/Qualifiers
     source          1..6307
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(98..117)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      911..1499
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1400..1419
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     1653..1670
                     /label=L4440
                     /note="L4440 vector, forward primer"
     promoter        1743..1820
                     /gene="lacI"
                     /label=lacI promoter
     CDS             1821..2903
                     /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(1840..1859)
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        join(2952..2957,2958..2975,2976..2982)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2990..3006
                     /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     3014..3030
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     3014..3030
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             3026..3553
                     /codon_start=1
                     /gene="lacZ fragment"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /label=lacZ-alpha
                     /translation="MTMITDSLAVVLQRRDWENPGVTQLNRLAAHPPFASWRNSEEART
                     DRPSQQLRSLNGEWRFAWFPAPEAVPESWLECDLPEADTVVVPSNWQMHGYDAPIYTNV
                     TYPITVNPPFVPTENPTGCYSLTFNVDESWLQEGQTRIIFDGVGISLSTARCTNASGVR
                     QPSEAVVWLCRS"
     primer_bind     complement(3046..3063)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(3046..3062)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3055..3077)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     promoter        join(3655..3660,3661..3676,3677..3683)
                     /label=tac promoter
                     /note="strong E. coli promoter; hybrid between the trp and 
                     lac UV5 promoters"
     protein_bind    3691..3707
                     /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             3730..4383
                     /codon_start=1
                     /product="glutathione S-transferase from Schistosoma 
                     japonicum"
                     /label=GST
                     /translation="MSPILGYWKIKGLVQPTRLLLEYLEEKYEEHLYERDEGDKWRNKK
                     FELGLEFPNLPYYIDGDVKLTQSMAIIRYIADKHNMLGGCPKERAEISMLEGAVLDIRY
                     GVSRIAYSKDFETLKVDFLSKLPEMLKMFEDRLCHKTYLNGDHVTHPDFMLYDALDVVL
                     YMDPMCLDAFPKLVCFKKRIEAIPQIDKYLKSSKYIAWPLQGWQATFGGGDHPPK"
     primer_bind     4341..4363
                     /label=pGEX 5'
                     /note="pGEX vectors, Glutathione-S-transferase, forward 
                     primer"
     CDS             4390..4413
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     primer_bind     complement(5829..5851)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(5996..6014)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        6082..6186
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6187..6255,6256..6307,1..740)
                     /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"
ORIGIN
        1 gagtgggtta catcgaactg gatctcaaca gcggtaagat ccttgagagt tttcgccccg
       61 aagaacgttt tccaatgatg agcactttta aagttctgct atgtggcgcg gtattatccc
      121 gtgttgacgc cgggcaagag caactcggtc gccgcataca ctattctcag aatgacttgg
      181 ttgagtactc accagtcaca gaaaagcatc ttacggatgg catgacagta agagaattat
      241 gcagtgctgc cataaccatg agtgataaca ctgcggccaa cttacttctg acaacgatcg
      301 gaggaccgaa ggagctaacc gcttttttgc acaacatggg ggatcatgta actcgccttg
      361 atcgttggga accggagctg aatgaagcca taccaaacga cgagcgtgac accacgatgc
      421 ctgcagcaat ggcaacaacg ttgcgcaaac tattaactgg cgaactactt actctagctt
      481 cccggcaaca attaatagac tggatggagg cggataaagt tgcaggacca cttctgcgct
      541 cggcccttcc ggctggctgg tttattgctg ataaatctgg agccggtgag cgtgggtctc
      601 gcggtatcat tgcagcactg gggccagatg gtaagccctc ccgtatcgta gttatctaca
      661 cgacggggag tcaggcaact atggatgaac gaaatagaca gatcgctgag ataggtgcct
      721 cactgattaa gcattggtaa ctgtcagacc aagtttactc atatatactt tagattgatt
      781 taaaacttca tttttaattt aaaaggatct aggtgaagat cctttttgat aatctcatga
      841 ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc agaccccgta gaaaagatca
      901 aaggatcttc ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac
      961 caccgctacc agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg
     1021 taactggctt cagcagagcg cagataccaa atactgtcct tctagtgtag ccgtagttag
     1081 gccaccactt caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac
     1141 cagtggctgc tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt
     1201 taccggataa ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg
     1261 agcgaacgac ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc
     1321 ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc
     1381 gcacgaggga gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc
     1441 acctctgact tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa
     1501 acgccagcaa cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgt
     1561 tctttcctgc gttatcccct gattctgtgg ataaccgtat taccgccttt gagtgagctg
     1621 ataccgctcg ccgcagccga acgaccgagc gcagcgagtc agtgagcgag gaagcggaag
     1681 agcgcctgat gcggtatttt ctccttacgc atctgtgcgg tatttcacac cgcataaatt
     1741 ccgacaccat cgaatggcgc aaaacctttc gcggtatggc atgatagcgc ccggaagaga
     1801 gtcaattcag ggtggtgaat gtgaaaccag taacgttata cgatgtcgca gagtatgccg
     1861 gtgtctctta tcagaccgtt tcccgcgtgg tgaaccaggc cagccacgtt tctgcgaaaa
     1921 cgcgggaaaa agtggaagcg gcgatggcgg agctgaatta cattcccaac cgcgtggcac
     1981 aacaactggc gggcaaacag tcgttgctga ttggcgttgc cacctccagt ctggccctgc
     2041 acgcgccgtc gcaaattgtc gcggcgatta aatctcgcgc cgatcaactg ggtgccagcg
     2101 tggtggtgtc gatggtagaa cgaagcggcg tcgaagcctg taaagcggcg gtgcacaatc
     2161 ttctcgcgca acgcgtcagt gggctgatca ttaactatcc gctggatgac caggatgcca
     2221 ttgctgtgga agctgcctgc actaatgttc cggcgttatt tcttgatgtc tctgaccaga
     2281 cacccatcaa cagtattatt ttctcccatg aagacggtac gcgactgggc gtggagcatc
     2341 tggtcgcatt gggtcaccag caaatcgcgc tgttagcggg cccattaagt tctgtctcgg
     2401 cgcgtctgcg tctggctggc tggcataaat atctcactcg caatcaaatt cagccgatag
     2461 cggaacggga aggcgactgg agtgccatgt ccggttttca acaaaccatg caaatgctga
     2521 atgagggcat cgttcccact gcgatgctgg ttgccaacga tcagatggcg ctgggcgcaa
     2581 tgcgcgccat taccgagtcc gggctgcgcg ttggtgcgga tatctcggta gtgggatacg
     2641 acgataccga agacagctca tgttatatcc cgccgttaac caccatcaaa caggattttc
     2701 gcctgctggg gcaaaccagc gtggaccgct tgctgcaact ctctcagggc caggcggtga
     2761 agggcaatca gctgttgccc gtctcactgg tgaaaagaaa aaccacgcta gcgcccaata
     2821 cgcaaaccgc ctctccccgc gcgttggccg attcattaat gcagctggca cgacaggttt
     2881 cccgactgga aagcgggcag tgagcgcaac gcaattaatg taagttagct cactcattag
     2941 gcaccccagg ctttacactt tatgcttccg gctcgtatgt tgtgtggaat tgtgagcgga
     3001 taacaatttc acacaggaaa cagctatgac catgattacg gattcactgg ccgtcgtttt
     3061 acaacgtcgt gactgggaaa accctggcgt tacccaactt aatcgccttg cagcacatcc
     3121 ccctttcgcc agctggcgta atagcgaaga ggcccgcacc gatcgccctt cccaacagtt
     3181 gcgcagcctg aatggcgaat ggcgctttgc ctggtttccg gcaccagaag cggtgccgga
     3241 aagctggctg gagtgcgatc ttcctgaggc cgatactgtc gtcgtcccct caaactggca
     3301 gatgcacggt tacgatgcgc ccatctacac caacgtgacc tatcccatta cggtcaatcc
     3361 gccgtttgtt cccacggaga atccgacggg ttgttactcg ctcacattta atgttgatga
     3421 aagctggcta caggaaggcc agacgcgaat tatttttgat ggcgttggaa ttagcttatc
     3481 gactgcacgg tgcaccaatg cttctggcgt caggcagcca tcggaagctg tggtatggct
     3541 gtgcaggtcg taaatcactg cataattcgt gtcgctcaag gcgcactccc gttctggata
     3601 atgttttttg cgccgacatc ataacggttc tggcaaatat tctgaaatga gctgttgaca
     3661 attaatcatc ggctcgtata atgtgtggaa ttgtgagcgg ataacaattt cacacaggaa
     3721 acagtattca tgtcccctat actaggttat tggaaaatta agggccttgt gcaacccact
     3781 cgacttcttt tggaatatct tgaagaaaaa tatgaagagc atttgtatga gcgcgatgaa
     3841 ggtgataaat ggcgaaacaa aaagtttgaa ttgggtttgg agtttcccaa tcttccttat
     3901 tatattgatg gtgatgttaa attaacacag tctatggcca tcatacgtta tatagctgac
     3961 aagcacaaca tgttgggtgg ttgtccaaaa gagcgtgcag agatttcaat gcttgaagga
     4021 gcggttttgg atattagata cggtgtttcg agaattgcat atagtaaaga ctttgaaact
     4081 ctcaaagttg attttcttag caagctacct gaaatgctga aaatgttcga agatcgttta
     4141 tgtcataaaa catatttaaa tggtgatcat gtaacccatc ctgacttcat gttgtatgac
     4201 gctcttgatg ttgttttata catggaccca atgtgcctgg atgcgttccc aaaattagtt
     4261 tgttttaaaa aacgtattga agctatccca caaattgata agtacttgaa atccagcaag
     4321 tatatagcat ggcctttgca gggctggcaa gccacgtttg gtggtggcga ccatcctcca
     4381 aaatcggatc tggaagttct gttccagggg cccctgggat ccatgtgtaa ggactatgta
     4441 tatgacaaag acattgagca aattgccaaa gaagagcaag gagaagcttt gaaattacaa
     4501 gcctccacct caacagaggt ttctcaccag cagtgttcag tgccaggcct tggtgagaaa
     4561 ttcccaacct gggaaacaac caaaccagaa ttagaactgc tggggcacaa cccgaggaga
     4621 agaagaatca cctccagctt tacgatcggt ttaagaggac tcatcaatct tggcaacacg
     4681 gcctttatga actgcattgt ccaggccctc acccacacgc cgatactgag agatttcttt
     4741 ctctctgaca ggcaccgatg tgagatgccg agtcccgagt tgtgtctggt ctgtgagatg
     4801 tcgtcgctgt ttcgggagtt gtattctgga aacccgtctc ctcatgtgcc ctataagtta
     4861 ctgcacctgg tgtggataca tgcccgccat ttagcagggt acaggcaaca ggatgcccac
     4921 gagttcctca ttgcagcgtt agatgtcctg cacaggcact gcaaaggtga tgatgtcggg
     4981 aaggcggcca acaatcccaa ccactgtaac tgcatcatag accaaatctt cacaggtggc
     5041 ctgcagtctg atgtcacctg tcaagcctgc catggcgtct ccaccacgat agacccatgc
     5101 tgggacatta gtttggactt gcctggctct tgcacctcct tctggcccat gagcccaggg
     5161 agggagagca gtgtgaacgg ggaaagccac ataccaggaa tcaccaccct cacggactgc
     5221 ttgcggaggt ttacgaggcc agagcactta ggaagcagtg ccaaaatcaa atgtggtagt
     5281 tgccaaagct accaggaatc taccaaacag ctcacaatga ataaattacc tgtcgttgcc
     5341 tgttttcatt tcaaacggtt tgaacattca gcgaaacaga ggcgcaagat cactacatac
     5401 atttcctttc ctctggagct ggatatgacg ccgtttatgg cctcaagtaa agagagcaga
     5461 atgaatggac aattgcagct gccaaccaat agtggaaaca acgaaaataa gtattccttg
     5521 tttgctgtgg ttaatcacca aggaaccttg gagagtggcc actataccag cttcatccgg
     5581 caccacaagg accagtggtt caagtgtgat gatgccgtca tcactaaggc cagtattaag
     5641 gacgtactgg acagtgaagg gtatttactg ttctatcaca aacaggtgct agaacatgag
     5701 tcagaaaaag tgaaagaaat gaacacacaa gcctactgag gatccccgga attcccgggt
     5761 cgactcgagc ggccgcatcg tgactgactg acgatctgcc tcgcgcgttt cggtgatgac
     5821 ggtgaaaacc tctgacacat gcagctcccg gagacggtca cagcttgtct gtaagcggat
     5881 gccgggagca gacaagcccg tcagggcgcg tcagcgggtg ttggcgggtg tcggggcgca
     5941 gccatgaccc agtcacgtag cgatagcgga gtgtataatt cttgaagacg aaagggcctc
     6001 gtgatacgcc tatttttata ggttaatgtc atgataataa tggtttctta gacgtcaggt
     6061 ggcacttttc ggggaaatgt gcgcggaacc cctatttgtt tatttttcta aatacattca
     6121 aatatgtatc cgctcatgag acaataaccc tgataaatgc ttcaataata ttgaaaaagg
     6181 aagagtatga gtattcaaca tttccgtgtc gcccttattc ccttttttgc ggcattttgc
     6241 cttcctgttt ttgctcaccc agaaacgctg gtgaaagtaa aagatgctga agatcagttg
     6301 ggtgcac
//
