LOCUS       sequence-445582-        6278 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Bacterial expression of USP27X (Y144C) with GST tag.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6278)
  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 6278)
  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 Y144C
FEATURES             Location/Qualifiers
     source          1..6278
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(119..138)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      932..1520
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1421..1440
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     1674..1691
                     /label=L4440
                     /note="L4440 vector, forward primer"
     promoter        1764..1841
                     /gene="lacI"
                     /label=lacI promoter
     CDS             1842..2924
                     /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(1861..1880)
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        join(2973..2978,2979..2996,2997..3003)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3011..3027
                     /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     3035..3051
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     3035..3051
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             3047..3574
                     /codon_start=1
                     /gene="lacZ fragment"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /label=lacZ-alpha
                     /translation="MTMITDSLAVVLQRRDWENPGVTQLNRLAAHPPFASWRNSEEART
                     DRPSQQLRSLNGEWRFAWFPAPEAVPESWLECDLPEADTVVVPSNWQMHGYDAPIYTNV
                     TYPITVNPPFVPTENPTGCYSLTFNVDESWLQEGQTRIIFDGVGISLSTARCTNASGVR
                     QPSEAVVWLCRS"
     primer_bind     complement(3067..3084)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(3067..3083)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3076..3098)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     promoter        join(3676..3681,3682..3697,3698..3704)
                     /label=tac promoter
                     /note="strong E. coli promoter; hybrid between the trp and 
                     lac UV5 promoters"
     protein_bind    3712..3728
                     /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             3751..4404
                     /codon_start=1
                     /product="glutathione S-transferase from Schistosoma 
                     japonicum"
                     /label=GST
                     /translation="MSPILGYWKIKGLVQPTRLLLEYLEEKYEEHLYERDEGDKWRNKK
                     FELGLEFPNLPYYIDGDVKLTQSMAIIRYIADKHNMLGGCPKERAEISMLEGAVLDIRY
                     GVSRIAYSKDFETLKVDFLSKLPEMLKMFEDRLCHKTYLNGDHVTHPDFMLYDALDVVL
                     YMDPMCLDAFPKLVCFKKRIEAIPQIDKYLKSSKYIAWPLQGWQATFGGGDHPPK"
     primer_bind     4362..4384
                     /label=pGEX 5'
                     /note="pGEX vectors, Glutathione-S-transferase, forward 
                     primer"
     CDS             4411..4434
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     primer_bind     complement(5821..5843)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(5988..6006)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        6074..6178
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6179..6247,6248..6278,1..761)
                     /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 ctgaagatca gttgggtgca cgagtgggtt acatcgaact ggatctcaac agcggtaaga
       61 tccttgagag ttttcgcccc gaagaacgtt ttccaatgat gagcactttt aaagttctgc
      121 tatgtggcgc ggtattatcc cgtgttgacg ccgggcaaga gcaactcggt cgccgcatac
      181 actattctca gaatgacttg gttgagtact caccagtcac agaaaagcat cttacggatg
      241 gcatgacagt aagagaatta tgcagtgctg ccataaccat gagtgataac actgcggcca
      301 acttacttct gacaacgatc ggaggaccga aggagctaac cgcttttttg cacaacatgg
      361 gggatcatgt aactcgcctt gatcgttggg aaccggagct gaatgaagcc ataccaaacg
      421 acgagcgtga caccacgatg cctgcagcaa tggcaacaac gttgcgcaaa ctattaactg
      481 gcgaactact tactctagct tcccggcaac aattaataga ctggatggag gcggataaag
      541 ttgcaggacc acttctgcgc tcggcccttc cggctggctg gtttattgct gataaatctg
      601 gagccggtga gcgtgggtct cgcggtatca ttgcagcact ggggccagat ggtaagccct
      661 cccgtatcgt agttatctac acgacgggga gtcaggcaac tatggatgaa cgaaatagac
      721 agatcgctga gataggtgcc tcactgatta agcattggta actgtcagac caagtttact
      781 catatatact ttagattgat ttaaaacttc atttttaatt taaaaggatc taggtgaaga
      841 tcctttttga taatctcatg accaaaatcc cttaacgtga gttttcgttc cactgagcgt
      901 cagaccccgt agaaaagatc aaaggatctt cttgagatcc tttttttctg cgcgtaatct
      961 gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg gatcaagagc
     1021 taccaactct ttttccgaag gtaactggct tcagcagagc gcagatacca aatactgtcc
     1081 ttctagtgta gccgtagtta ggccaccact tcaagaactc tgtagcaccg cctacatacc
     1141 tcgctctgct aatcctgtta ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg
     1201 ggttggactc aagacgatag ttaccggata aggcgcagcg gtcgggctga acggggggtt
     1261 cgtgcacaca gcccagcttg gagcgaacga cctacaccga actgagatac ctacagcgtg
     1321 agctatgaga aagcgccacg cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg
     1381 gcagggtcgg aacaggagag cgcacgaggg agcttccagg gggaaacgcc tggtatcttt
     1441 atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg atttttgtga tgctcgtcag
     1501 gggggcggag cctatggaaa aacgccagca acgcggcctt tttacggttc ctggcctttt
     1561 gctggccttt tgctcacatg ttctttcctg cgttatcccc tgattctgtg gataaccgta
     1621 ttaccgcctt tgagtgagct gataccgctc gccgcagccg aacgaccgag cgcagcgagt
     1681 cagtgagcga ggaagcggaa gagcgcctga tgcggtattt tctccttacg catctgtgcg
     1741 gtatttcaca ccgcataaat tccgacacca tcgaatggcg caaaaccttt cgcggtatgg
     1801 catgatagcg cccggaagag agtcaattca gggtggtgaa tgtgaaacca gtaacgttat
     1861 acgatgtcgc agagtatgcc ggtgtctctt atcagaccgt ttcccgcgtg gtgaaccagg
     1921 ccagccacgt ttctgcgaaa acgcgggaaa aagtggaagc ggcgatggcg gagctgaatt
     1981 acattcccaa ccgcgtggca caacaactgg cgggcaaaca gtcgttgctg attggcgttg
     2041 ccacctccag tctggccctg cacgcgccgt cgcaaattgt cgcggcgatt aaatctcgcg
     2101 ccgatcaact gggtgccagc gtggtggtgt cgatggtaga acgaagcggc gtcgaagcct
     2161 gtaaagcggc ggtgcacaat cttctcgcgc aacgcgtcag tgggctgatc attaactatc
     2221 cgctggatga ccaggatgcc attgctgtgg aagctgcctg cactaatgtt ccggcgttat
     2281 ttcttgatgt ctctgaccag acacccatca acagtattat tttctcccat gaagacggta
     2341 cgcgactggg cgtggagcat ctggtcgcat tgggtcacca gcaaatcgcg ctgttagcgg
     2401 gcccattaag ttctgtctcg gcgcgtctgc gtctggctgg ctggcataaa tatctcactc
     2461 gcaatcaaat tcagccgata gcggaacggg aaggcgactg gagtgccatg tccggttttc
     2521 aacaaaccat gcaaatgctg aatgagggca tcgttcccac tgcgatgctg gttgccaacg
     2581 atcagatggc gctgggcgca atgcgcgcca ttaccgagtc cgggctgcgc gttggtgcgg
     2641 atatctcggt agtgggatac gacgataccg aagacagctc atgttatatc ccgccgttaa
     2701 ccaccatcaa acaggatttt cgcctgctgg ggcaaaccag cgtggaccgc ttgctgcaac
     2761 tctctcaggg ccaggcggtg aagggcaatc agctgttgcc cgtctcactg gtgaaaagaa
     2821 aaaccacgct agcgcccaat acgcaaaccg cctctccccg cgcgttggcc gattcattaa
     2881 tgcagctggc acgacaggtt tcccgactgg aaagcgggca gtgagcgcaa cgcaattaat
     2941 gtaagttagc tcactcatta ggcaccccag gctttacact ttatgcttcc ggctcgtatg
     3001 ttgtgtggaa ttgtgagcgg ataacaattt cacacaggaa acagctatga ccatgattac
     3061 ggattcactg gccgtcgttt tacaacgtcg tgactgggaa aaccctggcg ttacccaact
     3121 taatcgcctt gcagcacatc cccctttcgc cagctggcgt aatagcgaag aggcccgcac
     3181 cgatcgccct tcccaacagt tgcgcagcct gaatggcgaa tggcgctttg cctggtttcc
     3241 ggcaccagaa gcggtgccgg aaagctggct ggagtgcgat cttcctgagg ccgatactgt
     3301 cgtcgtcccc tcaaactggc agatgcacgg ttacgatgcg cccatctaca ccaacgtgac
     3361 ctatcccatt acggtcaatc cgccgtttgt tcccacggag aatccgacgg gttgttactc
     3421 gctcacattt aatgttgatg aaagctggct acaggaaggc cagacgcgaa ttatttttga
     3481 tggcgttgga attagcttat cgactgcacg gtgcaccaat gcttctggcg tcaggcagcc
     3541 atcggaagct gtggtatggc tgtgcaggtc gtaaatcact gcataattcg tgtcgctcaa
     3601 ggcgcactcc cgttctggat aatgtttttt gcgccgacat cataacggtt ctggcaaata
     3661 ttctgaaatg agctgttgac aattaatcat cggctcgtat aatgtgtgga attgtgagcg
     3721 gataacaatt tcacacagga aacagtattc atgtccccta tactaggtta ttggaaaatt
     3781 aagggccttg tgcaacccac tcgacttctt ttggaatatc ttgaagaaaa atatgaagag
     3841 catttgtatg agcgcgatga aggtgataaa tggcgaaaca aaaagtttga attgggtttg
     3901 gagtttccca atcttcctta ttatattgat ggtgatgtta aattaacaca gtctatggcc
     3961 atcatacgtt atatagctga caagcacaac atgttgggtg gttgtccaaa agagcgtgca
     4021 gagatttcaa tgcttgaagg agcggttttg gatattagat acggtgtttc gagaattgca
     4081 tatagtaaag actttgaaac tctcaaagtt gattttctta gcaagctacc tgaaatgctg
     4141 aaaatgttcg aagatcgttt atgtcataaa acatatttaa atggtgatca tgtaacccat
     4201 cctgacttca tgttgtatga cgctcttgat gttgttttat acatggaccc aatgtgcctg
     4261 gatgcgttcc caaaattagt ttgttttaaa aaacgtattg aagctatccc acaaattgat
     4321 aagtacttga aatccagcaa gtatatagca tggcctttgc agggctggca agccacgttt
     4381 ggtggtggcg accatcctcc aaaatcggat ctggaagttc tgttccaggg gcccctggga
     4441 tccatgtgta aggactatgt atatgacaaa gacattgagc aaattgccaa agaagagcaa
     4501 ggagaagctt tgaaattaca agcctccacc tcaacagagg tttctcacca gcagtgttca
     4561 gtgccaggcc ttggtgagaa attcccaacc tgggaaacaa ccaaaccaga attagaactg
     4621 ctggggcaca acccgaggag aagaagaatc acctccagct ttacgatcgg tttaagagga
     4681 ctcatcaatc ttggcaacac gtgctttatg aactgcattg tccaggccct cacccacacg
     4741 ccgatactga gagatttctt tctctctgac aggcaccgat gtgagatgcc gagtcccgag
     4801 ttgtgtctgg tctgtgagat gtcgtcgctg tttcgggagt tgtattctgg aaacccgtct
     4861 cctcatgtgc cctgtaagtt actgcacctg gtgtggatac atgcccgcca tttagcaggg
     4921 tacaggcaac aggatgccca cgagttcctc attgcagcgt tagatgtcct gcacaggcac
     4981 tgcaaaggtg atgatgtcgg gaaggcggcc aacaatccca accactgtaa ctgcatcata
     5041 gaccaaatct tcacaggtgg cctgcagtct gatgtcacct gtcaagcctg ccatggcgtc
     5101 tccaccacga tagacccatg ctgggacatt agtttggact tgcctggctc ttgcacctcc
     5161 ttctggccca tgagcccagg gagggagagc agtgtgaacg gggaaagcca cataccagga
     5221 atcaccaccc tcacggactg cttgcggagg tttacgaggc cagagcactt aggaagcagt
     5281 gccaaaatca aatgtggtag ttgccaaagc taccaggaat ctaccaaaca gctcacaatg
     5341 aataaattac ctgtcgttgc ctgttttcat ttcaaacggt ttgaacattc agcgaaacag
     5401 aggcgcaaga tcactacata catttccttt cctctggagc tggatatgac gccgtttatg
     5461 gcctcaagta aagagagcag aatgaatgga caattgcagc tgccaaccaa tagtggaaac
     5521 aacgaaaata agtattcctt gtttgctgtg gttaatcacc aaggaacctt ggagagtggc
     5581 cactatacca gcttcatccg gcaccacaag gaccagtggt tcaagtgtga tgatgccgtc
     5641 atcactaagg ccagtattaa ggacgtactg gacagtgaag ggtatttact gttctatcac
     5701 aaacaggtgc tagaacatga gtcagaaaaa gtgaaagaaa tgaacacaca agcctactga
     5761 gcggccgcat cgtgactgac tgacgatctg cctcgcgcgt ttcggtgatg acggtgaaaa
     5821 cctctgacac atgcagctcc cggagacggt cacagcttgt ctgtaagcgg atgccgggag
     5881 cagacaagcc cgtcagggcg cgtcagcggg tgttggcggg tgtcggggcg cagccatgac
     5941 ccagtcacgt agcgatagcg gagtgtataa ttcttgaaga cgaaagggcc tcgtgatacg
     6001 cctattttta taggttaatg tcatgataat aatggtttct tagacgtcag gtggcacttt
     6061 tcggggaaat gtgcgcggaa cccctatttg tttatttttc taaatacatt caaatatgta
     6121 tccgctcatg agacaataac cctgataaat gcttcaataa tattgaaaaa ggaagagtat
     6181 gagtattcaa catttccgtg tcgcccttat tccctttttt gcggcatttt gccttcctgt
     6241 ttttgctcac ccagaaacgc tggtgaaagt aaaagatg
//
