LOCUS       sequence-445583-        6278 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Bacterial expression of USP27X (F313V) 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 F313V
FEATURES             Location/Qualifiers
     source          1..6278
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(106..125)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      919..1507
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1408..1427
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     1661..1678
                     /label=L4440
                     /note="L4440 vector, forward primer"
     promoter        1751..1828
                     /gene="lacI"
                     /label=lacI promoter
     CDS             1829..2911
                     /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(1848..1867)
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        join(2960..2965,2966..2983,2984..2990)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2998..3014
                     /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     3022..3038
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     3022..3038
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             3034..3561
                     /codon_start=1
                     /gene="lacZ fragment"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /label=lacZ-alpha
                     /translation="MTMITDSLAVVLQRRDWENPGVTQLNRLAAHPPFASWRNSEEART
                     DRPSQQLRSLNGEWRFAWFPAPEAVPESWLECDLPEADTVVVPSNWQMHGYDAPIYTNV
                     TYPITVNPPFVPTENPTGCYSLTFNVDESWLQEGQTRIIFDGVGISLSTARCTNASGVR
                     QPSEAVVWLCRS"
     primer_bind     complement(3054..3071)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(3054..3070)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3063..3085)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     promoter        join(3663..3668,3669..3684,3685..3691)
                     /label=tac promoter
                     /note="strong E. coli promoter; hybrid between the trp and 
                     lac UV5 promoters"
     protein_bind    3699..3715
                     /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             3738..4391
                     /codon_start=1
                     /product="glutathione S-transferase from Schistosoma 
                     japonicum"
                     /label=GST
                     /translation="MSPILGYWKIKGLVQPTRLLLEYLEEKYEEHLYERDEGDKWRNKK
                     FELGLEFPNLPYYIDGDVKLTQSMAIIRYIADKHNMLGGCPKERAEISMLEGAVLDIRY
                     GVSRIAYSKDFETLKVDFLSKLPEMLKMFEDRLCHKTYLNGDHVTHPDFMLYDALDVVL
                     YMDPMCLDAFPKLVCFKKRIEAIPQIDKYLKSSKYIAWPLQGWQATFGGGDHPPK"
     primer_bind     4349..4371
                     /label=pGEX 5'
                     /note="pGEX vectors, Glutathione-S-transferase, forward 
                     primer"
     CDS             4398..4421
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     primer_bind     complement(5808..5830)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(5975..5993)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        6061..6165
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6166..6234,6235..6278,1..748)
                     /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 gggtgcacga gtgggttaca tcgaactgga tctcaacagc ggtaagatcc ttgagagttt
       61 tcgccccgaa gaacgttttc caatgatgag cacttttaaa gttctgctat gtggcgcggt
      121 attatcccgt gttgacgccg ggcaagagca actcggtcgc cgcatacact attctcagaa
      181 tgacttggtt gagtactcac cagtcacaga aaagcatctt acggatggca tgacagtaag
      241 agaattatgc agtgctgcca taaccatgag tgataacact gcggccaact tacttctgac
      301 aacgatcgga ggaccgaagg agctaaccgc ttttttgcac aacatggggg atcatgtaac
      361 tcgccttgat cgttgggaac cggagctgaa tgaagccata ccaaacgacg agcgtgacac
      421 cacgatgcct gcagcaatgg caacaacgtt gcgcaaacta ttaactggcg aactacttac
      481 tctagcttcc cggcaacaat taatagactg gatggaggcg gataaagttg caggaccact
      541 tctgcgctcg gcccttccgg ctggctggtt tattgctgat aaatctggag ccggtgagcg
      601 tgggtctcgc ggtatcattg cagcactggg gccagatggt aagccctccc gtatcgtagt
      661 tatctacacg acggggagtc aggcaactat ggatgaacga aatagacaga tcgctgagat
      721 aggtgcctca ctgattaagc attggtaact gtcagaccaa gtttactcat atatacttta
      781 gattgattta aaacttcatt tttaatttaa aaggatctag gtgaagatcc tttttgataa
      841 tctcatgacc aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag accccgtaga
      901 aaagatcaaa ggatcttctt gagatccttt ttttctgcgc gtaatctgct gcttgcaaac
      961 aaaaaaacca ccgctaccag cggtggtttg tttgccggat caagagctac caactctttt
     1021 tccgaaggta actggcttca gcagagcgca gataccaaat actgtccttc tagtgtagcc
     1081 gtagttaggc caccacttca agaactctgt agcaccgcct acatacctcg ctctgctaat
     1141 cctgttacca gtggctgctg ccagtggcga taagtcgtgt cttaccgggt tggactcaag
     1201 acgatagtta ccggataagg cgcagcggtc gggctgaacg gggggttcgt gcacacagcc
     1261 cagcttggag cgaacgacct acaccgaact gagataccta cagcgtgagc tatgagaaag
     1321 cgccacgctt cccgaaggga gaaaggcgga caggtatccg gtaagcggca gggtcggaac
     1381 aggagagcgc acgagggagc ttccaggggg aaacgcctgg tatctttata gtcctgtcgg
     1441 gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct
     1501 atggaaaaac gccagcaacg cggccttttt acggttcctg gccttttgct ggccttttgc
     1561 tcacatgttc tttcctgcgt tatcccctga ttctgtggat aaccgtatta ccgcctttga
     1621 gtgagctgat accgctcgcc gcagccgaac gaccgagcgc agcgagtcag tgagcgagga
     1681 agcggaagag cgcctgatgc ggtattttct ccttacgcat ctgtgcggta tttcacaccg
     1741 cataaattcc gacaccatcg aatggcgcaa aacctttcgc ggtatggcat gatagcgccc
     1801 ggaagagagt caattcaggg tggtgaatgt gaaaccagta acgttatacg atgtcgcaga
     1861 gtatgccggt gtctcttatc agaccgtttc ccgcgtggtg aaccaggcca gccacgtttc
     1921 tgcgaaaacg cgggaaaaag tggaagcggc gatggcggag ctgaattaca ttcccaaccg
     1981 cgtggcacaa caactggcgg gcaaacagtc gttgctgatt ggcgttgcca cctccagtct
     2041 ggccctgcac gcgccgtcgc aaattgtcgc ggcgattaaa tctcgcgccg atcaactggg
     2101 tgccagcgtg gtggtgtcga tggtagaacg aagcggcgtc gaagcctgta aagcggcggt
     2161 gcacaatctt ctcgcgcaac gcgtcagtgg gctgatcatt aactatccgc tggatgacca
     2221 ggatgccatt gctgtggaag ctgcctgcac taatgttccg gcgttatttc ttgatgtctc
     2281 tgaccagaca cccatcaaca gtattatttt ctcccatgaa gacggtacgc gactgggcgt
     2341 ggagcatctg gtcgcattgg gtcaccagca aatcgcgctg ttagcgggcc cattaagttc
     2401 tgtctcggcg cgtctgcgtc tggctggctg gcataaatat ctcactcgca atcaaattca
     2461 gccgatagcg gaacgggaag gcgactggag tgccatgtcc ggttttcaac aaaccatgca
     2521 aatgctgaat gagggcatcg ttcccactgc gatgctggtt gccaacgatc agatggcgct
     2581 gggcgcaatg cgcgccatta ccgagtccgg gctgcgcgtt ggtgcggata tctcggtagt
     2641 gggatacgac gataccgaag acagctcatg ttatatcccg ccgttaacca ccatcaaaca
     2701 ggattttcgc ctgctggggc aaaccagcgt ggaccgcttg ctgcaactct ctcagggcca
     2761 ggcggtgaag ggcaatcagc tgttgcccgt ctcactggtg aaaagaaaaa ccacgctagc
     2821 gcccaatacg caaaccgcct ctccccgcgc gttggccgat tcattaatgc agctggcacg
     2881 acaggtttcc cgactggaaa gcgggcagtg agcgcaacgc aattaatgta agttagctca
     2941 ctcattaggc accccaggct ttacacttta tgcttccggc tcgtatgttg tgtggaattg
     3001 tgagcggata acaatttcac acaggaaaca gctatgacca tgattacgga ttcactggcc
     3061 gtcgttttac aacgtcgtga ctgggaaaac cctggcgtta cccaacttaa tcgccttgca
     3121 gcacatcccc ctttcgccag ctggcgtaat agcgaagagg cccgcaccga tcgcccttcc
     3181 caacagttgc gcagcctgaa tggcgaatgg cgctttgcct ggtttccggc accagaagcg
     3241 gtgccggaaa gctggctgga gtgcgatctt cctgaggccg atactgtcgt cgtcccctca
     3301 aactggcaga tgcacggtta cgatgcgccc atctacacca acgtgaccta tcccattacg
     3361 gtcaatccgc cgtttgttcc cacggagaat ccgacgggtt gttactcgct cacatttaat
     3421 gttgatgaaa gctggctaca ggaaggccag acgcgaatta tttttgatgg cgttggaatt
     3481 agcttatcga ctgcacggtg caccaatgct tctggcgtca ggcagccatc ggaagctgtg
     3541 gtatggctgt gcaggtcgta aatcactgca taattcgtgt cgctcaaggc gcactcccgt
     3601 tctggataat gttttttgcg ccgacatcat aacggttctg gcaaatattc tgaaatgagc
     3661 tgttgacaat taatcatcgg ctcgtataat gtgtggaatt gtgagcggat aacaatttca
     3721 cacaggaaac agtattcatg tcccctatac taggttattg gaaaattaag ggccttgtgc
     3781 aacccactcg acttcttttg gaatatcttg aagaaaaata tgaagagcat ttgtatgagc
     3841 gcgatgaagg tgataaatgg cgaaacaaaa agtttgaatt gggtttggag tttcccaatc
     3901 ttccttatta tattgatggt gatgttaaat taacacagtc tatggccatc atacgttata
     3961 tagctgacaa gcacaacatg ttgggtggtt gtccaaaaga gcgtgcagag atttcaatgc
     4021 ttgaaggagc ggttttggat attagatacg gtgtttcgag aattgcatat agtaaagact
     4081 ttgaaactct caaagttgat tttcttagca agctacctga aatgctgaaa atgttcgaag
     4141 atcgtttatg tcataaaaca tatttaaatg gtgatcatgt aacccatcct gacttcatgt
     4201 tgtatgacgc tcttgatgtt gttttataca tggacccaat gtgcctggat gcgttcccaa
     4261 aattagtttg ttttaaaaaa cgtattgaag ctatcccaca aattgataag tacttgaaat
     4321 ccagcaagta tatagcatgg cctttgcagg gctggcaagc cacgtttggt ggtggcgacc
     4381 atcctccaaa atcggatctg gaagttctgt tccaggggcc cctgggatcc atgtgtaagg
     4441 actatgtata tgacaaagac attgagcaaa ttgccaaaga agagcaagga gaagctttga
     4501 aattacaagc ctccacctca acagaggttt ctcaccagca gtgttcagtg ccaggccttg
     4561 gtgagaaatt cccaacctgg gaaacaacca aaccagaatt agaactgctg gggcacaacc
     4621 cgaggagaag aagaatcacc tccagcttta cgatcggttt aagaggactc atcaatcttg
     4681 gcaacacgtg ctttatgaac tgcattgtcc aggccctcac ccacacgccg atactgagag
     4741 atttctttct ctctgacagg caccgatgtg agatgccgag tcccgagttg tgtctggtct
     4801 gtgagatgtc gtcgctgttt cgggagttgt attctggaaa cccgtctcct catgtgccct
     4861 ataagttact gcacctggtg tggatacatg cccgccattt agcagggtac aggcaacagg
     4921 atgcccacga gttcctcatt gcagcgttag atgtcctgca caggcactgc aaaggtgatg
     4981 atgtcgggaa ggcggccaac aatcccaacc actgtaactg catcatagac caaatcttca
     5041 caggtggcct gcagtctgat gtcacctgtc aagcctgcca tggcgtctcc accacgatag
     5101 acccatgctg ggacattagt ttggacttgc ctggctcttg cacctccttc tggcccatga
     5161 gcccagggag ggagagcagt gtgaacgggg aaagccacat accaggaatc accaccctca
     5221 cggactgctt gcggaggttt acgaggccag agcacttagg aagcagtgcc aaaatcaaat
     5281 gtggtagttg ccaaagctac caggaatcta ccaaacagct cacaatgaat aaattacctg
     5341 tcgttgcctg ttttcatttc aaacgggttg aacattcagc gaaacagagg cgcaagatca
     5401 ctacatacat ttcctttcct ctggagctgg atatgacgcc gtttatggcc tcaagtaaag
     5461 agagcagaat gaatggacaa ttgcagctgc caaccaatag tggaaacaac gaaaataagt
     5521 attccttgtt tgctgtggtt aatcaccaag gaaccttgga gagtggccac tataccagct
     5581 tcatccggca ccacaaggac cagtggttca agtgtgatga tgccgtcatc actaaggcca
     5641 gtattaagga cgtactggac agtgaagggt atttactgtt ctatcacaaa caggtgctag
     5701 aacatgagtc agaaaaagtg aaagaaatga acacacaagc ctactgagcg gccgcatcgt
     5761 gactgactga cgatctgcct cgcgcgtttc ggtgatgacg gtgaaaacct ctgacacatg
     5821 cagctcccgg agacggtcac agcttgtctg taagcggatg ccgggagcag acaagcccgt
     5881 cagggcgcgt cagcgggtgt tggcgggtgt cggggcgcag ccatgaccca gtcacgtagc
     5941 gatagcggag tgtataattc ttgaagacga aagggcctcg tgatacgcct atttttatag
     6001 gttaatgtca tgataataat ggtttcttag acgtcaggtg gcacttttcg gggaaatgtg
     6061 cgcggaaccc ctatttgttt atttttctaa atacattcaa atatgtatcc gctcatgaga
     6121 caataaccct gataaatgct tcaataatat tgaaaaagga agagtatgag tattcaacat
     6181 ttccgtgtcg cccttattcc cttttttgcg gcattttgcc ttcctgtttt tgctcaccca
     6241 gaaacgctgg tgaaagtaaa agatgctgaa gatcagtt
//
