LOCUS       sequence-445597-        7878 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Transfection of USP27X (Y144C) with HA tag in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7878)
  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 7878)
  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: pCAGGS puro HA-hUSP27X Y144C
FEATURES             Location/Qualifiers
     source          1..7878
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     polyA_signal    111..245
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(148..167)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     202..221
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     promoter        250..749
                     /label=PGK promoter
                     /note="mouse phosphoglycerate kinase 1 promoter"
     primer_bind     complement(285..308)
                     /label=MSCV-rev
                     /note="Murine stem cell virus, reverse primer"
     primer_bind     682..701
                     /label=mPGK-F
                     /note="Mouse PGK promoter, forward primer"
     CDS             765..1364
                     /codon_start=1
                     /gene="pac from Streptomyces alboniger"
                     /product="puromycin N-acetyltransferase"
                     /label=PuroR
                     /note="confers resistance to puromycin"
                     /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
                     VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
                     AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
                     APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
     primer_bind     complement(765..784)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     1261..1281
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     primer_bind     complement(1700..1717)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(1700..1716)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1709..1731)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      complement(1857..2312)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1994..2015)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2206..2225
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        2339..2443
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2444..2512,2513..3304)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(2662..2681)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3475..4063
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3964..3983
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4217..4234
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    4351..4372
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(4387..4392,4393..4410,4411..4417)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4425..4441
                     /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     4430..4452
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     4449..4465
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     4449..4465
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     4495..4514
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        4495..4513
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     enhancer        4523..4902
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     intron          5199..6215
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and 
                     rabbit beta-globin"
     primer_bind     6139..6161
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     6223..6242
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     CDS             6280..6306
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
ORIGIN
        1 catgacatca accatatcag caaaagtgat acgggtatta tttttgccgc tatttctctg
       61 ttctcgctat tattccaacc gctgtttggt ctgctttctg acaaactcgg aacttgttta
      121 ttgcagctta taatggttac aaataaagca atagcatcac aaatttcaca aataaagcat
      181 ttttttcact gcattctagt tgtggtttgt ccaaactcat caatgtatct tatcatgtct
      241 ggatctaccg ggtaggggag gcgcttttcc caaggcagtc tggagcatgc gctttagcag
      301 ccccgctggg cacttggcgc tacacaagtg gcctctggcc tcgcacacat tccacatcca
      361 ccggtaggcg ccaaccggct ccgttctttg gtggcccctt cgcgccacct tctactcctc
      421 ccctagtcag gaagttcccc cccgccccgc agctcgcgtc gtgcaggacg tgacaaatgg
      481 aagtagcacg tctcactagt ctcgtgcaga tggacagcac cgctgagcaa tggaagcggg
      541 taggcctttg gggcagcggc caatagcagc tttgctcctt cgctttctgg gctcagaggc
      601 tgggaagggg tgggtccggg ggcgggctca ggggcgggct caggggcggg gcgggcgccc
      661 gaaggtcctc cggaggcccg gcattctgca cgcttcaaaa gcgcacgtct gccgcgctgt
      721 tctcctcttc ctcatctccg ggcctttcga caagctagct taccatgacc gagtacaagc
      781 ccacggtgcg cctcgccacc cgcgacgacg tccccagggc cgtacgcacc ctcgccgccg
      841 cgttcgccga ctaccccgcc acgcgccaca ccgtcgatcc ggaccgccac atcgagcggg
      901 tcaccgagct gcaagaactc ttcctcacgc gcgtcgggct cgacatcggc aaggtgtggg
      961 tcgcggacga cggcgccgcg gtggcggtct ggaccacgcc ggagagcgtc gaagcggggg
     1021 cggtgttcgc cgagatcggc ccgcgcatgg ccgagttgag cggttcccgg ctggccgcgc
     1081 agcaacagat ggaaggcctc ctggcgccgc accggcccaa ggagcccgcg tggttcctgg
     1141 ccaccgtcgg cgtctcgccc gaccaccagg gcaagggtct gggcagcgcc gtcgtgctcc
     1201 ccggagtgga ggcggccgag cgcgccgggg tgcccgcctt cctggagacc tccgcgcccc
     1261 gcaacctccc cttctacgag cggctcggct tcaccgtcac cgccgacgtc gaggtgcccg
     1321 aaggaccgcg cacctggtgc atgacccgca agcccggtgc ctgacgcccg ccccacgacc
     1381 cgcagcgccc gaccgaaagg agcgcacgac cccatgcatc gatttgccag ccatctgttg
     1441 tttgcccctc ccccgtgcct tccttgaccc tggaaggtgc cactcccact gtcctttcct
     1501 aataaaatga ggaaattgca tcgcattgtc tgagtaggtg tcattctatt ctggggggtg
     1561 gggtggggca ggacagcaag ggggaggatt gggaagacaa tagcaggcat gctggggatg
     1621 cggtgggctc tatggcttct gaggcggaaa gaaccagctg gggctcgacg cggccggccg
     1681 ctgaccaagt cagcttggca ctggccgtcg ttttacaacg tcgtgactgg gaaaaccctg
     1741 gcgttaccca acttaatcgc cttgcagcac atcccccttt cgccagctgg cgtaatagcg
     1801 aagaggcccg caccgatcgc ccttcccaac agttgcgcag cctgaatggc gaatggaaat
     1861 tgtaagcgtt aatattttgt taaaattcgc gttaaatttt tgttaaatca gctcattttt
     1921 taaccaatag gccgaaatcg gcaaaatccc ttataaatca aaagaataga ccgagatagg
     1981 gttgagtgtt gttccagttt ggaacaagag tccactatta aagaacgtgg actccaacgt
     2041 caaagggcga aaaaccgtct atcagggcga tggcccacta cgtgaaccat caccctaatc
     2101 aagttttttg gggtcgaggt gccgtaaagc actaaatcgg aaccctaaag ggagcccccg
     2161 atttagagct tgacggggaa agccggcgaa cgtggcgaga aaggaaggga agaaagcgaa
     2221 aggagcgggc gctagggcgc tggcaagtgt agcggtcacg ctgcgcgtaa ccaccacacc
     2281 cgccgcgctt aatgcgccgc tacagggcgc gtcaggtggc acttttcggg gaaatgtgcg
     2341 cggaacccct atttgtttat ttttctaaat acattcaaat atgtatccgc tcatgagaca
     2401 ataaccctga taaatgcttc aataatattg aaaaaggaag agtatgagta ttcaacattt
     2461 ccgtgtcgcc cttattccct tttttgcggc attttgcctt cctgtttttg ctcacccaga
     2521 aacgctggtg aaagtaaaag atgctgaaga tcagttgggt gcacgagtgg gttacatcga
     2581 actggatctc aacagcggta agatccttga gagttttcgc cccgaagaac gttttccaat
     2641 gatgagcact tttaaagttc tgctatgtgg cgcggtatta tcccgtattg acgccgggca
     2701 agagcaactc ggtcgccgca tacactattc tcagaatgac ttggttgagt actcaccagt
     2761 cacagaaaag catcttacgg atggcatgac agtaagagaa ttatgcagtg ctgccataac
     2821 catgagtgat aacactgcgg ccaacttact tctgacaacg atcggaggac cgaaggagct
     2881 aaccgctttt ttgcacaaca tgggggatca tgtaactcgc cttgatcgtt gggaaccgga
     2941 gctgaatgaa gccataccaa acgacgagcg tgacaccacg atgcctgtag caatggcaac
     3001 aacgttgcgc aaactattaa ctggcgaact acttactcta gcttcccggc aacaattaat
     3061 agactggatg gaggcggata aagttgcagg accacttctg cgctcggccc ttccggctgg
     3121 ctggtttatt gctgataaat ctggagccgg tgagcgtggg tctcgcggta tcattgcagc
     3181 actggggcca gatggtaagc cctcccgtat cgtagttatc tacacgacgg ggagtcaggc
     3241 aactatggat gaacgaaata gacagatcgc tgagataggt gcctcactga ttaagcattg
     3301 gtaactgtca gaccaagttt actcatatat actttagatt gatttaaaac ttcattttta
     3361 atttaaaagg atctaggtga agatcctttt tgataatctc atgaccaaaa tcccttaacg
     3421 tgagttttcg ttccactgag cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga
     3481 tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt
     3541 ggtttgtttg ccggatcaag agctaccaac tctttttccg aaggtaactg gcttcagcag
     3601 agcgcagata ccaaatactg ttcttctagt gtagccgtag ttaggccacc acttcaagaa
     3661 ctctgtagca ccgcctacat acctcgctct gctaatcctg ttaccagtgg ctgctgccag
     3721 tggcgataag tcgtgtctta ccgggttgga ctcaagacga tagttaccgg ataaggcgca
     3781 gcggtcgggc tgaacggggg gttcgtgcac acagcccagc ttggagcgaa cgacctacac
     3841 cgaactgaga tacctacagc gtgagctatg agaaagcgcc acgcttcccg aagggagaaa
     3901 ggcggacagg tatccggtaa gcggcagggt cggaacagga gagcgcacga gggagcttcc
     3961 agggggaaac gcctggtatc tttatagtcc tgtcgggttt cgccacctct gacttgagcg
     4021 tcgatttttg tgatgctcgt caggggggcg gagcctatgg aaaaacgcca gcaacgcggc
     4081 ctttttacgg ttcctggcct tttgctggcc ttttgctcac atgttctttc ctgcgttatc
     4141 ccctgattct gtggataacc gtattaccgc ctttgagtga gctgataccg ctcgccgcag
     4201 ccgaacgacc gagcgcagcg agtcagtgag cgaggaagcg gaagagcgcc caatacgcaa
     4261 accgcctctc cccgcgcgtt ggccgattca ttaatgcagc tggcacgaca ggtttcccga
     4321 ctggaaagcg ggcagtgagc gcaacgcaat taatgtgagt tagctcactc attaggcacc
     4381 ccaggcttta cactttatgc ttccggctcg tatgttgtgt ggaattgtga gcggataaca
     4441 atttcacaca ggaaacagct atgacatgat tacgaatttg gccaagtcgg cctctaatac
     4501 gactcactat agggagctcg tcgacattga ttattgacta gttattaata gtaatcaatt
     4561 acggggtcat tagttcatag cccatatatg gagttccgcg ttacataact tacggtaaat
     4621 ggcccgcctg gctgaccgcc caacgacccc cgcccattga cgtcaataat gacgtatgtt
     4681 cccatagtaa cgccaatagg gactttccat tgacgtcaat gggtggagta tttacggtaa
     4741 actgcccact tggcagtaca tcaagtgtat catatgccaa gtacgccccc tattgacgtc
     4801 aatgacggta aatggcccgc ctggcattat gcccagtaca tgaccttatg ggactttcct
     4861 acttggcagt acatctacgt attagtcatc gctattacca tggtcgaggt gagccccacg
     4921 ttctgcttca ctctccccat ctcccccccc tccccacccc caattttgta tttatttatt
     4981 ttttaattat tttgtgcagc gatgggggcg gggggggggg gggggggggg gggggggggg
     5041 gggggggggg gggggggggg gggggggggg gggggggggg cggggcgagg cggagaggtg
     5101 cggcggcagc caatcagagc ggcgcgctcc gaaagtttcc ttttatggcg aggcggcggc
     5161 ggcggcggcc ctataaaaag cgaagcgcgc ggcgggcggg agtcgctgcg cgctgccttc
     5221 gccccgtgcc ccgctccgcc gccgcctcgc gccgcccgcc ccggctctga ctgaccgcgt
     5281 tactcccaca ggtgagcggg cgggacggcc cttctcctcc gggctgtaat tagcgcttgg
     5341 tttaatgacg gcttgtttct tttctgtggc tgcgtgaaag ccttgagggg ctccgggagg
     5401 gccctttgtg cggggggagc ggctcggggg gtgcgtgcgt gtgtgtgtgc gtggggagcg
     5461 ccgcgtgcgg ctccgcgctg cccggcggct gtgagcgctg cgggcgcggc gcggggcttt
     5521 gtgcgctccg cagtgtgcgc gaggggagcg cggccggggg cggtgccccg cggtgcgggg
     5581 ggggctgcga ggggaacaaa ggctgcgtgc ggggtgtgtg cgtggggggg tgagcagggg
     5641 gtgtgggcgc gtcggtcggg ctgcaacccc ccctgcaccc ccctccccga gttgctgagc
     5701 acggcccggc ttcgggtgcg gggctccgta cggggcgtgg cgcggggctc gccgtgccgg
     5761 gcggggggtg gcggcaggtg ggggtgccgg gcggggcggg gccgcctcgg gccggggagg
     5821 gctcggggga ggggcgcggc ggcccccgga gcgccggcgg ctgtcgaggc gcggcgagcc
     5881 gcagccattg ccttttatgg taatcgtgcg agagggcgca gggacttcct ttgtcccaaa
     5941 tctgtgcgga gccgaaatct gggaggcgcc gccgcacccc ctctagcggg cgcggggcga
     6001 agcggtgcgg cgccggcagg aaggaaatgg gcggggaggg ccttcgtgcg tcgccgcgcc
     6061 gccgtcccct tctccctctc cagcctcggg gctgtccgcg gggggacggc tgccttcggg
     6121 ggggacgggg cagggcgggg ttcggcttct ggcgtgtgac cggcggctct agagcctctg
     6181 ctaaccatgt tcatgccttc ttctttttcc tacagctcct gggcaacgtg ctggttattg
     6241 tgctgtctca tcattttggc aaagaattca actcgaatgt acccatacga tgtgccagat
     6301 tacgccggat ccatgtgtaa ggactatgta tatgacaaag acattgagca aattgccaaa
     6361 gaagagcaag gagaagcttt gaaattacaa gcctccacct caacagaggt ttctcaccag
     6421 cagtgttcag tgccaggcct tggtgagaaa ttcccaacct gggaaacaac caaaccagaa
     6481 ttagaactgc tggggcacaa cccgaggaga agaagaatca cctccagctt tacgatcggt
     6541 ttaagaggac tcatcaatct tggcaacacg tgctttatga actgcattgt ccaggccctc
     6601 acccacacgc cgatactgag agatttcttt ctctctgaca ggcaccgatg tgagatgccg
     6661 agtcccgagt tgtgtctggt ctgtgagatg tcgtcgctgt ttcgggagtt gtattctgga
     6721 aacccgtctc ctcatgtgcc ctgtaagtta ctgcacctgg tgtggataca tgcccgccat
     6781 ttagcagggt acaggcaaca ggatgcccac gagttcctca ttgcagcgtt agatgtcctg
     6841 cacaggcact gcaaaggtga tgatgtcggg aaggcggcca acaatcccaa ccactgtaac
     6901 tgcatcatag accaaatctt cacaggtggc ctgcagtctg atgtcacctg tcaagcctgc
     6961 catggcgtct ccaccacgat agacccatgc tgggacatta gtttggactt gcctggctct
     7021 tgcacctcct tctggcccat gagcccaggg agggagagca gtgtgaacgg ggaaagccac
     7081 ataccaggaa tcaccaccct cacggactgc ttgcggaggt ttacgaggcc agagcactta
     7141 ggaagcagtg ccaaaatcaa atgtggtagt tgccaaagct accaggaatc taccaaacag
     7201 ctcacaatga ataaattacc tgtcgttgcc tgttttcatt tcaaacggtt tgaacattca
     7261 gcgaaacaga ggcgcaagat cactacatac atttcctttc ctctggagct ggatatgacg
     7321 ccgtttatgg cctcaagtaa agagagcaga atgaatggac aattgcagct gccaaccaat
     7381 agtggaaaca acgaaaataa gtattccttg tttgctgtgg ttaatcacca aggaaccttg
     7441 gagagtggcc actataccag cttcatccgg caccacaagg accagtggtt caagtgtgat
     7501 gatgccgtca tcactaaggc cagtattaag gacgtactgg acagtgaagg gtatttactg
     7561 ttctatcaca aacaggtgct agaacatgag tcagaaaaag tgaaagaaat gaacacacaa
     7621 gcctactgat cgagccttaa ttaagggcta gcaaatgcat gcggccgcca attccagctg
     7681 agcgccggtc gctaccatta ccagttggtc tggtgtcaaa aataataata accgggcagg
     7741 ccatgtctgc ccgtatttcg cgtaaggaaa tccattatgt actatttaaa aaacacaaac
     7801 ttttggatgt tcggtttatt ctttttcttt tactttttta tcatgggagc ctacttcccg
     7861 tttttcccga tttggcta
//
