LOCUS       sequence-445604-        8003 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Bacterial expression of USP27X with HA tag.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8003)
  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 8003)
  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
FEATURES             Location/Qualifiers
     source          1..8003
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      784..1372
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1273..1292
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     1526..1543
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    1660..1681
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(1696..1701,1702..1719,1720..1726)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1734..1750
                     /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     1739..1761
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     1758..1774
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     1758..1774
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     1804..1823
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1804..1822
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     enhancer        1832..2211
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     intron          2633..3649
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and 
                     rabbit beta-globin"
     primer_bind     3573..3595
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     3657..3676
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     CDS             3714..3740
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     polyA_signal    5423..5557
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5460..5479)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5514..5533
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     promoter        5562..6061
                     /label=PGK promoter
                     /note="mouse phosphoglycerate kinase 1 promoter"
     primer_bind     complement(5597..5620)
                     /label=MSCV-rev
                     /note="Murine stem cell virus, reverse primer"
     primer_bind     5994..6013
                     /label=mPGK-F
                     /note="Mouse PGK promoter, forward primer"
     CDS             6077..6676
                     /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(6077..6096)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     6573..6593
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     primer_bind     complement(7012..7029)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(7012..7028)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(7021..7043)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      complement(7169..7624)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(7306..7327)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     7518..7537
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        7651..7755
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(7756..7824,7825..8003,1..613)
                     /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(7974..7993)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
ORIGIN
        1 cgccgggcaa gagcaactcg gtcgccgcat acactattct cagaatgact tggttgagta
       61 ctcaccagtc acagaaaagc atcttacgga tggcatgaca gtaagagaat tatgcagtgc
      121 tgccataacc atgagtgata acactgcggc caacttactt ctgacaacga tcggaggacc
      181 gaaggagcta accgcttttt tgcacaacat gggggatcat gtaactcgcc ttgatcgttg
      241 ggaaccggag ctgaatgaag ccataccaaa cgacgagcgt gacaccacga tgcctgtagc
      301 aatggcaaca acgttgcgca aactattaac tggcgaacta cttactctag cttcccggca
      361 acaattaata gactggatgg aggcggataa agttgcagga ccacttctgc gctcggccct
      421 tccggctggc tggtttattg ctgataaatc tggagccggt gagcgtgggt ctcgcggtat
      481 cattgcagca ctggggccag atggtaagcc ctcccgtatc gtagttatct acacgacggg
      541 gagtcaggca actatggatg aacgaaatag acagatcgct gagataggtg cctcactgat
      601 taagcattgg taactgtcag accaagttta ctcatatata ctttagattg atttaaaact
      661 tcatttttaa tttaaaagga tctaggtgaa gatccttttt gataatctca tgaccaaaat
      721 cccttaacgt gagttttcgt tccactgagc gtcagacccc gtagaaaaga tcaaaggatc
      781 ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct
      841 accagcggtg gtttgtttgc cggatcaaga gctaccaact ctttttccga aggtaactgg
      901 cttcagcaga gcgcagatac caaatactgt tcttctagtg tagccgtagt taggccacca
      961 cttcaagaac tctgtagcac cgcctacata cctcgctctg ctaatcctgt taccagtggc
     1021 tgctgccagt ggcgataagt cgtgtcttac cgggttggac tcaagacgat agttaccgga
     1081 taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca cagcccagct tggagcgaac
     1141 gacctacacc gaactgagat acctacagcg tgagctatga gaaagcgcca cgcttcccga
     1201 agggagaaag gcggacaggt atccggtaag cggcagggtc ggaacaggag agcgcacgag
     1261 ggagcttcca gggggaaacg cctggtatct ttatagtcct gtcgggtttc gccacctctg
     1321 acttgagcgt cgatttttgt gatgctcgtc aggggggcgg agcctatgga aaaacgccag
     1381 caacgcggcc tttttacggt tcctggcctt ttgctggcct tttgctcaca tgttctttcc
     1441 tgcgttatcc cctgattctg tggataaccg tattaccgcc tttgagtgag ctgataccgc
     1501 tcgccgcagc cgaacgaccg agcgcagcga gtcagtgagc gaggaagcgg aagagcgccc
     1561 aatacgcaaa ccgcctctcc ccgcgcgttg gccgattcat taatgcagct ggcacgacag
     1621 gtttcccgac tggaaagcgg gcagtgagcg caacgcaatt aatgtgagtt agctcactca
     1681 ttaggcaccc caggctttac actttatgct tccggctcgt atgttgtgtg gaattgtgag
     1741 cggataacaa tttcacacag gaaacagcta tgacatgatt acgaatttgg ccaagtcggc
     1801 ctctaatacg actcactata gggagctcgt cgacattgat tattgactag ttattaatag
     1861 taatcaatta cggggtcatt agttcatagc ccatatatgg agttccgcgt tacataactt
     1921 acggtaaatg gcccgcctgg ctgaccgccc aacgaccccc gcccattgac gtcaataatg
     1981 acgtatgttc ccatagtaac gccaataggg actttccatt gacgtcaatg ggtggagtat
     2041 ttacggtaaa ctgcccactt ggcagtacat caagtgtatc atatgccaag tacgccccct
     2101 attgacgtca atgacggtaa atggcccgcc tggcattatg cccagtacat gaccttatgg
     2161 gactttccta cttggcagta catctacgta ttagtcatcg ctattaccat ggtcgaggtg
     2221 agccccacgt tctgcttcac tctccccatc tcccccccct ccccaccccc aattttgtat
     2281 ttatttattt tttaattatt ttgtgcagcg atgggggcgg gggggggggg gggggcgggc
     2341 ccgggggggg gggggggggg gggggggggg gcgggggggg gggggggggg gggggggggc
     2401 cccataacag gcgggcgccc cccactatct tctttttttc ggggggcggg cggggggggg
     2461 gggggggggg ggggggggcg gccggggggg gggggggggg gggggggggg gggggggggc
     2521 ggggcggaga ggtgcggcgg cagccaatca gagcggcgcg ctccgaaagt ttccttttat
     2581 ggcggggcgg cggcggcggc ggccctataa aaagcgaagc gcgcggcggg cgggagtcgc
     2641 tgcgcgctgc cttcgccccg tgccccgctc cgccgccgcc tcgcgccgcc cgccccggct
     2701 ctgactgacc gcgttactcc cacaggtgag cgggcgggac ggcccttctc ctccgggctg
     2761 taattagcgc ttggtttaat gacggcttgt ttcttttctg tggctgcgtg aaagccttga
     2821 ggggctccgg gagggccctt tgtgcggggg gagcggctcg gggggtgcgt gcgtgtgtgt
     2881 gtgcgtgggg agcgccgcgt gcggctccgc gctgcccggc ggctgtgagc gctgcgggcg
     2941 cggcgcgggg ctttgtgcgc tccgcagtgt gcgcgagggg agcgcggccg ggggcggtgc
     3001 cccgcggtgc ggggggggct gcgaggggaa caaaggctgc gtgcggggtg tgtgcgtggg
     3061 ggggtgagca gggggtgtgg gcgcgtcggt cgggctgcaa ccccccctgc acccccctcc
     3121 ccgagttgct gagcacggcc cggcttcggg tgcggggctc cgtacggggc gtggcgcggg
     3181 gctcgccgtg ccgggcgggg ggtggcggca ggtgggggtg ccgggcgggg cggggccgcc
     3241 tcgggccggg gagggctcgg gggaggggcg cggcggcccc cggagcgccg gcggctgtcg
     3301 aggcgcggcg agccgcagcc attgcctttt atggtaatcg tgcgagaggg cgcagggact
     3361 tcctttgtcc caaatctgtg cggagccgaa atctgggagg cgccgccgca ccccctctag
     3421 cgggcgcggg gcgaagcggt gcggcgccgg caggaaggaa atgggcgggg agggccttcg
     3481 tgcgtcgccg cgccgccgtc cccttctccc tctccagcct cggggctgtc cgcgggggga
     3541 cggctgcctt cgggggggac ggggcagggc ggggttcggc ttctggcgtg tgaccggcgg
     3601 ctctagagcc tctgctaacc atgttcatgc cttcttcttt ttcctacagc tcctgggcaa
     3661 cgtgctggtt attgtgctgt ctcatcattt tggcaaagaa ttcaactcga atgtacccat
     3721 acgatgtgcc agattacgcc ggatccatgt gtaaggacta tgtatatgac aaagacattg
     3781 agcaaattgc caaagaagag caaggagaag ctttgaaatt acaagcctcc acctcaacag
     3841 aggtttctca ccagcagtgt tcagtgccag gccttggtga gaaattccca acctgggaaa
     3901 caaccaaacc agaattagaa ctgctggggc acaacccgag gagaagaaga atcacctcca
     3961 gctttacgat cggtttaaga ggactcatca atcttggcaa cacgtgcttt atgaactgca
     4021 ttgtccaggc cctcacccac acgccgatac tgagagattt ctttctctct gacaggcacc
     4081 gatgtgagat gccgagtccc gagttgtgtc tggtctgtga gatgtcgtcg ctgtttcggg
     4141 agttgtattc tggaaacccg tctcctcatg tgccctataa gttactgcac ctggtgtgga
     4201 tacatgcccg ccatttagca gggtacaggc aacaggatgc ccacgagttc ctcattgcag
     4261 cgttagatgt cctgcacagg cactgcaaag gtgatgatgt cgggaaggcg gccaacaatc
     4321 ccaaccactg taactgcatc atagaccaaa tcttcacagg tggcctgcag tctgatgtca
     4381 cctgtcaagc ctgccatggc gtctccacca cgatagaccc atgctgggac attagtttgg
     4441 acttgcctgg ctcttgcacc tccttctggc ccatgagccc agggagggag agcagtgtga
     4501 acggggaaag ccacatacca ggaatcacca ccctcacgga ctgcttgcgg aggtttacga
     4561 ggccagagca cttaggaagc agtgccaaaa tcaaatgtgg tagttgccaa agctaccagg
     4621 aatctaccaa acagctcaca atgaataaat tacctgtcgt tgcctgtttt catttcaaac
     4681 ggtttgaaca ttcagcgaaa cagaggcgca agatcactac atacatttcc tttcctctgg
     4741 agctggatat gacgccgttt atggcctcaa gtaaagagag cagaatgaat ggacaattgc
     4801 agctgccaac caatagtgga aacaacgaaa ataagtattc cttgtttgct gtggttaatc
     4861 accaaggaac cttggagagt ggccactata ccagcttcat ccggcaccac aaggaccagt
     4921 ggttcaagtg tgatgatgcc gtcatcacta aggccagtat taaggacgta ctggacagtg
     4981 aagggtattt actgttctat cacaaacagg tgctagaaca tgagtcagaa aaagtgaaag
     5041 aaatgaacac acaagcctac tgatcgagcc ttaattaagg gctagcaaat gcatgcggcc
     5101 gccaattcca gctgagcgcc ggtcgctacc attaccagtt ggtctggtgt caaaaataat
     5161 aataaccggg caggccatgt ctgcccgtat ttcgcgtaag gaaatccatt atgtactatt
     5221 taaaaaacac aaacttttgg atgttcggtt tattcttttt cttttacttt tttatcatgg
     5281 gagcctactt cccgtttttc ccgatttggc tacatgacat caaccatatc agcaaaagtg
     5341 atacgggtat tatttttgcc gctatttctc tgttctcgct attattccaa ccgctgtttg
     5401 gtctgctttc tgacaaactc ggaacttgtt tattgcagct tataatggtt acaaataaag
     5461 caatagcatc acaaatttca caaataaagc atttttttca ctgcattcta gttgtggttt
     5521 gtccaaactc atcaatgtat cttatcatgt ctggatctac cgggtagggg aggcgctttt
     5581 cccaaggcag tctggagcat gcgctttagc agccccgctg ggcacttggc gctacacaag
     5641 tggcctctgg cctcgcacac attccacatc caccggtagg cgccaaccgg ctccgttctt
     5701 tggtggcccc ttcgcgccac cttctactcc tcccctagtc aggaagttcc cccccgcccc
     5761 gcagctcgcg tcgtgcagga cgtgacaaat ggaagtagca cgtctcacta gtctcgtgca
     5821 gatggacagc accgctgagc aatggaagcg ggtaggcctt tggggcagcg gccaatagca
     5881 gctttgctcc ttcgctttct gggctcagag gctgggaagg ggtgggtccg ggggcgggct
     5941 caggggcggg ctcaggggcg gggcgggcgc ccgaaggtcc tccggaggcc cggcattctg
     6001 cacgcttcaa aagcgcacgt ctgccgcgct gttctcctct tcctcatctc cgggcctttc
     6061 gacaagctag cttaccatga ccgagtacaa gcccacggtg cgcctcgcca cccgcgacga
     6121 cgtccccagg gccgtacgca ccctcgccgc cgcgttcgcc gactaccccg ccacgcgcca
     6181 caccgtcgat ccggaccgcc acatcgagcg ggtcaccgag ctgcaagaac tcttcctcac
     6241 gcgcgtcggg ctcgacatcg gcaaggtgtg ggtcgcggac gacggcgccg cggtggcggt
     6301 ctggaccacg ccggagagcg tcgaagcggg ggcggtgttc gccgagatcg gcccgcgcat
     6361 ggccgagttg agcggttccc ggctggccgc gcagcaacag atggaaggcc tcctggcgcc
     6421 gcaccggccc aaggagcccg cgtggttcct ggccaccgtc ggcgtctcgc ccgaccacca
     6481 gggcaagggt ctgggcagcg ccgtcgtgct ccccggagtg gaggcggccg agcgcgccgg
     6541 ggtgcccgcc ttcctggaga cctccgcgcc ccgcaacctc cccttctacg agcggctcgg
     6601 cttcaccgtc accgccgacg tcgaggtgcc cgaaggaccg cgcacctggt gcatgacccg
     6661 caagcccggt gcctgacgcc cgccccacga cccgcagcgc ccgaccgaaa ggagcgcacg
     6721 accccatgca tcgatttgcc agccatctgt tgtttgcccc tcccccgtgc cttccttgac
     6781 cctggaaggt gccactccca ctgtcctttc ctaataaaat gaggaaattg catcgcattg
     6841 tctgagtagg tgtcattcta ttctgggggg tggggtgggg caggacagca agggggagga
     6901 ttgggaagac aatagcaggc atgctgggga tgcggtgggc tctatggctt ctgaggcgga
     6961 aagaaccagc tggggctcga cgcggccggc cgctgaccaa gtcagcttgg cactggccgt
     7021 cgttttacaa cgtcgtgact gggaaaaccc tggcgttacc caacttaatc gccttgcagc
     7081 acatccccct ttcgccagct ggcgtaatag cgaagaggcc cgcaccgatc gcccttccca
     7141 acagttgcgc agcctgaatg gcgaatggaa attgtaagcg ttaatatttt gttaaaattc
     7201 gcgttaaatt tttgttaaat cagctcattt tttaaccaat aggccgaaat cggcaaaatc
     7261 ccttataaat caaaagaata gaccgagata gggttgagtg ttgttccagt ttggaacaag
     7321 agtccactat taaagaacgt ggactccaac gtcaaagggc gaaaaaccgt ctatcagggc
     7381 gatggcccac tacgtgaacc atcaccctaa tcaagttttt tggggtcgag gtgccgtaaa
     7441 gcactaaatc ggaaccctaa agggagcccc cgatttagag cttgacgggg aaagccggcg
     7501 aacgtggcga gaaaggaagg gaagaaagcg aaaggagcgg gcgctagggc gctggcaagt
     7561 gtagcggtca cgctgcgcgt aaccaccaca cccgccgcgc ttaatgcgcc gctacagggc
     7621 gcgtcaggtg gcacttttcg gggaaatgtg cgcggaaccc ctatttgttt atttttctaa
     7681 atacattcaa atatgtatcc gctcatgaga caataaccct gataaatgct tcaataatat
     7741 tgaaaaagga agagtatgag tattcaacat ttccgtgtcg cccttattcc cttttttgcg
     7801 gcattttgcc ttcctgtttt tgctcaccca gaaacgctgg tgaaagtaaa agatgctgaa
     7861 gatcagttgg gtgcacgagt gggttacatc gaactggatc tcaacagcgg taagatcctt
     7921 gagagttttc gccccgaaga acgttttcca atgatgagca cttttaaagt tctgctatgt
     7981 ggcgcggtat tatcccgtat tga
//
