LOCUS       sequence-445601-        8008 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Transfection of USP27X (S404N) with HA tag in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8008)
  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 8008)
  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 S404N
FEATURES             Location/Qualifiers
     source          1..8008
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      775..1363
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1264..1283
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     1517..1534
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    1651..1672
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(1687..1692,1693..1710,1711..1717)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1725..1741
                     /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     1730..1752
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     1749..1765
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     1749..1765
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     1795..1814
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1795..1813
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     enhancer        1823..2202
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     intron          2629..3645
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and 
                     rabbit beta-globin"
     primer_bind     3569..3591
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     3653..3672
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     CDS             3710..3736
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     polyA_signal    5419..5553
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5456..5475)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5510..5529
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     promoter        5558..6057
                     /label=PGK promoter
                     /note="mouse phosphoglycerate kinase 1 promoter"
     primer_bind     complement(5593..5616)
                     /label=MSCV-rev
                     /note="Murine stem cell virus, reverse primer"
     primer_bind     5990..6009
                     /label=mPGK-F
                     /note="Mouse PGK promoter, forward primer"
     CDS             6073..6672
                     /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(6073..6092)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     6569..6589
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     primer_bind     complement(7008..7025)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(7008..7024)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(7017..7039)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      complement(7165..7620)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(7302..7323)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     7514..7533
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        7647..7751
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(7752..7820,7821..8008,1..604)
                     /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(7970..7989)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
ORIGIN
        1 agagcaactc ggtcgccgca tacactattc tcagaatgac ttggttgagt actcaccagt
       61 cacagaaaag catcttacgg atggcatgac agtaagagaa ttatgcagtg ctgccataac
      121 catgagtgat aacactgcgg ccaacttact tctgacaacg atcggaggac cgaaggagct
      181 aaccgctttt ttgcacaaca tgggggatca tgtaactcgc cttgatcgtt gggaaccgga
      241 gctgaatgaa gccataccaa acgacgagcg tgacaccacg atgcctgtag caatggcaac
      301 aacgttgcgc aaactattaa ctggcgaact acttactcta gcttcccggc aacaattaat
      361 agactggatg gaggcggata aagttgcagg accacttctg cgctcggccc ttccggctgg
      421 ctggtttatt gctgataaat ctggagccgg tgagcgtggg tctcgcggta tcattgcagc
      481 actggggcca gatggtaagc cctcccgtat cgtagttatc tacacgacgg ggagtcaggc
      541 aactatggat gaacgaaata gacagatcgc tgagataggt gcctcactga ttaagcattg
      601 gtaactgtca gaccaagttt actcatatat actttagatt gatttaaaac ttcattttta
      661 atttaaaagg atctaggtga agatcctttt tgataatctc atgaccaaaa tcccttaacg
      721 tgagttttcg ttccactgag cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga
      781 tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt
      841 ggtttgtttg ccggatcaag agctaccaac tctttttccg aaggtaactg gcttcagcag
      901 agcgcagata ccaaatactg ttcttctagt gtagccgtag ttaggccacc acttcaagaa
      961 ctctgtagca ccgcctacat acctcgctct gctaatcctg ttaccagtgg ctgctgccag
     1021 tggcgataag tcgtgtctta ccgggttgga ctcaagacga tagttaccgg ataaggcgca
     1081 gcggtcgggc tgaacggggg gttcgtgcac acagcccagc ttggagcgaa cgacctacac
     1141 cgaactgaga tacctacagc gtgagctatg agaaagcgcc acgcttcccg aagggagaaa
     1201 ggcggacagg tatccggtaa gcggcagggt cggaacagga gagcgcacga gggagcttcc
     1261 agggggaaac gcctggtatc tttatagtcc tgtcgggttt cgccacctct gacttgagcg
     1321 tcgatttttg tgatgctcgt caggggggcg gagcctatgg aaaaacgcca gcaacgcggc
     1381 ctttttacgg ttcctggcct tttgctggcc ttttgctcac atgttctttc ctgcgttatc
     1441 ccctgattct gtggataacc gtattaccgc ctttgagtga gctgataccg ctcgccgcag
     1501 ccgaacgacc gagcgcagcg agtcagtgag cgaggaagcg gaagagcgcc caatacgcaa
     1561 accgcctctc cccgcgcgtt ggccgattca ttaatgcagc tggcacgaca ggtttcccga
     1621 ctggaaagcg ggcagtgagc gcaacgcaat taatgtgagt tagctcactc attaggcacc
     1681 ccaggcttta cactttatgc ttccggctcg tatgttgtgt ggaattgtga gcggataaca
     1741 atttcacaca ggaaacagct atgacatgat tacgaatttg gccaagtcgg cctctaatac
     1801 gactcactat agggagctcg tcgacattga ttattgacta gttattaata gtaatcaatt
     1861 acggggtcat tagttcatag cccatatatg gagttccgcg ttacataact tacggtaaat
     1921 ggcccgcctg gctgaccgcc caacgacccc cgcccattga cgtcaataat gacgtatgtt
     1981 cccatagtaa cgccaatagg gactttccat tgacgtcaat gggtggagta tttacggtaa
     2041 actgcccact tggcagtaca tcaagtgtat catatgccaa gtacgccccc tattgacgtc
     2101 aatgacggta aatggcccgc ctggcattat gcccagtaca tgaccttatg ggactttcct
     2161 acttggcagt acatctacgt attagtcatc gctattacca tggtcgaggt gagccccacg
     2221 ttctgcttca ctctccccat ctcccccccc tccccacccc caattttgta tttatttatt
     2281 ttttaattat tttgtgcagc gatgggggcg gggggggggg ggggggggcg ccgggggggg
     2341 gggggggggg gggggggggg gggggggggg gggggggggg gggggggggg ggaaaaagaa
     2401 ggggggggcc ccggaagttt tttttttttg gggggggggg gggggggggg cgcgcaaaaa
     2461 aaaaaggggg gggggggggg gggcggcggg gggggggggg gggggcgggg cggggcgagg
     2521 cggagaggtg cggcggcagc caatcagagc ggcgcgctcc gaaagtttcc ttttatggcg
     2581 aggcggcggc ggcggcggcc ctataaaaag cgaagcgcgc ggcgggcggg agtcgctgcg
     2641 cgctgccttc gccccgtgcc ccgctccgcc gccgcctcgc gccgcccgcc ccggctctga
     2701 ctgaccgcgt tactcccaca ggtgagcggg cgggacggcc cttctcctcc gggctgtaat
     2761 tagcgcttgg tttaatgacg gcttgtttct tttctgtggc tgcgtgaaag ccttgagggg
     2821 ctccgggagg gccctttgtg cggggggagc ggctcggggg gtgcgtgcgt gtgtgtgtgc
     2881 gtggggagcg ccgcgtgcgg ctccgcgctg cccggcggct gtgagcgctg cgggcgcggc
     2941 gcggggcttt gtgcgctccg cagtgtgcgc gaggggagcg cggccggggg cggtgccccg
     3001 cggtgcgggg ggggctgcga ggggaacaaa ggctgcgtgc ggggtgtgtg cgtggggggg
     3061 tgagcagggg gtgtgggcgc gtcggtcggg ctgcaacccc ccctgcaccc ccctccccga
     3121 gttgctgagc acggcccggc ttcgggtgcg gggctccgta cggggcgtgg cgcggggctc
     3181 gccgtgccgg gcggggggtg gcggcaggtg ggggtgccgg gcggggcggg gccgcctcgg
     3241 gccggggagg gctcggggga ggggcgcggc ggcccccgga gcgccggcgg ctgtcgaggc
     3301 gcggcgagcc gcagccattg ccttttatgg taatcgtgcg agagggcgca gggacttcct
     3361 ttgtcccaaa tctgtgcgga gccgaaatct gggaggcgcc gccgcacccc ctctagcggg
     3421 cgcggggcga agcggtgcgg cgccggcagg aaggaaatgg gcggggaggg ccttcgtgcg
     3481 tcgccgcgcc gccgtcccct tctccctctc cagcctcggg gctgtccgcg gggggacggc
     3541 tgccttcggg ggggacgggg cagggcgggg ttcggcttct ggcgtgtgac cggcggctct
     3601 agagcctctg ctaaccatgt tcatgccttc ttctttttcc tacagctcct gggcaacgtg
     3661 ctggttattg tgctgtctca tcattttggc aaagaattca actcgaatgt acccatacga
     3721 tgtgccagat tacgccggat ccatgtgtaa ggactatgta tatgacaaag acattgagca
     3781 aattgccaaa gaagagcaag gagaagcttt gaaattacaa gcctccacct caacagaggt
     3841 ttctcaccag cagtgttcag tgccaggcct tggtgagaaa ttcccaacct gggaaacaac
     3901 caaaccagaa ttagaactgc tggggcacaa cccgaggaga agaagaatca cctccagctt
     3961 tacgatcggt ttaagaggac tcatcaatct tggcaacacg tgctttatga actgcattgt
     4021 ccaggccctc acccacacgc cgatactgag agatttcttt ctctctgaca ggcaccgatg
     4081 tgagatgccg agtcccgagt tgtgtctggt ctgtgagatg tcgtcgctgt ttcgggagtt
     4141 gtattctgga aacccgtctc ctcatgtgcc ctataagtta ctgcacctgg tgtggataca
     4201 tgcccgccat ttagcagggt acaggcaaca ggatgcccac gagttcctca ttgcagcgtt
     4261 agatgtcctg cacaggcact gcaaaggtga tgatgtcggg aaggcggcca acaatcccaa
     4321 ccactgtaac tgcatcatag accaaatctt cacaggtggc ctgcagtctg atgtcacctg
     4381 tcaagcctgc catggcgtct ccaccacgat agacccatgc tgggacatta gtttggactt
     4441 gcctggctct tgcacctcct tctggcccat gagcccaggg agggagagca gtgtgaacgg
     4501 ggaaagccac ataccaggaa tcaccaccct cacggactgc ttgcggaggt ttacgaggcc
     4561 agagcactta ggaagcagtg ccaaaatcaa atgtggtagt tgccaaagct accaggaatc
     4621 taccaaacag ctcacaatga ataaattacc tgtcgttgcc tgttttcatt tcaaacggtt
     4681 tgaacattca gcgaaacaga ggcgcaagat cactacatac atttcctttc ctctggagct
     4741 ggatatgacg ccgtttatgg cctcaagtaa agagagcaga atgaatggac aattgcagct
     4801 gccaaccaat agtggaaaca acgaaaataa gtattccttg tttgctgtgg ttaatcacca
     4861 aggaaccttg gagagtggcc actataccag cttcatccgg caccacaagg accagtggtt
     4921 caagtgtgat gatgccgtca tcactaaggc caatattaag gacgtactgg acagtgaagg
     4981 gtatttactg ttctatcaca aacaggtgct agaacatgag tcagaaaaag tgaaagaaat
     5041 gaacacacaa gcctactgat cgagccttaa ttaagggcta gcaaatgcat gcggccgcca
     5101 attccagctg agcgccggtc gctaccatta ccagttggtc tggtgtcaaa aataataata
     5161 accgggcagg ccatgtctgc ccgtatttcg cgtaaggaaa tccattatgt actatttaaa
     5221 aaacacaaac ttttggatgt tcggtttatt ctttttcttt tactttttta tcatgggagc
     5281 ctacttcccg tttttcccga tttggctaca tgacatcaac catatcagca aaagtgatac
     5341 gggtattatt tttgccgcta tttctctgtt ctcgctatta ttccaaccgc tgtttggtct
     5401 gctttctgac aaactcggaa cttgtttatt gcagcttata atggttacaa ataaagcaat
     5461 agcatcacaa atttcacaaa taaagcattt ttttcactgc attctagttg tggtttgtcc
     5521 aaactcatca atgtatctta tcatgtctgg atctaccggg taggggaggc gcttttccca
     5581 aggcagtctg gagcatgcgc tttagcagcc ccgctgggca cttggcgcta cacaagtggc
     5641 ctctggcctc gcacacattc cacatccacc ggtaggcgcc aaccggctcc gttctttggt
     5701 ggccccttcg cgccaccttc tactcctccc ctagtcagga agttcccccc cgccccgcag
     5761 ctcgcgtcgt gcaggacgtg acaaatggaa gtagcacgtc tcactagtct cgtgcagatg
     5821 gacagcaccg ctgagcaatg gaagcgggta ggcctttggg gcagcggcca atagcagctt
     5881 tgctccttcg ctttctgggc tcagaggctg ggaaggggtg ggtccggggg cgggctcagg
     5941 ggcgggctca ggggcggggc gggcgcccga aggtcctccg gaggcccggc attctgcacg
     6001 cttcaaaagc gcacgtctgc cgcgctgttc tcctcttcct catctccggg cctttcgaca
     6061 agctagctta ccatgaccga gtacaagccc acggtgcgcc tcgccacccg cgacgacgtc
     6121 cccagggccg tacgcaccct cgccgccgcg ttcgccgact accccgccac gcgccacacc
     6181 gtcgatccgg accgccacat cgagcgggtc accgagctgc aagaactctt cctcacgcgc
     6241 gtcgggctcg acatcggcaa ggtgtgggtc gcggacgacg gcgccgcggt ggcggtctgg
     6301 accacgccgg agagcgtcga agcgggggcg gtgttcgccg agatcggccc gcgcatggcc
     6361 gagttgagcg gttcccggct ggccgcgcag caacagatgg aaggcctcct ggcgccgcac
     6421 cggcccaagg agcccgcgtg gttcctggcc accgtcggcg tctcgcccga ccaccagggc
     6481 aagggtctgg gcagcgccgt cgtgctcccc ggagtggagg cggccgagcg cgccggggtg
     6541 cccgccttcc tggagacctc cgcgccccgc aacctcccct tctacgagcg gctcggcttc
     6601 accgtcaccg ccgacgtcga ggtgcccgaa ggaccgcgca cctggtgcat gacccgcaag
     6661 cccggtgcct gacgcccgcc ccacgacccg cagcgcccga ccgaaaggag cgcacgaccc
     6721 catgcatcga tttgccagcc atctgttgtt tgcccctccc ccgtgccttc cttgaccctg
     6781 gaaggtgcca ctcccactgt cctttcctaa taaaatgagg aaattgcatc gcattgtctg
     6841 agtaggtgtc attctattct ggggggtggg gtggggcagg acagcaaggg ggaggattgg
     6901 gaagacaata gcaggcatgc tggggatgcg gtgggctcta tggcttctga ggcggaaaga
     6961 accagctggg gctcgacgcg gccggccgct gaccaagtca gcttggcact ggccgtcgtt
     7021 ttacaacgtc gtgactggga aaaccctggc gttacccaac ttaatcgcct tgcagcacat
     7081 ccccctttcg ccagctggcg taatagcgaa gaggcccgca ccgatcgccc ttcccaacag
     7141 ttgcgcagcc tgaatggcga atggaaattg taagcgttaa tattttgtta aaattcgcgt
     7201 taaatttttg ttaaatcagc tcatttttta accaataggc cgaaatcggc aaaatccctt
     7261 ataaatcaaa agaatagacc gagatagggt tgagtgttgt tccagtttgg aacaagagtc
     7321 cactattaaa gaacgtggac tccaacgtca aagggcgaaa aaccgtctat cagggcgatg
     7381 gcccactacg tgaaccatca ccctaatcaa gttttttggg gtcgaggtgc cgtaaagcac
     7441 taaatcggaa ccctaaaggg agcccccgat ttagagcttg acggggaaag ccggcgaacg
     7501 tggcgagaaa ggaagggaag aaagcgaaag gagcgggcgc tagggcgctg gcaagtgtag
     7561 cggtcacgct gcgcgtaacc accacacccg ccgcgcttaa tgcgccgcta cagggcgcgt
     7621 caggtggcac ttttcgggga aatgtgcgcg gaacccctat ttgtttattt ttctaaatac
     7681 attcaaatat gtatccgctc atgagacaat aaccctgata aatgcttcaa taatattgaa
     7741 aaaggaagag tatgagtatt caacatttcc gtgtcgccct tattcccttt tttgcggcat
     7801 tttgccttcc tgtttttgct cacccagaaa cgctggtgaa agtaaaagat gctgaagatc
     7861 agttgggtgc acgagtgggt tacatcgaac tggatctcaa cagcggtaag atccttgaga
     7921 gttttcgccc cgaagaacgt tttccaatga tgagcacttt taaagttctg ctatgtggcg
     7981 cggtattatc ccgtattgac gccgggca
//
