LOCUS       sequence-445599-        7861 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Transfection of USP27X (F313V) with HA tag in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7861)
  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 7861)
  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 F313V
FEATURES             Location/Qualifiers
     source          1..7861
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     439..458
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(677..781)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      808..1263
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(895..914)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1105..1126
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     1389..1411
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     1403..1420
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     1404..1420
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     CDS             complement(1756..2355)
                     /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(1839..1859)
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     primer_bind     2336..2355
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     promoter        complement(2371..2870)
                     /label=PGK promoter
                     /note="mouse phosphoglycerate kinase 1 promoter"
     primer_bind     complement(2419..2438)
                     /label=mPGK-F
                     /note="Mouse PGK promoter, forward primer"
     primer_bind     2812..2835
                     /label=MSCV-rev
                     /note="Murine stem cell virus, reverse primer"
     polyA_signal    2875..3009
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2899..2918)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2953..2972
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     CDS             complement(4692..4718)
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     primer_bind     complement(4756..4775)
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     intron          4783..5799
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and 
                     rabbit beta-globin"
     primer_bind     complement(4837..4859)
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     enhancer        6079..6458
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     primer_bind     complement(6467..6486)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(6468..6486)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(6516..6532)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6516..6532)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(6529..6551)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    6540..6556
                     /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)."
     promoter        complement(join(6564..6570,6571..6588,6589..6594))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6609..6630
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(6747..6764)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6918..7506)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6998..7017)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7677..7861,1..607,608..676))
                     /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"
ORIGIN
        1 cagccggaag ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc atccagtcta
       61 ttaattgttg ccgggaagct agagtaagta gttcgccagt taatagtttg cgcaacgttg
      121 ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct tcattcagct
      181 ccggttccca acgatcaagg cgagttacat gatcccccat gttgtgcaaa aaagcggtta
      241 gctccttcgg tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg
      301 ttatggcagc actgcataat tctcttactg tcatgccatc cgtaagatgc ttttctgtga
      361 ctggtgagta ctcaaccaag tcattctgag aatagtgtat gcggcgaccg agttgctctt
      421 gcccggcgtc aatacgggat aataccgcgc cacatagcag aactttaaaa gtgctcatca
      481 ttggaaaacg ttcttcgggg cgaaaactct caaggatctt accgctgttg agatccagtt
      541 cgatgtaacc cactcgtgca cccaactgat cttcagcatc ttttactttc accagcgttt
      601 ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga
      661 aatgttgaat actcatactc ttcctttttc aatattattg aagcatttat cagggttatt
      721 gtctcatgag cggatacata tttgaatgta tttagaaaaa taaacaaata ggggttccgc
      781 gcacatttcc ccgaaaagtg ccacctgacg cgccctgtag cggcgcatta agcgcggcgg
      841 gtgtggtggt tacgcgcagc gtgaccgcta cacttgccag cgccctagcg cccgctcctt
      901 tcgctttctt cccttccttt ctcgccacgt tcgccggctt tccccgtcaa gctctaaatc
      961 gggggctccc tttagggttc cgatttagtg ctttacggca cctcgacccc aaaaaacttg
     1021 attagggtga tggttcacgt agtgggccat cgccctgata gacggttttt cgccctttga
     1081 cgttggagtc cacgttcttt aatagtggac tcttgttcca aactggaaca acactcaacc
     1141 ctatctcggt ctattctttt gatttataag ggattttgcc gatttcggcc tattggttaa
     1201 aaaatgagct gatttaacaa aaatttaacg cgaattttaa caaaatatta acgcttacaa
     1261 tttccattcg ccattcaggc tgcgcaactg ttgggaaggg cgatcggtgc gggcctcttc
     1321 gctattacgc cagctggcga aagggggatg tgctgcaagg cgattaagtt gggtaacgcc
     1381 agggttttcc cagtcacgac gttgtaaaac gacggccagt gccaagctga cttggtcagc
     1441 ggccggccgc gtcgagcccc agctggttct ttccgcctca gaagccatag agcccaccgc
     1501 atccccagca tgcctgctat tgtcttccca atcctccccc ttgctgtcct gccccacccc
     1561 accccccaga atagaatgac acctactcag acaatgcgat gcaatttcct cattttatta
     1621 ggaaaggaca gtgggagtgg caccttccag ggtcaaggaa ggcacggggg aggggcaaac
     1681 aacagatggc tggcaaatcg atgcatgggg tcgtgcgctc ctttcggtcg ggcgctgcgg
     1741 gtcgtggggc gggcgtcagg caccgggctt gcgggtcatg caccaggtgc gcggtccttc
     1801 gggcacctcg acgtcggcgg tgacggtgaa gccgagccgc tcgtagaagg ggaggttgcg
     1861 gggcgcggag gtctccagga aggcgggcac cccggcgcgc tcggccgcct ccactccggg
     1921 gagcacgacg gcgctgccca gacccttgcc ctggtggtcg ggcgagacgc cgacggtggc
     1981 caggaaccac gcgggctcct tgggccggtg cggcgccagg aggccttcca tctgttgctg
     2041 cgcggccagc cgggaaccgc tcaactcggc catgcgcggg ccgatctcgg cgaacaccgc
     2101 ccccgcttcg acgctctccg gcgtggtcca gaccgccacc gcggcgccgt cgtccgcgac
     2161 ccacaccttg ccgatgtcga gcccgacgcg cgtgaggaag agttcttgca gctcggtgac
     2221 ccgctcgatg tggcggtccg gatcgacggt gtggcgcgtg gcggggtagt cggcgaacgc
     2281 ggcggcgagg gtgcgtacgg ccctggggac gtcgtcgcgg gtggcgaggc gcaccgtggg
     2341 cttgtactcg gtcatggtaa gctagcttgt cgaaaggccc ggagatgagg aagaggagaa
     2401 cagcgcggca gacgtgcgct tttgaagcgt gcagaatgcc gggcctccgg aggaccttcg
     2461 ggcgcccgcc ccgcccctga gcccgcccct gagcccgccc ccggacccac cccttcccag
     2521 cctctgagcc cagaaagcga aggagcaaag ctgctattgg ccgctgcccc aaaggcctac
     2581 ccgcttccat tgctcagcgg tgctgtccat ctgcacgaga ctagtgagac gtgctacttc
     2641 catttgtcac gtcctgcacg acgcgagctg cggggcgggg gggaacttcc tgactagggg
     2701 aggagtagaa ggtggcgcga aggggccacc aaagaacgga gccggttggc gcctaccggt
     2761 ggatgtggaa tgtgtgcgag gccagaggcc acttgtgtag cgccaagtgc ccagcggggc
     2821 tgctaaagcg catgctccag actgccttgg gaaaagcgcc tcccctaccc ggtagatcca
     2881 gacatgataa gatacattga tgagtttgga caaaccacaa ctagaatgca gtgaaaaaaa
     2941 tgctttattt gtgaaatttg tgatgctatt gctttatttg taaccattat aagctgcaat
     3001 aaacaagttc cgagtttgtc agaaagcaga ccaaacagcg gttggaataa tagcgagaac
     3061 agagaaatag cggcaaaaat aatacccgta tcacttttgc tgatatggtt gatgtcatgt
     3121 agccaaatcg ggaaaaacgg gaagtaggct cccatgataa aaaagtaaaa gaaaaagaat
     3181 aaaccgaaca tccaaaagtt tgtgtttttt aaatagtaca taatggattt ccttacgcga
     3241 aatacgggca gacatggcct gcccggttat tattattttt gacaccagac caactggtaa
     3301 tggtagcgac cggcgctcag ctggaattgg cggccgcatg catttgctag cccttaatta
     3361 aggctcgatc agtaggcttg tgtgttcatt tctttcactt tttctgactc atgttctagc
     3421 acctgtttgt gatagaacag taaataccct tcactgtcca gtacgtcctt aatactggcc
     3481 ttagtgatga cggcatcatc acacttgaac cactggtcct tgtggtgccg gatgaagctg
     3541 gtatagtggc cactctccaa ggttccttgg tgattaacca cagcaaacaa ggaatactta
     3601 ttttcgttgt ttccactatt ggttggcagc tgcaattgtc cattcattct gctctcttta
     3661 cttgaggcca taaacggcgt catatccagc tccagaggaa aggaaatgta tgtagtgatc
     3721 ttgcgcctct gtttcgctga atgttcaacc cgtttgaaat gaaaacaggc aacgacaggt
     3781 aatttattca ttgtgagctg tttggtagat tcctggtagc tttggcaact accacatttg
     3841 attttggcac tgcttcctaa gtgctctggc ctcgtaaacc tccgcaagca gtccgtgagg
     3901 gtggtgattc ctggtatgtg gctttccccg ttcacactgc tctccctccc tgggctcatg
     3961 ggccagaagg aggtgcaaga gccaggcaag tccaaactaa tgtcccagca tgggtctatc
     4021 gtggtggaga cgccatggca ggcttgacag gtgacatcag actgcaggcc acctgtgaag
     4081 atttggtcta tgatgcagtt acagtggttg ggattgttgg ccgccttccc gacatcatca
     4141 cctttgcagt gcctgtgcag gacatctaac gctgcaatga ggaactcgtg ggcatcctgt
     4201 tgcctgtacc ctgctaaatg gcgggcatgt atccacacca ggtgcagtaa cttatagggc
     4261 acatgaggag acgggtttcc agaatacaac tcccgaaaca gcgacgacat ctcacagacc
     4321 agacacaact cgggactcgg catctcacat cggtgcctgt cagagagaaa gaaatctctc
     4381 agtatcggcg tgtgggtgag ggcctggaca atgcagttca taaagcacgt gttgccaaga
     4441 ttgatgagtc ctcttaaacc gatcgtaaag ctggaggtga ttcttcttct cctcgggttg
     4501 tgccccagca gttctaattc tggtttggtt gtttcccagg ttgggaattt ctcaccaagg
     4561 cctggcactg aacactgctg gtgagaaacc tctgttgagg tggaggcttg taatttcaaa
     4621 gcttctcctt gctcttcttt ggcaatttgc tcaatgtctt tgtcatatac atagtcctta
     4681 cacatggatc cggcgtaatc tggcacatcg tatgggtaca ttcgagttga attctttgcc
     4741 aaaatgatga gacagcacaa taaccagcac gttgcccagg agctgtagga aaaagaagaa
     4801 ggcatgaaca tggttagcag aggctctaga gccgccggtc acacgccaga agccgaaccc
     4861 cgccctgccc cgtccccccc gaaggcagcc gtccccccgc ggacagcccc gaggctggag
     4921 agggagaagg ggacggcggc gcggcgacgc acgaaggccc tccccgccca tttccttcct
     4981 gccggcgccg caccgcttcg ccccgcgccc gctagagggg gtgcggcggc gcctcccaga
     5041 tttcggctcc gcacagattt gggacaaagg aagtccctgc gccctctcgc acgattacca
     5101 taaaaggcaa tggctgcggc tcgccgcgcc tcgacagccg ccggcgctcc gggggccgcc
     5161 gcgcccctcc cccgagccct ccccggcccg aggcggcccc gccccgcccg gcacccccac
     5221 ctgccgccac cccccgcccg gcacggcgag ccccgcgcca cgccccgtac ggagccccgc
     5281 acccgaagcc gggccgtgct cagcaactcg gggagggggg tgcagggggg gttgcagccc
     5341 gaccgacgcg cccacacccc ctgctcaccc ccccacgcac acaccccgca cgcagccttt
     5401 gttcccctcg cagccccccc cgcaccgcgg ggcaccgccc ccggccgcgc tcccctcgcg
     5461 cacactgcgg agcgcacaaa gccccgcgcc gcgcccgcag cgctcacagc cgccgggcag
     5521 cgcggagccg cacgcggcgc tccccacgca cacacacacg cacgcacccc ccgagccgct
     5581 ccccccgcac aaagggccct cccggagccc ctcaaggctt tcacgcagcc acagaaaaga
     5641 aacaagccgt cattaaacca agcgctaatt acagcccgga ggagaagggc cgtcccgccc
     5701 gctcacctgt gggagtaacg cggtcagtca gagccggggc gggcggcgcg aggcggcggc
     5761 ggagcggggc acggggcgaa ggcagcgcgc agcgactccc gcccgccgcg cgcttcgctt
     5821 tttatagggc cgccgccgcc gccgcctcgc cataaaagga aactttcgga gcgcgccgct
     5881 ctgattggct gccgccgcac ctctccgcct cgcccccccc cccccccccc cccccccccc
     5941 ccccccccgg cccccccccc cccccccccc cgcccccatc gctgcacaaa ataattaaaa
     6001 aataaataaa tacaaaattg ggggtgggga ggggggggag atggggagag tgaagcagaa
     6061 cgtggggctc acctcgacca tggtaatagc gatgactaat acgtagatgt actgccaagt
     6121 aggaaagtcc cataaggtca tgtactgggc ataatgccag gcgggccatt taccgtcatt
     6181 gacgtcaata gggggcgtac ttggcatatg atacacttga tgtactgcca agtgggcagt
     6241 ttaccgtaaa tactccaccc attgacgtca atggaaagtc cctattggcg ttactatggg
     6301 aacatacgtc attattgacg tcaatgggcg ggggtcgttg ggcggtcagc caggcgggcc
     6361 atttaccgta agttatgtaa cgcggaactc catatatggg ctatgaacta atgaccccgt
     6421 aattgattac tattaataac tagtcaataa tcaatgtcga cgagctccct atagtgagtc
     6481 gtattagagg ccgacttggc caaattcgta atcatgtcat agctgtttcc tgtgtgaaat
     6541 tgttatccgc tcacaattcc acacaacata cgagccggaa gcataaagtg taaagcctgg
     6601 ggtgcctaat gagtgagcta actcacatta attgcgttgc gctcactgcc cgctttccag
     6661 tcgggaaacc tgtcgtgcca gctgcattaa tgaatcggcc aacgcgcggg gagaggcggt
     6721 ttgcgtattg ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg
     6781 ctgcggcgag cggtatcagc tcactcaaag gcggtaatac ggttatccac agaatcaggg
     6841 gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag
     6901 gccgcgttgc tggcgttttt ccataggctc cgcccccctg acgagcatca caaaaatcga
     6961 cgctcaagtc agaggtggcg aaacccgaca ggactataaa gataccaggc gtttccccct
     7021 ggaagctccc tcgtgcgctc tcctgttccg accctgccgc ttaccggata cctgtccgcc
     7081 tttctccctt cgggaagcgt ggcgctttct catagctcac gctgtaggta tctcagttcg
     7141 gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac cccccgttca gcccgaccgc
     7201 tgcgccttat ccggtaacta tcgtcttgag tccaacccgg taagacacga cttatcgcca
     7261 ctggcagcag ccactggtaa caggattagc agagcgaggt atgtaggcgg tgctacagag
     7321 ttcttgaagt ggtggcctaa ctacggctac actagaagaa cagtatttgg tatctgcgct
     7381 ctgctgaagc cagttacctt cggaaaaaga gttggtagct cttgatccgg caaacaaacc
     7441 accgctggta gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga
     7501 tctcaagaag atcctttgat cttttctacg gggtctgacg ctcagtggaa cgaaaactca
     7561 cgttaaggga ttttggtcat gagattatca aaaaggatct tcacctagat ccttttaaat
     7621 taaaaatgaa gttttaaatc aatctaaagt atatatgagt aaacttggtc tgacagttac
     7681 caatgcttaa tcagtgaggc acctatctca gcgatctgtc tatttcgttc atccatagtt
     7741 gcctgactcc ccgtcgtgta gataactacg atacgggagg gcttaccatc tggccccagt
     7801 gctgcaatga taccgcgaga cccacgctca ccggctccag atttatcagc aataaaccag
     7861 c
//
