LOCUS       sequence-92067-e        8086 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Plasmid for constitutive or doxycycline-inducible expression of 
            G641E mutant of human NAT10 resistant to a siRNA. Confers resistance
            to puromycin. Use T-REx cells for doxycycline-inducible expression..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8086)
  AUTHORS   Larrieu D, Britton S, Demir M, Rodriguez R, Jackson SP
  TITLE     Chemical inhibition of NAT10 corrects defects of laminopathic cells.
  JOURNAL   Science. 2014 May 2;344(6183):527-32. doi: 10.1126/science.1252651.
  PUBMED    24786082
REFERENCE   2  (bases 1 to 8086)
  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"; doi: "10.1126/science"; 
            journalName: "Science"; date: "2014-05-2- 2"; volume: "344"; issue: 
            "6183"; pages: "527-32"
COMMENT     Sequence Label: pICE-FLAG-NAT10-siR-G641E
FEATURES             Location/Qualifiers
     source          1..8086
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     regulatory      16..25
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             25..48
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     regulatory      694..703
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     primer_bind     complement(3167..3184)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3173..3397
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      3443..3871
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3530..3549)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3740..3761
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3880..3900)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3885..4214
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      4065..4200
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     4127..4146
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             4266..4865
                     /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(4266..4285)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     4762..4782
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     polyA_signal    4995..5116
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5032..5051)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5086..5105
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5166..5182)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5166..5182)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5179..5201)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5190..5206
                     /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(5214..5220,5221..5238,5239..5244))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5259..5280
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5397..5414)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5568..6153)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5648..5667)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6324..7115,7116..7184))
                     /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     6947..6966
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7185..7289)
                     /gene="bla"
                     /label=AmpR promoter
     enhancer        7361..7740
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        7741..7944
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     7895..7915
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     protein_bind    7946..7964
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    7967..7985
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     primer_bind     7995..8019
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
ORIGIN
        1 aagcttgcgg ccgccgccac catggattac aaggatgacg acgataaggg atccatgcat
       61 cggaaaaagg tggataaccg aatccggatt ctcattgaga atggagtagc tgagcggcaa
      121 agatctctct ttgttgtagt tggggatcga ggaaaagatc aggtggtaat acttcatcac
      181 atgttatcca aagcaactgt gaaggctcgg ccttcagtgc tgtggtgtta taagaaagag
      241 ctggggttta gcagtcaccg gaagaaaaga atgcgacagc tgcagaagaa aataaagaat
      301 ggaacactga acataaagca ggacgacccc tttgaactct tcatagcagc cacaaacatt
      361 cgctactgct actacaacga gacccacaag atcctgggca ataccttcgg catgtgtgtg
      421 ctgcaggatt ttgaagcctt aactccaaac ttgctggcca ggactgtaga aacagtggaa
      481 ggtggtgggc tagtggtcat cctcctacgg accatgaact cactcaagca attgtacaca
      541 gtgactatgg atgtgcattc caggtacaga actgaggccc atcaggatgt ggtgggaaga
      601 tttaatgaaa ggtttattct gtctctggcc tcttgtaaga agtgtctcgt cattgatgac
      661 cagctcaaca tcctgcccat ctcctcccac gttgccacca tggaggccct gcctccccag
      721 actccggatg agagtcttgg tccttctgat ctggagctga gggagttgaa ggagagcttg
      781 caggacaccc agcctgtggg tgtgttggtg gactgctgta agactctaga ccaggccaaa
      841 gctgtcttga aatttatcga gggcatctct gaaaagaccc tgaggagtac tgttgcactc
      901 acagctgctc gaggacgggg aaaatctgca gccctgggat tggcgattgc tggggcggtg
      961 gcatttgggt actccaatat ctttgttacc tccccaagcc ctgataacct ccatactctg
     1021 tttgaatttg tatttaaagg atttgatgct ctgcaatatc aggaacatct ggattatgag
     1081 attatccagt ctctaaatcc tgaatttaac aaagcagtga tcagagtgaa tgtatttcga
     1141 gaacacaggc agactattca gtatatacat cctgcagatg ctgtgaagct gggccaggct
     1201 gaactagttg tgattgatga agctgccgcc atccccctcc ccttggtgaa gagcctactt
     1261 ggcccctacc ttgttttcat ggcatccacc atcaatggct atgagggcac tggccggtca
     1321 ctgtccctca agctaattca gcagctccgt caacagagcg cccagagcca ggtcagcacc
     1381 actgctgaga ataagaccac gacgacagcc agattggcat cagcgcggac actgcatgag
     1441 gtttccctcc aggagtcaat ccgatacgcc cctggggatg cagtggagaa gtggctgaat
     1501 gacttgctgt gcctggattg cctcaacatc actcggatag tctcaggctg ccccttgcct
     1561 gaagcttgtg aactgtacta tgttaataga gataccctct tttgctacca caaggcctct
     1621 gaagttttcc tccaacggct tatggccctc tacgtggctt ctcactacaa gaactctccc
     1681 aatgatctcc agatgctctc cgatgcacct gctcaccatc tcttctgcct tctgcctcct
     1741 gtgcccccca cccagaatgc ccttccagaa gtgcttgctg ttatccaggt gtgccttgaa
     1801 ggggagattt ctcgccagtc catcttgaac agtctgtctc gaggcaagaa ggcttcaggg
     1861 gacctgattc catggacagt gtcagaacag ttccaagatc cagactttgg tggtctgtct
     1921 ggtggaaggg tcgttcgcat tgctgttcac ccagattatc aagggatggg ctatgagagc
     1981 cgtgctctgc agctgctgca gatgtactat gaaggcaggt ttccttgtct ggaggaaaag
     2041 gtccttgaga caccacagga aattcacacc gtaagcagcg aggctgtcag cttgttggaa
     2101 gaggtcatca ctccccggaa ggacctgcct cctttactcc tcaaattgaa tgagaggcct
     2161 gccgaacgcc tggattacct gggtgtttcc tatggcttga cccccaggct cctcaagttc
     2221 tggaaacgag ctggatttgt tcctgtttat ctgagacaga ccccgaatga cctgaccgga
     2281 gagcactcgt gcatcatgct gaagacgctc actgatgagg atgaggctga ccagggaggc
     2341 tggcttgcag ccttctggaa agatttccga cggcggttcc tagccttgct ctcctaccag
     2401 ttcagtacct tctctccttc cctggctctg aacatcattc agaacaggaa catggggaag
     2461 ccagcccagc ctgccctgag ccgggaggag ctggaagcac tcttcctccc ctatgacctg
     2521 aagcggctgg agatgtattc acggaatatg gtggactatc acctcatcat ggacatgatc
     2581 ccggccatct ctcgcatcta tttcctgaac cagctggggg acctggccct gtctgcggct
     2641 cagtcggctc ttctcttggg gattggcctg cagcataagt ctgtggacca gctggaaaag
     2701 gagattgagc tgccctcggg ccagttgatg ggacttttca accggatcat ccgcaaagtt
     2761 gtgaagctat ttaatgaagt tcaggaaaag gccattgagg agcagatggt ggcagcgaag
     2821 gatgtggtca tggagcccac gatgaagacc ctcagtgacg acctagatga agcagcaaag
     2881 gaatttcagg agaaacacaa gaaggaagta gggaagctga aatcaatgga tttgagtgaa
     2941 tatattatcc gtggggacga tgaagagtgg aatgaagttt tgaacaaagc tgggccgaac
     3001 gcctcgatca tcagcctgaa aagtgacaag aaaaggaagt tagaggccaa acaagaaccc
     3061 aaacagagca agaagttgaa gaacagagag acaaagaaca aaaaagatat gaaactgaag
     3121 cggaagaaat agacgcgtct gcagtctaga taaacccgct gatcagcctc gactgtgcct
     3181 tctagttgcc agccatctgt tgtttgcccc tcccccgtgc cttccttgac cctggaaggt
     3241 gccactccca ctgtcctttc ctaataaaat gaggaaattg catcgcattg tctgagtagg
     3301 tgtcattcta ttctgggggg tggggtgggg caggacagca agggggagga ttgggaagac
     3361 aatagcaggc atgctgggga tgcggtgggc tctatggctt ctgaggcgga aagaaccagc
     3421 tggggctcta gggggtatcc ccacgcgccc tgtagcggcg cattaagcgc ggcgggtgtg
     3481 gtggttacgc gcagcgtgac cgctacactt gccagcgccc tagcgcccgc tcctttcgct
     3541 ttcttccctt cctttctcgc cacgttcgcc ggctttcccc gtcaagctct aaatcggggg
     3601 ctccctttag ggttccgatt tagtgcttta cggcacctcg accccaaaaa acttgattag
     3661 ggtgatggtt cacgtagtgg gccatcgccc tgatagacgg tttttcgccc tttgacgttg
     3721 gagtccacgt tctttaatag tggactcttg ttccaaactg gaacaacact caaccctatc
     3781 tcggtctatt cttttgattt ataagggatt ttgccgattt cggcctattg gttaaaaaat
     3841 gagctgattt aacaaaaatt taacgcgaat taattctgtg gaatgtgtgt cagttagggt
     3901 gtggaaagtc cccaggctcc ccagcaggca gaagtatgca aagcatgcat ctcaattagt
     3961 cagcaaccag gtgtggaaag tccccaggct ccccagcagg cagaagtatg caaagcatgc
     4021 atctcaatta gtcagcaacc atagtcccgc ccctaactcc gcccatcccg cccctaactc
     4081 cgcccagttc cgcccattct ccgccccatg gctgactaat tttttttatt tatgcagagg
     4141 ccgaggccgc ctctgcctct gagctattcc agaagtagtg aggaggcttt tttggaggcc
     4201 taggcttttg caaaaagctc ccgggagctt gtatatccat tttcggatct gatcagcacg
     4261 tgaccatgac cgagtacaag cccacggtgc gcctcgccac ccgcgacgac gtccccaggg
     4321 ccgtacgcac cctcgccgcc gcgttcgccg actaccccgc cacgcgccac accgtcgatc
     4381 cggaccgcca catcgagcgg gtcaccgagc tgcaagaact cttcctcacg cgcgtcgggc
     4441 tcgacatcgg caaggtgtgg gtcgcggacg acggcgccgc ggtggcggtc tggaccacgc
     4501 cggagagcgt cgaagcgggg gcggtgttcg ccgagatcgg cccgcgcatg gccgagttga
     4561 gcggttcccg gctggccgcg cagcaacaga tggaaggcct cctggcgccg caccggccca
     4621 aggagcccgc gtggttcctg gccaccgtcg gcgtctcgcc cgaccaccag ggcaagggtc
     4681 tgggcagcgc cgtcgtgctc cccggagtgg aggcggccga gcgcgccggg gtgcccgcct
     4741 tcctggagac ctccgcgccc cgcaacctcc ccttctacga gcggctcggc ttcaccgtca
     4801 ccgccgacgt cgaggtgccc gaaggaccgc gcacctggtg catgacccgc aagcccggtg
     4861 cctgacacgt gctacgagat ttcgattcca ccgccgcctt ctatgaaagg ttgggcttcg
     4921 gaatcgtttt ccgggacgcc ggctggatga tcctccagcg cggggatctc atgctggagt
     4981 tcttcgccca ccccaacttg tttattgcag cttataatgg ttacaaataa agcaatagca
     5041 tcacaaattt cacaaataaa gcattttttt cactgcattc tagttgtggt ttgtccaaac
     5101 tcatcaatgt atcttatcat gtctggctag ccgtcgacct ctagctagag cttggcgtaa
     5161 tcatggtcat agctgtttcc tgtgtgaaat tgttatccgc tcacaattcc acacaacata
     5221 cgagccggaa gcataaagtg taaagcctgg ggtgcctaat gagtgagcta actcacatta
     5281 attgcgttgc gctcactgcc cgctttccag tcgggaaacc tgtcgtgcca gctgcattaa
     5341 tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg ggcgctcttc cgcttcctcg
     5401 ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag
     5461 gcggtaatac ggttatccac agaatcaggg gataacgcag gaaagaacat gtgagcaaaa
     5521 ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc
     5581 cgcccccctg acgagcatca caaaaatcga cgctcaagtc agaggtggcg aaacccgaca
     5641 ggactataaa gataccaggc gtttccccct ggaagctccc tcgtgcgctc tcctgttccg
     5701 accctgccgc ttaccggata cctgtccgcc tttctccctt cgggaagcgt ggcgctttct
     5761 catagctcac gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt
     5821 gtgcacgaac cccccgttca gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag
     5881 tccaacccgg taagacacga cttatcgcca ctggcagcag ccactggtaa caggattagc
     5941 agagcgaggt atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa ctacggctac
     6001 actagaagaa cagtatttgg tatctgcgct ctgctgaagc cagttacctt cggaaaaaga
     6061 gttggtagct cttgatccgg caaacaaacc accgctggta gcggtttttt tgtttgcaag
     6121 cagcagatta cgcgcagaaa aaaaggatct caagaagatc ctttgatctt ttctacgggg
     6181 tctgacgctc agtggaacga aaactcacgt taagggattt tggtcatgag attatcaaaa
     6241 aggatcttca cctagatcct tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata
     6301 tatgagtaaa cttggtctga cagttaccaa tgcttaatca gtgaggcacc tatctcagcg
     6361 atctgtctat ttcgttcatc catagttgcc tgactccccg tcgtgtagat aactacgata
     6421 cgggagggct taccatctgg ccccagtgct gcaatgatac cgcgagaccc acgctcaccg
     6481 gctccagatt tatcagcaat aaaccagcca gccggaaggg ccgagcgcag aagtggtcct
     6541 gcaactttat ccgcctccat ccagtctatt aattgttgcc gggaagctag agtaagtagt
     6601 tcgccagtta atagtttgcg caacgttgtt gccattgcta caggcatcgt ggtgtcacgc
     6661 tcgtcgtttg gtatggcttc attcagctcc ggttcccaac gatcaaggcg agttacatga
     6721 tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt tgtcagaagt
     6781 aagttggccg cagtgttatc actcatggtt atggcagcac tgcataattc tcttactgtc
     6841 atgccatccg taagatgctt ttctgtgact ggtgagtact caaccaagtc attctgagaa
     6901 tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca
     6961 catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg aaaactctca
     7021 aggatcttac cgctgttgag atccagttcg atgtaaccca ctcgtgcacc caactgatct
     7081 tcagcatctt ttactttcac cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc
     7141 gcaaaaaagg gaataagggc gacacggaaa tgttgaatac tcatactctt cctttttcaa
     7201 tattattgaa gcatttatca gggttattgt ctcatgagcg gatacatatt tgaatgtatt
     7261 tagaaaaata aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc acctgacgtc
     7321 gacggatcgg gagatcctgt acgggccaga tatccgcgtt gacattgatt attgactagt
     7381 tattaatagt aatcaattac ggggtcatta gttcatagcc catatatgga gttccgcgtt
     7441 acataactta cggtaaatgg cccgcctggc tgaccgccca acgacccccg cccattgacg
     7501 tcaataatga cgtatgttcc catagtaacg ccaataggga ctttccattg acgtcaatgg
     7561 gtggagtatt tacggtaaac tgcccacttg gcagtacatc aagtgtatca tatgccaagt
     7621 acgcccccta ttgacgtcaa tgacggtaaa tggcccgcct ggcattatgc ccagtacatg
     7681 accttatggg actttcctac ttggcagtac atctacgtat tagtcatcgc tattaccatg
     7741 gtgatgcggt tttggcagta catcaatggg cgtggatagc ggtttgactc acggggattt
     7801 ccaagtctcc accccattga cgtcaatggg agtttgtttt ggcaccaaaa tcaacgggac
     7861 tttccaaaat gtcgtaacaa ctccgcccca ttgacgcaaa tgggcggtag gcgtgtacgg
     7921 tgggaggtct atataagcag agctctccct atcagtgata gagatctccc tatcagtgat
     7981 agagatcgtc gacgagctcg tttagtgaac cgtcagatcg cctggagacg ccatccacgc
     8041 tgttttgacc tccatagaag acaccgggac cgatccagcc tccggc
//
