LOCUS       sequence-150753-        7519 bp    DNA     circular SYN 14-APR-2025
DEFINITION  bacterial expression of wild type human PRMT9.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7519)
  AUTHORS   Hadjikyriacou A, Yang Y, Espejo A, Bedford MT, Clarke SG
  TITLE     Unique Features of Human Protein Arginine Methyltransferase 9 
            (PRMT9) and its Substrate RNA Splicing Factor SF3B2.
  JOURNAL   J Biol Chem. 2015 May 15. pii: jbc.M115.659433.
  PUBMED    25979344
REFERENCE   2  (bases 1 to 7519)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 14, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "J Biol 
            Chem. 2015 May 15. pii: jbc.M115.659433."
COMMENT     Sequence Label: GST-PRMT9 WT
FEATURES             Location/Qualifiers
     source          1..7519
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        join(183..188,189..204,205..211)
                     /label=tac promoter
                     /note="strong E. coli promoter; hybrid between the trp and 
                     lac UV5 promoters"
     protein_bind    219..235
                     /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)."
     CDS             258..911
                     /codon_start=1
                     /product="glutathione S-transferase from Schistosoma 
                     japonicum"
                     /label=GST
                     /translation="MSPILGYWKIKGLVQPTRLLLEYLEEKYEEHLYERDEGDKWRNKK
                     FELGLEFPNLPYYIDGDVKLTQSMAIIRYIADKHNMLGGCPKERAEISMLEGAVLDIRY
                     GVSRIAYSKDFETLKVDFLSKLPEMLKMFEDRLCHKTYLNGDHVTHPDFMLYDALDVVL
                     YMDPMCLDAFPKLVCFKKRIEAIPQIDKYLKSSKYIAWPLQGWQATFGGGDHPPK"
     primer_bind     869..891
                     /label=pGEX 5'
                     /note="pGEX vectors, Glutathione-S-transferase, forward 
                     primer"
     CDS             918..941
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     primer_bind     complement(3569..3591)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(3736..3754)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3822..3926
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3927..3995,3996..4787)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(4145..4164)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4958..5546
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5447..5466
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5700..5717
                     /label=L4440
                     /note="L4440 vector, forward primer"
     promoter        5790..5867
                     /gene="lacI (mutant)"
                     /label=lacIq promoter
                     /note="In the lacIq allele, a single base change in the 
                     promoter boosts expression of the lacI gene about 10-fold."
     CDS             5868..6950
                     /codon_start=1
                     /gene="lacI"
                     /product="lac repressor"
                     /label=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)."
                     /translation="MKPVTLYDVAEYAGVSYQTVSRVVNQASHVSAKTREKVEAAMAEL
                     NYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSRADQLGASVVVSMVERSGV
                     EACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSIIFSH
                     EDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREGDWSA
                     MSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTEDSSC
                     YIPPSTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR
                     ALADSLMQLARQVSRLESGQ"
     primer_bind     complement(5887..5906)
                     /label=LacI-R
                     /note="LacI, reverse primer"
     protein_bind    6963..6984
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(6999..7004,7005..7022,7023..7029)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7037..7053
                     /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     7061..7077
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     7061..7077
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             join(7073..7519,1..81)
                     /codon_start=1
                     /gene="lacZ fragment"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /label=lacZ-alpha
                     /translation="MTMITDSLAVVLQRRDWENPGVTQLNRLAAHPPFASWRNSEEART
                     DRPSQQLRSLNGEWRFAWFPAPEAVPESWLECDLPEADTVVVPSNWQMHGYDAPIYTNV
                     TYPITVNPPFVPTENPTGCYSLTFNVDESWLQEGQTRIIFDGVGITLSTARCTNASGVR
                     QPSEAVVWLCRS"
     primer_bind     complement(7093..7110)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(7093..7109)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(7102..7124)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
ORIGIN
        1 acgttatcga ctgcacggtg caccaatgct tctggcgtca ggcagccatc ggaagctgtg
       61 gtatggctgt gcaggtcgta aatcactgca taattcgtgt cgctcaaggc gcactcccgt
      121 tctggataat gttttttgcg ccgacatcat aacggttctg gcaaatattc tgaaatgagc
      181 tgttgacaat taatcatcgg ctcgtataat gtgtggaatt gtgagcggat aacaatttca
      241 cacaggaaac agtattcatg tcccctatac taggttattg gaaaattaag ggccttgtgc
      301 aacccactcg acttcttttg gaatatcttg aagaaaaata tgaagagcat ttgtatgagc
      361 gcgatgaagg tgataaatgg cgaaacaaaa agtttgaatt gggtttggag tttcccaatc
      421 ttccttatta tattgatggt gatgttaaat taacacagtc tatggccatc atacgttata
      481 tagctgacaa gcacaacatg ttgggtggtt gtccaaaaga gcgtgcagag atttcaatgc
      541 ttgaaggagc ggttttggat attagatacg gtgtttcgag aattgcatat agtaaagact
      601 ttgaaactct caaagttgat tttcttagca agctacctga aatgctgaaa atgttcgaag
      661 atcgtttatg tcataaaaca tatttaaatg gtgatcatgt aacccatcct gacttcatgt
      721 tgtatgacgc tcttgatgtt gttttataca tggacccaat gtgcctggat gcgttcccaa
      781 aattagtttg ttttaaaaaa cgtattgaag ctatcccaca aattgataag tacttgaaat
      841 ccagcaagta tatagcatgg cctttgcagg gctggcaagc cacgtttggt ggtggcgacc
      901 atcctccaaa atcggatctg gaagttctgt tccaggggcc cctgggatcc atgtcgaact
      961 cgcggcccag gtcccgccga gacgccgggg gtggcgctgg ggcagccggc cgggacgagc
     1021 tggtgtcgcg gtccttgcag agcgcagagc actgtctggg cgtccaggac ttcggcactg
     1081 cctatgccca ctacctcctc gtgctcagcc tggcgccgga gctgaaacac gacgtgaagg
     1141 aaacttttca gtacacactt ttcagatggg ctgaagagct tgatgctctc agtcggatac
     1201 aagacttact tggttgctat gagcaggcct tggaactgtt tcctgatgat gaagtgattt
     1261 gcaatagtat gggggagcat ctcttcagaa tgggctttag ggatgaagca gctgggtatt
     1321 ttcataaagc agtgaagcta aaccctgatt tcagtgatgc aaaggagaat ttttatcgtg
     1381 ttgcaaactg gttggtggag cgctggcact ttatcatgct taatgacacc aagaggaata
     1441 caatttataa tgcagcaatc caaaaggcag tttgtttggg gtccaaaagt gttttggaca
     1501 ttggagcagg aactggaata ctaagcatgt ttgctaaaaa agctggagca cattccgtgt
     1561 atgcctgtga gttatccaag accatgtatg aacttgcctg tgatgtcgtg gcagcaaaca
     1621 agatggaagc agggatcaaa ctcttacata cgaagtcact tgacatagag attccaaaac
     1681 atattcccga aagagtgtcc ctagttgtaa cagaaactgt cgatgcaggt ttatttggag
     1741 aaggaattgt ggagagtttg attcatgcat gggagcattt acttttacag ccaaagacca
     1801 aaggtgaaag tgctaattgt gaaaagtatg ggaaagttat accagcaagt gctgttatat
     1861 ttgggatggc agtagaatgt gcagagataa gaagacatca tagagtgggt attaaggaca
     1921 ttgctggtat ccatttgcca acaaatgtga aatttcagag tccggcttat tcttctgtag
     1981 atactgaaga aacaattgaa ccttatacaa ctgaaaagat gagtcgagtt cctggaggat
     2041 atttggcttt gacagagtgc tttgaaatta tgacagtaga tttcaacaac cttcaggaat
     2101 taaaaagtct tgcaactaaa aagcctgata agattggtat tcctgttatt aaagaaggca
     2161 tactagatgc tattatggtt tggtttgtgc tccagcttga tgatgaacat agtttatcca
     2221 caagtcctag tgaggaaaca tgttgggaac aggctgtcta ccccgtacag gaccttgcag
     2281 actactggat aaagcctgga gaccatgtga tgatggaagt atcttgtcaa gactgttact
     2341 taagaatcca gagtattagt gtcttgggtt tggaatgtga aatggatgtt gcaaaaagtt
     2401 ttacccagaa taaagacttg ttatcgttag gaaatgaggc tgaactttgt agtgccctcg
     2461 ctaaccttca gaccagtaaa ccagatgctg tagagcagac atgtatattg gaatctacag
     2521 aaattgcttt gcttaacaac atcccatatc atgaaggctt taaaatggca atgagcaaag
     2581 ttttgtcttc actgactcca gagaaactgt atcagaccat ggatactcac tgtcagaatg
     2641 agatgagctc tggaactgga cagagtaata ctgtacagaa catccttgaa cctttctacg
     2701 tgttagatgt gtccgaaggc ttctctgttc tgcctgttat tgctggcaca cttgggcagg
     2761 ttaaaccata cagttctgtg gagaaagacc agcatcgtat tgctctggac ctcatatctg
     2821 aagccaatca ctttcctaaa gaaacacttg agttttggct gagacatgtg gaggatgaat
     2881 ctgctatgtt acaaaggcca aaatcagaca agttatggag cataattata ttggatgtca
     2941 ttgagccatc tgggctcatt cagcaggaaa taatggaaaa agctgcaata tccaggtgtt
     3001 tactacaatc tggaggcaag atctttcctc agtatgtgct gatgtttggg ttgcttgtgg
     3061 aatcacagac actcctagag gagaatgctg ttcaaggaac agaacgtact cttggattaa
     3121 atatagcacc ttttattaac cagtttcagg tacctatacg tgtatttttg gacctatcct
     3181 cattgccctg tataccttta agcaagccag tggaactctt aagactagat ttaatgactc
     3241 cgtatttgaa cacctctaac agagaagtaa aggtatacgt ttgtaaatct ggaagactga
     3301 ctgctattcc attttggtat catatgtacc ttgatgaaga gattaggttg gatacttcaa
     3361 gtgaagcctc ccactggaaa caagctgcag ttgttttaga taatcccatc caggttgaaa
     3421 tgggagagga acttgtactc agcattcagc atcacaaaag caatgtcagc atcacagtaa
     3481 agcaatgaga attcccgggt cgactcgagc ggccgcatcg tgactgactg acgatctgcc
     3541 tcgcgcgttt cggtgatgac ggtgaaaacc tctgacacat gcagctcccg gagacggtca
     3601 cagcttgtct gtaagcggat gccgggagca gacaagcccg tcagggcgcg tcagcgggtg
     3661 ttggcgggtg tcggggcgca gccatgaccc agtcacgtag cgatagcgga gtgtataatt
     3721 cttgaagacg aaagggcctc gtgatacgcc tatttttata ggttaatgtc atgataataa
     3781 tggtttctta gacgtcaggt ggcacttttc ggggaaatgt gcgcggaacc cctatttgtt
     3841 tatttttcta aatacattca aatatgtatc cgctcatgag acaataaccc tgataaatgc
     3901 ttcaataata ttgaaaaagg aagagtatga gtattcaaca tttccgtgtc gcccttattc
     3961 ccttttttgc ggcattttgc cttcctgttt ttgctcaccc agaaacgctg gtgaaagtaa
     4021 aagatgctga agatcagttg ggtgcacgag tgggttacat cgaactggat ctcaacagcg
     4081 gtaagatcct tgagagtttt cgccccgaag aacgttttcc aatgatgagc acttttaaag
     4141 ttctgctatg tggcgcggta ttatcccgtg ttgacgccgg gcaagagcaa ctcggtcgcc
     4201 gcatacacta ttctcagaat gacttggttg agtactcacc agtcacagaa aagcatctta
     4261 cggatggcat gacagtaaga gaattatgca gtgctgccat aaccatgagt gataacactg
     4321 cggccaactt acttctgaca acgatcggag gaccgaagga gctaaccgct tttttgcaca
     4381 acatggggga tcatgtaact cgccttgatc gttgggaacc ggagctgaat gaagccatac
     4441 caaacgacga gcgtgacacc acgatgcctg cagcaatggc aacaacgttg cgcaaactat
     4501 taactggcga actacttact ctagcttccc ggcaacaatt aatagactgg atggaggcgg
     4561 ataaagttgc aggaccactt ctgcgctcgg cccttccggc tggctggttt attgctgata
     4621 aatctggagc cggtgagcgt gggtctcgcg gtatcattgc agcactgggg ccagatggta
     4681 agccctcccg tatcgtagtt atctacacga cggggagtca ggcaactatg gatgaacgaa
     4741 atagacagat cgctgagata ggtgcctcac tgattaagca ttggtaactg tcagaccaag
     4801 tttactcata tatactttag attgatttaa aacttcattt ttaatttaaa aggatctagg
     4861 tgaagatcct ttttgataat ctcatgacca aaatccctta acgtgagttt tcgttccact
     4921 gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg agatcctttt tttctgcgcg
     4981 taatctgctg cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc
     5041 aagagctacc aactcttttt ccgaaggtaa ctggcttcag cagagcgcag ataccaaata
     5101 ctgtccttct agtgtagccg tagttaggcc accacttcaa gaactctgta gcaccgccta
     5161 catacctcgc tctgctaatc ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc
     5221 ttaccgggtt ggactcaaga cgatagttac cggataaggc gcagcggtcg ggctgaacgg
     5281 ggggttcgtg cacacagccc agcttggagc gaacgaccta caccgaactg agatacctac
     5341 agcgtgagct atgagaaagc gccacgcttc ccgaagggag aaaggcggac aggtatccgg
     5401 taagcggcag ggtcggaaca ggagagcgca cgagggagct tccaggggga aacgcctggt
     5461 atctttatag tcctgtcggg tttcgccacc tctgacttga gcgtcgattt ttgtgatgct
     5521 cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc ggccttttta cggttcctgg
     5581 ccttttgctg gccttttgct cacatgttct ttcctgcgtt atcccctgat tctgtggata
     5641 accgtattac cgcctttgag tgagctgata ccgctcgccg cagccgaacg accgagcgca
     5701 gcgagtcagt gagcgaggaa gcggaagagc gcctgatgcg gtattttctc cttacgcatc
     5761 tgtgcggtat ttcacaccgc ataaattccg acaccatcga atggtgcaaa acctttcgcg
     5821 gtatggcatg atagcgcccg gaagagagtc aattcagggt ggtgaatgtg aaaccagtaa
     5881 cgttatacga tgtcgcagag tatgccggtg tctcttatca gaccgtttcc cgcgtggtga
     5941 accaggccag ccacgtttct gcgaaaacgc gggaaaaagt ggaagcggcg atggcggagc
     6001 tgaattacat tcccaaccgc gtggcacaac aactggcggg caaacagtcg ttgctgattg
     6061 gcgttgccac ctccagtctg gccctgcacg cgccgtcgca aattgtcgcg gcgattaaat
     6121 ctcgcgccga tcaactgggt gccagcgtgg tggtgtcgat ggtagaacga agcggcgtcg
     6181 aagcctgtaa agcggcggtg cacaatcttc tcgcgcaacg cgtcagtggg ctgatcatta
     6241 actatccgct ggatgaccag gatgccattg ctgtggaagc tgcctgcact aatgttccgg
     6301 cgttatttct tgatgtctct gaccagacac ccatcaacag tattattttc tcccatgaag
     6361 acggtacgcg actgggcgtg gagcatctgg tcgcattggg tcaccagcaa atcgcgctgt
     6421 tagcgggccc attaagttct gtctcggcgc gtctgcgtct ggctggctgg cataaatatc
     6481 tcactcgcaa tcaaattcag ccgatagcgg aacgggaagg cgactggagt gccatgtccg
     6541 gttttcaaca aaccatgcaa atgctgaatg agggcatcgt tcccactgcg atgctggttg
     6601 ccaacgatca gatggcgctg ggcgcaatgc gcgccattac cgagtccggg ctgcgcgttg
     6661 gtgcggatat ctcggtagtg ggatacgacg ataccgaaga cagctcatgt tatatcccgc
     6721 cgtcaaccac catcaaacag gattttcgcc tgctggggca aaccagcgtg gaccgcttgc
     6781 tgcaactctc tcagggccag gcggtgaagg gcaatcagct gttgcccgtc tcactggtga
     6841 aaagaaaaac caccctggcg cccaatacgc aaaccgcctc tccccgcgcg ttggccgatt
     6901 cattaatgca gctggcacga caggtttccc gactggaaag cgggcagtga gcgcaacgca
     6961 attaatgtga gttagctcac tcattaggca ccccaggctt tacactttat gcttccggct
     7021 cgtatgttgt gtggaattgt gagcggataa caatttcaca caggaaacag ctatgaccat
     7081 gattacggat tcactggccg tcgttttaca acgtcgtgac tgggaaaacc ctggcgttac
     7141 ccaacttaat cgccttgcag cacatccccc tttcgccagc tggcgtaata gcgaagaggc
     7201 ccgcaccgat cgcccttccc aacagttgcg cagcctgaat ggcgaatggc gctttgcctg
     7261 gtttccggca ccagaagcgg tgccggaaag ctggctggag tgcgatcttc ctgaggccga
     7321 tactgtcgtc gtcccctcaa actggcagat gcacggttac gatgcgccca tctacaccaa
     7381 cgtaacctat cccattacgg tcaatccgcc gtttgttccc acggagaatc cgacgggttg
     7441 ttactcgctc acatttaatg ttgatgaaag ctggctacag gaaggccaga cgcgaattat
     7501 ttttgatggc gttggaatt
//
