LOCUS       sequence-217041-        6383 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses N-terminally GST-tagged human RNMT in bacterial cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6383)
  AUTHORS   Trotman JB, Giltmier AJ, Mukherjee C, Schoenberg DR
  TITLE     RNA guanine-7 methyltransferase catalyzes the methylation of 
            cytoplasmically recapped RNAs.
  JOURNAL   Nucleic Acids Res. 2017 Oct 13;45(18):10726-10739. doi: 
            10.1093/nar/gkx801.
  PUBMED    28981715
REFERENCE   2  (bases 1 to 6383)
  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"; journalName: 
            "Nucleic Acids Res"; date: "2017-10-13- 13"; volume: "45"; issue: 
            "18"; pages: "10726-10739"
COMMENT     Sequence Label: pGEX-2T-GST-RNMT
FEATURES             Location/Qualifiers
     source          1..6383
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        join(1..6,7..22,23..29)
                     /label=tac promoter
                     /note="strong E. coli promoter; hybrid between the trp and 
                     lac UV5 promoters"
     protein_bind    37..53
                     /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             76..729
                     /codon_start=1
                     /product="glutathione S-transferase from Schistosoma 
                     japonicum"
                     /label=GST
                     /translation="MSPILGYWKIKGLVQPTRLLLEYLEEKYEEHLYERDEGDKWRNKK
                     FELGLEFPNLPYYIDGDVKLTQSMAIIRYIADKHNMLGGCPKERAEISMLEGAVLDIRY
                     GVSRIAYSKDFETLKVDFLSKLPEMLKMFEDRLCHKTYLNGDHVTHPDFMLYDALDVVL
                     YMDPMCLDAFPKLVCFKKRIEAIPQIDKYLKSSKYIAWPLQGWQATFGGGDHPPK"
     primer_bind     687..709
                     /label=pGEX 5'
                     /note="pGEX vectors, Glutathione-S-transferase, forward 
                     primer"
     CDS             736..753
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
     primer_bind     complement(2251..2273)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(2418..2436)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        2504..2608
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2609..2677,2678..3469)
                     /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(2827..2846)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3640..4228
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4129..4148
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4382..4399
                     /label=L4440
                     /note="L4440 vector, forward primer"
     promoter        4472..4549
                     /gene="lacI"
                     /label=lacI promoter
     CDS             4550..5632
                     /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
                     YIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR
                     ALADSLMQLARQVSRLESGQ"
     primer_bind     complement(4569..4588)
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        join(5681..5686,5687..5704,5705..5711)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5719..5735
                     /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     5743..5759
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     5743..5759
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             5755..6282
                     /codon_start=1
                     /gene="lacZ fragment"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /label=lacZ-alpha
                     /translation="MTMITDSLAVVLQRRDWENPGVTQLNRLAAHPPFASWRNSEEART
                     DRPSQQLRSLNGEWRFAWFPAPEAVPESWLECDLPEADTVVVPSNWQMHGYDAPIYTNV
                     TYPITVNPPFVPTENPTGCYSLTFNVDESWLQEGQTRIIFDGVGISLSTARCTNASGVR
                     QPSEAVVWLCRS"
     primer_bind     complement(5775..5792)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(5775..5791)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5784..5806)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
ORIGIN
        1 ttgacaatta atcatcggct cgtataatgt gtggaattgt gagcggataa caatttcaca
       61 caggaaacag tattcatgtc ccctatacta ggttattgga aaattaaggg ccttgtgcaa
      121 cccactcgac ttcttttgga atatcttgaa gaaaaatatg aagagcattt gtatgagcgc
      181 gatgaaggtg ataaatggcg aaacaaaaag tttgaattgg gtttggagtt tcccaatctt
      241 ccttattata ttgatggtga tgttaaatta acacagtcta tggccatcat acgttatata
      301 gctgacaagc acaacatgtt gggtggttgt ccaaaagagc gtgcagagat ttcaatgctt
      361 gaaggagcgg ttttggatat tagatacggt gtttcgagaa ttgcatatag taaagacttt
      421 gaaactctca aagttgattt tcttagcaag ctacctgaaa tgctgaaaat gttcgaagat
      481 cgtttatgtc ataaaacata tttaaatggt gatcatgtaa cccatcctga cttcatgttg
      541 tatgacgctc ttgatgttgt tttatacatg gacccaatgt gcctggatgc gttcccaaaa
      601 ttagtttgtt ttaaaaaacg tattgaagct atcccacaaa ttgataagta cttgaaatcc
      661 agcaagtata tagcatggcc tttgcagggc tggcaagcca cgtttggtgg tggcgaccat
      721 cctccaaaat cggatctggt tccgcgtgga tccccgggaa ttcatgcaaa ttctgcaaaa
      781 gcagaagaat atgaaaagat gtctcttgaa caggcaaaag cgtcagtgaa ttctgaaaca
      841 gagtcttcat tcaatattaa tgaaaacaca acagcttctg ggactgggct ttctgaaaag
      901 acttctgtct gtaggcaagt agacatagca agaaagagaa aagagtttga agatgatctt
      961 gtaaaggaaa gttctagttg tgggaaagac actccatcca agaagagaaa acttgatcct
     1021 gaaattgtcc cagaggaaaa agattgtggt gatgctgaag gcaattcaaa gaaaagaaaa
     1081 agagaaactg aggatgttcc aaaagataaa tcttctactg gagatggcac tcaaaataag
     1141 agaaaaatag cacttgagga tgttcctgaa aagcagaaaa atctggaaga aggacacagc
     1201 tcaacagtgg ctgcccatta caatgaactt caggaagttg gtttggagaa gcgtagtcaa
     1261 agtcgtattt tttacctaag aaactttaat aattggatga aaagtgttct cattggagaa
     1321 tttttggaaa aggtacgaca gaagaaaaaa cgtgatatca ctgttttgga cctgggatgt
     1381 ggtaaaggtg gagatttgct gaaatggaaa aaaggaagaa ttaacaagct agtttgtact
     1441 gatattgccg atgtttctgt caaacagtgt cagcagcggt atgaggacat gaaaaatcgt
     1501 cgtgatagtg aatatatttt cagtgcagaa tttataactg ctgacagctc aaaggaactt
     1561 ctgattgaca aatttcgtga cccacaaatg tgttttgaca tctgcagttg tcagtttgtc
     1621 tgtcattact catttgagtc ttatgagcag gctgacatga tgctgagaaa tgcgtgtgag
     1681 agacttagcc ctgggggcta ttttattggt actactccca atagctttga attgataaga
     1741 cgccttgaag cttcagaaac agaatcattt ggaaatgaaa tatatactgt gaaatttcag
     1801 aagaaaggag attatccttt atttggctgc aaatatgact tcaacttgga aggtgttgtg
     1861 gatgttcctg aattcttggt ctattttcca ttgctaaatg aaatggcaaa gaagtacaat
     1921 atgaaactag tctacaaaaa aacatttctg gaattctacg aagaaaagat taagaacaat
     1981 gaaaataaaa tgctcttaaa acgaatgcag gccttggagc catatcctgc aaatgagagt
     2041 tctaaacttg tctctgagaa ggtggatgac tatgaacatg cagcaaagta catgaagaac
     2101 agtcaagtaa ggttaccttt gggaacctta agtaaatcag aatgggaagc tacaagtatt
     2161 tacttggtgt ttgcctttga gaaacagcag tgaaattcat cgtgactgac tgacgatctg
     2221 cctcgcgcgt ttcggtgatg acggtgaaaa cctctgacac atgcagctcc cggagacggt
     2281 cacagcttgt ctgtaagcgg atgccgggag cagacaagcc cgtcagggcg cgtcagcggg
     2341 tgttggcggg tgtcggggcg cagccatgac ccagtcacgt agcgatagcg gagtgtataa
     2401 ttcttgaaga cgaaagggcc tcgtgatacg cctattttta taggttaatg tcatgataat
     2461 aatggtttct tagacgtcag gtggcacttt tcggggaaat gtgcgcggaa cccctatttg
     2521 tttatttttc taaatacatt caaatatgta tccgctcatg agacaataac cctgataaat
     2581 gcttcaataa tattgaaaaa ggaagagtat gagtattcaa catttccgtg tcgcccttat
     2641 tccctttttt gcggcatttt gccttcctgt ttttgctcac ccagaaacgc tggtgaaagt
     2701 aaaagatgct gaagatcagt tgggtgcacg agtgggttac atcgaactgg atctcaacag
     2761 cggtaagatc cttgagagtt ttcgccccga agaacgtttt ccaatgatga gcacttttaa
     2821 agttctgcta tgtggcgcgg tattatcccg tgttgacgcc gggcaagagc aactcggtcg
     2881 ccgcatacac tattctcaga atgacttggt tgagtactca ccagtcacag aaaagcatct
     2941 tacggatggc atgacagtaa gagaattatg cagtgctgcc ataaccatga gtgataacac
     3001 tgcggccaac ttacttctga caacgatcgg aggaccgaag gagctaaccg cttttttgca
     3061 caacatgggg gatcatgtaa ctcgccttga tcgttgggaa ccggagctga atgaagccat
     3121 accaaacgac gagcgtgaca ccacgatgcc tgcagcaatg gcaacaacgt tgcgcaaact
     3181 attaactggc gaactactta ctctagcttc ccggcaacaa ttaatagact ggatggaggc
     3241 ggataaagtt gcaggaccac ttctgcgctc ggcccttccg gctggctggt ttattgctga
     3301 taaatctgga gccggtgagc gtgggtctcg cggtatcatt gcagcactgg ggccagatgg
     3361 taagccctcc cgtatcgtag ttatctacac gacggggagt caggcaacta tggatgaacg
     3421 aaatagacag atcgctgaga taggtgcctc actgattaag cattggtaac tgtcagacca
     3481 agtttactca tatatacttt agattgattt aaaacttcat ttttaattta aaaggatcta
     3541 ggtgaagatc ctttttgata atctcatgac caaaatccct taacgtgagt tttcgttcca
     3601 ctgagcgtca gaccccgtag aaaagatcaa aggatcttct tgagatcctt tttttctgcg
     3661 cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca gcggtggttt gtttgccgga
     3721 tcaagagcta ccaactcttt ttccgaaggt aactggcttc agcagagcgc agataccaaa
     3781 tactgtcctt ctagtgtagc cgtagttagg ccaccacttc aagaactctg tagcaccgcc
     3841 tacatacctc gctctgctaa tcctgttacc agtggctgct gccagtggcg ataagtcgtg
     3901 tcttaccggg ttggactcaa gacgatagtt accggataag gcgcagcggt cgggctgaac
     3961 ggggggttcg tgcacacagc ccagcttgga gcgaacgacc tacaccgaac tgagatacct
     4021 acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc
     4081 ggtaagcggc agggtcggaa caggagagcg cacgagggag cttccagggg gaaacgcctg
     4141 gtatctttat agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg
     4201 ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac gcggcctttt tacggttcct
     4261 ggccttttgc tggccttttg ctcacatgtt ctttcctgcg ttatcccctg attctgtgga
     4321 taaccgtatt accgcctttg agtgagctga taccgctcgc cgcagccgaa cgaccgagcg
     4381 cagcgagtca gtgagcgagg aagcggaaga gcgcctgatg cggtattttc tccttacgca
     4441 tctgtgcggt atttcacacc gcataaattc cgacaccatc gaatggcgca aaacctttcg
     4501 cggtatggca tgatagcgcc cggaagagag tcaattcagg gtggtgaatg tgaaaccagt
     4561 aacgttatac gatgtcgcag agtatgccgg tgtctcttat cagaccgttt cccgcgtggt
     4621 gaaccaggcc agccacgttt ctgcgaaaac gcgggaaaaa gtggaagcgg cgatggcgga
     4681 gctgaattac attcccaacc gcgtggcaca acaactggcg ggcaaacagt cgttgctgat
     4741 tggcgttgcc acctccagtc tggccctgca cgcgccgtcg caaattgtcg cggcgattaa
     4801 atctcgcgcc gatcaactgg gtgccagcgt ggtggtgtcg atggtagaac gaagcggcgt
     4861 cgaagcctgt aaagcggcgg tgcacaatct tctcgcgcaa cgcgtcagtg ggctgatcat
     4921 taactatccg ctggatgacc aggatgccat tgctgtggaa gctgcctgca ctaatgttcc
     4981 ggcgttattt cttgatgtct ctgaccagac acccatcaac agtattattt tctcccatga
     5041 agacggtacg cgactgggcg tggagcatct ggtcgcattg ggtcaccagc aaatcgcgct
     5101 gttagcgggc ccattaagtt ctgtctcggc gcgtctgcgt ctggctggct ggcataaata
     5161 tctcactcgc aatcaaattc agccgatagc ggaacgggaa ggcgactgga gtgccatgtc
     5221 cggttttcaa caaaccatgc aaatgctgaa tgagggcatc gttcccactg cgatgctggt
     5281 tgccaacgat cagatggcgc tgggcgcaat gcgcgccatt accgagtccg ggctgcgcgt
     5341 tggtgcggat atctcggtag tgggatacga cgataccgaa gacagctcat gttatatccc
     5401 gccgttaacc accatcaaac aggattttcg cctgctgggg caaaccagcg tggaccgctt
     5461 gctgcaactc tctcagggcc aggcggtgaa gggcaatcag ctgttgcccg tctcactggt
     5521 gaaaagaaaa accaccctgg cgcccaatac gcaaaccgcc tctccccgcg cgttggccga
     5581 ttcattaatg cagctggcac gacaggtttc ccgactggaa agcgggcagt gagcgcaacg
     5641 caattaatgt aagttagctc actcattagg caccccaggc tttacacttt atgcttccgg
     5701 ctcgtatgtt gtgtggaatt gtgagcggat aacaatttca cacaggaaac agctatgacc
     5761 atgattacgg attcactggc cgtcgtttta caacgtcgtg actgggaaaa ccctggcgtt
     5821 acccaactta atcgccttgc agcacatccc cctttcgcca gctggcgtaa tagcgaagag
     5881 gcccgcaccg atcgcccttc ccaacagttg cgcagcctga atggcgaatg gcgctttgcc
     5941 tggtttccgg caccagaagc ggtgccggaa agctggctgg agtgcgatct tcctgaggcc
     6001 gatactgtcg tcgtcccctc aaactggcag atgcacggtt acgatgcgcc catctacacc
     6061 aacgtgacct atcccattac ggtcaatccg ccgtttgttc ccacggagaa tccgacgggt
     6121 tgttactcgc tcacatttaa tgttgatgaa agctggctac aggaaggcca gacgcgaatt
     6181 atttttgatg gcgttggaat tagcttatcg actgcacggt gcaccaatgc ttctggcgtc
     6241 aggcagccat cggaagctgt ggtatggctg tgcaggtcgt aaatcactgc ataattcgtg
     6301 tcgctcaagg cgcactcccg ttctggataa tgttttttgc gccgacatca taacggttct
     6361 ggcaaatatt ctgaaatgag ctg
//
