LOCUS       sequence-369236-        7642 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Bacterial expression plasmid for human NSUN2; for activity testing.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7642)
  AUTHORS   Schwickert M, Fischer TR, Zimmermann RA, Hoba SN, Meidner JL, Weber 
            M, Weber M, Stark MM, Koch J, Jung N, Kersten C, Windbergs M, Lyko 
            F, Helm M, Schirmeister T
  TITLE     Discovery of Inhibitors of DNA Methyltransferase 2, an 
            Epitranscriptomic Modulator and Potential Target for Cancer 
            Treatment.
  JOURNAL   J Med Chem. 2022 Jul 18. doi: 10.1021/acs.jmedchem.2c00388.
  PUBMED    35849534
REFERENCE   2  (bases 1 to 7642)
  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: "J Med Chem.
            2022 Jul 18. doi: 10.1021/acs.jmedchem.2c00388."
COMMENT     Sequence Label: NSUN2
FEATURES             Location/Qualifiers
     source          1..7642
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     965..984
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(1004..1081)
                     /gene="lacI"
                     /label=lacI promoter
     primer_bind     1287..1306
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     1390..1409
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1390..1408
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    1409..1433
                     /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)."
     RBS             1448..1470
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             1489..1506
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             1516..1533
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
     CDS             1537..1569
                     /codon_start=1
                     /product="leader peptide from bacteriophage T7 gene 10"
                     /label=T7 tag (gene 10 leader)
                     /note="promotes efficient translation in E. coli"
                     /translation="MASMTGGQQMG"
     CDS             3892..3909
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(3962..3980)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      3976..4023
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      4060..4515
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4147..4166)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4357..4378
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(4608..5423)
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     CDS             complement(4608..5423)
                     /codon_start=1
                     /gene="aph(3')-Ia"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     5331..5350
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     5415..5434
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     rep_origin      5545..6133
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6034..6053
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6287..6304
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    6319..6461
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(6387..6406)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     6547..6569
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(6563..6754)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(join(7563..7642,1..1003))
                     /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"
ORIGIN
        1 gcgtattggg cgccagggtg gtttttcttt tcaccagtga gacgggcaac agctgattgc
       61 ccttcaccgc ctggccctga gagagttgca gcaagcggtc cacgctggtt tgccccagca
      121 ggcgaaaatc ctgtttgatg gtggttaacg gcgggatata acatgagctg tcttcggtat
      181 cgtcgtatcc cactaccgag atatccgcac caacgcgcag cccggactcg gtaatggcgc
      241 gcattgcgcc cagcgccatc tgatcgttgg caaccagcat cgcagtggga acgatgccct
      301 cattcagcat ttgcatggtt tgttgaaaac cggacatggc actccagtcg ccttcccgtt
      361 ccgctatcgg ctgaatttga ttgcgagtga gatatttatg ccagccagcc agacgcagac
      421 gcgccgagac agaacttaat gggcccgcta acagcgcgat ttgctggtga cccaatgcga
      481 ccagatgctc cacgcccagt cgcgtaccgt cttcatggga gaaaataata ctgttgatgg
      541 gtgtctggtc agagacatca agaaataacg ccggaacatt agtgcaggca gcttccacag
      601 caatggcatc ctggtcatcc agcggatagt taatgatcag cccactgacg cgttgcgcga
      661 gaagattgtg caccgccgct ttacaggctt cgacgccgct tcgttctacc atcgacacca
      721 ccacgctggc acccagttga tcggcgcgag atttaatcgc cgcgacaatt tgcgacggcg
      781 cgtgcagggc cagactggag gtggcaacgc caatcagcaa cgactgtttg cccgccagtt
      841 gttgtgccac gcggttggga atgtaattca gctccgccat cgccgcttcc actttttccc
      901 gcgttttcgc agaaacgtgg ctggcctggt tcaccacgcg ggaaacggtc tgataagaga
      961 caccggcata ctctgcgaca tcgtataacg ttactggttt cacattcacc accctgaatt
     1021 gactctcttc cgggcgctat catgccatac cgcgaaaggt tttgcgccat tcgatggtgt
     1081 ccgggatctc gacgctctcc cttatgcgac tcctgcatta ggaagcagcc cagtagtagg
     1141 ttgaggccgt tgagcaccgc cgccgcaagg aatggtgcat gcaaggagat ggcgcccaac
     1201 agtcccccgg ccacggggcc tgccaccata cccacgccga aacaagcgct catgagcccg
     1261 aagtggcgag cccgatcttc cccatcggtg atgtcggcga tataggcgcc agcaaccgca
     1321 cctgtggcgc cggtgatgcc ggccacgatg cgtccggcgt agaggatcga gatctcgatc
     1381 ccgcgaaatt aatacgactc actatagggg aattgtgagc ggataacaat tcccctctag
     1441 aaataatttt gtttaacttt aagaaggaga tataccatgg gcagcagcca tcatcatcat
     1501 catcacagca gcggcctggt gccgcgcggc agccatatgg ctagcatgac tggtggacag
     1561 caaatgggtc gcggatccga attcatgggt cgtcgtagcc gtggtcgtcg tctgcaacaa
     1621 caacaacgcc cggaagatgc ggaggatggt gcggaaggtg gtggtaaacg cggtgaagcg
     1681 ggttgggaag gtggctaccc ggagatcgtt aaagaaaaca agctgttcga acactactat
     1741 caggagctga agattgtgcc ggagggtgaa tggggccaat ttatggatgc gctgcgtgag
     1801 ccgctgccgg cgaccctgcg tatcaccggt tacaaaagcc acgcgaagga gattctgcac
     1861 tgcctgaaaa acaagtattt caaggagctg gaagacctgg aagttgatgg ccagaaagtt
     1921 gaggtgccgc aaccgctgag ctggtatccg gaggaactgg cgtggcacac caacctgagc
     1981 cgtaaaatcc tgcgtaagag cccgcacctg gaaaaattcc accagtttct ggtgagcgaa
     2041 accgaaagcg gtaacatcag ccgtcaagag gcggttagca tgattccgcc gctgctgctg
     2101 aacgtgcgtc cgcaccacaa gatcctggat atgtgcgcgg cgccgggtag caaaaccacc
     2161 caactgattg aaatgctgca cgcggacatg aacgttccgt tcccggaggg ctttgttatc
     2221 gcgaacgacg tggataacaa gcgttgctac ctgctggtgc accaagcgaa acgtctgagc
     2281 agcccgtgca ttatggtggt taaccacgac gcgagcagca tcccgcgtct gcagattgac
     2341 gttgatggtc gtaaggaaat cctgttctat gaccgtattc tgtgcgatgt tccgtgcagc
     2401 ggtgacggca ccatgcgtaa aaacatcgat gtgtggaaga aatggaccac cctgaacagc
     2461 ctgcagctgc acggtctgca actgcgtatt gcgacccgtg gcgcggagca gctggcggaa
     2521 ggtggccgta tggtttacag cacctgcagc ctgaacccga tcgaggatga agcggtgatt
     2581 gcgagcctgc tggagaagag cgaaggtgcg ctggaactgg cggacgtgag caacgagctg
     2641 ccgggtctga aatggatgcc gggcatcacc cagtggaaag ttatgaccaa ggatggccaa
     2701 tggtttaccg actgggatgc ggtgccgcac agccgtcaca cccagatccg tccgaccatg
     2761 tttccgccga aagacccgga aaagctgcaa gcgatgcacc tggagcgttg cctgcgtatt
     2821 ctgccgcacc accagaacac cggtggcttc tttgtggcgg ttctggtgaa gaaaagcagc
     2881 atgccgtgga acaaacgtca gccgaagctg caaggtaaaa gcgcggaaac ccgtgagagc
     2941 acccaactga gcccggcgga cctgaccgaa ggcaagccga ccgatccgag caaactggag
     3001 agcccgagct tcaccggcac cggtgacacc gaaattgcgc atgcgaccga ggatctggaa
     3061 aacaacggta gcaagaaaga cggtgtttgc ggtccgccgc cgagcaagaa aatgaagctg
     3121 ttcggcttta aagaagatcc gttcgtgttt atcccggagg acgatccgct gttcccgccg
     3181 attgaaaagt tttacgcgct ggacccgagc tttccgcgta tgaacctgct gacccgtacc
     3241 accgagggta agaaacgtca actgtatatg gttagcaagg aactgcgtaa cgtgctgctg
     3301 aacaacagcg agaaaatgaa ggttatcaac accggtatta aagtgtggtg ccgtaacaac
     3361 agcggcgagg agttcgattg cgcgtttcgt ctggcgcagg aaggtatcta caccctgtat
     3421 ccgttcatta acagccgtat cattaccgtt agcatggaag atgtgaagat cctgctgacc
     3481 caagagaacc cgttctttcg taaactgagc agcgaaacct acagccaggc gaaggacctg
     3541 gcgaaaggca gcattgttct gaaatatgaa ccggatagcg cgaacccgga tgcgctgcag
     3601 tgcccgatcg ttctgtgcgg ttggcgtggc aaggcgagca ttcgtacctt tgtgccgaaa
     3661 aacgaacgtc tgcactacct gcgtatgatg ggtctggaag tgctgggcga gaagaagaaa
     3721 gagggtgtga tcctgaccaa cgagagcgcg gcgagcaccg gtcagccgga caacgatgtg
     3781 accgagggtc agcgtgcggg tgaaccgaac agcccggacg cggaggaagc gaatagcccg
     3841 gacgttaccg cgggttgcga tccggcgggt gttcatccgc cgcgtctcga gcaccaccac
     3901 caccaccact gagatccggc tgctaacaaa gcccgaaagg aagctgagtt ggctgctgcc
     3961 accgctgagc aataactagc ataacccctt ggggcctcta aacgggtctt gaggggtttt
     4021 ttgctgaaag gaggaactat atccggattg gcgaatggga cgcgccctgt agcggcgcat
     4081 taagcgcggc gggtgtggtg gttacgcgca gcgtgaccgc tacacttgcc agcgccctag
     4141 cgcccgctcc tttcgctttc ttcccttcct ttctcgccac gttcgccggc tttccccgtc
     4201 aagctctaaa tcgggggctc cctttagggt tccgatttag tgctttacgg cacctcgacc
     4261 ccaaaaaact tgattagggt gatggttcac gtagtgggcc atcgccctga tagacggttt
     4321 ttcgcccttt gacgttggag tccacgttct ttaatagtgg actcttgttc caaactggaa
     4381 caacactcaa ccctatctcg gtctattctt ttgatttata agggattttg ccgatttcgg
     4441 cctattggtt aaaaaatgag ctgatttaac aaaaatttaa cgcgaatttt aacaaaatat
     4501 taacgtttac aatttcaggt ggcacttttc ggggaaatgt gcgcggaacc cctatttgtt
     4561 tatttttcta aatacattca aatatgtatc cgctcatgaa ttaattctta gaaaaactca
     4621 tcgagcatca aatgaaactg caatttattc atatcaggat tatcaatacc atatttttga
     4681 aaaagccgtt tctgtaatga aggagaaaac tcaccgaggc agttccatag gatggcaaga
     4741 tcctggtatc ggtctgcgat tccgactcgt ccaacatcaa tacaacctat taatttcccc
     4801 tcgtcaaaaa taaggttatc aagtgagaaa tcaccatgag tgacgactga atccggtgag
     4861 aatggcaaaa gtttatgcat ttctttccag acttgttcaa caggccagcc attacgctcg
     4921 tcatcaaaat cactcgcatc aaccaaaccg ttattcattc gtgattgcgc ctgagcgaga
     4981 cgaaatacgc gatcgctgtt aaaaggacaa ttacaaacag gaatcgaatg caaccggcgc
     5041 aggaacactg ccagcgcatc aacaatattt tcacctgaat caggatattc ttctaatacc
     5101 tggaatgctg ttttcccggg gatcgcagtg gtgagtaacc atgcatcatc aggagtacgg
     5161 ataaaatgct tgatggtcgg aagaggcata aattccgtca gccagtttag tctgaccatc
     5221 tcatctgtaa catcattggc aacgctacct ttgccatgtt tcagaaacaa ctctggcgca
     5281 tcgggcttcc catacaatcg atagattgtc gcacctgatt gcccgacatt atcgcgagcc
     5341 catttatacc catataaatc agcatccatg ttggaattta atcgcggcct agagcaagac
     5401 gtttcccgtt gaatatggct cataacaccc cttgtattac tgtttatgta agcagacagt
     5461 tttattgttc atgaccaaaa tcccttaacg tgagttttcg ttccactgag cgtcagaccc
     5521 cgtagaaaag atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt
     5581 gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg ccggatcaag agctaccaac
     5641 tctttttccg aaggtaactg gcttcagcag agcgcagata ccaaatactg tccttctagt
     5701 gtagccgtag ttaggccacc acttcaagaa ctctgtagca ccgcctacat acctcgctct
     5761 gctaatcctg ttaccagtgg ctgctgccag tggcgataag tcgtgtctta ccgggttgga
     5821 ctcaagacga tagttaccgg ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac
     5881 acagcccagc ttggagcgaa cgacctacac cgaactgaga tacctacagc gtgagctatg
     5941 agaaagcgcc acgcttcccg aagggagaaa ggcggacagg tatccggtaa gcggcagggt
     6001 cggaacagga gagcgcacga gggagcttcc agggggaaac gcctggtatc tttatagtcc
     6061 tgtcgggttt cgccacctct gacttgagcg tcgatttttg tgatgctcgt caggggggcg
     6121 gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct tttgctggcc
     6181 ttttgctcac atgttctttc ctgcgttatc ccctgattct gtggataacc gtattaccgc
     6241 ctttgagtga gctgataccg ctcgccgcag ccgaacgacc gagcgcagcg agtcagtgag
     6301 cgaggaagcg gaagagcgcc tgatgcggta ttttctcctt acgcatctgt gcggtatttc
     6361 acaccgcata tatggtgcac tctcagtaca atctgctctg atgccgcata gttaagccag
     6421 tatacactcc gctatcgcta cgtgactggg tcatggctgc gccccgacac ccgccaacac
     6481 ccgctgacgc gccctgacgg gcttgtctgc tcccggcatc cgcttacaga caagctgtga
     6541 ccgtctccgg gagctgcatg tgtcagaggt tttcaccgtc atcaccgaaa cgcgcgaggc
     6601 agctgcggta aagctcatca gcgtggtcgt gaagcgattc acagatgtct gcctgttcat
     6661 ccgcgtccag ctcgttgagt ttctccagaa gcgttaatgt ctggcttctg ataaagcggg
     6721 ccatgttaag ggcggttttt tcctgtttgg tcactgatgc ctccgtgtaa gggggatttc
     6781 tgttcatggg ggtaatgata ccgatgaaac gagagaggat gctcacgata cgggttactg
     6841 atgatgaaca tgcccggtta ctggaacgtt gtgagggtaa acaactggcg gtatggatgc
     6901 ggcgggacca gagaaaaatc actcagggtc aatgccagcg cttcgttaat acagatgtag
     6961 gtgttccaca gggtagccag cagcatcctg cgatgcagat ccggaacata atggtgcagg
     7021 gcgctgactt ccgcgtttcc agactttacg aaacacggaa accgaagacc attcatgttg
     7081 ttgctcaggt cgcagacgtt ttgcagcagc agtcgcttca cgttcgctcg cgtatcggtg
     7141 attcattctg ctaaccagta aggcaacccc gccagcctag ccgggtcctc aacgacagga
     7201 gcacgatcat gcgcacccgt ggggccgcca tgccggcgat aatggcctgc ttctcgccga
     7261 aacgtttggt ggcgggacca gtgacgaagg cttgagcgag ggcgtgcaag attccgaata
     7321 ccgcaagcga caggccgatc atcgtcgcgc tccagcgaaa gcggtcctcg ccgaaaatga
     7381 cccagagcgc tgccggcacc tgtcctacga gttgcatgat aaagaagaca gtcataagtg
     7441 cggcgacgat agtcatgccc cgcgcccacc ggaaggagct gactgggttg aaggctctca
     7501 agggcatcgg tcgagatccc ggtgcctaat gagtgagcta acttacatta attgcgttgc
     7561 gctcactgcc cgctttccag tcgggaaacc tgtcgtgcca gctgcattaa tgaatcggcc
     7621 aacgcgcggg gagaggcggt tt
//
