LOCUS       sequence-369237-        6770 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Bacterial expression plasmid for human NSUN6; for activity testing.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6770)
  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 6770)
  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: NSUN6
FEATURES             Location/Qualifiers
     source          1..6770
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(79..98)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     289..310
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        473..577
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(578..646,647..1438)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(796..815)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      1609..2197
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2098..2117
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2351..2368
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    2383..2522
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(2448..2467)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     2608..2630
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(2624..2815)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(3624..4706)
                     /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     4668..4687
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(4707..4784)
                     /gene="lacI"
                     /label=lacI promoter
     primer_bind     4990..5009
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     5093..5112
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        5093..5111
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    5112..5136
                     /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             5151..5173
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             6594..6611
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(6664..6682)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      6678..6725
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      join(6762..6770,1..447)
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
ORIGIN
        1 gtagcggcgc attaagcgcg gcgggtgtgg tggttacgcg cagcgtgacc gctacacttg
       61 ccagcgccct agcgcccgct cctttcgctt tcttcccttc ctttctcgcc acgttcgccg
      121 gctttccccg tcaagctcta aatcgggggc tccctttagg gttccgattt agtgctttac
      181 ggcacctcga ccccaaaaaa cttgattagg gtgatggttc acgtagtggg ccatcgccct
      241 gatagacggt ttttcgccct ttgacgttgg agtccacgtt ctttaatagt ggactcttgt
      301 tccaaactgg aacaacactc aaccctatct cggtctattc ttttgattta taagggattt
      361 tgccgatttc ggcctattgg ttaaaaaatg agctgattta acaaaaattt aacgcgaatt
      421 ttaacaaaat attaacgctt acaatttagg tggcactttt cggggaaatg tgcgcggaac
      481 ccctatttgt ttatttttct aaatacattc aaatatgtat ccgctcatga gacaataacc
      541 ctgataaatg cttcaataat attgaaaaag gaagagtatg agtattcaac atttccgtgt
      601 cgcccttatt cccttttttg cggcattttg ccttcctgtt tttgctcacc cagaaacgct
      661 ggtgaaagta aaagatgctg aagatcagtt gggtgcacga gtgggttaca tcgaactgga
      721 tctcaacagc ggtaagatcc ttgagagttt tcgccccgaa gaacgttttc caatgatgag
      781 cacttttaaa gttctgctat gtggcgcggt attatcccgt attgacgccg ggcaagagca
      841 actcggtcgc cgcatacact attctcagaa tgacttggtt gagtactcac cagtcacaga
      901 aaagcatctt acggatggca tgacagtaag agaattatgc agtgctgcca taaccatgag
      961 tgataacact gcggccaact tacttctgac aacgatcgga ggaccgaagg agctaaccgc
     1021 ttttttgcac aacatggggg atcatgtaac tcgccttgat cgttgggaac cggagctgaa
     1081 tgaagccata ccaaacgacg agcgtgacac cacgatgcct gcagcaatgg caacaacgtt
     1141 gcgcaaacta ttaactggcg aactacttac tctagcttcc cggcaacaat taatagactg
     1201 gatggaggcg gataaagttg caggaccact tctgcgctcg gcccttccgg ctggctggtt
     1261 tattgctgat aaatctggag ccggtgagcg tgggtctcgc ggtatcattg cagcactggg
     1321 gccagatggt aagccctccc gtatcgtagt tatctacacg acggggagtc aggcaactat
     1381 ggatgaacga aatagacaga tcgctgagat aggtgcctca ctgattaagc attggtaact
     1441 gtcagaccaa gtttactcat atatacttta gattgattta aaacttcatt tttaatttaa
     1501 aaggatctag gtgaagatcc tttttgataa tctcatgacc aaaatccctt aacgtgagtt
     1561 ttcgttccac tgagcgtcag accccgtaga aaagatcaaa ggatcttctt gagatccttt
     1621 ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg
     1681 tttgccggat caagagctac caactctttt tccgaaggta actggcttca gcagagcgca
     1741 gataccaaat actgtccttc tagtgtagcc gtagttaggc caccacttca agaactctgt
     1801 agcaccgcct acatacctcg ctctgctaat cctgttacca gtggctgctg ccagtggcga
     1861 taagtcgtgt cttaccgggt tggactcaag acgatagtta ccggataagg cgcagcggtc
     1921 gggctgaacg gggggttcgt gcacacagcc cagcttggag cgaacgacct acaccgaact
     1981 gagataccta cagcgtgagc tatgagaaag cgccacgctt cccgaaggga gaaaggcgga
     2041 caggtatccg gtaagcggca gggtcggaac aggagagcgc acgagggagc ttccaggggg
     2101 aaacgcctgg tatctttata gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt
     2161 tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac gccagcaacg cggccttttt
     2221 acggttcctg gccttttgct ggccttttgc tcacatgttc tttcctgcgt tatcccctga
     2281 ttctgtggat aaccgtatta ccgcctttga gtgagctgat accgctcgcc gcagccgaac
     2341 gaccgagcgc agcgagtcag tgagcgagga agcggaagag cgcctgatgc ggtattttct
     2401 ccttacgcat ctgtgcggta tttcacaccg caatggtgca ctctcagtac aatctgctct
     2461 gatgccgcat agttaagcca gtatacactc cgctatcgct acgtgactgg gtcatggctg
     2521 cgccccgaca cccgccaaca cccgctgacg cgccctgacg ggcttgtctg ctcccggcat
     2581 ccgcttacag acaagctgtg accgtctccg ggagctgcat gtgtcagagg ttttcaccgt
     2641 catcaccgaa acgcgcgagg cagctgcggt aaagctcatc agcgtggtcg tgaagcgatt
     2701 cacagatgtc tgcctgttca tccgcgtcca gctcgttgag tttctccaga agcgttaatg
     2761 tctggcttct gataaagcgg gccatgttaa gggcggtttt ttcctgtttg gtcactgatg
     2821 cctccgtgta agggggattt ctgttcatgg gggtaatgat accgatgaaa cgagagagga
     2881 tgctcacgat acgggttact gatgatgaac atgcccggtt actggaacgt tgtgagggta
     2941 aacaactggc ggtatggatg cggcgggacc agagaaaaat cactcagggt caatgccagc
     3001 gcttcgttaa tacagatgta ggtgttccac agggtagcca gcagcatcct gcgatgcaga
     3061 tccggaacat aatggtgcag ggcgctgact tccgcgtttc cagactttac gaaacacgga
     3121 aaccgaagac cattcatgtt gttgctcagg tcgcagacgt tttgcagcag cagtcgcttc
     3181 acgttcgctc gcgtatcggt gattcattct gctaaccagt aaggcaaccc cgccagccta
     3241 gccgggtcct caacgacagg agcacgatca tgcgcacccg tggggccgcc atgccggcga
     3301 taatggcctg cttctcgccg aaacgtttgg tggcgggacc agtgacgaag gcttgagcga
     3361 gggcgtgcaa gattccgaat accgcaagcg acaggccgat catcgtcgcg ctccagcgaa
     3421 agcggtcctc gccgaaaatg acccagagcg ctgccggcac ctgtcctacg agttgcatga
     3481 taaagaagac agtcataagt gcggcgacga tagtcatgcc ccgcgcccac cggaaggagc
     3541 tgactgggtt gaaggctctc aagggcatcg gtcgagatcc cggtgcctaa tgagtgagct
     3601 aacttacatt aattgcgttg cgctcactgc ccgctttcca gtcgggaaac ctgtcgtgcc
     3661 agctgcatta atgaatcggc caacgcgcgg ggagaggcgg tttgcgtatt gggcgccagg
     3721 gtggtttttc ttttcaccag tgagacgggc aacagctgat tgcccttcac cgcctggccc
     3781 tgagagagtt gcagcaagcg gtccacgctg gtttgcccca gcaggcgaaa atcctgtttg
     3841 atggtggtta acggcgggat ataacatgag ctgtcttcgg tatcgtcgta tcccactacc
     3901 gagatatccg caccaacgcg cagcccggac tcggtaatgg cgcgcattgc gcccagcgcc
     3961 atctgatcgt tggcaaccag catcgcagtg ggaacgatgc cctcattcag catttgcatg
     4021 gtttgttgaa aaccggacat ggcactccag tcgccttccc gttccgctat cggctgaatt
     4081 tgattgcgag tgagatattt atgccagcca gccagacgca gacgcgccga gacagaactt
     4141 aatgggcccg ctaacagcgc gatttgctgg tgacccaatg cgaccagatg ctccacgccc
     4201 agtcgcgtac cgtcttcatg ggagaaaata atactgttga tgggtgtctg gtcagagaca
     4261 tcaagaaata acgccggaac attagtgcag gcagcttcca cagcaatggc atcctggtca
     4321 tccagcggat agttaatgat cagcccactg acgcgttgcg cgagaagatt gtgcaccgcc
     4381 gctttacagg cttcgacgcc gcttcgttct accatcgaca ccaccacgct ggcacccagt
     4441 tgatcggcgc gagatttaat cgccgcgaca atttgcgacg gcgcgtgcag ggccagactg
     4501 gaggtggcaa cgccaatcag caacgactgt ttgcccgcca gttgttgtgc cacgcggttg
     4561 ggaatgtaat tcagctccgc catcgccgct tccacttttt cccgcgtttt cgcagaaacg
     4621 tggctggcct ggttcaccac gcgggaaacg gtctgataag agacaccggc atactctgcg
     4681 acatcgtata acgttactgg tttcacattc accaccctga attgactctc ttccgggcgc
     4741 tatcatgcca taccgcgaaa ggttttgcgc cattcgatgg tgtccgggat ctcgacgctc
     4801 tcccttatgc gactcctgca ttaggaagca gcccagtagt aggttgaggc cgttgagcac
     4861 cgccgccgca aggaatggtg catgcaagga gatggcgccc aacagtcccc cggccacggg
     4921 gcctgccacc atacccacgc cgaaacaagc gctcatgagc ccgaagtggc gagcccgatc
     4981 ttccccatcg gtgatgtcgg cgatataggc gccagcaacc gcacctgtgg cgccggtgat
     5041 gccggccacg atgcgtccgg cgtagaggat cgagatctcg atcccgcgaa attaatacga
     5101 ctcactatag gggaattgtg agcggataac aattcccctc tagaaataat tttgtttaac
     5161 tttaagaagg agatatacat atgagcattt tcccgaagat tagcctgcgc ccggaagtgg
     5221 agaattacct gaaagagggt tttatgaaca aggagatcgt taccgcgctg ggcaagcaag
     5281 aggcggaacg taaattcgaa accctgctga agcacctgag ccacccgccg agctttacca
     5341 ccgtgcgtgt taacacccac ctggcgagcg tgcagcacgt taaaaacctg ctgctggatg
     5401 aactgcagaa gcaattcaac ggtctgagcg ttccgatcct gcaacacccg gatctgcagg
     5461 acgtgctgct gatcccggtt attggcccgc gtaaaaacat caagaaacag caatgcgagg
     5521 cgattgttgg tgcgcaatgc ggtaacgcgg ttctgcgtgg tgcgcatgtt tatgcgccgg
     5581 gcattgtgag cgcgagccag ttcatgaagg cgggtgatgt gatcagcgtt tatagcgaca
     5641 ttaagggtaa atgcaagaaa ggcgcgaaag aattcgacgg taccaaggtt tttctgggta
     5701 acggcatcag cgagctgagc cgtaaagaaa tttttagcgg cctgccggag ctgaagggta
     5761 tgggcatccg tatgaccgaa ccggtttacc tgagcccgag cttcgatagc gtgctgccgc
     5821 gttacctgtt cctgcaaaac ctgccgagcg cgctggtgag ccacgttctg aacccgcagc
     5881 cgggcgagaa aattctggac ctgtgcgcgg cgccgggtgg caagaccacc cacattgcgg
     5941 cgctgatgca cgatcaaggc gaggttatcg cgctggacaa gatcttcaac aaggtggaaa
     6001 agatcaagca gaacgcgctg ctgctgggtc tgaacagcat tcgtgcgttc tgctttgatg
     6061 gcaccaaggc ggtgaaactg gatatggttg aggacaccga gggtgaaccg ccgttcctgc
     6121 cggaaagctt tgatcgtatc ctgctggatg cgccgtgcag cggtatgggt cagcgtccga
     6181 acatggcgtg cacctggagc gtgaaagagg ttgcgagcta ccaaccgctg cagcgtaaac
     6241 tgtttaccgc ggcggttcaa ctgctgaagc cggagggtgt gctggtttat agcacctgca
     6301 ccatcaccct ggcggaaaac gaggaacagg tggcgtgggc gctgaccaaa tttccgtgcc
     6361 tgcagctgca accgcaggag ccgcaaattg gtggcgaagg catgcgtggt gcgggcctga
     6421 gctgcgaaca gctgaagcaa ctgcagcgtt ttgatccgag cgcggtgccg ctgccggaca
     6481 ccgatatgga cagcctgcgt gaggcgcgtc gtgaagacat gctgcgtctg gcgaataagg
     6541 acagcattgg ctttttcatt gcgaagtttg ttaagtgcaa gagcaccctc gagcaccacc
     6601 accaccacca ctgagatccg gctgctaaca aagcccgaaa ggaagctgag ttggctgctg
     6661 ccaccgctga gcaataacta gcataacccc ttggggcctc taaacgggtc ttgaggggtt
     6721 ttttgctgaa aggaggaact atatccggat tggcgaatgg gacgcgccct
//
