LOCUS       sequence-436606-        5716 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses NSD2 in bacterial cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5716)
  AUTHORS   Weirich S, Kusevic D, Schnee P, Reiter J, Pleiss J, Jeltsch A
  TITLE     Discovery of NSD2 non-histone substrates and design of a 
            super-substrate.
  JOURNAL   Commun Biol. 2024 Jun 8;7(1):707. doi: 10.1038/s42003-024-06395-z.
  PUBMED    38851815
REFERENCE   2  (bases 1 to 5716)
  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: "Commun 
            Biol."; date: "2024-06-8"; volume: "7(1):707. doi"; pages: " 
            10.1038/s42003-024-06395-z"
COMMENT     Sequence Label: pGex-6p2_NSD2
FEATURES             Location/Qualifiers
     source          1..5716
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     743..762
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(782..859)
                     /gene="lacI"
                     /label=lacI promoter
     primer_bind     complement(932..949)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1103..1691)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1183..1202)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(1862..2653,2654..2722))
                     /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     2485..2504
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(2723..2827)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     2895..2913
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     3058..3080
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(3905..3928)
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     CDS             complement(3935..4588)
                     /codon_start=1
                     /product="glutathione S-transferase from Schistosoma 
                     japonicum"
                     /label=GST
                     /translation="MSPILGYWKIKGLVQPTRLLLEYLEEKYEEHLYERDEGDKWRNKK
                     FELGLEFPNLPYYIDGDVKLTQSMAIIRYIADKHNMLGGCPKERAEISMLEGAVLDIRY
                     GVSRIAYSKDFETLKVDFLSKLPEMLKMFEDRLCHKTYLNGDHVTHPDFMLYDALDVVL
                     YMDPMCLDAFPKLVCFKKRIEAIPQIDKYLKSSKYIAWPLQGWQATFGGGDHPPK"
     primer_bind     complement(3955..3977)
                     /label=pGEX 5'
                     /note="pGEX vectors, Glutathione-S-transferase, forward 
                     primer"
     protein_bind    4611..4627
                     /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(4635..4641,4642..4657,4658..4663))
                     /label=tac promoter
                     /note="strong E. coli promoter; hybrid between the trp and 
                     lac UV5 promoters"
     CDS             complement(4765..5292)
                     /codon_start=1
                     /gene="lacZ fragment"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /label=lacZ-alpha
                     /translation="MTMITDSLAVVLQRRDWENPGVTQLNRLAAHPPFASWRNSEEART
                     DRPSQQLRSLNGEWRFAWFPAPEAVPESWLECDLPEADTVVVPSNWQMHGYDAPIYTNV
                     TYPITVNPPFVPTENPTGCYSLTFNVDESWLQEGQTRIIFDGVGISLSTARCTNASGVR
                     QPSEAVVWLCRS"
     primer_bind     5241..5263
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     5255..5272
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     5256..5272
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5288..5304)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5288..5304)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     protein_bind    5312..5328
                     /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(5336..5342,5343..5360,5361..5366))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     CDS             complement(join(5415..5716,1..781))
                     /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 cggactcggt aatggcgcgc attgcgccca gcgccatctg atcgttggca accagcatcg
       61 cagtgggaac gatgccctca ttcagcattt gcatggtttg ttgaaaaccg gacatggcac
      121 tccagtcgcc ttcccgttcc gctatcggct gaatttgatt gcgagtgaga tatttatgcc
      181 agccagccag acgcagacgc gccgagacag aacttaatgg gcccgctaac agcgcgattt
      241 gctggtgacc caatgcgacc agatgctcca cgcccagtcg cgtaccgtct tcatgggaga
      301 aaataatact gttgatgggt gtctggtcag agacatcaag aaataacgcc ggaacattag
      361 tgcaggcagc ttccacagca atggcatcct ggtcatccag cggatagtta atgatcagcc
      421 cactgacgcg ttgcgcgaga agattgtgca ccgccgcttt acaggcttcg acgccgcttc
      481 gttctaccat cgacaccacc acgctggcac ccagttgatc ggcgcgagat ttaatcgccg
      541 cgacaatttg cgacggcgcg tgcagggcca gactggaggt ggcaacgcca atcagcaacg
      601 actgtttgcc cgccagttgt tgtgccacgc ggttgggaat gtaattcagc tccgccatcg
      661 ccgcttccac tttttcccgc gttttcgcag aaacgtggct ggcctggttc accacgcggg
      721 aaacggtctg ataagagaca ccggcatact ctgcgacatc gtataacgtt actggtttca
      781 cattcaccac cctgaattga ctctcttccg ggcgctatca tgccataccg cgaaaggttt
      841 tgcgccattc gatggtgtcg gaatttatgc ggtgtgaaat accgcacaga tgcgtaagga
      901 gaaaataccg catcaggcgc tcttccgctt cctcgctcac tgactcgctg cgctcggtcg
      961 ttcggctgcg gcgagcggta tcagctcact caaaggcggt aatacggtta tccacagaat
     1021 caggggataa cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta
     1081 aaaaggccgc gttgctggcg tttttccata ggctccgccc ccctgacgag catcacaaaa
     1141 atcgacgctc aagtcagagg tggcgaaacc cgacaggact ataaagatac caggcgtttc
     1201 cccctggaag ctccctcgtg cgctctcctg ttccgaccct gccgcttacc ggatacctgt
     1261 ccgcctttct cccttcggga agcgtggcgc tttctcatag ctcacgctgt aggtatctca
     1321 gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc gttcagcccg
     1381 accgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggtaaga cacgacttat
     1441 cgccactggc agcagccact ggtaacagga ttagcagagc gaggtatgta ggcggtgcta
     1501 cagagttctt gaagtggtgg cctaactacg gctacactag aaggacagta tttggtatct
     1561 gcgctctgct gaagccagtt accttcggaa aaagagttgg tagctcttga tccggcaaac
     1621 aaaccaccgc tggtagcggt ggtttttttg tttgcaagca gcagattacg cgcagaaaaa
     1681 aaggatctca agaagatcct ttgatctttt ctacggggtc tgacgctcag tggaacgaaa
     1741 actcacgtta agggattttg gtcatgagat tatcaaaaag gatcttcacc tagatccttt
     1801 taaattaaaa atgaagtttt aaatcaatct aaagtatata tgagtaaact tggtctgaca
     1861 gttaccaatg cttaatcagt gaggcaccta tctcagcgat ctgtctattt cgttcatcca
     1921 tagttgcctg actccccgtc gtgtagataa ctacgatacg ggagggctta ccatctggcc
     1981 ccagtgctgc aatgataccg cgagacccac gctcaccggc tccagattta tcagcaataa
     2041 accagccagc cggaagggcc gagcgcagaa gtggtcctgc aactttatcc gcctccatcc
     2101 agtctattaa ttgttgccgg gaagctagag taagtagttc gccagttaat agtttgcgca
     2161 acgttgttgc cattgctgca ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat
     2221 tcagctccgg ttcccaacga tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag
     2281 cggttagctc cttcggtcct ccgatcgttg tcagaagtaa gttggccgca gtgttatcac
     2341 tcatggttat ggcagcactg cataattctc ttactgtcat gccatccgta agatgctttt
     2401 ctgtgactgg tgagtactca accaagtcat tctgagaata gtgtatgcgg cgaccgagtt
     2461 gctcttgccc ggcgtcaaca cgggataata ccgcgccaca tagcagaact ttaaaagtgc
     2521 tcatcattgg aaaacgttct tcggggcgaa aactctcaag gatcttaccg ctgttgagat
     2581 ccagttcgat gtaacccact cgtgcaccca actgatcttc agcatctttt actttcacca
     2641 gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga ataagggcga
     2701 cacggaaatg ttgaatactc atactcttcc tttttcaata ttattgaagc atttatcagg
     2761 gttattgtct catgagcgga tacatatttg aatgtattta gaaaaataaa caaatagggg
     2821 ttccgcgcac atttccccga aaagtgccac ctgacgtcta agaaaccatt attatcatga
     2881 cattaaccta taaaaatagg cgtatcacga ggccctttcg tcttcaagaa ttatacactc
     2941 cgctatcgct acgtgactgg gtcatggctg cgccccgaca cccgccaaca cccgctgacg
     3001 cgccctgacg ggcttgtctg ctcccggcat ccgcttacag acaagctgtg accgtctccg
     3061 ggagctgcat gtgtcagagg ttttcaccgt catcaccgaa acgcgcgagg cagatcgtca
     3121 gtcagtcacg atgcggccgc tcgaggtcct ctgactgcct cttcccttcc ccttttgctc
     3181 tgcgccgcct cgttttcttc ttggtctttt tgcccttttc ctctgatgaa agggtcgtcg
     3241 aggtctttgg tctatccccg aggaatccac tgcaattgga ggctccacac cggcagaccg
     3301 ttttttcatt gcccagacaa tcgaggttgt agttaaaagt cagctccgtc cctgcaggaa
     3361 tgtcacagac ggcaaacagg cccacacgag tgtccccatt cactgtccac ttgagggtct
     3421 cacagttggg ctggcagctg tgattcataa atcgagagta gtttcctttg gggccagcgt
     3481 ctattatacg gtccttgtct atagtgagca tgtagaagtg ggtgatgtcg ttctcgtgtg
     3541 cgtgcttgat tctcgccatg cactcctcct cgtcgatcag ctccccaacg tactcgttaa
     3601 caaattctcc ctttctgatg tccctcttgg cgaccaggcc ccaccctttg ccatctgtct
     3661 tgatgatctt ggtctctggg tactggcgct tggtgaagca ctggttctgg cagaactcgc
     3721 ccgcgggaca cacctgcggg tggcactcaa acatcagcat cctgttcaga cactccgaat
     3781 caaagccaca aggattctca tctgtgggct tgcagttgca cttagggatt tctgaaatat
     3841 ccgctgtgta gatctggact ttcccgtaag gcttattcac cttgatgtgc ttgtaggatc
     3901 ccaggggccc ctggaacaga acttccagat ccgattttgg aggatggtcg ccaccaccaa
     3961 acgtggcttg ccagccctgc aaaggccatg ctatatactt gctggatttc aagtacttat
     4021 caatttgtgg gatagcttca atacgttttt taaaacaaac taattttggg aacgcatcca
     4081 ggcacattgg gtccatgtat aaaacaacat caagagcgtc atacaacatg aagtcaggat
     4141 gggttacatg atcaccattt aaatatgttt tatgacataa acgatcttcg aacattttca
     4201 gcatttcagg tagcttgcta agaaaatcaa ctttgagagt ttcaaagtct ttactatatg
     4261 caattctcga aacaccgtat ctaatatcca aaaccgctcc ttcaagcatt gaaatctctg
     4321 cacgctcttt tggacaacca cccaacatgt tgtgcttgtc agctatataa cgtatgatgg
     4381 ccatagactg tgttaattta acatcaccat caatataata aggaagattg ggaaactcca
     4441 aacccaattc aaactttttg tttcgccatt tatcaccttc atcgcgctca tacaaatgct
     4501 cttcatattt ttcttcaaga tattccaaaa gaagtcgagt gggttgcaca aggcccttaa
     4561 ttttccaata acctagtata ggggacatga atactgtttc ctgtgtgaaa ttgttatccg
     4621 ctcacaattc cacacattat acgagccgat gattaattgt caacagctca tttcagaata
     4681 tttgccagaa ccgttatgat gtcggcgcaa aaaacattat ccagaacggg agtgcgcctt
     4741 gagcgacacg aattatgcag tgatttacga cctgcacagc cataccacag cttccgatgg
     4801 ctgcctgacg ccagaagcat tggtgcaccg tgcagtcgat aagctaattc caacgccatc
     4861 aaaaataatt cgcgtctggc cttcctgtag ccagctttca tcaacattaa atgtgagcga
     4921 gtaacaaccc gtcggattct ccgtgggaac aaacggcgga ttgaccgtaa tgggataggt
     4981 cacgttggtg tagatgggcg catcgtaacc gtgcatctgc cagtttgagg ggacgacgac
     5041 agtatcggcc tcaggaagat cgcactccag ccagctttcc ggcaccgctt ctggtgccgg
     5101 aaaccaggca aagcgccatt cgccattcag gctgcgcaac tgttgggaag ggcgatcggt
     5161 gcgggcctct tcgctattac gccagctggc gaaaggggga tgtgctgcaa ggcgattaag
     5221 ttgggtaacg ccagggtttt cccagtcacg acgttgtaaa acgacggcca gtgaatccgt
     5281 aatcatggtc atagctgttt cctgtgtgaa attgttatcc gctcacaatt ccacacaaca
     5341 tacgagccgg aagcataaag tgtaaagcct ggggtgccta atgagtgagc taacttacat
     5401 taattgcgtt gcgctcactg cccgctttcc agtcgggaaa cctgtcgtgc cagctgcatt
     5461 aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat tgggcgctag cgtggttttt
     5521 cttttcacca gtgagacggg caacagctga ttgcccttca ccgcctggcc ctgagagagt
     5581 tgcagcaagc ggtccacgct ggtttgcccc agcaggcgaa aatcctgttt gatggtggtt
     5641 aacggcggga tataacatga gctgtcttcg gtatcgtcgt atcccactac cgagatatcc
     5701 gcaccaacgc gcagcc
//
