LOCUS       sequence-436975-        8829 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses NSD2 in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8829)
  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 8829)
  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: pECFP_C1_NSD2 FL
FEATURES             Location/Qualifiers
     source          1..8829
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        61..364
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        365..568
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     519..539
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      607..616
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(613..615,616..618,619..1329)
                     /codon_start=1
                     /product="enhanced CFP"
                     /label=ECFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTWGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYISHNVYITADKQKNGIK
                     ANFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDERYK"
     primer_bind     complement(658..679)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(919..938)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1266..1287
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     CDS             4014..4025
                     /codon_start=1
                     /product="WELQut protease recognition and cleavage site"
                     /label=WELQut site
                     /translation="WELQ"
     polyA_signal    5617..5738
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5654..5673)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5708..5727
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(5745..6200)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5882..5903)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     6094..6113
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        6227..6331
                     /gene="bla"
                     /label=AmpR promoter
     promoter        6333..6690
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        6333..6524
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(6356..6376)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      6541..6676
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             6725..7519
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     CDS             6725..7519
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(6779..6798)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     7389..7408
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(7707..7726)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    7751..7798
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      8127..8715
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     8616..8635
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata tggagttccg
       61 cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt
      121 gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca
      181 atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc
      241 aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta
      301 catgacctta tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac
      361 catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg actcacgggg
      421 atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg
      481 ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg gtaggcgtgt
      541 acggtgggag gtctatataa gcagagctgg tttagtgaac cgtcagatcc gctagcgcta
      601 ccggtcgcca ccatggtgag caagggcgag gagctgttca ccggggtggt gcccatcctg
      661 gtcgagctgg acggcgacgt aaacggccac aagttcagcg tgtccggcga gggcgagggc
      721 gatgccacct acggcaagct gaccctgaag ttcatctgca ccaccggcaa gctgcccgtg
      781 ccctggccca ccctcgtgac caccctgacc tggggcgtgc agtgcttcag ccgctacccc
      841 gaccacatga agcagcacga cttcttcaag tccgccatgc ccgaaggcta cgtccaggag
      901 cgcaccatct tcttcaagga cgacggcaac tacaagaccc gcgccgaggt gaagttcgag
      961 ggcgacaccc tggtgaaccg catcgagctg aagggcatcg acttcaagga ggacggcaac
     1021 atcctggggc acaagctgga gtacaactac atcagccaca acgtctatat caccgccgac
     1081 aagcagaaga acggcatcaa ggccaacttc aagatccgcc acaacatcga ggacggcagc
     1141 gtgcagctcg ccgaccacta ccagcagaac acccccatcg gcgacggccc cgtgctgctg
     1201 cccgacaacc actacctgag cacccagtcc gccctgagca aagaccccaa cgagaagcgc
     1261 gatcacatgg tcctgctgga gttcgtgacc gccgccggga tcactctcgg catggacgag
     1321 cggtacaagt ccggactcag atctcgagct atggaattta gcatcaagca gagtcccctt
     1381 tctgttcaga gtgttgtaaa gtgcataaag atgaagcagg caccagaaat cctcggcagt
     1441 gccaacggga agactccgag ctgcgaggtg aaccgcgagt gttctgtgtt cctcagcaaa
     1501 gcccagctct ccagtagcct gcaggagggg gtcatgcaga agtttaacgg ccacgacgcc
     1561 ctgcccttta ttccagccga caagctgaaa gatcttactt cccgggtgtt taatggagaa
     1621 cccggcgcac acgatgccaa actgcgtttt gagtcccagg aaatgaaagg gattgggaca
     1681 ccccctaaca ctacccctat caaaaatggc tctccagaaa ttaagctgaa aatcaccaaa
     1741 acatacatga atgggaagcc tctctttgaa tcttccattt gtggtgacag tgctgctgat
     1801 gtgtctcagt cagaagaaaa tggacaaaaa ccagaaaaca aggcgagaag gaacaggaag
     1861 aggagcataa aatatgactc cttgctggag cagggccttg tcgaagcagc tcttgtgtct
     1921 aagatctcaa gtccttcaga taaaaagatt ccagctaaga aagagtcttg tccaaacact
     1981 ggaagagaca aagaccacct gttgaaatac aacgttggtg atttggtgtg gtccaaagtg
     2041 tcgggttacc cttggtggcc ttgcatggtt tctgcagatc cactccttca cagctatacc
     2101 aaacttaaag gtcagaaaaa gagtgcacgc cagtatcacg tacagttctt tggtgacgcc
     2161 ccagaaagag cttggatatt tgagaagagc ctcgtagctt ttgaaggaga aggacagttt
     2221 gaaaaattat gccaggaaag tgccaagcag gcacccacga aagctgagaa aattaagcta
     2281 ttgaaaccaa tttcagggaa attgagggcc cagtgggaaa tgggcattgt tcaagcagaa
     2341 gaagctgcaa gcatgtcagt ggaggagcgg aaagccaagt tcacctttct ctatgtgggg
     2401 gaccagcttc atctcaaccc tcaagtagcc aaggaggctg gcattgctgc agagtctttg
     2461 ggagaaatgg cagaatcctc aggagtcagt gaagaagctg ctgaaaaccc caagtctgtg
     2521 agagaagagt gcattcccat gaagagaagg cggagggcca aactgtgtag ctctgcagag
     2581 accctggaga gtcaccccga catagggaag agtactcctc aaaagacggc agaggctgac
     2641 cccagaagag gagtagggtc tcctcctggg aggaagaaga ccacagtctc catgccacga
     2701 agcaggaagg gagatgcagc atcccagttt ttggtcttct gtcaaaaaca cagggatgag
     2761 gtggtagctg agcacccaga tgcttcaggt gaggagattg aagagctgct caggtcacag
     2821 tggagtctgc tgagtgagaa gcagagagca cgctacaaca ccaagtttgc cctggtggcc
     2881 cctgtccagg ctgaagaaga ctctggtaat gtaaatggga aaaaaagaaa ccacacaaag
     2941 aggatacagg accctacaga agatgctgaa gctgaggaca cacccaggaa aagactcagg
     3001 acggacaagc acagtcttcg gaagagagac acaatcactg acaaaacggc cagaacaagc
     3061 tcttacaagg ccatggaggc agcctcctcg ctcaagagcc aggcagcaac gaaaaatctg
     3121 tctgatgcat gtaaaccact gaagaagcga aatcgggctt ccacggcagc atcttcagct
     3181 cttgggttta gcaaaagttc atctccttct gcatccttaa ctgagaatga ggtctcggac
     3241 agcccgggag acgagccctc ggagtcccca tacgaaagtg cagacgaaac acaaactgaa
     3301 gtatctgtct catccaaaaa gtctgagcga ggagtgactg ccaaaaagga gtatgtgtgc
     3361 cagctgtgtg agaagccggg cagcctcctg ctctgtgaag gaccctgctg cggagctttc
     3421 cacctcgcct gccttgggct ttcccggagg ccagaaggga ggttcacctg cagcgagtgt
     3481 gcctcaggga ttcactcatg tttcgtgtgt aaagagagca agacagatgt taagcgctgt
     3541 gtggtaactc agtgtggaaa attttaccat gaggcttgtg tgaaaaaata ccctctgact
     3601 gtatttgaga gccgaggttt ccgctgcccc ctccacagct gtgtgagctg ccatgcttcc
     3661 aacccttcaa acccaaggcc gtcaaaaggt aaaatgatgc ggtgtgtccg ctgccccgtt
     3721 gcctatcaca gcggggatgc ttgtctggca gcaggatgct cagtgatcgc ctccaacagc
     3781 atcatctgca ctgcccactt cactgctcgg aaggggaagc gacaccacgc ccacgtcaac
     3841 gtgagctggt gcttcgtgtg ctccaaaggg gggagccttc tgtgctgtga gtcctgccca
     3901 gcggccttcc accctgactg cctgaacatc gagatgcctg acggcagctg gttctgcaat
     3961 gactgcaggg ctgggaagaa gctgcacttc caggatatca tttgggtgaa acttgggaac
     4021 tacagatggt ggccggcaga agtttgccat cccaaaaatg ttcccccaaa tattcagaaa
     4081 atgaagcacg agattggaga attccctgtg tttttctttg ggtctaaaga ttattactgg
     4141 acgcatcagg cgcgagtgtt cccgtacatg gagggggacc ggggcagccg ctaccagggg
     4201 gtcagaggga tcggaagagt cttcaaaaac gcactgcaag aagctgaagc tcgttttcgt
     4261 gaaattaagc ttcagaggga agcccgagaa acacaggaga gcgagcgcaa gcccccacca
     4321 tacaagcaca tcaaggtgaa taagccttac gggaaagtcc agatctacac agcggatatt
     4381 tcagaaatcc ctaagtgcaa ctgcaagccc acagatgaga atccttgtgg ctttgattcg
     4441 gagtgtctga acaggatgct gatgtttgag tgccacccgc aggtgtgtcc cgcgggcgag
     4501 ttctgccaga accagtgctt caccaagcgc cagtacccag agaccaagat catcaagaca
     4561 gatggcaaag ggtggggcct ggtcgccaag agggacatca gaaagggaga atttgttaac
     4621 gagtacgttg gggagctgat cgacgaggag gagtgcatgg cgagaatcaa gcacgcacac
     4681 gagaacgaca tcacccactt ctacatgctc actatagaca aggaccgtat aatagacgct
     4741 ggccccaaag gaaactactc tcgatttatg aatcacagct gccagcccaa ctgtgagacc
     4801 ctcaagtgga cagtgaatgg ggacactcgt gtgggcctgt ttgccgtctg tgacattcct
     4861 gcagggacgg agctgacttt taactacaac ctcgattgtc tgggcaatga aaaaacggtc
     4921 tgccggtgtg gagcctccaa ttgcagtgga ttcctcgggg atagaccaaa gacctcgacg
     4981 accctttcat cagaggaaaa gggcaaaaag accaagaaga aaacgaggcg gcgcagagca
     5041 aaaggggaag ggaagaggca gtcagaggac gagtgcttcc gctgcggtga tggcgggcag
     5101 ctggtgctgt gtgaccgcaa gttctgcacc aaggcctacc acctgtcctg cctgggcctt
     5161 ggcaagcggc ccttcgggaa gtgggaatgt ccttggcatc attgtgacgt gtgtggcaaa
     5221 ccttcgactt cattttgcca cctctgcccc aattcgttct gtaaggagca ccaggacggg
     5281 acagccttca gctgcacccc ggacgggcgg tcctactgct gtgagcatga cttaggggcg
     5341 gcatcggtca gaagcaccaa gactgagaag ccccccccag agccagggaa gccgaagggg
     5401 aagaggcggc ggcggagggg ctggcggaga gtcacagagg gcaaataaca agcttcgaat
     5461 tctgcagtcg acgataccgc gggcccggga tccaccggat ctagataact gatcataatc
     5521 agccatacca catttgtaga ggttttactt gctttaaaaa acctcccaca cctccccctg
     5581 aacctgaaac ataaaatgaa tgcaattgtt gttgttaact tgtttattgc agcttataat
     5641 ggttacaaat aaagcaatag catcacaaat ttcacaaata aagcattttt ttcactgcat
     5701 tctagttgtg gtttgtccaa actcatcaat gtatcttaag gcgtaaattg taagcgttaa
     5761 tattttgtta aaattcgcgt taaatttttg ttaaatcagc tcatttttta accaataggc
     5821 cgaaatcggc aaaatccctt ataaatcaaa agaatagacc gagatagggt tgagtgttgt
     5881 tccagtttgg aacaagagtc cactattaaa gaacgtggac tccaacgtca aagggcgaaa
     5941 aaccgtctat cagggcgatg gcccactacg tgaaccatca ccctaatcaa gttttttggg
     6001 gtcgaggtgc cgtaaagcac taaatcggaa ccctaaaggg agcccccgat ttagagcttg
     6061 acggggaaag ccggcgaacg tggcgagaaa ggaagggaag aaagcgaaag gagcgggcgc
     6121 tagggcgctg gcaagtgtag cggtcacgct gcgcgtaacc accacacccg ccgcgcttaa
     6181 tgcgccgcta cagggcgcgt caggtggcac ttttcgggga aatgtgcgcg gaacccctat
     6241 ttgtttattt ttctaaatac attcaaatat gtatccgctc atgagacaat aaccctgata
     6301 aatgcttcaa taatattgaa aaaggaagag tcctgaggcg gaaagaacca gctgtggaat
     6361 gtgtgtcagt tagggtgtgg aaagtcccca ggctccccag caggcagaag tatgcaaagc
     6421 atgcatctca attagtcagc aaccaggtgt ggaaagtccc caggctcccc agcaggcaga
     6481 agtatgcaaa gcatgcatct caattagtca gcaaccatag tcccgcccct aactccgccc
     6541 atcccgcccc taactccgcc cagttccgcc cattctccgc cccatggctg actaattttt
     6601 tttatttatg cagaggccga ggccgcctcg gcctctgagc tattccagaa gtagtgagga
     6661 ggcttttttg gaggcctagg cttttgcaaa gatcgatcaa gagacaggat gaggatcgtt
     6721 tcgcatgatt gaacaagatg gattgcacgc aggttctccg gccgcttggg tggagaggct
     6781 attcggctat gactgggcac aacagacaat cggctgctct gatgccgccg tgttccggct
     6841 gtcagcgcag gggcgcccgg ttctttttgt caagaccgac ctgtccggtg ccctgaatga
     6901 actgcaagac gaggcagcgc ggctatcgtg gctggccacg acgggcgttc cttgcgcagc
     6961 tgtgctcgac gttgtcactg aagcgggaag ggactggctg ctattgggcg aagtgccggg
     7021 gcaggatctc ctgtcatctc accttgctcc tgccgagaaa gtatccatca tggctgatgc
     7081 aatgcggcgg ctgcatacgc ttgatccggc tacctgccca ttcgaccacc aagcgaaaca
     7141 tcgcatcgag cgagcacgta ctcggatgga agccggtctt gtcgatcagg atgatctgga
     7201 cgaagagcat caggggctcg cgccagccga actgttcgcc aggctcaagg cgagcatgcc
     7261 cgacggcgag gatctcgtcg tgacccatgg cgatgcctgc ttgccgaata tcatggtgga
     7321 aaatggccgc ttttctggat tcatcgactg tggccggctg ggtgtggcgg accgctatca
     7381 ggacatagcg ttggctaccc gtgatattgc tgaagagctt ggcggcgaat gggctgaccg
     7441 cttcctcgtg ctttacggta tcgccgctcc cgattcgcag cgcatcgcct tctatcgcct
     7501 tcttgacgag ttcttctgag cgggactctg gggttcgaaa tgaccgacca agcgacgccc
     7561 aacctgccat cacgagattt cgattccacc gccgccttct atgaaaggtt gggcttcgga
     7621 atcgttttcc gggacgccgg ctggatgatc ctccagcgcg gggatctcat gctggagttc
     7681 ttcgcccacc ctagggggag gctaactgaa acacggaagg agacaatacc ggaaggaacc
     7741 cgcgctatga cggcaataaa aagacagaat aaaacgcacg gtgttgggtc gtttgttcat
     7801 aaacgcgggg ttcggtccca gggctggcac tctgtcgata ccccaccgag accccattgg
     7861 ggccaatacg cccgcgtttc ttccttttcc ccaccccacc ccccaagttc gggtgaaggc
     7921 ccagggctcg cagccaacgt cggggcggca ggccctgcca tagcctcagg ttactcatat
     7981 atactttaga ttgatttaaa acttcatttt taatttaaaa ggatctaggt gaagatcctt
     8041 tttgataatc tcatgaccaa aatcccttaa cgtgagtttt cgttccactg agcgtcagac
     8101 cccgtagaaa agatcaaagg atcttcttga gatccttttt ttctgcgcgt aatctgctgc
     8161 ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt tgccggatca agagctacca
     8221 actctttttc cgaaggtaac tggcttcagc agagcgcaga taccaaatac tgttcttcta
     8281 gtgtagccgt agttaggcca ccacttcaag aactctgtat caccgcctac atacctcgct
     8341 ctgctaatcc tgttaccagt ggctgctgcc agtggcgata agtcgtgtct taccgggttg
     8401 gactcaagac gatagttacc ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc
     8461 acacagccca gcttggagcg aacgacctac accgaactga gatacctaca gcgtgagcta
     8521 tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca ggtatccggt aagcggcagg
     8581 gtcggaacag gagagcgcac gagggagctt ccagggggaa acgcctggta tctttatagt
     8641 cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg
     8701 cggagcctat ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc cttttgctgg
     8761 ccttttgctc acatgttctt tcctgcgtta tcccctgatt ctgtggataa ccgtattacc
     8821 gccatgcat
//
