LOCUS       sequence-469844-        6126 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  expresses His-tagged human SET in E coli.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6126)
  AUTHORS   Seibert M, Kruger M, Watson NA, Sen O, Daum JR, Slotman JA, Braun T,
            Houtsmuller AB, Gorbsky GJ, Jacob R, Kracht M, Higgins JMG, Schmitz 
            ML
  TITLE     CDK1-mediated phosphorylation at H2B serine 6 is required for 
            mitotic chromosome segregation.
  JOURNAL   J Cell Biol. 2019 Apr 1;218(4):1164-1181. doi: 
            10.1083/jcb.201806057. Epub 2019 Feb 14.
  PUBMED    30765437
REFERENCE   2  (bases 1 to 6126)
  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: 
            "10.1083/jcb.201806057"; journalName: "J Cell Biol"; date: 
            "2019-04-1- 1"; volume: "218"; issue: "4"; pages: "1164-1181"
COMMENT     Sequence Label: pET28a-SET
FEATURES             Location/Qualifiers
     source          1..6126
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      complement(26..73)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     primer_bind     69..87
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     CDS             complement(140..157)
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             complement(1004..1021)
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
     CDS             complement(1031..1048)
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     RBS             1067..1089
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     protein_bind    1104..1128
                     /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     complement(1128..1147)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1129..1147)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1231..1250)
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     promoter        1456..1533
                     /gene="lacI"
                     /label=lacI promoter
     CDS             1534..2616
                     /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(1553..1572)
                     /label=LacI-R
                     /note="LacI, reverse primer"
     CDS             3425..3616
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     primer_bind     complement(3610..3632)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     misc_feature    3718..3857
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     3773..3792
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(3872..3889)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4043..4631)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4123..4142)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     complement(4742..4761)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             4753..5568
                     /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             4753..5568
                     /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     complement(4826..4845)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      complement(5660..6115)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5797..5818)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     6009..6028
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
ORIGIN
        1 atccggatat agttcctcct ttcagcaaaa aacccctcaa gacccgttta gaggccccaa
       61 ggggttatgc tagttattgc tcagcggtgg cagcagccaa ctcagcttcc tttcgggctt
      121 tgttagcagc cggatctcag tggtggtggt ggtggtgctc gagttagtca tcttctcctt
      181 catcctcctc tccttcctcc ccttcatcat catcttcatc ttcttcacct tcatcctcat
      241 ccccttcttc gtcaatatct tctaatcctt cctcctcttc atcatcatca tcatcttctt
      301 ctccttctcc ttcttcatca tccatatcgg gaaccaagta gtactgtaat gggtttggcc
      361 aaatatcatc tttgatgacc tctcctaact catcagcacc tgcatcagaa tggtcagtaa
      421 accaggtaaa gaagctctct ggttcctcat gctgcctctt cctgctggct ttattctgcg
      481 tttgactcga acgtttcgtc aaatcctttc cagatttcca tttgatttcg gtggacttcg
      541 aagatggatc accactctca ttcagatgaa attctttgga gagaacttta ttttcaaagt
      601 aaggattttc atcaaaataa aaatctattc tgtaacctga tttaatatct tcaaattctg
      661 tcacttcaac tctggtcaaa taatgcagtg cctcttcatc ttcctcccca agcagtgcag
      721 acacttgtgg atggttgaca aatgttgtta cccaaaaatt tgggattttg gcgatcaatt
      781 ctgacctctt ctgaaaaaat ggttggcgga gtttgttata tttctgttct actttcaaaa
      841 tctcctcact ggcttgttca ttaagtctgt ctatttcatt ttgtacttca tcaatgtgtt
      901 caatcgcttc ttgctgttct ttttctgagg tctcgtcggc cccgtcgtgg ttggagttga
      961 gctccttttt actgactttg gccgccggcg ccgacatcat atggctgccg cgcggcacca
     1021 ggccgctgct gtgatgatga tgatgatggc tgctgcccat ggtatatctc cttcttaaag
     1081 ttaaacaaaa ttatttctag aggggaattg ttatccgctc acaattcccc tatagtgagt
     1141 cgtattaatt tcgcgggatc gagatctcga tcctctacgc cggacgcatc gtggccggca
     1201 tcaccggcgc cacaggtgcg gttgctggcg cctatatcgc cgacatcacc gatggggaag
     1261 atcgggctcg ccacttcggg ctcatgagcg cttgtttcgg cgtgggtatg gtggcaggcc
     1321 ccgtggccgg gggactgttg ggcgccatct ccttgcatgc accattcctt gcggcggcgg
     1381 tgctcaacgg cctcaaccta ctactgggct gcttcctaat gcaggagtcg cataagggag
     1441 agcgtcgaga tcccggacac catcgaatgg cgcaaaacct ttcgcggtat ggcatgatag
     1501 cgcccggaag agagtcaatt cagggtggtg aatgtgaaac cagtaacgtt atacgatgtc
     1561 gcagagtatg ccggtgtctc ttatcagacc gtttcccgcg tggtgaacca ggccagccac
     1621 gtttctgcga aaacgcggga aaaagtggaa gcggcgatgg cggagctgaa ttacattccc
     1681 aaccgcgtgg cacaacaact ggcgggcaaa cagtcgttgc tgattggcgt tgccacctcc
     1741 agtctggccc tgcacgcgcc gtcgcaaatt gtcgcggcga ttaaatctcg cgccgatcaa
     1801 ctgggtgcca gcgtggtggt gtcgatggta gaacgaagcg gcgtcgaagc ctgtaaagcg
     1861 gcggtgcaca atcttctcgc gcaacgcgtc agtgggctga tcattaacta tccgctggat
     1921 gaccaggatg ccattgctgt ggaagctgcc tgcactaatg ttccggcgtt atttcttgat
     1981 gtctctgacc agacacccat caacagtatt attttctccc atgaagacgg tacgcgactg
     2041 ggcgtggagc atctggtcgc attgggtcac cagcaaatcg cgctgttagc gggcccatta
     2101 agttctgtct cggcgcgtct gcgtctggct ggctggcata aatatctcac tcgcaatcaa
     2161 attcagccga tagcggaacg ggaaggcgac tggagtgcca tgtccggttt tcaacaaacc
     2221 atgcaaatgc tgaatgaggg catcgttccc actgcgatgc tggttgccaa cgatcagatg
     2281 gcgctgggcg caatgcgcgc cattaccgag tccgggctgc gcgttggtgc ggatatctcg
     2341 gtagtgggat acgacgatac cgaagacagc tcatgttata tcccgccgtt aaccaccatc
     2401 aaacaggatt ttcgcctgct ggggcaaacc agcgtggacc gcttgctgca actctctcag
     2461 ggccaggcgg tgaagggcaa tcagctgttg cccgtctcac tggtgaaaag aaaaaccacc
     2521 ctggcgccca atacgcaaac cgcctctccc cgcgcgttgg ccgattcatt aatgcagctg
     2581 gcacgacagg tttcccgact ggaaagcggg cagtgagcgc aacgcaatta atgtaagtta
     2641 gctcactcat taggcaccgg gatctcgacc gatgcccttg agagccttca acccagtcag
     2701 ctccttccgg tgggcgcggg gcatgactat cgtcgccgca cttatgactg tcttctttat
     2761 catgcaactc gtaggacagg tgccggcagc gctctgggtc attttcggcg aggaccgctt
     2821 tcgctggagc gcgacgatga tcggcctgtc gcttgcggta ttcggaatct tgcacgccct
     2881 cgctcaagcc ttcgtcactg gtcccgccac caaacgtttc ggcgagaagc aggccattat
     2941 cgccggcatg gcggccccac gggtgcgcat gatcgtgctc ctgtcgttga ggacccggct
     3001 aggctggcgg ggttgcctta ctggttagca gaatgaatca ccgatacgcg agcgaacgtg
     3061 aagcgactgc tgctgcaaaa cgtctgcgac ctgagcaaca acatgaatgg tcttcggttt
     3121 ccgtgtttcg taaagtctgg aaacgcggaa gtcagcgccc tgcaccatta tgttccggat
     3181 ctgcatcgca ggatgctgct ggctaccctg tggaacacct acatctgtat taacgaagcg
     3241 ctggcattga ccctgagtga tttttctctg gtcccgccgc atccataccg ccagttgttt
     3301 accctcacaa cgttccagta accgggcatg ttcatcatca gtaacccgta tcgtgagcat
     3361 cctctctcgt ttcatcggta tcattacccc catgaacaga aatccccctt acacggaggc
     3421 atcagtgacc aaacaggaaa aaaccgccct taacatggcc cgctttatca gaagccagac
     3481 attaacgctt ctggagaaac tcaacgagct ggacgcggat gaacaggcag acatctgtga
     3541 atcgcttcac gaccacgctg atgagcttta ccgcagctgc ctcgcgcgtt tcggtgatga
     3601 cggtgaaaac ctctgacaca tgcagctccc ggagacggtc acagcttgtc tgtaagcgga
     3661 tgccgggagc agacaagccc gtcagggcgc gtcagcgggt gttggcgggt gtcggggcgc
     3721 agccatgacc cagtcacgta gcgatagcgg agtgtatact ggcttaacta tgcggcatca
     3781 gagcagattg tactgagagt gcaccattgc ggtgtgaaat accgcacaga tgcgtaagga
     3841 gaaaataccg catcaggcgc tcttccgctt cctcgctcac tgactcgctg cgctcggtcg
     3901 ttcggctgcg gcgagcggta tcagctcact caaaggcggt aatacggtta tccacagaat
     3961 caggggataa cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta
     4021 aaaaggccgc gttgctggcg tttttccata ggctccgccc ccctgacgag catcacaaaa
     4081 atcgacgctc aagtcagagg tggcgaaacc cgacaggact ataaagatac caggcgtttc
     4141 cccctggaag ctccctcgtg cgctctcctg ttccgaccct gccgcttacc ggatacctgt
     4201 ccgcctttct cccttcggga agcgtggcgc tttctcatag ctcacgctgt aggtatctca
     4261 gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc gttcagcccg
     4321 accgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggtaaga cacgacttat
     4381 cgccactggc agcagccact ggtaacagga ttagcagagc gaggtatgta ggcggtgcta
     4441 cagagttctt gaagtggtgg cctaactacg gctacactag aaggacagta tttggtatct
     4501 gcgctctgct gaagccagtt accttcggaa aaagagttgg tagctcttga tccggcaaac
     4561 aaaccaccgc tggtagcggt ggtttttttg tttgcaagca gcagattacg cgcagaaaaa
     4621 aaggatctca agaagatcct ttgatctttt ctacggggtc tgacgctcag tggaacgaaa
     4681 actcacgtta agggattttg gtcatgaaca ataaaactgt ctgcttacat aaacagtaat
     4741 acaaggggtg ttatgagcca tattcaacgg gaaacgtctt gctctaggcc gcgattaaat
     4801 tccaacatgg atgctgattt atatgggtat aaatgggctc gcgataatgt cgggcaatca
     4861 ggtgcgacaa tctatcgatt gtatgggaag cccgatgcgc cagagttgtt tctgaaacat
     4921 ggcaaaggta gcgttgccaa tgatgttaca gatgagatgg tcagactaaa ctggctgacg
     4981 gaatttatgc ctcttccgac catcaagcat tttatccgta ctcctgatga tgcatggtta
     5041 ctcaccactg cgatccccgg gaaaacagca ttccaggtat tagaagaata tcctgattca
     5101 ggtgaaaata ttgttgatgc gctggcagtg ttcctgcgcc ggttgcattc gattcctgtt
     5161 tgtaattgtc cttttaacag cgatcgcgta tttcgtctcg ctcaggcgca atcacgaatg
     5221 aataacggtt tggttgatgc gagtgatttt gatgacgagc gtaatggctg gcctgttgaa
     5281 caagtctgga aagaaatgca taaacttttg ccattctcac cggattcagt cgtcactcat
     5341 ggtgatttct cacttgataa ccttattttt gacgagggga aattaatagg ttgtattgat
     5401 gttggacgag tcggaatcgc agaccgatac caggatcttg ccatcctatg gaactgcctc
     5461 ggtgagtttt ctccttcatt acagaaacgg ctttttcaaa aatatggtat tgataatcct
     5521 gatatgaata aattgcagtt tcatttgatg ctcgatgagt ttttctaaga attaattcat
     5581 gagcggatac atatttgaat gtatttagaa aaataaacaa ataggggttc cgcgcacatt
     5641 tccccgaaaa gtgccaccta aattgtaagc gttaatattt tgttaaaatt cgcgttaaat
     5701 ttttgttaaa tcagctcatt ttttaaccaa taggccgaaa tcggcaaaat cccttataaa
     5761 tcaaaagaat agaccgagat agggttgagt gttgttccag tttggaacaa gagtccacta
     5821 ttaaagaacg tggactccaa cgtcaaaggg cgaaaaaccg tctatcaggg cgatggccca
     5881 ctacgtgaac catcacccta atcaagtttt ttggggtcga ggtgccgtaa agcactaaat
     5941 cggaacccta aagggagccc ccgatttaga gcttgacggg gaaagccggc gaacgtggcg
     6001 agaaaggaag ggaagaaagc gaaaggagcg ggcgctaggg cgctggcaag tgtagcggtc
     6061 acgctgcgcg taaccaccac acccgccgcg cttaatgcgc cgctacaggg cgcgtcccat
     6121 tcgcca
//
