LOCUS       sequence-305064-        6979 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Polycistronic coexpression of Xenopus laevis histones (H2A, H2B, H3 
            and H4).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6979)
  AUTHORS   Shim Y, Duan MR, Chen X, Smerdon MJ, Min JH
  TITLE     Polycistronic coexpression and nondenaturing purification of histone
            octamers.
  JOURNAL   Anal Biochem. 2012 Aug 15;427(2):190-2. doi: 
            10.1016/j.ab.2012.05.006. Epub 2012 May 19.
  PUBMED    22617796
REFERENCE   2  (bases 1 to 6979)
  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: 
            "10.1016/j.ab.2012.05.006"; journalName: "Anal Biochem"; date: 
            "2012-08-15- 15"; volume: "427"; issue: "2"; pages: "190-2"
COMMENT     Sequence Label: pET29a-YS14
FEATURES             Location/Qualifiers
     source          1..6979
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(165..182)
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
     CDS             complement(189..206)
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             complement(216..260)
                     /codon_start=1
                     /product="affinity and epitope tag derived from pancreatic 
                     ribonuclease A"
                     /label=S-Tag
                     /translation="KETAAAKFERQHMDS"
     RBS             271..293
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     protein_bind    308..332
                     /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(332..351)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(333..351)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(439..458)
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     promoter        664..741
                     /gene="lacI"
                     /label=lacI promoter
     CDS             742..1824
                     /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(761..780)
                     /label=LacI-R
                     /note="LacI, reverse primer"
     CDS             2633..2824
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     primer_bind     complement(2818..2840)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     misc_feature    2926..3065
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     2981..3000
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(3080..3097)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3251..3839)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3331..3350)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     complement(3950..3969)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             3961..4776
                     /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             3961..4776
                     /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(4034..4053)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      complement(4868..5323)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5005..5026)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     5217..5236
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     terminator      complement(5360..5407)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     primer_bind     5403..5421
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     CDS             complement(5474..5491)
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             complement(5498..5515)
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
     RBS             5841..5863
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     RBS             6296..6318
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     RBS             6710..6732
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
ORIGIN
        1 gccagataga ctggagctcc agctcccacc cgctctgcat aattgccttt ccttaacagc
       61 cggtgaacac ggccaacagg gaactgtagc ccagcccgag atgagcgagt cttggcctta
      121 gcgcgggttt taccgccttg tttgcctctt cctgacatac ccatggaacc gcgtggcacc
      181 agggtaccgt ggtggtggtg gtggtgcaga tctgggctgt ccatgtgctg gcgttcgaat
      241 ttagcagcag cggtttcttt catatgtata tctccttctt aaagttaaac aaaattattt
      301 ctagagggga attgttatcc gctcacaatt cccctatagt gagtcgtatt aatttcgcgg
      361 gatcgagatc gatctcgatc ctctacgccg gacgcatcgt ggccggcatc accggcgcca
      421 caggtgcggt tgctggcgcc tatatcgccg acatcaccga tggggaagat cgggctcgcc
      481 acttcgggct catgagcgct tgtttcggcg tgggtatggt ggcaggcccc gtggccgggg
      541 gactgttggg cgccatctcc ttgcatgcac cattccttgc ggcggcggtg ctcaacggcc
      601 tcaacctact actgggctgc ttcctaatgc aggagtcgca taagggagag cgtcgagatc
      661 ccggacacca tcgaatggcg caaaaccttt cgcggtatgg catgatagcg cccggaagag
      721 agtcaattca gggtggtgaa tgtgaaacca gtaacgttat acgatgtcgc agagtatgcc
      781 ggtgtctctt atcagaccgt ttcccgcgtg gtgaaccagg ccagccacgt ttctgcgaaa
      841 acgcgggaaa aagtggaagc ggcgatggcg gagctgaatt acattcccaa ccgcgtggca
      901 caacaactgg cgggcaaaca gtcgttgctg attggcgttg ccacctccag tctggccctg
      961 cacgcgccgt cgcaaattgt cgcggcgatt aaatctcgcg ccgatcaact gggtgccagc
     1021 gtggtggtgt cgatggtaga acgaagcggc gtcgaagcct gtaaagcggc ggtgcacaat
     1081 cttctcgcgc aacgcgtcag tgggctgatc attaactatc cgctggatga ccaggatgcc
     1141 attgctgtgg aagctgcctg cactaatgtt ccggcgttat ttcttgatgt ctctgaccag
     1201 acacccatca acagtattat tttctcccat gaagacggta cgcgactggg cgtggagcat
     1261 ctggtcgcat tgggtcacca gcaaatcgcg ctgttagcgg gcccattaag ttctgtctcg
     1321 gcgcgtctgc gtctggctgg ctggcataaa tatctcactc gcaatcaaat tcagccgata
     1381 gcggaacggg aaggcgactg gagtgccatg tccggttttc aacaaaccat gcaaatgctg
     1441 aatgagggca tcgttcccac tgcgatgctg gttgccaacg atcagatggc gctgggcgca
     1501 atgcgcgcca ttaccgagtc cgggctgcgc gttggtgcgg acatctcggt agtgggatac
     1561 gacgataccg aagacagctc atgttatatc ccgccgttaa ccaccatcaa acaggatttt
     1621 cgcctgctgg ggcaaaccag cgtggaccgc ttgctgcaac tctctcaggg ccaggcggtg
     1681 aagggcaatc agctgttgcc cgtctcactg gtgaaaagaa aaaccaccct ggcgcccaat
     1741 acgcaaaccg cctctccccg cgcgttggcc gattcattaa tgcagctggc acgacaggtt
     1801 tcccgactgg aaagcgggca gtgagcgcaa cgcaattaat gtaagttagc tcactcatta
     1861 ggcaccggga tctcgaccga tgcccttgag agccttcaac ccagtcagct ccttccggtg
     1921 ggcgcggggc atgactatcg tcgccgcact tatgactgtc ttctttatca tgcaactcgt
     1981 aggacaggtg ccggcagcgc tctgggtcat tttcggcgag gaccgctttc gctggagcgc
     2041 gacgatgatc ggcctgtcgc ttgcggtatt cggaatcttg cacgccctcg ctcaagcctt
     2101 cgtcactggt cccgccacca aacgtttcgg cgagaagcag gccattatcg ccggcatggc
     2161 ggccccacgg gtgcgcatga tcgtgctcct gtcgttgagg acccggctag gctggcgggg
     2221 ttgccttact ggttagcaga atgaatcacc gatacgcgag cgaacgtgaa gcgactgctg
     2281 ctgcaaaacg tctgcgacct gagcaacaac atgaatggtc ttcggtttcc gtgtttcgta
     2341 aagtctggaa acgcggaagt cagcgccctg caccattatg ttccggatct gcatcgcagg
     2401 atgctgctgg ctaccctgtg gaacacctac atctgtatta acgaagcgct ggcattgacc
     2461 ctgagtgatt tttctctggt cccgccgcat ccataccgcc agttgtttac cctcacaacg
     2521 ttccagtaac cgggcatgtt catcatcagt aacccgtatc gtgagcatcc tctctcgttt
     2581 catcggtatc attaccccca tgaacagaaa tcccccttac acggaggcat cagtgaccaa
     2641 acaggaaaaa accgccctta acatggcccg ctttatcaga agccagacat taacgcttct
     2701 ggagaaactc aacgagctgg acgcggatga acaggcagac atctgtgaat cgcttcacga
     2761 ccacgctgat gagctttacc gcagctgcct cgcgcgtttc ggtgatgacg gtgaaaacct
     2821 ctgacacatg cagctcccgg agacggtcac agcttgtctg taagcggatg ccgggagcag
     2881 acaagcccgt cagggcgcgt cagcgggtgt tggcgggtgt cggggcgcag ccatgaccca
     2941 gtcacgtagc gatagcggag tgtatactgg cttaactatg cggcatcaga gcagattgta
     3001 ctgagagtgc accattgcgg tgtgaaatac cgcacagatg cgtaaggaga aaataccgca
     3061 tcaggcgctc ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc
     3121 gagcggtatc agctcactca aaggcggtaa tacggttatc cacagaatca ggggataacg
     3181 caggaaagaa catgtgagca aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt
     3241 tgctggcgtt tttccatagg ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa
     3301 gtcagaggtg gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggaagct
     3361 ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc
     3421 cttcgggaag cgtggcgctt tctcatagct cacgctgtag gtatctcagt tcggtgtagg
     3481 tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct
     3541 tatccggtaa ctatcgtctt gagtccaacc cggtaagaca cgacttatcg ccactggcag
     3601 cagccactgg taacaggatt agcagagcga ggtatgtagg cggtgctaca gagttcttga
     3661 agtggtggcc taactacggc tacactagaa ggacagtatt tggtatctgc gctctgctga
     3721 agccagttac cttcggaaaa agagttggta gctcttgatc cggcaaacaa accaccgctg
     3781 gtagcggtgg tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag
     3841 aagatccttt gatcttttct acggggtctg acgctcagtg gaacgaaaac tcacgttaag
     3901 ggattttggt catgaacaat aaaactgtct gcttacataa acagtaatac aaggggtgtt
     3961 atgagccata ttcaacggga aacgtcttgc tctaggccgc gattaaattc caacatggat
     4021 gctgatttat atgggtataa atgggctcgc gataatgtcg ggcaatcagg tgcgacaatc
     4081 tatcgattgt atgggaagcc cgatgcgcca gagttgtttc tgaaacatgg caaaggtagc
     4141 gttgccaatg atgttacaga tgagatggtc agactaaact ggctgacgga atttatgcct
     4201 cttccgacca tcaagcattt tatccgtact cctgatgatg catggttact caccactgcg
     4261 atccccggga aaacagcatt ccaggtatta gaagaatatc ctgattcagg tgaaaatatt
     4321 gttgatgcgc tggcagtgtt cctgcgccgg ttgcattcga ttcctgtttg taattgtcct
     4381 tttaacagcg atcgcgtatt tcgtctcgct caggcgcaat cacgaatgaa taacggtttg
     4441 gttgatgcga gtgattttga tgacgagcgt aatggctggc ctgttgaaca agtctggaaa
     4501 gaaatgcata aacttttgcc attctcaccg gattcagtcg tcactcatgg tgatttctca
     4561 cttgataacc ttatttttga cgaggggaaa ttaataggtt gtattgatgt tggacgagtc
     4621 ggaatcgcag accgatacca ggatcttgcc atcctatgga actgcctcgg tgagttttct
     4681 ccttcattac agaaacggct ttttcaaaaa tatggtattg ataatcctga tatgaataaa
     4741 ttgcagtttc atttgatgct cgatgagttt ttctaagaat taattcatga gcggatacat
     4801 atttgaatgt atttagaaaa ataaacaaat aggggttccg cgcacatttc cccgaaaagt
     4861 gccacctaaa ttgtaagcgt taatattttg ttaaaattcg cgttaaattt ttgttaaatc
     4921 agctcatttt ttaaccaata ggccgaaatc ggcaaaatcc cttataaatc aaaagaatag
     4981 accgagatag ggttgagtgt tgttccagtt tggaacaaga gtccactatt aaagaacgtg
     5041 gactccaacg tcaaagggcg aaaaaccgtc tatcagggcg atggcccact acgtgaacca
     5101 tcaccctaat caagtttttt ggggtcgagg tgccgtaaag cactaaatcg gaaccctaaa
     5161 gggagccccc gatttagagc ttgacgggga aagccggcga acgtggcgag aaaggaaggg
     5221 aagaaagcga aaggagcggg cgctagggcg ctggcaagtg tagcggtcac gctgcgcgta
     5281 accaccacac ccgccgcgct taatgcgccg ctacagggcg cgtcccattc gccaatccgg
     5341 atatagttcc tcctttcagc aaaaaacccc tcaagacccg tttagaggcc ccaaggggtt
     5401 atgctagtta ttgctcagcg gtggcagcag ccaactcagc ttcctttcgg gctttgttag
     5461 cagccggatc tcagtggtgg tggtggtggt gctcgaggct gccgcgcggc accaggccgc
     5521 tgctaccacc gaaaccgtac agggtacgac cctgacgttt cagagcgtaa acaacgtcca
     5581 tagcggtaac ggttttacgt ttagcgtgtt cggtgtaggt aacagcgtca cggataacgt
     5641 tttccaggaa aactttcaga acaccgcggg tttcttcgta gatcagaccg gagatacgtt
     5701 taacaccacc acgacgagcc agacgacgga tagccggctt ggtgataccc tggatgttgt
     5761 cacgcagaac tttacggtga cgtttagcac cacctttacc cagaccttta ccacctttac
     5821 cacgaccaga catatgtata tctccttctt aaagttaaac aaaattattg cggccgccta
     5881 agccctctcg cctcggattc tgcgggccag ctggatgtcc ttgggcatga tggtgaccct
     5941 cttggcgtgg atggcacaca ggttggtgtc ctcaaagaga gcgaccagat aagcctcgct
     6001 ggcctcctgc agagccataa cggccgagct ctggaagcgc aggtcggtct tgaagtcctg
     6061 agcgatctcc cggaccaggc gctggaaagg cagtttgcgg atgagcagct cggtggattt
     6121 ctggtagcgg cggatctcgc ggagagcgac tgtgccgggc cggtaacggt gaggtttctt
     6181 gactccgccg gtagcaggag cggacttcct ggctgccttg gtggccagct gcttgcgggg
     6241 agccttccct ccggtggatt tacgggcggt ctgcttggta cgggccatat gtatatctcc
     6301 ttcttaaagt taaacaaaat tattgtcgac ttacttggcg ctggtgtact tggtgacagc
     6361 cttggtgccc tcggacacgg cgtgtttggc caactcccca ggcagcagca gtcggaccgc
     6421 ggtctggatc tcccgggagg tgatggtgga gcgcttgttg taatgagcca ggcgggaggc
     6481 ttcccctgcg atgcgctcaa acacatcgtt gacaaaggag ttcatgatgc tcatggcctt
     6541 ggacgagatg ccggtatcgg ggtgcacctg cttcagcacc ttgtacacgt aaatggcata
     6601 actctccttc cttgtcttcc tgcgcttttt cccgtctttc ttctgagtct tggtcaccgc
     6661 tttcttggag cctttcttcg gggctggagc ggacttggcc atatgtatat ctccttctta
     6721 aagttaaaca aaattattga attctcactt gctcttggcc gacttggaac tctcggtttt
     6781 cttgggcagc agcacggact ggatgttggg caggaccccg ccctgagcga tagtgactct
     6841 tccgagcagt ttgttcagtt cctcatcgtt gcgcacagcg agctgcaggt gtctggggat
     6901 aatgcgagtc ttcttgttat cacgggccgc attcccggcc aattccaaaa tctcagcggt
     6961 cagatactcc aacactgca
//
