LOCUS       sequence-386607-        6168 bp    DNA     circular SYN 15-APR-2025
DEFINITION  Human (hu) histones H2Ax and H2B inserted into a plasmid.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6168)
  TITLE     Preparation of recombinant nucleosomes
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 6168)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: 
            "Unpublished"
COMMENT     Sequence Label: His-huH2Ax-huH2B
FEATURES             Location/Qualifiers
     source          1..6168
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    3..27
                     /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             42..64
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             83..100
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             113..133
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     promoter        637..655
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    656..680
                     /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)."
     CDS             723..1103
                     /codon_start=1
                     /gene="HIST1H2BJ"
                     /product="human histone H2B"
                     /label=H2B
                     /translation="MPEPAKSAPAPKKGSKKAVTKAQKKDGKKRKRSRKESYSIYVYKV
                     LKQVHPDTGISSKAMGIMNSFVNDIFERIAGEASRLAHYNKRSTITSREIQTAVRLLLP
                     GELAKHAVSEGTKAVTKYTSAK"
     CDS             1117..1161
                     /codon_start=1
                     /product="affinity and epitope tag derived from pancreatic 
                     ribonuclease A"
                     /label=S-Tag
                     /translation="KETAAAKFERQHMDS"
     primer_bind     complement(1199..1217)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      1213..1260
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      1297..1752
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1384..1403)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1594..1615
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(join(1867..2658,2659..2727))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     2490..2509
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(2728..2820)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      2901..3489
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3390..3409
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3643..3660
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    3675..3814
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(3740..3759)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3900..3922
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(3916..4107)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(4679..5761)
                     /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     5723..5742
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(5762..5839)
                     /gene="lacI"
                     /label=lacI promoter
     primer_bind     6045..6064
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     promoter        join(6152..6168,1..2)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
ORIGIN
        1 ggggaattgt gagcggataa caattcccct ctagaaataa ttttgtttaa ctttaagaag
       61 gagatatacc atgggcagca gccatcacca tcatcaccac agccaggatc cagagaacct
      121 ctacttccaa ggcatgtcgg gccgcggcaa gactggcggc aaggcccgcg ccaaggccaa
      181 gtcgcgctcg tcgcgcgccg gcctccagtt cccagtgggc cgtgtacacc ggctgctgcg
      241 gaagggccac tacgccgagc gcgttggcgc cggcgcgcca gtgtacctgg cggcagtgct
      301 ggagtacctc accgctgaga tcctggagct ggcgggcaat gcggcccgcg acaacaagaa
      361 gacgcgaatc atcccccgcc acctgcagct ggccatccgc aacgacgagg agctcaacaa
      421 gctgctgggc ggcgtgacga tcgcccaggg aggcgtcctg cccaacatcc aggccgtgct
      481 gctgcccaag aagaccagcg ccaccgtggg gccgaaggcg ccctcgggcg gcaagaaggc
      541 cgagcaggac tccgaggagt actaaaagct tgcggccgca taatgcttaa gtcgaacaga
      601 aagtaatcgt attgtacacg gccgcataat cgaaattaat acgactcact ataggggaat
      661 tgtgagcgga taacaattcc ccatcttagt atattagtta agtataagaa ggagatatac
      721 atatgccaga gccagcgaag tctgctcccg ccccgaaaaa gggctccaag aaggcggtga
      781 ctaaggcgca gaagaaagac ggcaagaagc gcaagcgcag ccgcaaggag agctattcca
      841 tctatgtgta caaggttctg aagcaggtcc accctgacac cggcatttcg tccaaggcca
      901 tgggcatcat gaattcgttt gtgaacgaca ttttcgagcg catcgcaggt gaggcttccc
      961 gcctggcgca ttacaacaag cgctcgacca tcacctccag ggagatccag acggccgtgc
     1021 gcctgctgct gcctggggag ttggccaagc acgccgtgtc cgagggtact aaggccgtca
     1081 ccaagtacac cagcgctaag tagtctcgag tctggtaaag aaaccgctgc tgcgaaattt
     1141 gaacgccagc acatggactc gtctactagc gcagcttaat taacctaggc tgctgccacc
     1201 gctgagcaat aactagcata accccttggg gcctctaaac gggtcttgag gggttttttg
     1261 ctgaaaggag gaactatatc cggattggcg aatgggacgc gccctgtagc ggcgcattaa
     1321 gcgcggcggg tgtggtggtt acgcgcagcg tgaccgctac acttgccagc gccctagcgc
     1381 ccgctccttt cgctttcttc ccttcctttc tcgccacgtt cgccggcttt ccccgtcaag
     1441 ctctaaatcg ggggctccct ttagggttcc gatttagtgc tttacggcac ctcgacccca
     1501 aaaaacttga ttagggtgat ggttcacgta gtgggccatc gccctgatag acggtttttc
     1561 gccctttgac gttggagtcc acgttcttta atagtggact cttgttccaa actggaacaa
     1621 cactcaaccc tatctcggtc tattcttttg atttataagg gattttgccg atttcggcct
     1681 attggttaaa aaatgagctg atttaacaaa aatttaacgc gaattttaac aaaatattaa
     1741 cgtttacaat ttctggcggc acgatggcat gagattatca aaaaggatct tcacctagat
     1801 ccttttaaat taaaaatgaa gttttaaatc aatctaaagt atatatgagt aaacttggtc
     1861 tgacagttac caatgcttaa tcagtgaggc acctatctca gcgatctgtc tatttcgttc
     1921 atccatagtt gcctgactcc ccgtcgtgta gataactacg atacgggagg gcttaccatc
     1981 tggccccagt gctgcaatga taccgcgaga cccacgctca ccggctccag atttatcagc
     2041 aataaaccag ccagccggaa gggccgagcg cagaagtggt cctgcaactt tatccgcctc
     2101 catccagtct attaattgtt gccgggaagc tagagtaagt agttcgccag ttaatagttt
     2161 gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc
     2221 ttcattcagc tccggttccc aacgatcaag gcgagttaca tgatccccca tgttgtgcaa
     2281 aaaagcggtt agctccttcg gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt
     2341 atcactcatg gttatggcag cactgcataa ttctcttact gtcatgccat ccgtaagatg
     2401 cttttctgtg actggtgagt actcaaccaa gtcattctga gaatagtgta tgcggcgacc
     2461 gagttgctct tgcccggcgt caatacggga taataccgcg ccacatagca gaactttaaa
     2521 agtgctcatc attggaaaac gttcttcggg gcgaaaactc tcaaggatct taccgctgtt
     2581 gagatccagt tcgatgtaac ccactcgtgc acccaactga tcttcagcat cttttacttt
     2641 caccagcgtt tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa agggaataag
     2701 ggcgacacgg aaatgttgaa tactcatact cttccttttt caatcatgat tgaagcattt
     2761 atcagggtta ttgtctcatg agcggataca tatttgaatg tatttagaaa aataaacaaa
     2821 taggtcatga ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc agaccccgta
     2881 gaaaagatca aaggatcttc ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa
     2941 acaaaaaaac caccgctacc agcggtggtt tgtttgccgg atcaagagct accaactctt
     3001 tttccgaagg taactggctt cagcagagcg cagataccaa atactgtcct tctagtgtag
     3061 ccgtagttag gccaccactt caagaactct gtagcaccgc ctacatacct cgctctgcta
     3121 atcctgttac cagtggctgc tgccagtggc gataagtcgt gtcttaccgg gttggactca
     3181 agacgatagt taccggataa ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag
     3241 cccagcttgg agcgaacgac ctacaccgaa ctgagatacc tacagcgtga gctatgagaa
     3301 agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga
     3361 acaggagagc gcacgaggga gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc
     3421 gggtttcgcc acctctgact tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc
     3481 ctatggaaaa acgccagcaa cgcggccttt ttacggttcc tggccttttg ctggcctttt
     3541 gctcacatgt tctttcctgc gttatcccct gattctgtgg ataaccgtat taccgccttt
     3601 gagtgagctg ataccgctcg ccgcagccga acgaccgagc gcagcgagtc agtgagcgag
     3661 gaagcggaag agcgcctgat gcggtatttt ctccttacgc atctgtgcgg tatttcacac
     3721 cgcaatggtg cactctcagt acaatctgct ctgatgccgc atagttaagc cagtatacac
     3781 tccgctatcg ctacgtgact gggtcatggc tgcgccccga cacccgccaa cacccgctga
     3841 cgcgccctga cgggcttgtc tgctcccggc atccgcttac agacaagctg tgaccgtctc
     3901 cgggagctgc atgtgtcaga ggttttcacc gtcatcaccg aaacgcgcga ggcagctgcg
     3961 gtaaagctca tcagcgtggt cgtgaagcga ttcacagatg tctgcctgtt catccgcgtc
     4021 cagctcgttg agtttctcca gaagcgttaa tgtctggctt ctgataaagc gggccatgtt
     4081 aagggcggtt ttttcctgtt tggtcactga tgcctccgtg taagggggat ttctgttcat
     4141 gggggtaatg ataccgatga aacgagagag gatgctcacg atacgggtta ctgatgatga
     4201 acatgcccgg ttactggaac gttgtgaggg taaacaactg gcggtatgga tgcggcggga
     4261 ccagagaaaa atcactcagg gtcaatgcca gcgcttcgtt aatacagatg taggtgttcc
     4321 acagggtagc cagcagcatc ctgcgatgca gatccggaac ataatggtgc agggcgctga
     4381 cttccgcgtt tccagacttt acgaaacacg gaaaccgaag accattcatg ttgttgctca
     4441 ggtcgcagac gttttgcagc agcagtcgct tcacgttcgc tcgcgtatcg gtgattcatt
     4501 ctgctaacca gtaaggcaac cccgccagcc tagccgggtc ctcaacgaca ggagcacgat
     4561 catgctagtc atgccccgcg cccaccggaa ggagctgact gggttgaagg ctctcaaggg
     4621 catcggtcga gatcccggtg cctaatgagt gagctaactt acattaattg cgttgcgctc
     4681 actgcccgct ttccagtcgg gaaacctgtc gtgccagctg cattaatgaa tcggccaacg
     4741 cgcggggaga ggcggtttgc gtattgggcg ccagggtggt ttttcttttc accagtgaga
     4801 cgggcaacag ctgattgccc ttcaccgcct ggccctgaga gagttgcagc aagcggtcca
     4861 cgctggtttg ccccagcagg cgaaaatcct gtttgatggt ggttaacggc gggatataac
     4921 atgagctgtc ttcggtatcg tcgtatccca ctaccgagat gtccgcacca acgcgcagcc
     4981 cggactcggt aatggcgcgc attgcgccca gcgccatctg atcgttggca accagcatcg
     5041 cagtgggaac gatgccctca ttcagcattt gcatggtttg ttgaaaaccg gacatggcac
     5101 tccagtcgcc ttcccgttcc gctatcggct gaatttgatt gcgagtgaga tatttatgcc
     5161 agccagccag acgcagacgc gccgagacag aacttaatgg gcccgctaac agcgcgattt
     5221 gctggtgacc caatgcgacc agatgctcca cgcccagtcg cgtaccgtct tcatgggaga
     5281 aaataatact gttgatgggt gtctggtcag agacatcaag aaataacgcc ggaacattag
     5341 tgcaggcagc ttccacagca atggcatcct ggtcatccag cggatagtta atgatcagcc
     5401 cactgacgcg ttgcgcgaga agattgtgca ccgccgcttt acaggcttcg acgccgcttc
     5461 gttctaccat cgacaccacc acgctggcac ccagttgatc ggcgcgagat ttaatcgccg
     5521 cgacaatttg cgacggcgcg tgcagggcca gactggaggt ggcaacgcca atcagcaacg
     5581 actgtttgcc cgccagttgt tgtgccacgc ggttgggaat gtaattcagc tccgccatcg
     5641 ccgcttccac tttttcccgc gttttcgcag aaacgtggct ggcctggttc accacgcggg
     5701 aaacggtctg ataagagaca ccggcatact ctgcgacatc gtataacgtt actggtttca
     5761 cattcaccac cctgaattga ctctcttccg ggcgctatca tgccataccg cgaaaggttt
     5821 tgcgccattc gatggtgtcc gggatctcga cgctctccct tatgcgactc ctgcattagg
     5881 aagcagccca gtagtaggtt gaggccgttg agcaccgccg ccgcaaggaa tggtgcatgc
     5941 aaggagatgg cgcccaacag tcccccggcc acggggcctg ccaccatacc cacgccgaaa
     6001 caagcgctca tgagcccgaa gtggcgagcc cgatcttccc catcggtgat gtcggcgata
     6061 taggcgccag caaccgcacc tgtggcgccg gtgatgccgg ccacgatgcg tccggcgtag
     6121 aggatcgaga tcgatctcga tcccgcgaaa ttaatacgac tcactata
//
