LOCUS       sequence-204394-        6469 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Co-expression of Drosophila histones H3 and H4.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6469)
  AUTHORS   Khuong MT, Fei J, Cruz-Becerra G, Kadonaga JT
  TITLE     A simple and versatile system for the ATP-dependent assembly of 
            chromatin.
  JOURNAL   J Biol Chem. 2017 Nov 24;292(47):19478-19490. doi: 
            10.1074/jbc.M117.815365. Epub 2017 Oct 5.
  PUBMED    28982979
REFERENCE   2  (bases 1 to 6469)
  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: 
            "10.1074/jbc.M117.815365"; journalName: "J Biol Chem"; date: 
            "2017-11-24- 24"; volume: "292"; issue: "47"; pages: "19478-19490"
COMMENT     Sequence Label: pET11a-dH3-dH4
FEATURES             Location/Qualifiers
     source          1..6469
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     106..125
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     promoter        209..227
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    228..252
                     /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             267..289
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     promoter        728..746
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    747..771
                     /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             786..808
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     primer_bind     complement(1178..1196)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      1192..1239
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     promoter        complement(join(1414..1419,1420..1436,1437..1442))
                     /label=tet promoter
                     /note="E. coli promoter for tetracycline efflux protein 
                     gene"
     primer_bind     complement(1469..1487)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        1555..1659
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1660..1728,1729..2520)
                     /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     complement(1878..1897)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2691..3279
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3180..3199
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3433..3450
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    3465..3604
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(3530..3549)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3690..3712
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(3706..3897)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(5209..6291)
                     /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     6253..6272
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(6292..6369)
                     /gene="lacI"
                     /label=lacI promoter
ORIGIN
        1 caaggagatg gcgcccaaca gtcccccggc cacggggcct gccaccatac ccacgccgaa
       61 acaagcgctc atgagcccga agtggcgagc ccgatcttcc ccatcggtga tgtcggcgat
      121 ataggcgcca gcaaccgcac ctgtggcgcc ggtgatgccg gccacgatgc gtccggcgta
      181 gaggatcgag atctcgatcc cgcgaaatta atacgactca ctatagggga attgtgagcg
      241 gataacaatt cccctctaga aataattttg tttaacttta agaaggagat atacatatgg
      301 cgcgtaccaa acagactgcg cgtaaatcta ccggtggtaa agctccgcgt aaacagctgg
      361 ctaccaaagc agcgcgtaaa tccgctccgg ccactggcgg cgttaaaaag ccgcatcgtt
      421 atcgcccggg tacggttgct ctgcgcgaaa ttcgtcgtta ccagaagtct actgagctgc
      481 tgatccgtaa actgccgttc caacgtctgg tgcgtgaaat cgctcaggac tttaaaactg
      541 acctgcgttt tcaatcctcc gctgttatgg cgctgcagga agcgtctgag gcgtacctgg
      601 taggcctgtt cgaagacacc aatctgtgtg ctatccacgc gaaacgcgtg accatcatgc
      661 caaaagacat ccagctggct cgtcgtatcc gcggcgaacg tgcttaagga tctcgatccc
      721 gcgaaattaa tacgactcac tataggggaa ttgtgagcgg ataacaattc ccctctagaa
      781 ataattttgt ttaactttaa gaaggagata tacatatgac cggtcgtggc aaaggcggta
      841 aaggcctggg caaaggtggc gcaaaacgcc accgtaaggt gctgcgtgat aacattcagg
      901 gcatcaccaa accggccatc cgtcgcctgg cacgtcgcgg tggcgttaaa cgtattagcg
      961 gtctgatcta cgaagagact cgtggcgtac tgaaagtatt tctggaaaac gtgatccgcg
     1021 acgctgttac ttacactgag cacgcgaaac gtaaaacggt gaccgccctg gacgtagttt
     1081 acgcgctgaa acgtcagggt cgcaccctgt atggcttcgg tggctaagga tccggctgct
     1141 aacaaagccc gaaaggaagc tgagttggct gctgccaccg ctgagcaata actagcataa
     1201 ccccttgggg cctctaaacg ggtcttgagg ggttttttgc tgaaaggagg aactatatcc
     1261 ggatatcccg caagaggccc ggcagtaccg gcataaccaa gcctatgcct acagcatcca
     1321 gggtgacggt gccgaggatg acgatgagcg cattgttaga tttcatacac ggtgcctgac
     1381 tgcgttagca atttaactgt gataaactac cgcattaaag cttatcgatg ataagctgtc
     1441 aaacatgaga attcttgaag acgaaagggc ctcgtgatac gcctattttt ataggttaat
     1501 gtcatgataa taatggtttc ttagacgtca ggtggcactt ttcggggaaa tgtgcgcgga
     1561 acccctattt gtttattttt ctaaatacat tcaaatatgt atccgctcat gagacaataa
     1621 ccctgataaa tgcttcaata atattgaaaa aggaagagta tgagtattca acatttccgt
     1681 gtcgccctta ttcccttttt tgcggcattt tgccttcctg tttttgctca cccagaaacg
     1741 ctggtgaaag taaaagatgc tgaagatcag ttgggtgcac gagtgggtta catcgaactg
     1801 gatctcaaca gcggtaagat ccttgagagt tttcgccccg aagaacgttt tccaatgatg
     1861 agcactttta aagttctgct atgtggcgcg gtattatccc gtgttgacgc cgggcaagag
     1921 caactcggtc gccgcataca ctattctcag aatgacttgg ttgagtactc accagtcaca
     1981 gaaaagcatc ttacggatgg catgacagta agagaattat gcagtgctgc cataaccatg
     2041 agtgataaca ctgcggccaa cttacttctg acaacgatcg gaggaccgaa ggagctaacc
     2101 gcttttttgc acaacatggg ggatcatgta actcgccttg atcgttggga accggagctg
     2161 aatgaagcca taccaaacga cgagcgtgac accacgatgc ctgcagcaat ggcaacaacg
     2221 ttgcgcaaac tattaactgg cgaactactt actctagctt cccggcaaca attaatagac
     2281 tggatggagg cggataaagt tgcaggacca cttctgcgct cggcccttcc ggctggctgg
     2341 tttattgctg ataaatctgg agccggtgag cgtgggtctc gcggtatcat tgcagcactg
     2401 gggccagatg gtaagccctc ccgtatcgta gttatctaca cgacggggag tcaggcaact
     2461 atggatgaac gaaatagaca gatcgctgag ataggtgcct cactgattaa gcattggtaa
     2521 ctgtcagacc aagtttactc atatatactt tagattgatt taaaacttca tttttaattt
     2581 aaaaggatct aggtgaagat cctttttgat aatctcatga ccaaaatccc ttaacgtgag
     2641 ttttcgttcc actgagcgtc agaccccgta gaaaagatca aaggatcttc ttgagatcct
     2701 ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt
     2761 tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt cagcagagcg
     2821 cagataccaa atactgtcct tctagtgtag ccgtagttag gccaccactt caagaactct
     2881 gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc tgccagtggc
     2941 gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa ggcgcagcgg
     3001 tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac ctacaccgaa
     3061 ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg
     3121 gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga gcttccaggg
     3181 ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact tgagcgtcga
     3241 tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa cgcggccttt
     3301 ttacggttcc tggccttttg ctggcctttt gctcacatgt tctttcctgc gttatcccct
     3361 gattctgtgg ataaccgtat taccgccttt gagtgagctg ataccgctcg ccgcagccga
     3421 acgaccgagc gcagcgagtc agtgagcgag gaagcggaag agcgcctgat gcggtatttt
     3481 ctccttacgc atctgtgcgg tatttcacac cgcaatggtg cactctcagt acaatctgct
     3541 ctgatgccgc atagttaagc cagtatacac tccgctatcg ctacgtgact gggtcatggc
     3601 tgcgccccga cacccgccaa cacccgctga cgcgccctga cgggcttgtc tgctcccggc
     3661 atccgcttac agacaagctg tgaccgtctc cgggagctgc atgtgtcaga ggttttcacc
     3721 gtcatcaccg aaacgcgcga ggcagctgcg gtaaagctca tcagcgtggt cgtgaagcga
     3781 ttcacagatg tctgcctgtt catccgcgtc cagctcgttg agtttctcca gaagcgttaa
     3841 tgtctggctt ctgataaagc gggccatgtt aagggcggtt ttttcctgtt tggtcactga
     3901 tgcctccgtg taagggggat ttctgttcat gggggtaatg ataccgatga aacgagagag
     3961 gatgctcacg atacgggtta ctgatgatga acatgcccgg ttactggaac gttgtgaggg
     4021 taaacaactg gcggtatgga tgcggcggga ccagagaaaa atcactcagg gtcaatgcca
     4081 gcgcttcgtt aatacagatg taggtgttcc acagggtagc cagcagcatc ctgcgatgca
     4141 gatccggaac ataatggtgc agggcgctga cttccgcgtt tccagacttt acgaaacacg
     4201 gaaaccgaag accattcatg ttgttgctca ggtcgcagac gttttgcagc agcagtcgct
     4261 tcacgttcgc tcgcgtatcg gtgattcatt ctgctaacca gtaaggcaac cccgccagcc
     4321 tagccgggtc ctcaacgaca ggagcacgat catgcgcacc cgtggccagg acccaacgct
     4381 gcccgagatg cgccgcgtgc ggctgctgga gatggcggac gcgatggata tgttctgcca
     4441 agggttggtt tgcgcattca cagttctccg caagaattga ttggctccaa ttcttggagt
     4501 ggtgaatccg ttagcgaggt gccgccggct tccattcagg tcgaggtggc ccggctccat
     4561 gcaccgcgac gcaacgcggg gaggcagaca aggtataggg cggcgcctac aatccatgcc
     4621 aacccgttcc atgtgctcgc cgaggcggca taaatcgccg tgacgatcag cggtccaatg
     4681 atcgaagtta ggctggtaag agccgcgagc gatccttgaa gctgtccctg atggtcgtca
     4741 tctacctgcc tggacagcat ggcctgcaac gcgggcatcc cgatgccgcc ggaagcgaga
     4801 agaatcataa tggggaaggc catccagcct cgcgtcgcga acgccagcaa gacgtagccc
     4861 agcgcgtcgg ccgccatgcc ggcgataatg gcctgcttct cgccgaaacg tttggtggcg
     4921 ggaccagtga cgaaggcttg agcgagggcg tgcaagattc cgaataccgc aagcgacagg
     4981 ccgatcatcg tcgcgctcca gcgaaagcgg tcctcgccga aaatgaccca gagcgctgcc
     5041 ggcacctgtc ctacgagttg catgataaag aagacagtca taagtgcggc gacgatagtc
     5101 atgccccgcg cccaccggaa ggagctgact gggttgaagg ctctcaaggg catcggtcga
     5161 gatcccggtg cctaatgagt gagctaactt acattaattg cgttgcgctc actgcccgct
     5221 ttccagtcgg gaaacctgtc gtgccagctg cattaatgaa tcggccaacg cgcggggaga
     5281 ggcggtttgc gtattgggcg ccagggtggt ttttcttttc accagtgaga cgggcaacag
     5341 ctgattgccc ttcaccgcct ggccctgaga gagttgcagc aagcggtcca cgctggtttg
     5401 ccccagcagg cgaaaatcct gtttgatggt ggttaacggc gggatataac atgagctgtc
     5461 ttcggtatcg tcgtatccca ctaccgagat atccgcacca acgcgcagcc cggactcggt
     5521 aatggcgcgc attgcgccca gcgccatctg atcgttggca accagcatcg cagtgggaac
     5581 gatgccctca ttcagcattt gcatggtttg ttgaaaaccg gacatggcac tccagtcgcc
     5641 ttcccgttcc gctatcggct gaatttgatt gcgagtgaga tatttatgcc agccagccag
     5701 acgcagacgc gccgagacag aacttaatgg gcccgctaac agcgcgattt gctggtgacc
     5761 caatgcgacc agatgctcca cgcccagtcg cgtaccgtct tcatgggaga aaataatact
     5821 gttgatgggt gtctggtcag agacatcaag aaataacgcc ggaacattag tgcaggcagc
     5881 ttccacagca atggcatcct ggtcatccag cggatagtta atgatcagcc cactgacgcg
     5941 ttgcgcgaga agattgtgca ccgccgcttt acaggcttcg acgccgcttc gttctaccat
     6001 cgacaccacc acgctggcac ccagttgatc ggcgcgagat ttaatcgccg cgacaatttg
     6061 cgacggcgcg tgcagggcca gactggaggt ggcaacgcca atcagcaacg actgtttgcc
     6121 cgccagttgt tgtgccacgc ggttgggaat gtaattcagc tccgccatcg ccgcttccac
     6181 tttttcccgc gttttcgcag aaacgtggct ggcctggttc accacgcggg aaacggtctg
     6241 ataagagaca ccggcatact ctgcgacatc gtataacgtt actggtttca cattcaccac
     6301 cctgaattga ctctcttccg ggcgctatca tgccataccg cgaaaggttt tgcgccattc
     6361 gatggtgtcc gggatctcga cgctctccct tatgcgactc ctgcattagg aagcagccca
     6421 gtagtaggtt gaggccgttg agcaccgccg ccgcaaggaa tggtgcatg
//
