LOCUS       sequence-204395-        6316 bp    DNA     circular SYN 14-APR-2025
DEFINITION  Expression of Drosophila histone H2B. An additional Ile codon was 
            inserted after the initiating Met codon to enhance the protein 
            expression level.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6316)
  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 6316)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 14, 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-dH2B
FEATURES             Location/Qualifiers
     source          1..6316
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     106..125
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     209..228
                     /label=T7
                     /note="T7 promoter, forward 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)"
     primer_bind     complement(1025..1043)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      1039..1086
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     promoter        complement(join(1261..1266,1267..1283,1284..1289))
                     /label=tet promoter
                     /note="E. coli promoter for tetracycline efflux protein 
                     gene"
     primer_bind     complement(1316..1334)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        1402..1506
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1507..1575,1576..2367)
                     /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(1725..1744)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2538..3126
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3027..3046
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3280..3297
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    3312..3451
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(3377..3396)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3537..3559
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(3553..3744)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(5056..6138)
                     /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     6100..6119
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(6139..6216)
                     /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 atacatatga
      301 ttccgccgaa aacctctgga gatatacata tgattccgcc gaaaacctct gggagatata
      361 catatgattc cgccgaaaac ctctgggaga tatacatatg attccgccga aaacctctgg
      421 gagatataca tatgattccg ccgaaaacct ctggatatac atatgattcc gccgaaaacc
      481 tctgatatac atatgattcc gccgaaaacc tctgggagat atacatatga ttccgccgaa
      541 aacctctggg agatatacat atgattccgc cgaaaacctc tggagatata catatgattc
      601 cgccgaaaac ctctggtaaa gcggctaaaa aagcgggtaa agctcagaaa aacatcacca
      661 aaactgacaa gaaaaagaaa cgcaaacgta aagaaagcta cgcaatctac atttacaagg
      721 ttctgaaaca ggttcacccg gacaccggca tctcttccaa agctatgagc attatgaact
      781 ctttcgttaa cgatatcttc gaacgtattg cggctgaagc gtcccgtctg gctcactaca
      841 acaaacgtag caccattact tctcgtgaaa tccaaacggc tgtgcgtctg ctgctgccgg
      901 gcgaactggc gaagcacgcg gtgtctgaag gtaccaaagc ggttacgaaa tacactagct
      961 ctaaataatg atagggatcc ggctgctaac aaagcccgaa aggaagctga gttggctgct
     1021 gccaccgctg agcaataact agcataaccc cttggggcct ctaaacgggt cttgaggggt
     1081 tttttgctga aaggaggaac tatatccgga tatcccgcaa gaggcccggc agtaccggca
     1141 taaccaagcc tatgcctaca gcatccaggg tgacggtgcc gaggatgacg atgagcgcat
     1201 tgttagattt catacacggt gcctgactgc gttagcaatt taactgtgat aaactaccgc
     1261 attaaagctt atcgatgata agctgtcaaa catgagaatt cttgaagacg aaagggcctc
     1321 gtgatacgcc tatttttata ggttaatgtc atgataataa tggtttctta gacgtcaggt
     1381 ggcacttttc ggggaaatgt gcgcggaacc cctatttgtt tatttttcta aatacattca
     1441 aatatgtatc cgctcatgag acaataaccc tgataaatgc ttcaataata ttgaaaaagg
     1501 aagagtatga gtattcaaca tttccgtgtc gcccttattc ccttttttgc ggcattttgc
     1561 cttcctgttt ttgctcaccc agaaacgctg gtgaaagtaa aagatgctga agatcagttg
     1621 ggtgcacgag tgggttacat cgaactggat ctcaacagcg gtaagatcct tgagagtttt
     1681 cgccccgaag aacgttttcc aatgatgagc acttttaaag ttctgctatg tggcgcggta
     1741 ttatcccgtg ttgacgccgg gcaagagcaa ctcggtcgcc gcatacacta ttctcagaat
     1801 gacttggttg agtactcacc agtcacagaa aagcatctta cggatggcat gacagtaaga
     1861 gaattatgca gtgctgccat aaccatgagt gataacactg cggccaactt acttctgaca
     1921 acgatcggag gaccgaagga gctaaccgct tttttgcaca acatggggga tcatgtaact
     1981 cgccttgatc gttgggaacc ggagctgaat gaagccatac caaacgacga gcgtgacacc
     2041 acgatgcctg cagcaatggc aacaacgttg cgcaaactat taactggcga actacttact
     2101 ctagcttccc ggcaacaatt aatagactgg atggaggcgg ataaagttgc aggaccactt
     2161 ctgcgctcgg cccttccggc tggctggttt attgctgata aatctggagc cggtgagcgt
     2221 gggtctcgcg gtatcattgc agcactgggg ccagatggta agccctcccg tatcgtagtt
     2281 atctacacga cggggagtca ggcaactatg gatgaacgaa atagacagat cgctgagata
     2341 ggtgcctcac tgattaagca ttggtaactg tcagaccaag tttactcata tatactttag
     2401 attgatttaa aacttcattt ttaatttaaa aggatctagg tgaagatcct ttttgataat
     2461 ctcatgacca aaatccctta acgtgagttt tcgttccact gagcgtcaga ccccgtagaa
     2521 aagatcaaag gatcttcttg agatcctttt tttctgcgcg taatctgctg cttgcaaaca
     2581 aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc aagagctacc aactcttttt
     2641 ccgaaggtaa ctggcttcag cagagcgcag ataccaaata ctgtccttct agtgtagccg
     2701 tagttaggcc accacttcaa gaactctgta gcaccgccta catacctcgc tctgctaatc
     2761 ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga
     2821 cgatagttac cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc
     2881 agcttggagc gaacgaccta caccgaactg agatacctac agcgtgagct atgagaaagc
     2941 gccacgcttc ccgaagggag aaaggcggac aggtatccgg taagcggcag ggtcggaaca
     3001 ggagagcgca cgagggagct tccaggggga aacgcctggt atctttatag tcctgtcggg
     3061 tttcgccacc tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta
     3121 tggaaaaacg ccagcaacgc ggccttttta cggttcctgg ccttttgctg gccttttgct
     3181 cacatgttct ttcctgcgtt atcccctgat tctgtggata accgtattac cgcctttgag
     3241 tgagctgata ccgctcgccg cagccgaacg accgagcgca gcgagtcagt gagcgaggaa
     3301 gcggaagagc gcctgatgcg gtattttctc cttacgcatc tgtgcggtat ttcacaccgc
     3361 aatggtgcac tctcagtaca atctgctctg atgccgcata gttaagccag tatacactcc
     3421 gctatcgcta cgtgactggg tcatggctgc gccccgacac ccgccaacac ccgctgacgc
     3481 gccctgacgg gcttgtctgc tcccggcatc cgcttacaga caagctgtga ccgtctccgg
     3541 gagctgcatg tgtcagaggt tttcaccgtc atcaccgaaa cgcgcgaggc agctgcggta
     3601 aagctcatca gcgtggtcgt gaagcgattc acagatgtct gcctgttcat ccgcgtccag
     3661 ctcgttgagt ttctccagaa gcgttaatgt ctggcttctg ataaagcggg ccatgttaag
     3721 ggcggttttt tcctgtttgg tcactgatgc ctccgtgtaa gggggatttc tgttcatggg
     3781 ggtaatgata ccgatgaaac gagagaggat gctcacgata cgggttactg atgatgaaca
     3841 tgcccggtta ctggaacgtt gtgagggtaa acaactggcg gtatggatgc ggcgggacca
     3901 gagaaaaatc actcagggtc aatgccagcg cttcgttaat acagatgtag gtgttccaca
     3961 gggtagccag cagcatcctg cgatgcagat ccggaacata atggtgcagg gcgctgactt
     4021 ccgcgtttcc agactttacg aaacacggaa accgaagacc attcatgttg ttgctcaggt
     4081 cgcagacgtt ttgcagcagc agtcgcttca cgttcgctcg cgtatcggtg attcattctg
     4141 ctaaccagta aggcaacccc gccagcctag ccgggtcctc aacgacagga gcacgatcat
     4201 gcgcacccgt ggccaggacc caacgctgcc cgagatgcgc cgcgtgcggc tgctggagat
     4261 ggcggacgcg atggatatgt tctgccaagg gttggtttgc gcattcacag ttctccgcaa
     4321 gaattgattg gctccaattc ttggagtggt gaatccgtta gcgaggtgcc gccggcttcc
     4381 attcaggtcg aggtggcccg gctccatgca ccgcgacgca acgcggggag gcagacaagg
     4441 tatagggcgg cgcctacaat ccatgccaac ccgttccatg tgctcgccga ggcggcataa
     4501 atcgccgtga cgatcagcgg tccaatgatc gaagttaggc tggtaagagc cgcgagcgat
     4561 ccttgaagct gtccctgatg gtcgtcatct acctgcctgg acagcatggc ctgcaacgcg
     4621 ggcatcccga tgccgccgga agcgagaaga atcataatgg ggaaggccat ccagcctcgc
     4681 gtcgcgaacg ccagcaagac gtagcccagc gcgtcggccg ccatgccggc gataatggcc
     4741 tgcttctcgc cgaaacgttt ggtggcggga ccagtgacga aggcttgagc gagggcgtgc
     4801 aagattccga ataccgcaag cgacaggccg atcatcgtcg cgctccagcg aaagcggtcc
     4861 tcgccgaaaa tgacccagag cgctgccggc acctgtccta cgagttgcat gataaagaag
     4921 acagtcataa gtgcggcgac gatagtcatg ccccgcgccc accggaagga gctgactggg
     4981 ttgaaggctc tcaagggcat cggtcgagat cccggtgcct aatgagtgag ctaacttaca
     5041 ttaattgcgt tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg ccagctgcat
     5101 taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta ttgggcgcca gggtggtttt
     5161 tcttttcacc agtgagacgg gcaacagctg attgcccttc accgcctggc cctgagagag
     5221 ttgcagcaag cggtccacgc tggtttgccc cagcaggcga aaatcctgtt tgatggtggt
     5281 taacggcggg atataacatg agctgtcttc ggtatcgtcg tatcccacta ccgagatatc
     5341 cgcaccaacg cgcagcccgg actcggtaat ggcgcgcatt gcgcccagcg ccatctgatc
     5401 gttggcaacc agcatcgcag tgggaacgat gccctcattc agcatttgca tggtttgttg
     5461 aaaaccggac atggcactcc agtcgccttc ccgttccgct atcggctgaa tttgattgcg
     5521 agtgagatat ttatgccagc cagccagacg cagacgcgcc gagacagaac ttaatgggcc
     5581 cgctaacagc gcgatttgct ggtgacccaa tgcgaccaga tgctccacgc ccagtcgcgt
     5641 accgtcttca tgggagaaaa taatactgtt gatgggtgtc tggtcagaga catcaagaaa
     5701 taacgccgga acattagtgc aggcagcttc cacagcaatg gcatcctggt catccagcgg
     5761 atagttaatg atcagcccac tgacgcgttg cgcgagaaga ttgtgcaccg ccgctttaca
     5821 ggcttcgacg ccgcttcgtt ctaccatcga caccaccacg ctggcaccca gttgatcggc
     5881 gcgagattta atcgccgcga caatttgcga cggcgcgtgc agggccagac tggaggtggc
     5941 aacgccaatc agcaacgact gtttgcccgc cagttgttgt gccacgcggt tgggaatgta
     6001 attcagctcc gccatcgccg cttccacttt ttcccgcgtt ttcgcagaaa cgtggctggc
     6061 ctggttcacc acgcgggaaa cggtctgata agagacaccg gcatactctg cgacatcgta
     6121 taacgttact ggtttcacat tcaccaccct gaattgactc tcttccgggc gctatcatgc
     6181 cataccgcga aaggttttgc gccattcgat ggtgtccggg atctcgacgc tctcccttat
     6241 gcgactcctg cattaggaag cagcccagta gtaggttgag gccgttgagc accgccgccg
     6301 caaggaatgg tgcatg
//
