LOCUS       sequence-204402-        6013 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of Drosophila histone H2A.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6013)
  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 6013)
  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-dH2A
FEATURES             Location/Qualifiers
     source          1..6013
                     /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(722..740)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      736..783
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     promoter        complement(join(958..963,964..980,981..986))
                     /label=tet promoter
                     /note="E. coli promoter for tetracycline efflux protein 
                     gene"
     primer_bind     complement(1013..1031)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        1099..1203
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1204..1272,1273..2064)
                     /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(1422..1441)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2235..2823
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2724..2743
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2977..2994
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    3009..3148
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(3074..3093)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3234..3256
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(3250..3441)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(4753..5835)
                     /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     5797..5816
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(5836..5913)
                     /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 atacatatgt
      301 ctggacgtgg aaaaggtggc aaagtgaagg gaaaggcaaa gtcccgctca aaccgtgccg
      361 gtcttcaatt ccctgtgggc cgtattcacc gtttgctccg gaagggaaac tacgcagagc
      421 gtgttggtgc aggcgctcca gtttacctag ctgccgtaat ggaatatctg gccgctgagg
      481 ttctcgagtt ggctggcaat gctgctcgtg acaacaagaa gactagaatt attccgcgtc
      541 atctgcaact ggccatccgc aacgacgagg agttaaacaa gctgctctcc ggcgtcacaa
      601 ttgcacaagg tggcgtgttg cctaatatac aggctgttct gttgcccaag aagaccgaga
      661 agaaggccta aggatccggc tgctaacaaa gcccgaaagg aagctgagtt ggctgctgcc
      721 accgctgagc aataactagc ataacccctt ggggcctcta aacgggtctt gaggggtttt
      781 ttgctgaaag gaggaactat atccggatat cccgcaagag gcccggcagt accggcataa
      841 ccaagcctat gcctacagca tccagggtga cggtgccgag gatgacgatg agcgcattgt
      901 tagatttcat acacggtgcc tgactgcgtt agcaatttaa ctgtgataaa ctaccgcatt
      961 aaagcttatc gatgataagc tgtcaaacat gagaattctt gaagacgaaa gggcctcgtg
     1021 atacgcctat ttttataggt taatgtcatg ataataatgg tttcttagac gtcaggtggc
     1081 acttttcggg gaaatgtgcg cggaacccct atttgtttat ttttctaaat acattcaaat
     1141 atgtatccgc tcatgagaca ataaccctga taaatgcttc aataatattg aaaaaggaag
     1201 agtatgagta ttcaacattt ccgtgtcgcc cttattccct tttttgcggc attttgcctt
     1261 cctgtttttg ctcacccaga aacgctggtg aaagtaaaag atgctgaaga tcagttgggt
     1321 gcacgagtgg gttacatcga actggatctc aacagcggta agatccttga gagttttcgc
     1381 cccgaagaac gttttccaat gatgagcact tttaaagttc tgctatgtgg cgcggtatta
     1441 tcccgtgttg acgccgggca agagcaactc ggtcgccgca tacactattc tcagaatgac
     1501 ttggttgagt actcaccagt cacagaaaag catcttacgg atggcatgac agtaagagaa
     1561 ttatgcagtg ctgccataac catgagtgat aacactgcgg ccaacttact tctgacaacg
     1621 atcggaggac cgaaggagct aaccgctttt ttgcacaaca tgggggatca tgtaactcgc
     1681 cttgatcgtt gggaaccgga gctgaatgaa gccataccaa acgacgagcg tgacaccacg
     1741 atgcctgcag caatggcaac aacgttgcgc aaactattaa ctggcgaact acttactcta
     1801 gcttcccggc aacaattaat agactggatg gaggcggata aagttgcagg accacttctg
     1861 cgctcggccc ttccggctgg ctggtttatt gctgataaat ctggagccgg tgagcgtggg
     1921 tctcgcggta tcattgcagc actggggcca gatggtaagc cctcccgtat cgtagttatc
     1981 tacacgacgg ggagtcaggc aactatggat gaacgaaata gacagatcgc tgagataggt
     2041 gcctcactga ttaagcattg gtaactgtca gaccaagttt actcatatat actttagatt
     2101 gatttaaaac ttcattttta atttaaaagg atctaggtga agatcctttt tgataatctc
     2161 atgaccaaaa tcccttaacg tgagttttcg ttccactgag cgtcagaccc cgtagaaaag
     2221 atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa
     2281 aaaccaccgc taccagcggt ggtttgtttg ccggatcaag agctaccaac tctttttccg
     2341 aaggtaactg gcttcagcag agcgcagata ccaaatactg tccttctagt gtagccgtag
     2401 ttaggccacc acttcaagaa ctctgtagca ccgcctacat acctcgctct gctaatcctg
     2461 ttaccagtgg ctgctgccag tggcgataag tcgtgtctta ccgggttgga ctcaagacga
     2521 tagttaccgg ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac acagcccagc
     2581 ttggagcgaa cgacctacac cgaactgaga tacctacagc gtgagctatg agaaagcgcc
     2641 acgcttcccg aagggagaaa ggcggacagg tatccggtaa gcggcagggt cggaacagga
     2701 gagcgcacga gggagcttcc agggggaaac gcctggtatc tttatagtcc tgtcgggttt
     2761 cgccacctct gacttgagcg tcgatttttg tgatgctcgt caggggggcg gagcctatgg
     2821 aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct tttgctggcc ttttgctcac
     2881 atgttctttc ctgcgttatc ccctgattct gtggataacc gtattaccgc ctttgagtga
     2941 gctgataccg ctcgccgcag ccgaacgacc gagcgcagcg agtcagtgag cgaggaagcg
     3001 gaagagcgcc tgatgcggta ttttctcctt acgcatctgt gcggtatttc acaccgcaat
     3061 ggtgcactct cagtacaatc tgctctgatg ccgcatagtt aagccagtat acactccgct
     3121 atcgctacgt gactgggtca tggctgcgcc ccgacacccg ccaacacccg ctgacgcgcc
     3181 ctgacgggct tgtctgctcc cggcatccgc ttacagacaa gctgtgaccg tctccgggag
     3241 ctgcatgtgt cagaggtttt caccgtcatc accgaaacgc gcgaggcagc tgcggtaaag
     3301 ctcatcagcg tggtcgtgaa gcgattcaca gatgtctgcc tgttcatccg cgtccagctc
     3361 gttgagtttc tccagaagcg ttaatgtctg gcttctgata aagcgggcca tgttaagggc
     3421 ggttttttcc tgtttggtca ctgatgcctc cgtgtaaggg ggatttctgt tcatgggggt
     3481 aatgataccg atgaaacgag agaggatgct cacgatacgg gttactgatg atgaacatgc
     3541 ccggttactg gaacgttgtg agggtaaaca actggcggta tggatgcggc gggaccagag
     3601 aaaaatcact cagggtcaat gccagcgctt cgttaataca gatgtaggtg ttccacaggg
     3661 tagccagcag catcctgcga tgcagatccg gaacataatg gtgcagggcg ctgacttccg
     3721 cgtttccaga ctttacgaaa cacggaaacc gaagaccatt catgttgttg ctcaggtcgc
     3781 agacgttttg cagcagcagt cgcttcacgt tcgctcgcgt atcggtgatt cattctgcta
     3841 accagtaagg caaccccgcc agcctagccg ggtcctcaac gacaggagca cgatcatgcg
     3901 cacccgtggc caggacccaa cgctgcccga gatgcgccgc gtgcggctgc tggagatggc
     3961 ggacgcgatg gatatgttct gccaagggtt ggtttgcgca ttcacagttc tccgcaagaa
     4021 ttgattggct ccaattcttg gagtggtgaa tccgttagcg aggtgccgcc ggcttccatt
     4081 caggtcgagg tggcccggct ccatgcaccg cgacgcaacg cggggaggca gacaaggtat
     4141 agggcggcgc ctacaatcca tgccaacccg ttccatgtgc tcgccgaggc ggcataaatc
     4201 gccgtgacga tcagcggtcc aatgatcgaa gttaggctgg taagagccgc gagcgatcct
     4261 tgaagctgtc cctgatggtc gtcatctacc tgcctggaca gcatggcctg caacgcgggc
     4321 atcccgatgc cgccggaagc gagaagaatc ataatgggga aggccatcca gcctcgcgtc
     4381 gcgaacgcca gcaagacgta gcccagcgcg tcggccgcca tgccggcgat aatggcctgc
     4441 ttctcgccga aacgtttggt ggcgggacca gtgacgaagg cttgagcgag ggcgtgcaag
     4501 attccgaata ccgcaagcga caggccgatc atcgtcgcgc tccagcgaaa gcggtcctcg
     4561 ccgaaaatga cccagagcgc tgccggcacc tgtcctacga gttgcatgat aaagaagaca
     4621 gtcataagtg cggcgacgat agtcatgccc cgcgcccacc ggaaggagct gactgggttg
     4681 aaggctctca agggcatcgg tcgagatccc ggtgcctaat gagtgagcta acttacatta
     4741 attgcgttgc gctcactgcc cgctttccag tcgggaaacc tgtcgtgcca gctgcattaa
     4801 tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg ggcgccaggg tggtttttct
     4861 tttcaccagt gagacgggca acagctgatt gcccttcacc gcctggccct gagagagttg
     4921 cagcaagcgg tccacgctgg tttgccccag caggcgaaaa tcctgtttga tggtggttaa
     4981 cggcgggata taacatgagc tgtcttcggt atcgtcgtat cccactaccg agatatccgc
     5041 accaacgcgc agcccggact cggtaatggc gcgcattgcg cccagcgcca tctgatcgtt
     5101 ggcaaccagc atcgcagtgg gaacgatgcc ctcattcagc atttgcatgg tttgttgaaa
     5161 accggacatg gcactccagt cgccttcccg ttccgctatc ggctgaattt gattgcgagt
     5221 gagatattta tgccagccag ccagacgcag acgcgccgag acagaactta atgggcccgc
     5281 taacagcgcg atttgctggt gacccaatgc gaccagatgc tccacgccca gtcgcgtacc
     5341 gtcttcatgg gagaaaataa tactgttgat gggtgtctgg tcagagacat caagaaataa
     5401 cgccggaaca ttagtgcagg cagcttccac agcaatggca tcctggtcat ccagcggata
     5461 gttaatgatc agcccactga cgcgttgcgc gagaagattg tgcaccgccg ctttacaggc
     5521 ttcgacgccg cttcgttcta ccatcgacac caccacgctg gcacccagtt gatcggcgcg
     5581 agatttaatc gccgcgacaa tttgcgacgg cgcgtgcagg gccagactgg aggtggcaac
     5641 gccaatcagc aacgactgtt tgcccgccag ttgttgtgcc acgcggttgg gaatgtaatt
     5701 cagctccgcc atcgccgctt ccactttttc ccgcgttttc gcagaaacgt ggctggcctg
     5761 gttcaccacg cgggaaacgg tctgataaga gacaccggca tactctgcga catcgtataa
     5821 cgttactggt ttcacattca ccaccctgaa ttgactctct tccgggcgct atcatgccat
     5881 accgcgaaag gttttgcgcc attcgatggt gtccgggatc tcgacgctct cccttatgcg
     5941 actcctgcat taggaagcag cccagtagta ggttgaggcc gttgagcacc gccgccgcaa
     6001 ggaatggtgc atg
//
