LOCUS       sequence-388992-        4509 bp    DNA     circular SYN 15-APR-2025
DEFINITION  Human (hu) histones H3 and H4 inserted into a plasmid.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4509)
  TITLE     Preparation of recombinant nucleosomes
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 4509)
  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            "Unpublished"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-huH3-huH4
FEATURES             Location/Qualifiers
     source          1..4509
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     581..600
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(620..697)
                     /gene="lacI"
                     /label=lacI promoter
     promoter        743..761
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    762..786
                     /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             842..859
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             872..892
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     promoter        1375..1393
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    1394..1418
                     /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             1788..1832
                     /codon_start=1
                     /product="affinity and epitope tag derived from pancreatic 
                     ribonuclease A"
                     /label=S-Tag
                     /translation="KETAAAKFERQHMDS"
     primer_bind     complement(1870..1888)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      1884..1931
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     CDS             complement(2102..2893)
                     /codon_start=1
                     /gene="aadA"
                     /product="aminoglycoside adenylyltransferase (Murphy, 
                     1985)"
                     /label=SmR
                     /note="confers resistance to spectinomycin and 
                     streptomycin"
                     /translation="MREAVIAEVSTQLSEVVGVIERHLEPTLLAVHLYGSAVDGGLKPH
                     SDIDLLVTVTVRLDETTRRALINDLLETSASPGESEILRAVEVTIVVHDDIIPWRYPAK
                     RELQFGEWQRNDILAGIFEPATIDIDLAILLTKAREHSVALVGPAAEELFDPVPEQDLF
                     EALNETLTLWNSPPDWAGDERNVVLTLSRIWYSAVTGKIAPKDVAADWAMERLPAQYQP
                     VILEARQAYLGQEEDRLASRADQLEEFVHYVKGEITKVVGK"
     promoter        complement(2894..2985)
                     /gene="bla"
                     /label=AmpR promoter
     CDS             complement(join(4046..4509,1..619))
                     /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"
ORIGIN
        1 gagtgagata tttatgccag ccagccagac gcagacgcgc cgagacagaa cttaatgggc
       61 ccgctaacag cgcgatttgc tggtgaccca atgcgaccag atgctccacg cccagtcgcg
      121 taccgtcttc atgggagaaa ataatactgt tgatgggtgt ctggtcagag acatcaagaa
      181 ataacgccgg aacattagtg caggcagctt ccacagcaat ggcatcctgg tcatccagcg
      241 gatagttaat gatcagccca ctgacgcgtt gcgcgagaag attgtgcacc gccgctttac
      301 aggcttcgac gccgcttcgt tctaccatcg acaccaccac gctggcaccc agttgatcgg
      361 cgcgagattt aatcgccgcg acaatttgcg acggcgcgtg cagggccaga ctggaggtgg
      421 caacgccaat cagcaacgac tgtttgcccg ccagttgttg tgccacgcgg ttgggaatgt
      481 aattcagctc cgccatcgcc gcttccactt tttcccgcgt tttcgcagaa acgtggctgg
      541 cctggttcac cacgcgggaa acggtctgat aagagacacc ggcatactct gcgacatcgt
      601 ataacgttac tggtttcaca ttcaccaccc tgaattgact ctcttccggg cgctatcatg
      661 ccataccgcg aaaggttttg cgccattcga tggtgtccgg gatctcgacg ctctccctta
      721 tgcgactcct gcattaggaa attaatacga ctcactatag gggaattgtg agcggataac
      781 aattcccctg tagaaataat tttgtttaac tttaataagg agatatacca tgggcagcag
      841 ccatcaccat catcaccaca gccaggatcc agagaacctc tacttccaag gcatggctcg
      901 tactaaacag acagctcgga aatccaccgg cggtaaagcg ccacgcaagc agctggctac
      961 caaggctgct cgcaagagcg cgccggctac cggcggcgtg aaaaagcctc accgttaccg
     1021 cccgggcact gtggctctgc gcgagatccg ccgctaccaa aagtcgactg agttgctgat
     1081 tcggaagctg ccgttccagc gcctggtgcg agaaatcgcc caagacttca agaccgatct
     1141 tcgcttccag agctctgcgg tgatggcgct gcaggaggct tgtgaggcct acttggtagg
     1201 gctctttgag gacacaaacc tttgcgccat ccatgctaag cgagtgacta ttatgcccaa
     1261 agacatccag ctcgctcgcc gcattcgcgg agaaagagcg tagaagcttg cggccgcata
     1321 atgcttaagt cgaacagaaa gtaatcgtat tgtacacggc cgcataatcg aaattaatac
     1381 gactcactat aggggaattg tgagcggata acaattcccc atcttagtat attagttaag
     1441 tataagaagg agatatacat atgagcggcc gcggcaaagg tggcaaaggc ctgggcaagg
     1501 gtggcgcgaa acgccatcgt aaagtgctgc gtgataacat tcagggcatt accaaaccgg
     1561 ccattcgtcg cctggcgcgt cgtggcggtg tgaaacgcat tagcggtctg atttatgaag
     1621 aaacccgtgg cgtgctgaaa gtctttctgg aaaacgttat tcgtgatgcg gtgacctata
     1681 ccgaacatgc gaaacgcaag accgtgacgg cgatggatgt tgtgtatgca ctgaaacgcc
     1741 agggtcgcac cctgtatggt tttggcggtt aataactcga gtctggtaaa gaaaccgctg
     1801 ctgcgaaatt tgaacgccag cacatggact cgtctactag cgcagcttaa ttaacctagg
     1861 ctgctgccac cgctgagcaa taactagcat aaccccttgg ggcctctaaa cgggtcttga
     1921 ggggtttttt gctgaaacct caggcatttg agaagcacac ggtcacactg cttccggtag
     1981 tcaataaacc ggtaaaccag caatagacat aagcggctat ttaacgaccc tgccctgaac
     2041 cgacgaccgg gtcatcgtgg ccggatcttg cggcccctcg gcttgaacga attgttagac
     2101 attatttgcc gactaccttg gtgatctcgc ctttcacgta gtggacaaat tcttccaact
     2161 gatctgcgcg cgaggccaag cgatcttctt cttgtccaag ataagcctgt ctagcttcaa
     2221 gtatgacggg ctgatactgg gccggcaggc gctccattgc ccagtcggca gcgacatcct
     2281 tcggcgcgat tttgccggtt actgcgctgt accaaatgcg ggacaacgta agcactacat
     2341 ttcgctcatc gccagcccag tcgggcggcg agttccatag cgttaaggtt tcatttagcg
     2401 cctcaaatag atcctgttca ggaaccggat caaagagttc ctccgccgct ggacctacca
     2461 aggcaacgct atgttctctt gcttttgtca gcaagatagc cagatcaatg tcgatcgtgg
     2521 ctggctcgaa gatacctgca agaatgtcat tgcgctgcca ttctccaaat tgcagttcgc
     2581 gcttagctgg ataacgccac ggaatgatgt cgtcgtgcac aacaatggtg acttctacag
     2641 cgcggagaat ctcgctctct ccaggggaag ccgaagtttc caaaaggtcg ttgatcaaag
     2701 ctcgccgcgt tgtttcatca agccttacgg tcaccgtaac cagcaaatca atatcactgt
     2761 gtggcttcag gccgccatcc actgcggagc cgtacaaatg tacggccagc aacgtcggtt
     2821 cgagatggcg ctcgatgacg ccaactacct ctgatagttg agtcgatact tcggcgatca
     2881 ccgcttccct catactcttc ctttttcaat attattgaag catttatcag ggttattgtc
     2941 tcatgagcgg atacatattt gaatgtattt agaaaaataa acaaatagct agctcactcg
     3001 gtcgctacgc tccgggcgtg agactgcggc gggcgctgcg gacacataca aagttaccca
     3061 cagattccgt ggataagcag gggactaaca tgtgaggcaa aacagcaggg ccgcgccggt
     3121 ggcgtttttc cataggctcc gccctcctgc cagagttcac ataaacagac gcttttccgg
     3181 tgcatctgtg ggagccgtga ggctcaacca tgaatctgac agtacgggcg aaacccgaca
     3241 ggacttaaag atccccaccg tttccggcgg gtcgctccct cttgcgctct cctgttccga
     3301 ccctgccgtt taccggatac ctgttccgcc tttctccctt acgggaagtg tggcgctttc
     3361 tcatagctca cacactggta tctcggctcg gtgtaggtcg ttcgctccaa gctgggctgt
     3421 aagcaagaac tccccgttca gcccgactgc tgcgccttat ccggtaactg ttcacttgag
     3481 tccaacccgg aaaagcacgg taaaacgcca ctggcagcag ccattggtaa ctgggagttc
     3541 gcagaggatt tgtttagcta aacacgcggt tgctcttgaa gtgtgcgcca aagtccggct
     3601 acactggaag gacagatttg gttgctgtgc tctgcgaaag ccagttacca cggttaagca
     3661 gttccccaac tgacttaacc ttcgatcaaa ccacctcccc aggtggtttt ttcgaaggtt
     3721 aagtcagttg gggaactgct taaccttcga tcaaaccacc tccccaggtg gttttttcgt
     3781 ttacagggca aaagattacg cgcagaaaaa aaggatctca agaagatcct ttgatctttt
     3841 ctactgaacc gctctagatt tcagtgcaat ttatctcttc aaatgtagca cctgaagtca
     3901 gccccatacg atataagttg taattctcat gttagtcatg ccccgcgccc accggaagga
     3961 gctgactggg ttgaaggctc tcaagggcat cggtcgagat cccggtgcct aatgagtgag
     4021 ctaacttaca ttaattgcgt tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg
     4081 ccagctgcat taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta ttgggcgcca
     4141 gggtggtttt tcttttcacc agtgagacgg gcaacagctg attgcccttc accgcctggc
     4201 cctgagagag ttgcagcaag cggtccacgc tggtttgccc cagcaggcga aaatcctgtt
     4261 tgatggtggt taacggcggg atataacatg agctgtcttc ggtatcgtcg tatcccacta
     4321 ccgagatgtc cgcaccaacg cgcagcccgg actcggtaat ggcgcgcatt gcgcccagcg
     4381 ccatctgatc gttggcaacc agcatcgcag tgggaacgat gccctcattc agcatttgca
     4441 tggtttgttg aaaaccggac atggcactcc agtcgccttc ccgttccgct atcggctgaa
     4501 tttgattgc
//
