LOCUS       sequence-307839-        7398 bp    DNA     circular SYN 14-APR-2025
DEFINITION  Expression of human linker histone H1.4.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7398)
  AUTHORS   Gibson BA, Doolittle LK, Schneider MWG, Jensen LE, Gamarra N, Henry 
            L, Gerlich DW, Redding S, Rosen MK
  TITLE     Organization of Chromatin by Intrinsic and Regulated Phase 
            Separation.
  JOURNAL   Cell. 2019 Oct 3;179(2):470-484.e21. doi: 
            10.1016/j.cell.2019.08.037. Epub 2019 Sep 19.
  PUBMED    31543265
REFERENCE   2  (bases 1 to 7398)
  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: 
            "10.1016/j.cell.2019.08.037"; journalName: "Cell"; date: "2019-10-3-
            3"; volume: "179"; issue: "2"; pages: "470-484.e21"
COMMENT     Sequence Label: pMTTH+H1.4
FEATURES             Location/Qualifiers
     source          1..7398
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        join(99..104,105..120,121..127)
                     /label=tac promoter
                     /note="strong E. coli promoter; hybrid between the trp and 
                     lac UV5 promoters"
     protein_bind    135..151
                     /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)."
     primer_bind     140..162
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     CDS             221..1321
                     /codon_start=1
                     /gene="malE (mutated)"
                     /product="maltose binding protein from E. coli"
                     /label=MBP
                     /note="This version of the gene does not encode a signal 
                     sequence, so MBP will remain in the cytosol."
                     /translation="MKTEEGKLVIWINGDKGYNGLAEVGKKFEKDTGIKVTVEHPDKLE
                     EKFPQVAATGDGPDIIFWAHDRFGGYAQSGLLAEITPDKAFQDKLYPFTWDAVRYNGKL
                     IAYPIAVEALSLIYNKDLLPNPPKTWEEIPALDKELKAKGKSALMFNLQEPYFTWPLIA
                     ADGGYAFKYENGKYDIKDVGVDNAGAKAGLTFLVDLIKNKHMNADTDYSIAEAAFNKGE
                     TAMTINGPWAWSNIDTSKVNYGVTVLPTFKGQPSKPFVGVLSAGINAASPNKELAKEFL
                     ENYLLTDEGLEAVNKDKPLGAVALKSYEEELAKDPRIAATMENAQKGEIMPNIPQMSAF
                     WYAVRTAVINAASGRQTVDEALKDAQT"
     regulatory      1178..1187
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     primer_bind     1295..1318
                     /label=MBP-F
                     /note="Maltose binding protein, forward primer"
     CDS             1370..1381
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     CDS             1394..1414
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     CDS             2108..2128
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     CDS             2129..2146
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(2159..2176)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(2159..2175)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2168..2190)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     complement(2354..2371)
                     /label=pBAD Reverse
                     /note="For vectors with E. coli araBAD promoter, reverse 
                     primer"
     terminator      2524..2610
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      2702..2729
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     promoter        2748..2839
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2840..2908,2909..3700)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(3058..3077)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      complement(3742..4255)
                     /direction=LEFT
                     /label=M13 ori
                     /note="M13 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3937..3958)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4149..4168
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     rep_origin      4366..4954
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4855..4874
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5108..5125
                     /label=L4440
                     /note="L4440 vector, forward primer"
     primer_bind     complement(5231..5250)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5391..5413
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(5407..5598)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     promoter        6094..6171
                     /gene="lacI (mutant)"
                     /label=lacIq promoter
                     /note="In the lacIq allele, a single base change in the 
                     promoter boosts expression of the lacI gene about 10-fold."
     CDS             6172..7254
                     /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     complement(6191..6210)
                     /label=LacI-R
                     /note="LacI, reverse primer"
ORIGIN
        1 actgcataat tcgtgtcgct caaggcgcac tcccgttctg gataatgttt tttgcgccga
       61 catcataacg gttctggcaa atattctgaa atgagctgtt gacaattaat catcggctcg
      121 tataatgtgt ggaattgtga gcggataaca atttcacaca ggaaacagcc agtccgttta
      181 ggtgttttca cgagcacttc accaacaagg accatagatt atgaaaactg aagaaggtaa
      241 actggtaatc tggattaacg gcgataaagg ctataacggt ctcgctgaag tcggtaagaa
      301 attcgagaaa gataccggaa ttaaagtcac cgttgagcat ccggataaac tggaagagaa
      361 attcccacag gttgcggcaa ctggcgatgg ccctgacatt atcttctggg cacacgaccg
      421 ctttggtggc tacgctcaat ctggcctgtt ggctgaaatc accccggaca aagcgttcca
      481 ggacaagctg tatccgttta cctgggatgc cgtacgttac aacggcaagc tgattgctta
      541 cccgatcgct gttgaagcgt tatcgctgat ttataacaaa gatctgctgc cgaacccgcc
      601 aaaaacctgg gaagagatcc cggcgctgga taaagaactg aaagcgaaag gtaagagcgc
      661 gctgatgttc aacctgcaag aaccgtactt cacctggccg ctgattgctg ctgacggggg
      721 ttatgcgttc aagtatgaaa acggcaagta cgacattaaa gacgtgggcg tggataacgc
      781 tggcgcgaaa gcgggtctga ccttcctggt tgacctgatt aaaaacaaac acatgaatgc
      841 agacaccgat tactccatcg cagaagctgc ctttaataaa ggcgaaacag cgatgaccat
      901 caacggcccg tgggcatggt ccaacatcga caccagcaaa gtgaattatg gtgtaacggt
      961 actgccgacc ttcaagggtc aaccatccaa accgttcgtt ggcgtgctga gcgcaggtat
     1021 taacgccgcc agtccgaaca aagagctggc aaaagagttc ctcgaaaact atctgctgac
     1081 tgatgaaggt ctggaagcgg ttaataaaga caaaccgctg ggtgccgtag cgctgaagtc
     1141 ttacgaggaa gagttggcga aagatccacg tattgccgcc accatggaaa acgcccagaa
     1201 aggtgaaatc atgccgaaca tcccgcagat gtccgctttc tggtatgccg tgcgtactgc
     1261 ggtgatcaac gccgccagcg gtcgtcagac tgtcgatgaa gccctgaaag acgcgcagac
     1321 taattcgagc tcgaacaaca acaacaataa caataacaac aacctcggga tcgagggaag
     1381 gatttcagaa tttgaaaacc tgtattttca gggccatatg ggaggcagtg gtggatcatc
     1441 agaaactgca cctgcagcgc ctgctgctcc ggcacctgcc gagaaaactc ccgtgaagaa
     1501 gaaggctaga aagtctgcag gtgcggccaa gcgcaaagcg tctgggcccc cggtgtccga
     1561 gctcattact aaagctgttg ccgcctccaa ggagcgcagc ggcgtatctt tggccgctct
     1621 caagaaagcg ctggcagccg ctggctatga cgtggagaag aacaacagcc gcatcaagct
     1681 gggtctcaag agcctggtga gcaagggcac cctggtgcag accaagggca ccggcgcgtc
     1741 gggttccttc aaactcaaca agaaggcggc ctctggggaa gccaagccta aggctaaaaa
     1801 ggcaggtgca gctaaagcta agaagccagc aggagctgcg aagaagccga agaaggctac
     1861 tggagcggct acccctaaga agtcggctaa gaagacccca aagaaggcga agaagccggc
     1921 tgcagctgct ggagccaaaa aagcgaaaag cccgaaaaag gcgaaagcag ccaagccaaa
     1981 aaaggcgccc aagagcccag cgaaggccaa agcagttaaa cccaaggcgg ctaaaccaaa
     2041 gaccgccaag cccaaggcag caaagcctaa gaaggcggca gccaagaaaa agggctcgag
     2101 cggatccgaa aacctgtatt ttcagggcca tcatcatcat catcactgaa agcttggcac
     2161 tggccgtcgt tttacaacgt cgtgactggg aaaaccctgg cgttacccaa cttaatcgcc
     2221 ttgcagcaca tccccctttc gccagctggc gtaatagcga agaggcccgc accgatcgcc
     2281 cttcccaaca gttgcgcagc ctgaatggcg aatggcagct tggctgtttt ggcggatgag
     2341 ataagatttt cagcctgata cagattaaat cagaacgcag aagcggtctg ataaaacaga
     2401 atttgcctgg cggcagtagc gcggtggtcc cacctgaccc catgccgaac tcagaagtga
     2461 aacgccgtag cgccgatggt agtgtggggt ctccccatgc gagagtaggg aactgccagg
     2521 catcaaataa aacgaaaggc tcagtcgaaa gactgggcct ttcgttttat ctgttgtttg
     2581 tcggtgaacg ctctcctgag taggacaaat ccgccgggag cggatttgaa cgttgcgaag
     2641 caacggcccg gagggtggcg ggcaggacgc ccgccataaa ctgccaggca tcaaattaag
     2701 cagaaggcca tcctgacgga tggccttttt gcgtttctac aaactctttt gtttattttt
     2761 ctaaatacat tcaaatatgt atccgctcat gagacaataa ccctgataaa tgcttcaata
     2821 atattgaaaa aggaagagta tgagtattca acatttccgt gtcgccctta ttcccttttt
     2881 tgcggcattt tgccttcctg tttttgctca cccagaaacg ctggtgaaag taaaagatgc
     2941 tgaagatcag ttgggtgcac gagtgggtta catcgaactg gatctcaaca gcggtaagat
     3001 ccttgagagt tttcgccccg aagaacgttt cccaatgatg agcactttta aagttctgct
     3061 atgtggcgcg gtattatccc gtgttgacgc cgggcaagag caactcggtc gccgcataca
     3121 ctattctcag aatgacttgg ttgagtactc accagtcaca gaaaagcatc ttacggatgg
     3181 catgacagta agagaattat gcagtgctgc cataaccatg agtgataaca ctgcggccaa
     3241 cttacttctg acaacgatcg gaggaccgaa ggagctaacc gcttttttgc acaacatggg
     3301 ggatcatgta actcgccttg atcgttggga accggagctg aatgaagcca taccaaacga
     3361 cgagcgtgac accacgatgc ctgtagcaat ggcaacaacg ttgcgcaaac tattaactgg
     3421 cgaactactt actctagctt cccggcaaca attaatagac tggatggagg cggataaagt
     3481 tgcaggacca cttctgcgct cggcccttcc ggctggctgg tttattgctg ataaatctgg
     3541 agccggtgag cgtgggtctc gcggtatcat tgcagcactg gggccagatg gtaagccctc
     3601 ccgtatcgta gttatctaca cgacggggag tcaggcaact atggatgaac gaaatagaca
     3661 gatcgctgag ataggtgcct cactgattaa gcattggtaa ctgtcagacc aagtttactc
     3721 atatatactt tagattgatt taccccggtt gataatcaga aaagccccaa aaacaggaag
     3781 attgtataag caaatattta aattgtaaac gttaatattt tgttaaaatt cgcgttaaat
     3841 ttttgttaaa tcagctcatt ttttaaccaa taggccgaaa tcggcaaaat cccttataaa
     3901 tcaaaagaat agcccgagat agggttgagt gttgttccag tttggaacaa gagtccacta
     3961 ttaaagaacg tggactccaa cgtcaaaggg cgaaaaaccg tctatcaggg cgatggccca
     4021 ctacgtgaac catcacccaa atcaagtttt ttggggtcga ggtgccgtaa agcactaaat
     4081 cggaacccta aagggagccc ccgatttaga gcttgacggg gaaagccggc gaacgtggcg
     4141 agaaaggaag ggaagaaagc gaaaggagcg ggcgctaggg cgctggcaag tgtagcggtc
     4201 acgctgcgcg taaccaccac acccgccgcg cttaatgcgc cgctacaggg cgcgtaaaag
     4261 gatctaggtg aagatccttt ttgataatct catgaccaaa atcccttaac gtgagttttc
     4321 gttccactga gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag atcctttttt
     4381 tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt
     4441 gccggatcaa gagctaccaa ctctttttcc gaaggtaact ggcttcagca gagcgcagat
     4501 accaaatact gtccttctag tgtagccgta gttaggccac cacttcaaga actctgtagc
     4561 accgcctaca tacctcgctc tgctaatcct gttaccagtg gctgctgcca gtggcgataa
     4621 gtcgtgtctt accgggttgg actcaagacg atagttaccg gataaggcgc agcggtcggg
     4681 ctgaacgggg ggttcgtgca cacagcccag cttggagcga acgacctaca ccgaactgag
     4741 atacctacag cgtgagctat gagaaagcgc cacgcttccc gaagggagaa aggcggacag
     4801 gtatccggta agcggcaggg tcggaacagg agagcgcacg agggagcttc cagggggaaa
     4861 cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt
     4921 gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg
     4981 gttcctggcc ttttgctggc cttttgctca catgttcttt cctgcgttat cccctgattc
     5041 tgtggataac cgtattaccg cctttgagtg agctgatacc gctcgccgca gccgaacgac
     5101 cgagcgcagc gagtcagtga gcgaggaagc ggaagagcgc ctgatgcggt attttctcct
     5161 tacgcatctg tgcggtattt cacaccgcat gtggtgcact ctcagtacac cgcatgtggt
     5221 gcactctcag tacaatctgc tctgatgccg catagttaag ccagtataca ctccgctatc
     5281 gctacgtgac tgggtcatgg ctgcgccccg acacccgcca acacccgctg acgcgccctg
     5341 acgggcttgt ctgctcccgg catccgctta cagacaagct gtgaccgtct ccgggagctg
     5401 catgtgtcag aggttttcac cgtcatcacc gaaacgcgcg aggcagctgc ggtaaagctc
     5461 atcagcgtgg tcgtgcagcg attcacagat gtctgcctgt tcatccgcgt ccagctcgtt
     5521 gagtttctcc agaagcgtta atgtctggct tctgataaag cgggccatgt taagggcggt
     5581 tttttcctgt ttggtcactg atgcctccgt gtaaggggga tttctgttca tgggggtaat
     5641 gataccgatg aaacgagaga ggatgctcac gatacgggtt actgatgatg aacatgcccg
     5701 gttactggaa cgttgtgagg gtaaacaact ggcggtatgg atgcggcggg accagagaaa
     5761 aatcactcag ggtcaatgcc agcgcttcgt taatacagat gtaggtgttc cacagggtag
     5821 ccagcagcat cctgcgatgc agatccggaa cataatggtg cagggcgctg acttccgcgt
     5881 ttccagactt tacgaaacac ggaaaccgaa gaccattcat gttgttgctc aggtcgcaga
     5941 cgttttgcag cagcagtcgc ttcacgttcg ctcgcgtatc ggtgattcat tctgctaacc
     6001 agtaaggcaa ccccgccagc ctagccgggt cctcaacgac aggagcacga tcatgcgcac
     6061 ccgtggccag gacccaacgc tgcccgaaat tccgacacca tcgaatggtg caaaaccttt
     6121 cgcggtatgg catgatagcg cccggaagag agtcaattca gggtggtgaa tgtgaaacca
     6181 gtaacgttat acgatgtcgc agagtatgcc ggtgtctctt atcagaccgt ttcccgcgtg
     6241 gtgaaccagg ccagccacgt ttctgcgaaa acgcgggaaa aagtggaagc ggcgatggcg
     6301 gagctgaatt acattcccaa ccgcgtggca caacaactgg cgggcaaaca gtcgttgctg
     6361 attggcgttg ccacctccag tctggccctg cacgcgccgt cgcaaattgt cgcggcgatt
     6421 aaatctcgcg ccgatcaact gggtgccagc gtggtggtgt cgatggtaga acgaagcggc
     6481 gtcgaagcct gtaaagcggc ggtgcacaat cttctcgcgc aacgcgtcag tgggctgatc
     6541 attaactatc cgctggatga ccaggatgcc attgctgtgg aagctgcctg cactaatgtt
     6601 ccggcgttat ttcttgatgt ctctgaccag acacccatca acagtattat tttctcccat
     6661 gaagacggta cgcgactggg cgtggagcat ctggtcgcat tgggtcacca gcaaatcgcg
     6721 ctgttagcgg gcccattaag ttctgtctcg gcgcgtctgc gtctggctgg ctggcataaa
     6781 tatctcactc gcaatcaaat tcagccgata gcggaacggg aaggcgactg gagtgccatg
     6841 tccggttttc aacaaaccat gcaaatgctg aatgagggca tcgttcccac tgcgatgctg
     6901 gttgccaacg atcagatggc gctgggcgca atgcgcgcca ttaccgagtc cgggctgcgc
     6961 gttggtgcgg atatctcggt agtgggatac gacgataccg aagacagctc atgttatatc
     7021 ccgccgttaa ccaccatcaa acaggatttt cgcctgctgg ggcaaaccag cgtggaccgc
     7081 ttgctgcaac tctctcaggg ccaggcggtg aagggcaatc agctgttgcc cgtctcactg
     7141 gtgaaaagaa aaaccaccct ggcgcccaat acgcaaaccg cctctccccg cgcgttggcc
     7201 gattcattaa tgcagctggc acgacaggtt tcccgactgg aaagcgggca gtgagcgcaa
     7261 cgcaattaat gtaagttagc tcactcatta ggcacaattc tcatgtttga cagcttatca
     7321 tcgactgcac ggtgcaccaa tgcttctggc gtcaggcagc catcggaagc tgtggtatgg
     7381 ctgtgcaggt cgtaaatc
//
