LOCUS       sequence-307841-        7077 bp    DNA     circular SYN 14-APR-2025
DEFINITION  Expression of human linker histone H1.4 without c-terminal tail.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7077)
  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 7077)
  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-Delta-CTD
FEATURES             Location/Qualifiers
     source          1..7077
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        join(59..64,65..80,81..87)
                     /label=tac promoter
                     /note="strong E. coli promoter; hybrid between the trp and 
                     lac UV5 promoters"
     protein_bind    95..111
                     /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     100..122
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     CDS             181..1281
                     /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      1138..1147
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     primer_bind     1255..1278
                     /label=MBP-F
                     /note="Maltose binding protein, forward primer"
     CDS             1330..1341
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     CDS             1354..1374
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     CDS             1747..1767
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     CDS             1768..1785
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(1798..1815)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(1798..1814)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1807..1829)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     complement(1993..2010)
                     /label=pBAD Reverse
                     /note="For vectors with E. coli araBAD promoter, reverse 
                     primer"
     terminator      2163..2249
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      2341..2368
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     promoter        2387..2478
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2479..2547,2548..3339)
                     /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(2697..2716)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      complement(3381..3894)
                     /direction=LEFT
                     /label=M13 ori
                     /note="M13 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3576..3597)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3788..3807
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     rep_origin      4005..4593
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4494..4513
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4747..4764
                     /label=L4440
                     /note="L4440 vector, forward primer"
     primer_bind     complement(4870..4889)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5030..5052
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(5046..5237)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     promoter        5733..5810
                     /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             5811..6893
                     /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(5830..5849)
                     /label=LacI-R
                     /note="LacI, reverse primer"
ORIGIN
        1 gataatgttt tttgcgccga catcataacg gttctggcaa atattctgaa atgagctgtt
       61 gacaattaat catcggctcg tataatgtgt ggaattgtga gcggataaca atttcacaca
      121 ggaaacagcc agtccgttta ggtgttttca cgagcacttc accaacaagg accatagatt
      181 atgaaaactg aagaaggtaa actggtaatc tggattaacg gcgataaagg ctataacggt
      241 ctcgctgaag tcggtaagaa attcgagaaa gataccggaa ttaaagtcac cgttgagcat
      301 ccggataaac tggaagagaa attcccacag gttgcggcaa ctggcgatgg ccctgacatt
      361 atcttctggg cacacgaccg ctttggtggc tacgctcaat ctggcctgtt ggctgaaatc
      421 accccggaca aagcgttcca ggacaagctg tatccgttta cctgggatgc cgtacgttac
      481 aacggcaagc tgattgctta cccgatcgct gttgaagcgt tatcgctgat ttataacaaa
      541 gatctgctgc cgaacccgcc aaaaacctgg gaagagatcc cggcgctgga taaagaactg
      601 aaagcgaaag gtaagagcgc gctgatgttc aacctgcaag aaccgtactt cacctggccg
      661 ctgattgctg ctgacggggg ttatgcgttc aagtatgaaa acggcaagta cgacattaaa
      721 gacgtgggcg tggataacgc tggcgcgaaa gcgggtctga ccttcctggt tgacctgatt
      781 aaaaacaaac acatgaatgc agacaccgat tactccatcg cagaagctgc ctttaataaa
      841 ggcgaaacag cgatgaccat caacggcccg tgggcatggt ccaacatcga caccagcaaa
      901 gtgaattatg gtgtaacggt actgccgacc ttcaagggtc aaccatccaa accgttcgtt
      961 ggcgtgctga gcgcaggtat taacgccgcc agtccgaaca aagagctggc aaaagagttc
     1021 ctcgaaaact atctgctgac tgatgaaggt ctggaagcgg ttaataaaga caaaccgctg
     1081 ggtgccgtag cgctgaagtc ttacgaggaa gagttggcga aagatccacg tattgccgcc
     1141 accatggaaa acgcccagaa aggtgaaatc atgccgaaca tcccgcagat gtccgctttc
     1201 tggtatgccg tgcgtactgc ggtgatcaac gccgccagcg gtcgtcagac tgtcgatgaa
     1261 gccctgaaag acgcgcagac taattcgagc tcgaacaaca acaacaataa caataacaac
     1321 aacctcggga tcgagggaag gatttcagaa tttgaaaacc tgtattttca gggccatatg
     1381 ggaggcagtg gtggatcatc agaaactgca cctgcagcgc ctgctgctcc ggcacctgcc
     1441 gagaaaactc ccgtgaagaa gaaggctaga aagtctgcag gtgcggccaa gcgcaaagcg
     1501 tctgggcccc cggtgtccga gctcattact aaagctgttg ccgcctccaa ggagcgcagc
     1561 ggcgtatctt tggccgctct caagaaagcg ctggcagccg ctggctatga cgtggagaag
     1621 aacaacagcc gcatcaagct gggtctcaag agcctggtga gcaagggcac cctggtgcag
     1681 accaagggca ccggcgcgtc gggttccttc aaactcaaca agaaggcggc cggctcgagc
     1741 ggatccgaaa acctgtattt tcagggccat catcatcatc atcactgaaa gcttggcact
     1801 ggccgtcgtt ttacaacgtc gtgactggga aaaccctggc gttacccaac ttaatcgcct
     1861 tgcagcacat ccccctttcg ccagctggcg taatagcgaa gaggcccgca ccgatcgccc
     1921 ttcccaacag ttgcgcagcc tgaatggcga atggcagctt ggctgttttg gcggatgaga
     1981 taagattttc agcctgatac agattaaatc agaacgcaga agcggtctga taaaacagaa
     2041 tttgcctggc ggcagtagcg cggtggtccc acctgacccc atgccgaact cagaagtgaa
     2101 acgccgtagc gccgatggta gtgtggggtc tccccatgcg agagtaggga actgccaggc
     2161 atcaaataaa acgaaaggct cagtcgaaag actgggcctt tcgttttatc tgttgtttgt
     2221 cggtgaacgc tctcctgagt aggacaaatc cgccgggagc ggatttgaac gttgcgaagc
     2281 aacggcccgg agggtggcgg gcaggacgcc cgccataaac tgccaggcat caaattaagc
     2341 agaaggccat cctgacggat ggcctttttg cgtttctaca aactcttttg tttatttttc
     2401 taaatacatt caaatatgta tccgctcatg agacaataac cctgataaat gcttcaataa
     2461 tattgaaaaa ggaagagtat gagtattcaa catttccgtg tcgcccttat tccctttttt
     2521 gcggcatttt gccttcctgt ttttgctcac ccagaaacgc tggtgaaagt aaaagatgct
     2581 gaagatcagt tgggtgcacg agtgggttac atcgaactgg atctcaacag cggtaagatc
     2641 cttgagagtt ttcgccccga agaacgtttc ccaatgatga gcacttttaa agttctgcta
     2701 tgtggcgcgg tattatcccg tgttgacgcc gggcaagagc aactcggtcg ccgcatacac
     2761 tattctcaga atgacttggt tgagtactca ccagtcacag aaaagcatct tacggatggc
     2821 atgacagtaa gagaattatg cagtgctgcc ataaccatga gtgataacac tgcggccaac
     2881 ttacttctga caacgatcgg aggaccgaag gagctaaccg cttttttgca caacatgggg
     2941 gatcatgtaa ctcgccttga tcgttgggaa ccggagctga atgaagccat accaaacgac
     3001 gagcgtgaca ccacgatgcc tgtagcaatg gcaacaacgt tgcgcaaact attaactggc
     3061 gaactactta ctctagcttc ccggcaacaa ttaatagact ggatggaggc ggataaagtt
     3121 gcaggaccac ttctgcgctc ggcccttccg gctggctggt ttattgctga taaatctgga
     3181 gccggtgagc gtgggtctcg cggtatcatt gcagcactgg ggccagatgg taagccctcc
     3241 cgtatcgtag ttatctacac gacggggagt caggcaacta tggatgaacg aaatagacag
     3301 atcgctgaga taggtgcctc actgattaag cattggtaac tgtcagacca agtttactca
     3361 tatatacttt agattgattt accccggttg ataatcagaa aagccccaaa aacaggaaga
     3421 ttgtataagc aaatatttaa attgtaaacg ttaatatttt gttaaaattc gcgttaaatt
     3481 tttgttaaat cagctcattt tttaaccaat aggccgaaat cggcaaaatc ccttataaat
     3541 caaaagaata gcccgagata gggttgagtg ttgttccagt ttggaacaag agtccactat
     3601 taaagaacgt ggactccaac gtcaaagggc gaaaaaccgt ctatcagggc gatggcccac
     3661 tacgtgaacc atcacccaaa tcaagttttt tggggtcgag gtgccgtaaa gcactaaatc
     3721 ggaaccctaa agggagcccc cgatttagag cttgacgggg aaagccggcg aacgtggcga
     3781 gaaaggaagg gaagaaagcg aaaggagcgg gcgctagggc gctggcaagt gtagcggtca
     3841 cgctgcgcgt aaccaccaca cccgccgcgc ttaatgcgcc gctacagggc gcgtaaaagg
     3901 atctaggtga agatcctttt tgataatctc atgaccaaaa tcccttaacg tgagttttcg
     3961 ttccactgag cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga tccttttttt
     4021 ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg
     4081 ccggatcaag agctaccaac tctttttccg aaggtaactg gcttcagcag agcgcagata
     4141 ccaaatactg tccttctagt gtagccgtag ttaggccacc acttcaagaa ctctgtagca
     4201 ccgcctacat acctcgctct gctaatcctg ttaccagtgg ctgctgccag tggcgataag
     4261 tcgtgtctta ccgggttgga ctcaagacga tagttaccgg ataaggcgca gcggtcgggc
     4321 tgaacggggg gttcgtgcac acagcccagc ttggagcgaa cgacctacac cgaactgaga
     4381 tacctacagc gtgagctatg agaaagcgcc acgcttcccg aagggagaaa ggcggacagg
     4441 tatccggtaa gcggcagggt cggaacagga gagcgcacga gggagcttcc agggggaaac
     4501 gcctggtatc tttatagtcc tgtcgggttt cgccacctct gacttgagcg tcgatttttg
     4561 tgatgctcgt caggggggcg gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg
     4621 ttcctggcct tttgctggcc ttttgctcac atgttctttc ctgcgttatc ccctgattct
     4681 gtggataacc gtattaccgc ctttgagtga gctgataccg ctcgccgcag ccgaacgacc
     4741 gagcgcagcg agtcagtgag cgaggaagcg gaagagcgcc tgatgcggta ttttctcctt
     4801 acgcatctgt gcggtatttc acaccgcatg tggtgcactc tcagtacacc gcatgtggtg
     4861 cactctcagt acaatctgct ctgatgccgc atagttaagc cagtatacac tccgctatcg
     4921 ctacgtgact gggtcatggc tgcgccccga cacccgccaa cacccgctga cgcgccctga
     4981 cgggcttgtc tgctcccggc atccgcttac agacaagctg tgaccgtctc cgggagctgc
     5041 atgtgtcaga ggttttcacc gtcatcaccg aaacgcgcga ggcagctgcg gtaaagctca
     5101 tcagcgtggt cgtgcagcga ttcacagatg tctgcctgtt catccgcgtc cagctcgttg
     5161 agtttctcca gaagcgttaa tgtctggctt ctgataaagc gggccatgtt aagggcggtt
     5221 ttttcctgtt tggtcactga tgcctccgtg taagggggat ttctgttcat gggggtaatg
     5281 ataccgatga aacgagagag gatgctcacg atacgggtta ctgatgatga acatgcccgg
     5341 ttactggaac gttgtgaggg taaacaactg gcggtatgga tgcggcggga ccagagaaaa
     5401 atcactcagg gtcaatgcca gcgcttcgtt aatacagatg taggtgttcc acagggtagc
     5461 cagcagcatc ctgcgatgca gatccggaac ataatggtgc agggcgctga cttccgcgtt
     5521 tccagacttt acgaaacacg gaaaccgaag accattcatg ttgttgctca ggtcgcagac
     5581 gttttgcagc agcagtcgct tcacgttcgc tcgcgtatcg gtgattcatt ctgctaacca
     5641 gtaaggcaac cccgccagcc tagccgggtc ctcaacgaca ggagcacgat catgcgcacc
     5701 cgtggccagg acccaacgct gcccgaaatt ccgacaccat cgaatggtgc aaaacctttc
     5761 gcggtatggc atgatagcgc ccggaagaga gtcaattcag ggtggtgaat gtgaaaccag
     5821 taacgttata cgatgtcgca gagtatgccg gtgtctctta tcagaccgtt tcccgcgtgg
     5881 tgaaccaggc cagccacgtt tctgcgaaaa cgcgggaaaa agtggaagcg gcgatggcgg
     5941 agctgaatta cattcccaac cgcgtggcac aacaactggc gggcaaacag tcgttgctga
     6001 ttggcgttgc cacctccagt ctggccctgc acgcgccgtc gcaaattgtc gcggcgatta
     6061 aatctcgcgc cgatcaactg ggtgccagcg tggtggtgtc gatggtagaa cgaagcggcg
     6121 tcgaagcctg taaagcggcg gtgcacaatc ttctcgcgca acgcgtcagt gggctgatca
     6181 ttaactatcc gctggatgac caggatgcca ttgctgtgga agctgcctgc actaatgttc
     6241 cggcgttatt tcttgatgtc tctgaccaga cacccatcaa cagtattatt ttctcccatg
     6301 aagacggtac gcgactgggc gtggagcatc tggtcgcatt gggtcaccag caaatcgcgc
     6361 tgttagcggg cccattaagt tctgtctcgg cgcgtctgcg tctggctggc tggcataaat
     6421 atctcactcg caatcaaatt cagccgatag cggaacggga aggcgactgg agtgccatgt
     6481 ccggttttca acaaaccatg caaatgctga atgagggcat cgttcccact gcgatgctgg
     6541 ttgccaacga tcagatggcg ctgggcgcaa tgcgcgccat taccgagtcc gggctgcgcg
     6601 ttggtgcgga tatctcggta gtgggatacg acgataccga agacagctca tgttatatcc
     6661 cgccgttaac caccatcaaa caggattttc gcctgctggg gcaaaccagc gtggaccgct
     6721 tgctgcaact ctctcagggc caggcggtga agggcaatca gctgttgccc gtctcactgg
     6781 tgaaaagaaa aaccaccctg gcgcccaata cgcaaaccgc ctctccccgc gcgttggccg
     6841 attcattaat gcagctggca cgacaggttt cccgactgga aagcgggcag tgagcgcaac
     6901 gcaattaatg taagttagct cactcattag gcacaattct catgtttgac agcttatcat
     6961 cgactgcacg gtgcaccaat gcttctggcg tcaggcagcc atcggaagct gtggtatggc
     7021 tgtgcaggtc gtaaatcact gcataattcg tgtcgctcaa ggcgcactcc cgttctg
//
