LOCUS       sequence-388012-        5686 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of HMfB D14A K30A E34A in E. coli.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5686)
  AUTHORS   Erkelens AM, Henneman B, van der Valk RA, Kirolos NCS, Dame RT
  TITLE     Specific DNA binding of archaeal histones HMfA and HMfB.
  JOURNAL   Front Microbiol. 2023 Apr 18;14:1166608. doi: 
            10.3389/fmicb.2023.1166608. eCollection 2023.
  PUBMED    37143534
REFERENCE   2  (bases 1 to 5686)
  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"; journalName: "Front 
            Microbiol."; date: "2023-04-18"; volume: "14:1166608. doi"; pages: "
            10.3389/fmicb.2023.1166608. eCollection 2023"
COMMENT     Sequence Label: pRD324
FEATURES             Location/Qualifiers
     source          1..5686
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      complement(26..73)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     primer_bind     69..87
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     RBS             347..369
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     RBS             623..645
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     protein_bind    660..684
                     /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     complement(684..703)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(685..703)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(791..810)
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     promoter        1016..1093
                     /gene="lacI"
                     /label=lacI promoter
     CDS             1094..2176
                     /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(1113..1132)
                     /label=LacI-R
                     /note="LacI, reverse primer"
     CDS             2985..3176
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     primer_bind     complement(3170..3192)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     misc_feature    3278..3417
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     3333..3352
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(3432..3449)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3603..4191)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3683..3702)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     complement(4302..4321)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             4313..5128
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     CDS             4313..5128
                     /codon_start=1
                     /gene="aph(3')-Ia"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     complement(4386..4405)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      complement(5220..5675)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5357..5378)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     5569..5588
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
ORIGIN
        1 atccggatat agttcctcct ttcagcaaaa aacccctcaa gacccgttta gaggccccaa
       61 ggggttatgc tagttattgc tcagcggtgg cagcagccaa ctcagcttcc tttcgggctt
      121 tgttagcagt tatttcttaa atcttcgaac tgctagttca atgtcctctg cttttattgt
      181 tttccttccg gcgtgtcttg ctagttttat agcttctgaa gcaatgtctc tacccatcgc
      241 ttctaaaatc gctgctagag ttattcttgc atcatcactg actctttcgg ctcctgccgc
      301 ttttataatt ctaccaattg gtgctattgg taattccata tgtatatctc cttcttaaag
      361 ttaaacaaaa ttatttctag aggggctttc gggctttgtt agcagttatt tgaacatttt
      421 tcgagctaat tcaatgtctt ctgcctttat tgtttttctt cccgcatgtt ttgctaattt
      481 cacagcttct gaagctattt cttcgcccat cgcttctaaa accgctgcta atgcaattct
      541 tgcatcgtca ctaactcttt cagcaccagc gttttttatg attcttccga ttggggcaat
      601 tggtaactca cccatatgta tatctccttc ttaaagttaa acaaaattat ttctagaggg
      661 gaattgttat ccgctcacaa ttcccctata gtgagtcgta ttaatttcgc gggatcgaga
      721 tcgatctcga tcctctacgc cggacgcatc gtggccggca tcaccggcgc cacaggtgcg
      781 gttgctggcg cctatatcgc cgacatcacc gatggggaag atcgggctcg ccacttcggg
      841 ctcatgagcg cttgtttcgg cgtgggtatg gtggcaggcc ccgtggccgg gggactgttg
      901 ggcgccatct ccttgcatgc accattcctt gcggcggcgg tgctcaacgg cctcaaccta
      961 ctactgggct gcttcctaat gcaggagtcg cataagggag agcgtcgaga tcccggacac
     1021 catcgaatgg cgcaaaacct ttcgcggtat ggcatgatag cgcccggaag agagtcaatt
     1081 cagggtggtg aatgtgaaac cagtaacgtt atacgatgtc gcagagtatg ccggtgtctc
     1141 ttatcagacc gtttcccgcg tggtgaacca ggccagccac gtttctgcga aaacgcggga
     1201 aaaagtggaa gcggcgatgg cggagctgaa ttacattccc aaccgcgtgg cacaacaact
     1261 ggcgggcaaa cagtcgttgc tgattggcgt tgccacctcc agtctggccc tgcacgcgcc
     1321 gtcgcaaatt gtcgcggcga ttaaatctcg cgccgatcaa ctgggtgcca gcgtggtggt
     1381 gtcgatggta gaacgaagcg gcgtcgaagc ctgtaaagcg gcggtgcaca atcttctcgc
     1441 gcaacgcgtc agtgggctga tcattaacta tccgctggat gaccaggatg ccattgctgt
     1501 ggaagctgcc tgcactaatg ttccggcgtt atttcttgat gtctctgacc agacacccat
     1561 caacagtatt attttctccc atgaagacgg tacgcgactg ggcgtggagc atctggtcgc
     1621 attgggtcac cagcaaatcg cgctgttagc gggcccatta agttctgtct cggcgcgtct
     1681 gcgtctggct ggctggcata aatatctcac tcgcaatcaa attcagccga tagcggaacg
     1741 ggaaggcgac tggagtgcca tgtccggttt tcaacaaacc atgcaaatgc tgaatgaggg
     1801 catcgttccc actgcgatgc tggttgccaa cgatcagatg gcgctgggcg caatgcgcgc
     1861 cattaccgag tccgggctgc gcgttggtgc ggacatctcg gtagtgggat acgacgatac
     1921 cgaagacagc tcatgttata tcccgccgtt aaccaccatc aaacaggatt ttcgcctgct
     1981 ggggcaaacc agcgtggacc gcttgctgca actctctcag ggccaggcgg tgaagggcaa
     2041 tcagctgttg cccgtctcac tggtgaaaag aaaaaccacc ctggcgccca atacgcaaac
     2101 cgcctctccc cgcgcgttgg ccgattcatt aatgcagctg gcacgacagg tttcccgact
     2161 ggaaagcggg cagtgagcgc aacgcaatta atgtaagtta gctcactcat taggcaccgg
     2221 gatctcgacc gatgcccttg agagccttca acccagtcag ctccttccgg tgggcgcggg
     2281 gcatgactat cgtcgccgca cttatgactg tcttctttat catgcaactc gtaggacagg
     2341 tgccggcagc gctctgggtc attttcggcg aggaccgctt tcgctggagc gcgacgatga
     2401 tcggcctgtc gcttgcggta ttcggaatct tgcacgccct cgctcaagcc ttcgtcactg
     2461 gtcccgccac caaacgtttc ggcgagaagc aggccattat cgccggcatg gcggccccac
     2521 gggtgcgcat gatcgtgctc ctgtcgttga ggacccggct aggctggcgg ggttgcctta
     2581 ctggttagca gaatgaatca ccgatacgcg agcgaacgtg aagcgactgc tgctgcaaaa
     2641 cgtctgcgac ctgagcaaca acatgaatgg tcttcggttt ccgtgtttcg taaagtctgg
     2701 aaacgcggaa gtcagcgccc tgcaccatta tgttccggat ctgcatcgca ggatgctgct
     2761 ggctaccctg tggaacacct acatctgtat taacgaagcg ctggcattga ccctgagtga
     2821 tttttctctg gtcccgccgc atccataccg ccagttgttt accctcacaa cgttccagta
     2881 accgggcatg ttcatcatca gtaacccgta tcgtgagcat cctctctcgt ttcatcggta
     2941 tcattacccc catgaacaga aatccccctt acacggaggc atcagtgacc aaacaggaaa
     3001 aaaccgccct taacatggcc cgctttatca gaagccagac attaacgctt ctggagaaac
     3061 tcaacgagct ggacgcggat gaacaggcag acatctgtga atcgcttcac gaccacgctg
     3121 atgagcttta ccgcagctgc ctcgcgcgtt tcggtgatga cggtgaaaac ctctgacaca
     3181 tgcagctccc ggagacggtc acagcttgtc tgtaagcgga tgccgggagc agacaagccc
     3241 gtcagggcgc gtcagcgggt gttggcgggt gtcggggcgc agccatgacc cagtcacgta
     3301 gcgatagcgg agtgtatact ggcttaacta tgcggcatca gagcagattg tactgagagt
     3361 gcaccattgc ggtgtgaaat accgcacaga tgcgtaagga gaaaataccg catcaggcgc
     3421 tcttccgctt cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta
     3481 tcagctcact caaaggcggt aatacggtta tccacagaat caggggataa cgcaggaaag
     3541 aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg
     3601 tttttccata ggctccgccc ccctgacgag catcacaaaa atcgacgctc aagtcagagg
     3661 tggcgaaacc cgacaggact ataaagatac caggcgtttc cccctggaag ctccctcgtg
     3721 cgctctcctg ttccgaccct gccgcttacc ggatacctgt ccgcctttct cccttcggga
     3781 agcgtggcgc tttctcatag ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc
     3841 tccaagctgg gctgtgtgca cgaacccccc gttcagcccg accgctgcgc cttatccggt
     3901 aactatcgtc ttgagtccaa cccggtaaga cacgacttat cgccactggc agcagccact
     3961 ggtaacagga ttagcagagc gaggtatgta ggcggtgcta cagagttctt gaagtggtgg
     4021 cctaactacg gctacactag aaggacagta tttggtatct gcgctctgct gaagccagtt
     4081 accttcggaa aaagagttgg tagctcttga tccggcaaac aaaccaccgc tggtagcggt
     4141 ggtttttttg tttgcaagca gcagattacg cgcagaaaaa aaggatttca agaagatcct
     4201 ttgatctttt ctacggggtc tgacgctcag tggaacgaaa actcacgtta agggattttg
     4261 gtcatgaaca ataaaactgt ctgcttacat aaacagtaat acaaggggtg ttatgagcca
     4321 tattcaacgg gaaacgtctt gctctaggcc gcgattaaat tccaacatgg atgctgattt
     4381 atatgggtat aaatgggctc gcgataatgt cgggcaatca ggtgcgacaa tctatcgatt
     4441 gtatgggaag cccgatgcgc cagagttgtt tctgaaacat ggcaaaggta gcgttgccaa
     4501 tgatgttaca gatgagatgg tcagactaaa ctggctgacg gaatttatgc ctcttccgac
     4561 catcaagcat tttatccgta ctcctgatga tgcatggtta ctcaccactg cgatccccgg
     4621 gaaaacagca ttccaggtat tagaagaata tcctgattca ggtgaaaata ttgttgatgc
     4681 gctagcagtg ttcctgcgcc ggttgcattc gattcctgtt tgtaattgtc cttttaacag
     4741 cgatcgcgta tttcgtctcg ctcaggcgca atcacgaatg aataacggtt tggttgatgc
     4801 gagtgatttt gatgacgagc gtaatggctg gcctgttgaa caagtctgga aagaaatgca
     4861 taaacttttg ccattctcac cggattcagt cgtcactcat ggtgatttct cacttgataa
     4921 ccttattttt gacgagggga aattaatagg ttgtattgat gttggacgag tcggaatcgc
     4981 agaccgatac caggatcttg ccatcctatg gaactgcctc ggtgagtttt ctccttcatt
     5041 acagaaacgg ctttttcaaa aatatggtat tgataatcct gatatgaata aattgcagtt
     5101 tcatttgatg ctcgatgagt ttttctaaga attaattcat gagcggatac atatttgaat
     5161 gtatttagaa aaataaacaa ataggggttc cgcgcacatt tccccgaaaa gtgccaccta
     5221 aattgtaagc gttaatattt tgttaaaatt cgcgttaaat ttttgttaaa tcagctcatt
     5281 ttttaaccaa taggccgaaa tcggcaaaat cccttataaa tcaaaagaat agaccgagat
     5341 agggttgagt gttgttccag tttggaacaa gagtccacta ttaaagaacg tggactccaa
     5401 cgtcaaaggg cgaaaaaccg tctatcaggg cgatggccca ctacgtgaac catcacccta
     5461 atcaagtttt ttggggtcga ggtgccgtaa agcactaaat cggaacccta aagggagccc
     5521 ccgatttaga gcttgacggg gaaagccggc gaacgtggcg agaaaggaag ggaagaaagc
     5581 gaaaggagcg ggcgctaggg cgctggcaag tgtagcggtc acgctgcgcg taaccaccac
     5641 acccgccgcg cttaatgcgc cgctacaggg cgcgtcccat tcgcca
//
