LOCUS       sequence-380667-        4913 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expressed a H1-core mutant by introducing multiple 7SK elements 
            (PSE, spacer, and TATA box).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4913)
  AUTHORS   Gao Z, van der Velden YU, Fan M, van der Linden CA, Vink M, 
            Herrera-Carrillo E, Berkhout B
  TITLE     Engineered mini H1 promoters with dedicated RNA Polymerase II or III
            activity.
  JOURNAL   J Biol Chem. 2020 Nov 5. pii: S0021-9258(20)00012-5. doi: 
            10.1074/jbc.RA120.015386.
  PUBMED    33154168
REFERENCE   2  (bases 1 to 4913)
  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: "J Biol 
            Chem. 2020 Nov 5. pii: S0021-9258(20)00012-5. doi: 
            10.1074/jbc.RA120.015386."
COMMENT     Sequence Label: H1.M10.luc
FEATURES             Location/Qualifiers
     source          1..4913
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     47..66
                     /label=H1
                     /note="Human H1 promoter, forward primer"
     regulatory      186..195
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             192..1844
                     /codon_start=1
                     /gene="luc+"
                     /product="firefly luciferase"
                     /label=luciferase
                     /note="enhanced luc+ version of the luciferase gene"
                     /translation="MEDAKNIKKGPAPFYPLEDGTAGEQLHKAMKRYALVPGTIAFTDA
                     HIEVDITYAEYFEMSVRLAEAMKRYGLNTNHRIVVCSENSLQFFMPVLGALFIGVAVAP
                     ANDIYNERELLNSMGISQPTVVFVSKKGLQKILNVQKKLPIIQKIIIMDSKTDYQGFQS
                     MYTFVTSHLPPGFNEYDFVPESFDRDKTIALIMNSSGSTGLPKGVALPHRTACVRFSHA
                     RDPIFGNQIIPDTAILSVVPFHHGFGMFTTLGYLICGFRVVLMYRFEEELFLRSLQDYK
                     IQSALLVPTLFSFFAKSTLIDKYDLSNLHEIASGGAPLSKEVGEAVAKRFHLPGIRQGY
                     GLTETTSAILITPEGDDKPGAVGKVVPFFEAKVVDLDTGKTLGVNQRGELCVRGPMIMS
                     GYVNNPEATNALIDKDGWLHSGDIAYWDEDEHFFIVDRLKSLIKYKGYQVAPAELESIL
                     LQHPNIFDAGVAGLPDDDAGELPAAVVVLEHGKTMTEKEIVDYVASQVTTAKKLRGGVV
                     FVDEVPKGLTGKLDARKIREILIKAKKGGKIAV"
     primer_bind     complement(258..276)
                     /label=LucNrev
                     /note="Firefly luciferase, reverse primer"
     polyA_signal    1885..2006
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1896..1915)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     1950..1969
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(2254..2271)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2425..3013)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2505..2524)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3184..3975,3976..4044))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     3807..3826
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4045..4149)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      4176..4631
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4263..4282)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4473..4494
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     polyA_signal    4762..4810
                     /note="synthetic polyadenylation signal"
     misc_feature    join(4824..4913,1..2)
                     /label=pause site
                     /note="RNA polymerase II transcriptional pause signal from 
                     the human alpha-2 globin gene"
     primer_bind     4864..4883
                     /label=RVprimer3
                     /note="pGL3 vector, forward primer"
ORIGIN
        1 ctatcgatag gtaccgagct cttacgcgtg ctagcatatt tgcatgtcgc tatgtgttct
       61 gggaaacttg acctaagtgt aaagttgaga cttccttcag gtttatatag ttctgtatga
      121 gaccactttt tcccatactc gagtgtgaag gggcagtagt aatataccgg tgatcttttt
      181 tggaagccac catggaagac gccaaaaaca taaagaaagg cccggcgcca ttctatccgc
      241 tggaagatgg aaccgctgga gagcaactgc ataaggctat gaagagatac gccctggttc
      301 ctggaacaat tgcttttaca gatgcacata tcgaggtgga catcacttac gctgagtact
      361 tcgaaatgtc cgttcggttg gcagaagcta tgaaacgata tgggctgaat acaaatcaca
      421 gaatcgtcgt atgcagtgaa aactctcttc aattctttat gccggtgttg ggcgcgttat
      481 ttatcggagt tgcagttgcg cccgcgaacg acatttataa tgaacgtgaa ttgctcaaca
      541 gtatgggcat ttcgcagcct accgtggtgt tcgtttccaa aaaggggttg caaaaaattt
      601 tgaacgtgca aaaaaagctc ccaatcatcc aaaaaattat tatcatggat tctaaaacgg
      661 attaccaggg atttcagtcg atgtacacgt tcgtcacatc tcatctacct cccggtttta
      721 atgaatacga ttttgtgcca gagtccttcg atagggacaa gacaattgca ctgatcatga
      781 actcctctgg atctactggt ctgcctaaag gtgtcgctct gcctcataga actgcctgcg
      841 tgagattctc gcatgccaga gatcctattt ttggcaatca aatcattccg gatactgcga
      901 ttttaagtgt tgttccattc catcacggtt ttggaatgtt tactacactc ggatatttga
      961 tatgtggatt tcgagtcgtc ttaatgtata gatttgaaga agagctgttt ctgaggagcc
     1021 ttcaggatta caagattcaa agtgcgctgc tggtgccaac cctattctcc ttcttcgcca
     1081 aaagcactct gattgacaaa tacgatttat ctaatttaca cgaaattgct tctggtggcg
     1141 ctcccctctc taaggaagtc ggggaagcgg ttgccaagag gttccatctg ccaggtatca
     1201 ggcaaggata tgggctcact gagactacat cagctattct gattacaccc gagggggatg
     1261 ataaaccggg cgcggtcggt aaagttgttc cattttttga agcgaaggtt gtggatctgg
     1321 ataccgggaa aacgctgggc gttaatcaaa gaggcgaact gtgtgtgaga ggtcctatga
     1381 ttatgtccgg ttatgtaaac aatccggaag cgaccaacgc cttgattgac aaggatggat
     1441 ggctacattc tggagacata gcttactggg acgaagacga acacttcttc atcgttgacc
     1501 gcctgaagtc tctgattaag tacaaaggct atcaggtggc tcccgctgaa ttggaatcca
     1561 tcttgctcca acaccccaac atcttcgacg caggtgtcgc aggtcttccc gacgatgacg
     1621 ccggtgaact tcccgccgcc gttgttgttt tggagcacgg aaagacgatg acggaaaaag
     1681 agatcgtgga ttacgtcgcc agtcaagtaa caaccgcgaa aaagttgcgc ggaggagttg
     1741 tgtttgtgga cgaagtaccg aaaggtctta ccggaaaact cgacgcaaga aaaatcagag
     1801 agatcctcat aaaggccaag aagggcggaa agatcgccgt gtaattctag agtcggggcg
     1861 gccggccgct tcgagcagac atgataagat acattgatga gtttggacaa accacaacta
     1921 gaatgcagtg aaaaaaatgc tttatttgtg aaatttgtga tgctattgct ttatttgtaa
     1981 ccattataag ctgcaataaa caagttaaca acaacaattg cattcatttt atgtttcagg
     2041 ttcaggggga ggtgtgggag gttttttaaa gcaagtaaaa cctctacaaa tgtggtaaaa
     2101 tcgataagga tccgtcgacc gatgcccttg agagccttca acccagtcag ctccttccgg
     2161 tgggcgcggg gcatgactat cgtcgccgca cttatgactg tcttctttat catgcaactc
     2221 gtaggacagg tgccggcagc gctcttccgc ttcctcgctc actgactcgc tgcgctcggt
     2281 cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt tatccacaga
     2341 atcaggggat aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg
     2401 taaaaaggcc gcgttgctgg cgtttttcca taggctccgc ccccctgacg agcatcacaa
     2461 aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga ctataaagat accaggcgtt
     2521 tccccctgga agctccctcg tgcgctctcc tgttccgacc ctgccgctta ccggatacct
     2581 gtccgccttt ctcccttcgg gaagcgtggc gctttctcat agctcacgct gtaggtatct
     2641 cagttcggtg taggtcgttc gctccaagct gggctgtgtg cacgaacccc ccgttcagcc
     2701 cgaccgctgc gccttatccg gtaactatcg tcttgagtcc aacccggtaa gacacgactt
     2761 atcgccactg gcagcagcca ctggtaacag gattagcaga gcgaggtatg taggcggtgc
     2821 tacagagttc ttgaagtggt ggcctaacta cggctacact agaagaacag tatttggtat
     2881 ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa
     2941 acaaaccacc gctggtagcg gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa
     3001 aaaaggatct caagaagatc ctttgatctt ttctacgggg tctgacgctc agtggaacga
     3061 aaactcacgt taagggattt tggtcatgag attatcaaaa aggatcttca cctagatcct
     3121 tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa cttggtctga
     3181 cagttaccaa tgcttaatca gtgaggcacc tatctcagcg atctgtctat ttcgttcatc
     3241 catagttgcc tgactccccg tcgtgtagat aactacgata cgggagggct taccatctgg
     3301 ccccagtgct gcaatgatac cgcgagaccc acgctcaccg gctccagatt tatcagcaat
     3361 aaaccagcca gccggaaggg ccgagcgcag aagtggtcct gcaactttat ccgcctccat
     3421 ccagtctatt aattgttgcc gggaagctag agtaagtagt tcgccagtta atagtttgcg
     3481 caacgttgtt gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc
     3541 attcagctcc ggttcccaac gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa
     3601 agcggttagc tccttcggtc ctccgatcgt tgtcagaagt aagttggccg cagtgttatc
     3661 actcatggtt atggcagcac tgcataattc tcttactgtc atgccatccg taagatgctt
     3721 ttctgtgact ggtgagtact caaccaagtc attctgagaa tagtgtatgc ggcgaccgag
     3781 ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca catagcagaa ctttaaaagt
     3841 gctcatcatt ggaaaacgtt cttcggggcg aaaactctca aggatcttac cgctgttgag
     3901 atccagttcg atgtaaccca ctcgtgcacc caactgatct tcagcatctt ttactttcac
     3961 cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc
     4021 gacacggaaa tgttgaatac tcatactctt cctttttcaa tattattgaa gcatttatca
     4081 gggttattgt ctcatgagcg gatacatatt tgaatgtatt tagaaaaata aacaaatagg
     4141 ggttccgcgc acatttcccc gaaaagtgcc acctgacgcg ccctgtagcg gcgcattaag
     4201 cgcggcgggt gtggtggtta cgcgcagcgt gaccgctaca cttgccagcg ccctagcgcc
     4261 cgctcctttc gctttcttcc cttcctttct cgccacgttc gccggctttc cccgtcaagc
     4321 tctaaatcgg gggctccctt tagggttccg atttagtgct ttacggcacc tcgaccccaa
     4381 aaaacttgat tagggtgatg gttcacgtag tgggccatcg ccctgataga cggtttttcg
     4441 ccctttgacg ttggagtcca cgttctttaa tagtggactc ttgttccaaa ctggaacaac
     4501 actcaaccct atctcggtct attcttttga tttataaggg attttgccga tttcggccta
     4561 ttggttaaaa aatgagctga tttaacaaaa atttaacgcg aattttaaca aaatattaac
     4621 gcttacaatt tgccattcgc cattcaggct gcgcaactgt tgggaagggc gatcggtgcg
     4681 ggcctcttcg ctattacgcc agcccaagct accatgataa gtaagtaata ttaaggtacg
     4741 ggaggtactt ggagcggccg caataaaata tctttatttt cattacatct gtgtgttggt
     4801 tttttgtgtg aatcgatagt actaacatac gctctccatc aaaacaaaac gaaacaaaac
     4861 aaactagcaa aataggctgt ccccagtgca agtgcaggtg ccagaacatt tct
//
