LOCUS       sequence-374971-        5038 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses a truncated PolIII H1 promoter of 224 nucleotides (H1.224)
            with the TATA box mutated.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5038)
  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 5038)
  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: H1.224.M.luc
FEATURES             Location/Qualifiers
     source          1..5038
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        27..242
                     /label=H1 promoter
                     /note="human H1 RNA promoter"
     primer_bind     163..182
                     /label=H1
                     /note="Human H1 promoter, forward primer"
     regulatory      302..311
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             308..1960
                     /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(374..392)
                     /label=LucNrev
                     /note="Firefly luciferase, reverse primer"
     polyA_signal    2001..2122
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2012..2031)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2066..2085
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(2370..2387)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2541..3129)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2621..2640)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3300..4091,4092..4160))
                     /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     3923..3942
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4161..4265)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      4292..4747
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4379..4398)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4589..4610
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     polyA_signal    4878..4926
                     /note="synthetic polyadenylation signal"
     misc_feature    4940..5031
                     /label=pause site
                     /note="RNA polymerase II transcriptional pause signal from 
                     the human alpha-2 globin gene"
     primer_bind     4980..4999
                     /label=RVprimer3
                     /note="pGL3 vector, forward primer"
ORIGIN
        1 ggtaccgagc tcttacgcgt gctagcgaac gctgacgtca tcaacccgct ccaaggaatc
       61 gcgggcccag tgtcactagg cgggaacacc cagcgcgcgt gcgccctggc aggaagatgg
      121 ctgtgaggga caggggagtg gcgccctgca aatatttgca tgtcgctatg tgttctggga
      181 aatcaccata aacgtgaaat gtctttggat ttgggaatcc tcgaggttct gtatgagacc
      241 actctttccc atactcgagt gtgaaggggc agtagtaata taccggtgat cttttttgga
      301 agccaccatg gaagacgcca aaaacataaa gaaaggcccg gcgccattct atccgctgga
      361 agatggaacc gctggagagc aactgcataa ggctatgaag agatacgccc tggttcctgg
      421 aacaattgct tttacagatg cacatatcga ggtggacatc acttacgctg agtacttcga
      481 aatgtccgtt cggttggcag aagctatgaa acgatatggg ctgaatacaa atcacagaat
      541 cgtcgtatgc agtgaaaact ctcttcaatt ctttatgccg gtgttgggcg cgttatttat
      601 cggagttgca gttgcgcccg cgaacgacat ttataatgaa cgtgaattgc tcaacagtat
      661 gggcatttcg cagcctaccg tggtgttcgt ttccaaaaag gggttgcaaa aaattttgaa
      721 cgtgcaaaaa aagctcccaa tcatccaaaa aattattatc atggattcta aaacggatta
      781 ccagggattt cagtcgatgt acacgttcgt cacatctcat ctacctcccg gttttaatga
      841 atacgatttt gtgccagagt ccttcgatag ggacaagaca attgcactga tcatgaactc
      901 ctctggatct actggtctgc ctaaaggtgt cgctctgcct catagaactg cctgcgtgag
      961 attctcgcat gccagagatc ctatttttgg caatcaaatc attccggata ctgcgatttt
     1021 aagtgttgtt ccattccatc acggttttgg aatgtttact acactcggat atttgatatg
     1081 tggatttcga gtcgtcttaa tgtatagatt tgaagaagag ctgtttctga ggagccttca
     1141 ggattacaag attcaaagtg cgctgctggt gccaacccta ttctccttct tcgccaaaag
     1201 cactctgatt gacaaatacg atttatctaa tttacacgaa attgcttctg gtggcgctcc
     1261 cctctctaag gaagtcgggg aagcggttgc caagaggttc catctgccag gtatcaggca
     1321 aggatatggg ctcactgaga ctacatcagc tattctgatt acacccgagg gggatgataa
     1381 accgggcgcg gtcggtaaag ttgttccatt ttttgaagcg aaggttgtgg atctggatac
     1441 cgggaaaacg ctgggcgtta atcaaagagg cgaactgtgt gtgagaggtc ctatgattat
     1501 gtccggttat gtaaacaatc cggaagcgac caacgccttg attgacaagg atggatggct
     1561 acattctgga gacatagctt actgggacga agacgaacac ttcttcatcg ttgaccgcct
     1621 gaagtctctg attaagtaca aaggctatca ggtggctccc gctgaattgg aatccatctt
     1681 gctccaacac cccaacatct tcgacgcagg tgtcgcaggt cttcccgacg atgacgccgg
     1741 tgaacttccc gccgccgttg ttgttttgga gcacggaaag acgatgacgg aaaaagagat
     1801 cgtggattac gtcgccagtc aagtaacaac cgcgaaaaag ttgcgcggag gagttgtgtt
     1861 tgtggacgaa gtaccgaaag gtcttaccgg aaaactcgac gcaagaaaaa tcagagagat
     1921 cctcataaag gccaagaagg gcggaaagat cgccgtgtaa ttctagagtc ggggcggccg
     1981 gccgcttcga gcagacatga taagatacat tgatgagttt ggacaaacca caactagaat
     2041 gcagtgaaaa aaatgcttta tttgtgaaat ttgtgatgct attgctttat ttgtaaccat
     2101 tataagctgc aataaacaag ttaacaacaa caattgcatt cattttatgt ttcaggttca
     2161 gggggaggtg tgggaggttt tttaaagcaa gtaaaacctc tacaaatgtg gtaaaatcga
     2221 taaggatccg tcgaccgatg cccttgagag ccttcaaccc agtcagctcc ttccggtggg
     2281 cgcggggcat gactatcgtc gccgcactta tgactgtctt ctttatcatg caactcgtag
     2341 gacaggtgcc ggcagcgctc ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt
     2401 cggctgcggc gagcggtatc agctcactca aaggcggtaa tacggttatc cacagaatca
     2461 ggggataacg caggaaagaa catgtgagca aaaggccagc aaaaggccag gaaccgtaaa
     2521 aaggccgcgt tgctggcgtt tttccatagg ctccgccccc ctgacgagca tcacaaaaat
     2581 cgacgctcaa gtcagaggtg gcgaaacccg acaggactat aaagatacca ggcgtttccc
     2641 cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc
     2701 gcctttctcc cttcgggaag cgtggcgctt tctcatagct cacgctgtag gtatctcagt
     2761 tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt tcagcccgac
     2821 cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggtaagaca cgacttatcg
     2881 ccactggcag cagccactgg taacaggatt agcagagcga ggtatgtagg cggtgctaca
     2941 gagttcttga agtggtggcc taactacggc tacactagaa gaacagtatt tggtatctgc
     3001 gctctgctga agccagttac cttcggaaaa agagttggta gctcttgatc cggcaaacaa
     3061 accaccgctg gtagcggtgg tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa
     3121 ggatctcaag aagatccttt gatcttttct acggggtctg acgctcagtg gaacgaaaac
     3181 tcacgttaag ggattttggt catgagatta tcaaaaagga tcttcaccta gatcctttta
     3241 aattaaaaat gaagttttaa atcaatctaa agtatatatg agtaaacttg gtctgacagt
     3301 taccaatgct taatcagtga ggcacctatc tcagcgatct gtctatttcg ttcatccata
     3361 gttgcctgac tccccgtcgt gtagataact acgatacggg agggcttacc atctggcccc
     3421 agtgctgcaa tgataccgcg agacccacgc tcaccggctc cagatttatc agcaataaac
     3481 cagccagccg gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag
     3541 tctattaatt gttgccggga agctagagta agtagttcgc cagttaatag tttgcgcaac
     3601 gttgttgcca ttgctacagg catcgtggtg tcacgctcgt cgtttggtat ggcttcattc
     3661 agctccggtt cccaacgatc aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg
     3721 gttagctcct tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt gttatcactc
     3781 atggttatgg cagcactgca taattctctt actgtcatgc catccgtaag atgcttttct
     3841 gtgactggtg agtactcaac caagtcattc tgagaatagt gtatgcggcg accgagttgc
     3901 tcttgcccgg cgtcaatacg ggataatacc gcgccacata gcagaacttt aaaagtgctc
     3961 atcattggaa aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc
     4021 agttcgatgt aacccactcg tgcacccaac tgatcttcag catcttttac tttcaccagc
     4081 gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca
     4141 cggaaatgtt gaatactcat actcttcctt tttcaatatt attgaagcat ttatcagggt
     4201 tattgtctca tgagcggata catatttgaa tgtatttaga aaaataaaca aataggggtt
     4261 ccgcgcacat ttccccgaaa agtgccacct gacgcgccct gtagcggcgc attaagcgcg
     4321 gcgggtgtgg tggttacgcg cagcgtgacc gctacacttg ccagcgccct agcgcccgct
     4381 cctttcgctt tcttcccttc ctttctcgcc acgttcgccg gctttccccg tcaagctcta
     4441 aatcgggggc tccctttagg gttccgattt agtgctttac ggcacctcga ccccaaaaaa
     4501 cttgattagg gtgatggttc acgtagtggg ccatcgccct gatagacggt ttttcgccct
     4561 ttgacgttgg agtccacgtt ctttaatagt ggactcttgt tccaaactgg aacaacactc
     4621 aaccctatct cggtctattc ttttgattta taagggattt tgccgatttc ggcctattgg
     4681 ttaaaaaatg agctgattta acaaaaattt aacgcgaatt ttaacaaaat attaacgctt
     4741 acaatttgcc attcgccatt caggctgcgc aactgttggg aagggcgatc ggtgcgggcc
     4801 tcttcgctat tacgccagcc caagctacca tgataagtaa gtaatattaa ggtacgggag
     4861 gtacttggag cggccgcaat aaaatatctt tattttcatt acatctgtgt gttggttttt
     4921 tgtgtgaatc gatagtacta acatacgctc tccatcaaaa caaaacgaaa caaaacaaac
     4981 tagcaaaata ggctgtcccc agtgcaagtg caggtgccag aacatttctc tatcgata
//
