LOCUS       sequence-374918-        4913 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses a truncated Polymerase III H1 promoter of 99 nucleotides 
            (H1-99).
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: H1.99.luc
FEATURES             Location/Qualifiers
     source          1..4913
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        27..117
                     /label=H1 promoter
                     /note="human H1 RNA promoter"
     primer_bind     38..57
                     /label=H1
                     /note="Human H1 promoter, forward primer"
     regulatory      177..186
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             183..1835
                     /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(249..267)
                     /label=LucNrev
                     /note="Firefly luciferase, reverse primer"
     polyA_signal    1876..1997
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1887..1906)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     1941..1960
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(2245..2262)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2416..3004)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2496..2515)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3175..3966,3967..4035))
                     /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     3798..3817
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4036..4140)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      4167..4622
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4254..4273)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4464..4485
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     polyA_signal    4753..4801
                     /note="synthetic polyadenylation signal"
     misc_feature    4815..4906
                     /label=pause site
                     /note="RNA polymerase II transcriptional pause signal from 
                     the human alpha-2 globin gene"
     primer_bind     4855..4874
                     /label=RVprimer3
                     /note="pGL3 vector, forward primer"
ORIGIN
        1 ggtaccgagc tcttacgcgt gctagcatat ttgcatgtcg ctatgtgttc tgggaaatca
       61 ccataaacgt gaaatgtctt tggatttggg aatcttataa gttctgtatg agaccactct
      121 ttcccatact cgagtgtgaa ggggcagtag taatataccg gtgatctttt ttggaagcca
      181 ccatggaaga cgccaaaaac ataaagaaag gcccggcgcc attctatccg ctggaagatg
      241 gaaccgctgg agagcaactg cataaggcta tgaagagata cgccctggtt cctggaacaa
      301 ttgcttttac agatgcacat atcgaggtgg acatcactta cgctgagtac ttcgaaatgt
      361 ccgttcggtt ggcagaagct atgaaacgat atgggctgaa tacaaatcac agaatcgtcg
      421 tatgcagtga aaactctctt caattcttta tgccggtgtt gggcgcgtta tttatcggag
      481 ttgcagttgc gcccgcgaac gacatttata atgaacgtga attgctcaac agtatgggca
      541 tttcgcagcc taccgtggtg ttcgtttcca aaaaggggtt gcaaaaaatt ttgaacgtgc
      601 aaaaaaagct cccaatcatc caaaaaatta ttatcatgga ttctaaaacg gattaccagg
      661 gatttcagtc gatgtacacg ttcgtcacat ctcatctacc tcccggtttt aatgaatacg
      721 attttgtgcc agagtccttc gatagggaca agacaattgc actgatcatg aactcctctg
      781 gatctactgg tctgcctaaa ggtgtcgctc tgcctcatag aactgcctgc gtgagattct
      841 cgcatgccag agatcctatt tttggcaatc aaatcattcc ggatactgcg attttaagtg
      901 ttgttccatt ccatcacggt tttggaatgt ttactacact cggatatttg atatgtggat
      961 ttcgagtcgt cttaatgtat agatttgaag aagagctgtt tctgaggagc cttcaggatt
     1021 acaagattca aagtgcgctg ctggtgccaa ccctattctc cttcttcgcc aaaagcactc
     1081 tgattgacaa atacgattta tctaatttac acgaaattgc ttctggtggc gctcccctct
     1141 ctaaggaagt cggggaagcg gttgccaaga ggttccatct gccaggtatc aggcaaggat
     1201 atgggctcac tgagactaca tcagctattc tgattacacc cgagggggat gataaaccgg
     1261 gcgcggtcgg taaagttgtt ccattttttg aagcgaaggt tgtggatctg gataccggga
     1321 aaacgctggg cgttaatcaa agaggcgaac tgtgtgtgag aggtcctatg attatgtccg
     1381 gttatgtaaa caatccggaa gcgaccaacg ccttgattga caaggatgga tggctacatt
     1441 ctggagacat agcttactgg gacgaagacg aacacttctt catcgttgac cgcctgaagt
     1501 ctctgattaa gtacaaaggc tatcaggtgg ctcccgctga attggaatcc atcttgctcc
     1561 aacaccccaa catcttcgac gcaggtgtcg caggtcttcc cgacgatgac gccggtgaac
     1621 ttcccgccgc cgttgttgtt ttggagcacg gaaagacgat gacggaaaaa gagatcgtgg
     1681 attacgtcgc cagtcaagta acaaccgcga aaaagttgcg cggaggagtt gtgtttgtgg
     1741 acgaagtacc gaaaggtctt accggaaaac tcgacgcaag aaaaatcaga gagatcctca
     1801 taaaggccaa gaagggcgga aagatcgccg tgtaattcta gagtcggggc ggccggccgc
     1861 ttcgagcaga catgataaga tacattgatg agtttggaca aaccacaact agaatgcagt
     1921 gaaaaaaatg ctttatttgt gaaatttgtg atgctattgc tttatttgta accattataa
     1981 gctgcaataa acaagttaac aacaacaatt gcattcattt tatgtttcag gttcaggggg
     2041 aggtgtggga ggttttttaa agcaagtaaa acctctacaa atgtggtaaa atcgataagg
     2101 atccgtcgac cgatgccctt gagagccttc aacccagtca gctccttccg gtgggcgcgg
     2161 ggcatgacta tcgtcgccgc acttatgact gtcttcttta tcatgcaact cgtaggacag
     2221 gtgccggcag cgctcttccg cttcctcgct cactgactcg ctgcgctcgg tcgttcggct
     2281 gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg ttatccacag aatcagggga
     2341 taacgcagga aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc
     2401 cgcgttgctg gcgtttttcc ataggctccg cccccctgac gagcatcaca aaaatcgacg
     2461 ctcaagtcag aggtggcgaa acccgacagg actataaaga taccaggcgt ttccccctgg
     2521 aagctccctc gtgcgctctc ctgttccgac cctgccgctt accggatacc tgtccgcctt
     2581 tctcccttcg ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc tcagttcggt
     2641 gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg
     2701 cgccttatcc ggtaactatc gtcttgagtc caacccggta agacacgact tatcgccact
     2761 ggcagcagcc actggtaaca ggattagcag agcgaggtat gtaggcggtg ctacagagtt
     2821 cttgaagtgg tggcctaact acggctacac tagaagaaca gtatttggta tctgcgctct
     2881 gctgaagcca gttaccttcg gaaaaagagt tggtagctct tgatccggca aacaaaccac
     2941 cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc
     3001 tcaagaagat cctttgatct tttctacggg gtctgacgct cagtggaacg aaaactcacg
     3061 ttaagggatt ttggtcatga gattatcaaa aaggatcttc acctagatcc ttttaaatta
     3121 aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa acttggtctg acagttacca
     3181 atgcttaatc agtgaggcac ctatctcagc gatctgtcta tttcgttcat ccatagttgc
     3241 ctgactcccc gtcgtgtaga taactacgat acgggagggc ttaccatctg gccccagtgc
     3301 tgcaatgata ccgcgagacc cacgctcacc ggctccagat ttatcagcaa taaaccagcc
     3361 agccggaagg gccgagcgca gaagtggtcc tgcaacttta tccgcctcca tccagtctat
     3421 taattgttgc cgggaagcta gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt
     3481 tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc
     3541 cggttcccaa cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa aagcggttag
     3601 ctccttcggt cctccgatcg ttgtcagaag taagttggcc gcagtgttat cactcatggt
     3661 tatggcagca ctgcataatt ctcttactgt catgccatcc gtaagatgct tttctgtgac
     3721 tggtgagtac tcaaccaagt cattctgaga atagtgtatg cggcgaccga gttgctcttg
     3781 cccggcgtca atacgggata ataccgcgcc acatagcaga actttaaaag tgctcatcat
     3841 tggaaaacgt tcttcggggc gaaaactctc aaggatctta ccgctgttga gatccagttc
     3901 gatgtaaccc actcgtgcac ccaactgatc ttcagcatct tttactttca ccagcgtttc
     3961 tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa
     4021 atgttgaata ctcatactct tcctttttca atattattga agcatttatc agggttattg
     4081 tctcatgagc ggatacatat ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg
     4141 cacatttccc cgaaaagtgc cacctgacgc gccctgtagc ggcgcattaa gcgcggcggg
     4201 tgtggtggtt acgcgcagcg tgaccgctac acttgccagc gccctagcgc ccgctccttt
     4261 cgctttcttc ccttcctttc tcgccacgtt cgccggcttt ccccgtcaag ctctaaatcg
     4321 ggggctccct ttagggttcc gatttagtgc tttacggcac ctcgacccca aaaaacttga
     4381 ttagggtgat ggttcacgta gtgggccatc gccctgatag acggtttttc gccctttgac
     4441 gttggagtcc acgttcttta atagtggact cttgttccaa actggaacaa cactcaaccc
     4501 tatctcggtc tattcttttg atttataagg gattttgccg atttcggcct attggttaaa
     4561 aaatgagctg atttaacaaa aatttaacgc gaattttaac aaaatattaa cgcttacaat
     4621 ttgccattcg ccattcaggc tgcgcaactg ttgggaaggg cgatcggtgc gggcctcttc
     4681 gctattacgc cagcccaagc taccatgata agtaagtaat attaaggtac gggaggtact
     4741 tggagcggcc gcaataaaat atctttattt tcattacatc tgtgtgttgg ttttttgtgt
     4801 gaatcgatag tactaacata cgctctccat caaaacaaaa cgaaacaaaa caaactagca
     4861 aaataggctg tccccagtgc aagtgcaggt gccagaacat ttctctatcg ata
//
