LOCUS       sequence-380663-        4914 bp    DNA     circular SYN 15-APR-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 4914)
  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 4914)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 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..4914
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        44..134
                     /label=H1 promoter
                     /note="human H1 RNA promoter"
     primer_bind     55..74
                     /label=H1
                     /note="Human H1 promoter, forward primer"
     regulatory      178..187
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             184..1836
                     /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(250..268)
                     /label=LucNrev
                     /note="Firefly luciferase, reverse primer"
     polyA_signal    1877..1998
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1888..1907)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     1942..1961
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(2246..2263)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2417..3005)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2497..2516)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3176..3967,3968..4036))
                     /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     3799..3818
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4037..4141)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      4168..4623
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4255..4274)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4465..4486
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     polyA_signal    4754..4802
                     /note="synthetic polyadenylation signal"
     misc_feature    4816..4907
                     /label=pause site
                     /note="RNA polymerase II transcriptional pause signal from 
                     the human alpha-2 globin gene"
     primer_bind     4856..4875
                     /label=RVprimer3
                     /note="pGL3 vector, forward primer"
ORIGIN
        1 ggtaccgagc tcttacgcgt gctagcccgg gctcgagact agaatatttg catgtcgcta
       61 tgtgttctgg gaaatcacca taaacgtgaa atgtctttgg atttgggaat cttataagtt
      121 ctgtatgaga ccactctttc ccgatatcaa gcttggcatt ccggtactgt tggtaaagcc
      181 accatggaag acgccaaaaa cataaagaaa ggcccggcgc cattctatcc gctggaagat
      241 ggaaccgctg gagagcaact gcataaggct atgaagagat acgccctggt tcctggaaca
      301 attgctttta cagatgcaca tatcgaggtg gacatcactt acgctgagta cttcgaaatg
      361 tccgttcggt tggcagaagc tatgaaacga tatgggctga atacaaatca cagaatcgtc
      421 gtatgcagtg aaaactctct tcaattcttt atgccggtgt tgggcgcgtt atttatcgga
      481 gttgcagttg cgcccgcgaa cgacatttat aatgaacgtg aattgctcaa cagtatgggc
      541 atttcgcagc ctaccgtggt gttcgtttcc aaaaaggggt tgcaaaaaat tttgaacgtg
      601 caaaaaaagc tcccaatcat ccaaaaaatt attatcatgg attctaaaac ggattaccag
      661 ggatttcagt cgatgtacac gttcgtcaca tctcatctac ctcccggttt taatgaatac
      721 gattttgtgc cagagtcctt cgatagggac aagacaattg cactgatcat gaactcctct
      781 ggatctactg gtctgcctaa aggtgtcgct ctgcctcata gaactgcctg cgtgagattc
      841 tcgcatgcca gagatcctat ttttggcaat caaatcattc cggatactgc gattttaagt
      901 gttgttccat tccatcacgg ttttggaatg tttactacac tcggatattt gatatgtgga
      961 tttcgagtcg tcttaatgta tagatttgaa gaagagctgt ttctgaggag ccttcaggat
     1021 tacaagattc aaagtgcgct gctggtgcca accctattct ccttcttcgc caaaagcact
     1081 ctgattgaca aatacgattt atctaattta cacgaaattg cttctggtgg cgctcccctc
     1141 tctaaggaag tcggggaagc ggttgccaag aggttccatc tgccaggtat caggcaagga
     1201 tatgggctca ctgagactac atcagctatt ctgattacac ccgaggggga tgataaaccg
     1261 ggcgcggtcg gtaaagttgt tccatttttt gaagcgaagg ttgtggatct ggataccggg
     1321 aaaacgctgg gcgttaatca aagaggcgaa ctgtgtgtga gaggtcctat gattatgtcc
     1381 ggttatgtaa acaatccgga agcgaccaac gccttgattg acaaggatgg atggctacat
     1441 tctggagaca tagcttactg ggacgaagac gaacacttct tcatcgttga ccgcctgaag
     1501 tctctgatta agtacaaagg ctatcaggtg gctcccgctg aattggaatc catcttgctc
     1561 caacacccca acatcttcga cgcaggtgtc gcaggtcttc ccgacgatga cgccggtgaa
     1621 cttcccgccg ccgttgttgt tttggagcac ggaaagacga tgacggaaaa agagatcgtg
     1681 gattacgtcg ccagtcaagt aacaaccgcg aaaaagttgc gcggaggagt tgtgtttgtg
     1741 gacgaagtac cgaaaggtct taccggaaaa ctcgacgcaa gaaaaatcag agagatcctc
     1801 ataaaggcca agaagggcgg aaagatcgcc gtgtaattct agagtcgggg cggccggccg
     1861 cttcgagcag acatgataag atacattgat gagtttggac aaaccacaac tagaatgcag
     1921 tgaaaaaaat gctttatttg tgaaatttgt gatgctattg ctttatttgt aaccattata
     1981 agctgcaata aacaagttaa caacaacaat tgcattcatt ttatgtttca ggttcagggg
     2041 gaggtgtggg aggtttttta aagcaagtaa aacctctaca aatgtggtaa aatcgataag
     2101 gatccgtcga ccgatgccct tgagagcctt caacccagtc agctccttcc ggtgggcgcg
     2161 gggcatgact atcgtcgccg cacttatgac tgtcttcttt atcatgcaac tcgtaggaca
     2221 ggtgccggca gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc
     2281 tgcggcgagc ggtatcagct cactcaaagg cggtaatacg gttatccaca gaatcagggg
     2341 ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg
     2401 ccgcgttgct ggcgtttttc cataggctcc gcccccctga cgagcatcac aaaaatcgac
     2461 gctcaagtca gaggtggcga aacccgacag gactataaag ataccaggcg tttccccctg
     2521 gaagctccct cgtgcgctct cctgttccga ccctgccgct taccggatac ctgtccgcct
     2581 ttctcccttc gggaagcgtg gcgctttctc atagctcacg ctgtaggtat ctcagttcgg
     2641 tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct
     2701 gcgccttatc cggtaactat cgtcttgagt ccaacccggt aagacacgac ttatcgccac
     2761 tggcagcagc cactggtaac aggattagca gagcgaggta tgtaggcggt gctacagagt
     2821 tcttgaagtg gtggcctaac tacggctaca ctagaagaac agtatttggt atctgcgctc
     2881 tgctgaagcc agttaccttc ggaaaaagag ttggtagctc ttgatccggc aaacaaacca
     2941 ccgctggtag cggtggtttt tttgtttgca agcagcagat tacgcgcaga aaaaaaggat
     3001 ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc tcagtggaac gaaaactcac
     3061 gttaagggat tttggtcatg agattatcaa aaaggatctt cacctagatc cttttaaatt
     3121 aaaaatgaag ttttaaatca atctaaagta tatatgagta aacttggtct gacagttacc
     3181 aatgcttaat cagtgaggca cctatctcag cgatctgtct atttcgttca tccatagttg
     3241 cctgactccc cgtcgtgtag ataactacga tacgggaggg cttaccatct ggccccagtg
     3301 ctgcaatgat accgcgagac ccacgctcac cggctccaga tttatcagca ataaaccagc
     3361 cagccggaag ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc atccagtcta
     3421 ttaattgttg ccgggaagct agagtaagta gttcgccagt taatagtttg cgcaacgttg
     3481 ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct tcattcagct
     3541 ccggttccca acgatcaagg cgagttacat gatcccccat gttgtgcaaa aaagcggtta
     3601 gctccttcgg tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg
     3661 ttatggcagc actgcataat tctcttactg tcatgccatc cgtaagatgc ttttctgtga
     3721 ctggtgagta ctcaaccaag tcattctgag aatagtgtat gcggcgaccg agttgctctt
     3781 gcccggcgtc aatacgggat aataccgcgc cacatagcag aactttaaaa gtgctcatca
     3841 ttggaaaacg ttcttcgggg cgaaaactct caaggatctt accgctgttg agatccagtt
     3901 cgatgtaacc cactcgtgca cccaactgat cttcagcatc ttttactttc accagcgttt
     3961 ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga
     4021 aatgttgaat actcatactc ttcctttttc aatattattg aagcatttat cagggttatt
     4081 gtctcatgag cggatacata tttgaatgta tttagaaaaa taaacaaata ggggttccgc
     4141 gcacatttcc ccgaaaagtg ccacctgacg cgccctgtag cggcgcatta agcgcggcgg
     4201 gtgtggtggt tacgcgcagc gtgaccgcta cacttgccag cgccctagcg cccgctcctt
     4261 tcgctttctt cccttccttt ctcgccacgt tcgccggctt tccccgtcaa gctctaaatc
     4321 gggggctccc tttagggttc cgatttagtg ctttacggca cctcgacccc aaaaaacttg
     4381 attagggtga tggttcacgt agtgggccat cgccctgata gacggttttt cgccctttga
     4441 cgttggagtc cacgttcttt aatagtggac tcttgttcca aactggaaca acactcaacc
     4501 ctatctcggt ctattctttt gatttataag ggattttgcc gatttcggcc tattggttaa
     4561 aaaatgagct gatttaacaa aaatttaacg cgaattttaa caaaatatta acgcttacaa
     4621 tttgccattc gccattcagg ctgcgcaact gttgggaagg gcgatcggtg cgggcctctt
     4681 cgctattacg ccagcccaag ctaccatgat aagtaagtaa tattaaggta cgggaggtac
     4741 ttggagcggc cgcaataaaa tatctttatt ttcattacat ctgtgtgttg gttttttgtg
     4801 tgaatcgata gtactaacat acgctctcca tcaaaacaaa acgaaacaaa acaaactagc
     4861 aaaataggct gtccccagtg caagtgcagg tgccagaaca tttctctatc gata
//
