LOCUS       sequence-380666-        5039 bp    DNA     circular SYN 15-APR-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 5039)
  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 5039)
  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: H1.224.M.luc
FEATURES             Location/Qualifiers
     source          1..5039
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        64..278
                     /label=H1 promoter
                     /note="human H1 RNA promoter"
     primer_bind     199..218
                     /label=H1
                     /note="Human H1 promoter, forward primer"
     regulatory      322..331
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             328..1980
                     /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(394..412)
                     /label=LucNrev
                     /note="Firefly luciferase, reverse primer"
     polyA_signal    2021..2142
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2032..2051)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2086..2105
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(2390..2407)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2561..3149)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2641..2660)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3320..4111,4112..4180))
                     /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     3943..3962
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4181..4285)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      4312..4767
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4399..4418)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4609..4630
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     polyA_signal    4898..4946
                     /note="synthetic polyadenylation signal"
     misc_feature    join(4960..5039,1..12)
                     /label=pause site
                     /note="RNA polymerase II transcriptional pause signal from 
                     the human alpha-2 globin gene"
     primer_bind     5000..5019
                     /label=RVprimer3
                     /note="pGL3 vector, forward primer"
ORIGIN
        1 gaacatttct ctatcgatag gtaccgagct cttacgcgtg ctagcccggg ctcgagacta
       61 gacgaacgct gacgtcatca acccgctcca aggaatcgcg ggcccagtgt cactaggcgg
      121 gaacacccag cgcgcgtgcg ccctggcagg aagatggctg tgagggacag gggagtggcg
      181 ccctgcaata tttgcatgtc gctatgtgtt ctgggaaatc accataaacg tgaaatgtct
      241 ttggatttgg gaatcctcga ggttctgtat gagaccactc tttcccgata tcaagcttgg
      301 cattccggta ctgttggtaa agccaccatg gaagacgcca aaaacataaa gaaaggcccg
      361 gcgccattct atccgctgga agatggaacc gctggagagc aactgcataa ggctatgaag
      421 agatacgccc tggttcctgg aacaattgct tttacagatg cacatatcga ggtggacatc
      481 acttacgctg agtacttcga aatgtccgtt cggttggcag aagctatgaa acgatatggg
      541 ctgaatacaa atcacagaat cgtcgtatgc agtgaaaact ctcttcaatt ctttatgccg
      601 gtgttgggcg cgttatttat cggagttgca gttgcgcccg cgaacgacat ttataatgaa
      661 cgtgaattgc tcaacagtat gggcatttcg cagcctaccg tggtgttcgt ttccaaaaag
      721 gggttgcaaa aaattttgaa cgtgcaaaaa aagctcccaa tcatccaaaa aattattatc
      781 atggattcta aaacggatta ccagggattt cagtcgatgt acacgttcgt cacatctcat
      841 ctacctcccg gttttaatga atacgatttt gtgccagagt ccttcgatag ggacaagaca
      901 attgcactga tcatgaactc ctctggatct actggtctgc ctaaaggtgt cgctctgcct
      961 catagaactg cctgcgtgag attctcgcat gccagagatc ctatttttgg caatcaaatc
     1021 attccggata ctgcgatttt aagtgttgtt ccattccatc acggttttgg aatgtttact
     1081 acactcggat atttgatatg tggatttcga gtcgtcttaa tgtatagatt tgaagaagag
     1141 ctgtttctga ggagccttca ggattacaag attcaaagtg cgctgctggt gccaacccta
     1201 ttctccttct tcgccaaaag cactctgatt gacaaatacg atttatctaa tttacacgaa
     1261 attgcttctg gtggcgctcc cctctctaag gaagtcgggg aagcggttgc caagaggttc
     1321 catctgccag gtatcaggca aggatatggg ctcactgaga ctacatcagc tattctgatt
     1381 acacccgagg gggatgataa accgggcgcg gtcggtaaag ttgttccatt ttttgaagcg
     1441 aaggttgtgg atctggatac cgggaaaacg ctgggcgtta atcaaagagg cgaactgtgt
     1501 gtgagaggtc ctatgattat gtccggttat gtaaacaatc cggaagcgac caacgccttg
     1561 attgacaagg atggatggct acattctgga gacatagctt actgggacga agacgaacac
     1621 ttcttcatcg ttgaccgcct gaagtctctg attaagtaca aaggctatca ggtggctccc
     1681 gctgaattgg aatccatctt gctccaacac cccaacatct tcgacgcagg tgtcgcaggt
     1741 cttcccgacg atgacgccgg tgaacttccc gccgccgttg ttgttttgga gcacggaaag
     1801 acgatgacgg aaaaagagat cgtggattac gtcgccagtc aagtaacaac cgcgaaaaag
     1861 ttgcgcggag gagttgtgtt tgtggacgaa gtaccgaaag gtcttaccgg aaaactcgac
     1921 gcaagaaaaa tcagagagat cctcataaag gccaagaagg gcggaaagat cgccgtgtaa
     1981 ttctagagtc ggggcggccg gccgcttcga gcagacatga taagatacat tgatgagttt
     2041 ggacaaacca caactagaat gcagtgaaaa aaatgcttta tttgtgaaat ttgtgatgct
     2101 attgctttat ttgtaaccat tataagctgc aataaacaag ttaacaacaa caattgcatt
     2161 cattttatgt ttcaggttca gggggaggtg tgggaggttt tttaaagcaa gtaaaacctc
     2221 tacaaatgtg gtaaaatcga taaggatccg tcgaccgatg cccttgagag ccttcaaccc
     2281 agtcagctcc ttccggtggg cgcggggcat gactatcgtc gccgcactta tgactgtctt
     2341 ctttatcatg caactcgtag gacaggtgcc ggcagcgctc ttccgcttcc tcgctcactg
     2401 actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc agctcactca aaggcggtaa
     2461 tacggttatc cacagaatca ggggataacg caggaaagaa catgtgagca aaaggccagc
     2521 aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg ctccgccccc
     2581 ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg acaggactat
     2641 aaagatacca ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc
     2701 cgcttaccgg atacctgtcc gcctttctcc cttcgggaag cgtggcgctt tctcatagct
     2761 cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg
     2821 aaccccccgt tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc
     2881 cggtaagaca cgacttatcg ccactggcag cagccactgg taacaggatt agcagagcga
     2941 ggtatgtagg cggtgctaca gagttcttga agtggtggcc taactacggc tacactagaa
     3001 gaacagtatt tggtatctgc gctctgctga agccagttac cttcggaaaa agagttggta
     3061 gctcttgatc cggcaaacaa accaccgctg gtagcggtgg tttttttgtt tgcaagcagc
     3121 agattacgcg cagaaaaaaa ggatctcaag aagatccttt gatcttttct acggggtctg
     3181 acgctcagtg gaacgaaaac tcacgttaag ggattttggt catgagatta tcaaaaagga
     3241 tcttcaccta gatcctttta aattaaaaat gaagttttaa atcaatctaa agtatatatg
     3301 agtaaacttg gtctgacagt taccaatgct taatcagtga ggcacctatc tcagcgatct
     3361 gtctatttcg ttcatccata gttgcctgac tccccgtcgt gtagataact acgatacggg
     3421 agggcttacc atctggcccc agtgctgcaa tgataccgcg agacccacgc tcaccggctc
     3481 cagatttatc agcaataaac cagccagccg gaagggccga gcgcagaagt ggtcctgcaa
     3541 ctttatccgc ctccatccag tctattaatt gttgccggga agctagagta agtagttcgc
     3601 cagttaatag tttgcgcaac gttgttgcca ttgctacagg catcgtggtg tcacgctcgt
     3661 cgtttggtat ggcttcattc agctccggtt cccaacgatc aaggcgagtt acatgatccc
     3721 ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc agaagtaagt
     3781 tggccgcagt gttatcactc atggttatgg cagcactgca taattctctt actgtcatgc
     3841 catccgtaag atgcttttct gtgactggtg agtactcaac caagtcattc tgagaatagt
     3901 gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg ggataatacc gcgccacata
     3961 gcagaacttt aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa ctctcaagga
     4021 tcttaccgct gttgagatcc agttcgatgt aacccactcg tgcacccaac tgatcttcag
     4081 catcttttac tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa
     4141 aaaagggaat aagggcgaca cggaaatgtt gaatactcat actcttcctt tttcaatatt
     4201 attgaagcat ttatcagggt tattgtctca tgagcggata catatttgaa tgtatttaga
     4261 aaaataaaca aataggggtt ccgcgcacat ttccccgaaa agtgccacct gacgcgccct
     4321 gtagcggcgc attaagcgcg gcgggtgtgg tggttacgcg cagcgtgacc gctacacttg
     4381 ccagcgccct agcgcccgct cctttcgctt tcttcccttc ctttctcgcc acgttcgccg
     4441 gctttccccg tcaagctcta aatcgggggc tccctttagg gttccgattt agtgctttac
     4501 ggcacctcga ccccaaaaaa cttgattagg gtgatggttc acgtagtggg ccatcgccct
     4561 gatagacggt ttttcgccct ttgacgttgg agtccacgtt ctttaatagt ggactcttgt
     4621 tccaaactgg aacaacactc aaccctatct cggtctattc ttttgattta taagggattt
     4681 tgccgatttc ggcctattgg ttaaaaaatg agctgattta acaaaaattt aacgcgaatt
     4741 ttaacaaaat attaacgctt acaatttgcc attcgccatt caggctgcgc aactgttggg
     4801 aagggcgatc ggtgcgggcc tcttcgctat tacgccagcc caagctacca tgataagtaa
     4861 gtaatattaa ggtacgggag gtacttggag cggccgcaat aaaatatctt tattttcatt
     4921 acatctgtgt gttggttttt tgtgtgaatc gatagtacta acatacgctc tccatcaaaa
     4981 caaaacgaaa caaaacaaac tagcaaaata ggctgtcccc agtgcaagtg caggtgcca
//
