LOCUS       sequence-374974-        4914 bp    DNA     circular SYN 14-APR-2025
DEFINITION  Expressed a H1-core mutant by introducing multiple 7SK elements 
            (octamer, PSE, spacer, and TATA box).
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 14, 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.M11.luc
FEATURES             Location/Qualifiers
     source          1..4914
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    8..99
                     /label=pause site
                     /note="RNA polymerase II transcriptional pause signal from 
                     the human alpha-2 globin gene"
     primer_bind     complement(40..59)
                     /label=RVprimer3
                     /note="pGL3 vector, forward primer"
     polyA_signal    113..161
                     /note="synthetic polyadenylation signal"
     rep_origin      complement(292..747)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(429..450)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     641..660
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        774..878
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(879..947,948..1739)
                     /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     complement(1097..1116)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      1910..2498
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2399..2418
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2652..2669
                     /label=L4440
                     /note="L4440 vector, forward primer"
     polyA_signal    2917..3038
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2954..2973)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3008..3027
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     CDS             complement(3079..4731)
                     /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     4647..4665
                     /label=LucNrev
                     /note="Firefly luciferase, reverse primer"
     regulatory      4728..4737
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     primer_bind     complement(4857..4876)
                     /label=H1
                     /note="Human H1 promoter, forward primer"
ORIGIN
        1 tatcgataga gaaatgttct ggcacctgca cttgcactgg ggacagccta ttttgctagt
       61 ttgttttgtt tcgttttgtt ttgatggaga gcgtatgtta gtactatcga ttcacacaaa
      121 aaaccaacac acagatgtaa tgaaaataaa gatattttat tgcggccgct ccaagtacct
      181 cccgtacctt aatattactt acttatcatg gtagcttggg ctggcgtaat agcgaagagg
      241 cccgcaccga tcgcccttcc caacagttgc gcagcctgaa tggcgaatgg caaattgtaa
      301 gcgttaatat tttgttaaaa ttcgcgttaa atttttgtta aatcagctca ttttttaacc
      361 aataggccga aatcggcaaa atcccttata aatcaaaaga atagaccgag atagggttga
      421 gtgttgttcc agtttggaac aagagtccac tattaaagaa cgtggactcc aacgtcaaag
      481 ggcgaaaaac cgtctatcag ggcgatggcc cactacgtga accatcaccc taatcaagtt
      541 ttttggggtc gaggtgccgt aaagcactaa atcggaaccc taaagggagc ccccgattta
      601 gagcttgacg gggaaagccg gcgaacgtgg cgagaaagga agggaagaaa gcgaaaggag
      661 cgggcgctag ggcgctggca agtgtagcgg tcacgctgcg cgtaaccacc acacccgccg
      721 cgcttaatgc gccgctacag ggcgcgtcag gtggcacttt tcggggaaat gtgcgcggaa
      781 cccctatttg tttatttttc taaatacatt caaatatgta tccgctcatg agacaataac
      841 cctgataaat gcttcaataa tattgaaaaa ggaagagtat gagtattcaa catttccgtg
      901 tcgcccttat tccctttttt gcggcatttt gccttcctgt ttttgctcac ccagaaacgc
      961 tggtgaaagt aaaagatgct gaagatcagt tgggtgcacg agtgggttac atcgaactgg
     1021 atctcaacag cggtaagatc cttgagagtt ttcgccccga agaacgtttt ccaatgatga
     1081 gcacttttaa agttctgcta tgtggcgcgg tattatcccg tattgacgcc gggcaagagc
     1141 aactcggtcg ccgcatacac tattctcaga atgacttggt tgagtactca ccagtcacag
     1201 aaaagcatct tacggatggc atgacagtaa gagaattatg cagtgctgcc ataaccatga
     1261 gtgataacac tgcggccaac ttacttctga caacgatcgg aggaccgaag gagctaaccg
     1321 cttttttgca caacatgggg gatcatgtaa ctcgccttga tcgttgggaa ccggagctga
     1381 atgaagccat accaaacgac gagcgtgaca ccacgatgcc tgtagcaatg gcaacaacgt
     1441 tgcgcaaact attaactggc gaactactta ctctagcttc ccggcaacaa ttaatagact
     1501 ggatggaggc ggataaagtt gcaggaccac ttctgcgctc ggcccttccg gctggctggt
     1561 ttattgctga taaatctgga gccggtgagc gtgggtctcg cggtatcatt gcagcactgg
     1621 ggccagatgg taagccctcc cgtatcgtag ttatctacac gacggggagt caggcaacta
     1681 tggatgaacg aaatagacag atcgctgaga taggtgcctc actgattaag cattggtaac
     1741 tgtcagacca agtttactca tatatacttt agattgattt aaaacttcat ttttaattta
     1801 aaaggatcta ggtgaagatc ctttttgata atctcatgac caaaatccct taacgtgagt
     1861 tttcgttcca ctgagcgtca gaccccgtag aaaagatcaa aggatcttct tgagatcctt
     1921 tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca gcggtggttt
     1981 gtttgccgga tcaagagcta ccaactcttt ttccgaaggt aactggcttc agcagagcgc
     2041 agataccaaa tactgttctt ctagtgtagc cgtagttagg ccaccacttc aagaactctg
     2101 tagcaccgcc tacatacctc gctctgctaa tcctgttacc agtggctgct gccagtggcg
     2161 ataagtcgtg tcttaccggg ttggactcaa gacgatagtt accggataag gcgcagcggt
     2221 cgggctgaac ggggggttcg tgcacacagc ccagcttgga gcgaacgacc tacaccgaac
     2281 tgagatacct acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg
     2341 acaggtatcc ggtaagcggc agggtcggaa caggagagcg cacgagggag cttccagggg
     2401 gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat
     2461 ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac gcggcctttt
     2521 tacggttcct ggccttttgc tggccttttg ctcacatgtt ctttcctgcg ttatcccctg
     2581 attctgtgga taaccgtatt accgcctttg agtgagctga taccgctcgc cgcagccgaa
     2641 cgaccgagcg cagcgagtca gtgagcgagg aagcggaaga gcgctgccgg cacctgtcct
     2701 acgagttgca tgataaagaa gacagtcata agtgcggcga cgatagtcat gccccgcgcc
     2761 caccggaagg agctgactgg gttgaaggct ctcaagggca tcggtcgacg gatccttatc
     2821 gattttacca catttgtaga ggttttactt gctttaaaaa acctcccaca cctccccctg
     2881 aacctgaaac ataaaatgaa tgcaattgtt gttgttaact tgtttattgc agcttataat
     2941 ggttacaaat aaagcaatag catcacaaat ttcacaaata aagcattttt ttcactgcat
     3001 tctagttgtg gtttgtccaa actcatcaat gtatcttatc atgtctgctc gaagcggccg
     3061 gccgccccga ctctagaatt acacggcgat ctttccgccc ttcttggcct ttatgaggat
     3121 ctctctgatt tttcttgcgt cgagttttcc ggtaagacct ttcggtactt cgtccacaaa
     3181 cacaactcct ccgcgcaact ttttcgcggt tgttacttga ctggcgacgt aatccacgat
     3241 ctctttttcc gtcatcgtct ttccgtgctc caaaacaaca acggcggcgg gaagttcacc
     3301 ggcgtcatcg tcgggaagac ctgcgacacc tgcgtcgaag atgttggggt gttggagcaa
     3361 gatggattcc aattcagcgg gagccacctg atagcctttg tacttaatca gagacttcag
     3421 gcggtcaacg atgaagaagt gttcgtcttc gtcccagtaa gctatgtctc cagaatgtag
     3481 ccatccatcc ttgtcaatca aggcgttggt cgcttccgga ttgtttacat aaccggacat
     3541 aatcatagga cctctcacac acagttcgcc tctttgatta acgcccagcg ttttcccggt
     3601 atccagatcc acaaccttcg cttcaaaaaa tggaacaact ttaccgaccg cgcccggttt
     3661 atcatccccc tcgggtgtaa tcagaatagc tgatgtagtc tcagtgagcc catatccttg
     3721 cctgatacct ggcagatgga acctcttggc aaccgcttcc ccgacttcct tagagagggg
     3781 agcgccacca gaagcaattt cgtgtaaatt agataaatcg tatttgtcaa tcagagtgct
     3841 tttggcgaag aaggagaata gggttggcac cagcagcgca ctttgaatct tgtaatcctg
     3901 aaggctcctc agaaacagct cttcttcaaa tctatacatt aagacgactc gaaatccaca
     3961 tatcaaatat ccgagtgtag taaacattcc aaaaccgtga tggaatggaa caacacttaa
     4021 aatcgcagta tccggaatga tttgattgcc aaaaatagga tctctggcat gcgagaatct
     4081 cacgcaggca gttctatgag gcagagcgac acctttaggc agaccagtag atccagagga
     4141 gttcatgatc agtgcaattg tcttgtccct atcgaaggac tctggcacaa aatcgtattc
     4201 attaaaaccg ggaggtagat gagatgtgac gaacgtgtac atcgactgaa atccctggta
     4261 atccgtttta gaatccatga taataatttt ttggatgatt gggagctttt tttgcacgtt
     4321 caaaattttt tgcaacccct ttttggaaac gaacaccacg gtaggctgcg aaatgcccat
     4381 actgttgagc aattcacgtt cattataaat gtcgttcgcg ggcgcaactg caactccgat
     4441 aaataacgcg cccaacaccg gcataaagaa ttgaagagag ttttcactgc atacgacgat
     4501 tctgtgattt gtattcagcc catatcgttt catagcttct gccaaccgaa cggacatttc
     4561 gaagtactca gcgtaagtga tgtccacctc gatatgtgca tctgtaaaag caattgttcc
     4621 aggaaccagg gcgtatctct tcatagcctt atgcagttgc tctccagcgg ttccatcttc
     4681 cagcggatag aatggcgccg ggcctttctt tatgtttttg gcgtcttcca tggtggcttc
     4741 caaaaaagat caccggtata ttactactgc cccttcacac tcgagtatgg gaaagagtgg
     4801 tctcatacag aactatataa acctgaagga aatctcaact ttacacttag gtcaagtttc
     4861 ccagaacaca tagcgacatg ctaaatatgc tagcacgcgt aagagctcgg tacc
//
