LOCUS       sequence-193337-        4863 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  It contains 0.5 Kb of the miR-183-96-182 cluster promoter. This 
            fragment partially loses promoter activity in melanoma cells..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4863)
  AUTHORS   Segura MF, Jubierre L, Li S, Soriano A, Koetz L, Gaziel-Sovran A, 
            Masanas M, Kleffman K, Dankert JF, Walsh MJ, Hernando E
  TITLE     Kruppel-like factor 4 (KLF4) regulates the miR-183~96~182 cluster 
            under physiologic and pathologic conditions.
  JOURNAL   Oncotarget. 2017 Apr 18;8(16):26298-26311. doi: 
            10.18632/oncotarget.15459.
  PUBMED    28412746
REFERENCE   2  (bases 1 to 4863)
  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"; doi: 
            "10.18632/oncotarget"; journalName: "Oncotarget"; date: "2017-04-18-
            18"; volume: "8"; issue: "16"; pages: "26298-26311"
COMMENT     Sequence Label: pGL4 Cluster Promoter Delta 1.2
FEATURES             Location/Qualifiers
     source          1..4863
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     15..34
                     /label=RVprimer3
                     /note="pGL3 vector, forward primer"
     promoter        723..754
                     /label=minP
                     /note="minimal TATA-box promoter with low basal activity"
     regulatory      781..790
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             787..2439
                     /codon_start=1
                     /product="firefly luciferase"
                     /label=luciferase
                     /note="synthetic luc2 version of the luciferase gene"
                     /translation="MEDAKNIKKGPAPFYPLEDGTAGEQLHKAMKRYALVPGTIAFTDA
                     HIEVDITYAEYFEMSVRLAEAMKRYGLNTNHRIVVCSENSLQFFMPVLGALFIGVAVAP
                     ANDIYNERELLNSMGISQPTVVFVSKKGLQKILNVQKKLPIIQKIIIMDSKTDYQGFQS
                     MYTFVTSHLPPGFNEYDFVPESFDRDKTIALIMNSSGSTGLPKGVALPHRTACVRFSHA
                     RDPIFGNQIIPDTAILSVVPFHHGFGMFTTLGYLICGFRVVLMYRFEEELFLRSLQDYK
                     IQSALLVPTLFSFFAKSTLIDKYDLSNLHEIASGGAPLSKEVGEAVAKRFHLPGIRQGY
                     GLTETTSAILITPEGDDKPGAVGKVVPFFEAKVVDLDTGKTLGVNQRGELCVRGPMIMS
                     GYVNNPEATNALIDKDGWLHSGDIAYWDEDEHFFIVDRLKSLIKYKGYQVAPAELESIL
                     LQHPNIFDAGVAGLPDDDAGELPAAVVVLEHGKTMTEKEIVDYVASQVTTAKKLRGGVV
                     FVDEVPKGLTGKLDARKIREILIKAKKGGKIAV"
     polyA_signal    2483..2604
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2494..2513)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2548..2567
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(2852..2869)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3023..3611)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3103..3122)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3811..4602,4603..4671))
                     /codon_start=1
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     polyA_signal    4776..4824
                     /note="synthetic polyadenylation signal"
     misc_feature    join(4838..4863,1..66)
                     /label=pause site
                     /note="RNA polymerase II transcriptional pause signal from 
                     the human alpha-2 globin gene"
ORIGIN
        1 cgaaacaaaa caaactagca aaataggctg tccccagtgc aagtgcaggt gccagaacat
       61 ttctctggcc taactggccg gtaccgagct cggatccact agtaacggcc gccagtgtgc
      121 tggaattcgc ccttctcacc cccttctttt gctcgagccc aagacaaggt gttaacggtc
      181 ttcaagtgca agggttaggt ttggtgggag cataggctaa gcaccaccct cattttacag
      241 gggcttctag ggtgccagat ctgcgggggt cgattgggaa gaactagaag ctaaaaagga
      301 gaagggcgga ttaacgtttt attttaatct ggttttgagc tgccgttgcc gtctggggcg
      361 gccgggcggg gcggagctgg gactgccaag ggcagccaaa gcagcaggga gggagggggc
      421 gctccggtgc tcaggatgct ggggaagaag gaagaggcca ccccaacgct gacagcttgg
      481 aggacctagc ccctgccccc cggtcccttc tgaatgctcc gggtggtgtc agaccaaagg
      541 acggttgaga aggatctaat ccgcgtccgc gccccagccc cggctggccc cgccccctcc
      601 cgagggagcc actgtgtccg ggcagcccgc gccccctgtt cttccggtcc tcgggcccag
      661 gtacctgagc tcgctagcct cgaggatatc aagatctggc ctcggcggcc aagcttagac
      721 actagagggt atataatgga agctcgactt ccagcttggc aatccggtac tgttggtaaa
      781 gccaccatgg aagatgccaa aaacattaag aagggcccag cgccattcta cccactcgaa
      841 gacgggaccg ccggcgagca gctgcacaaa gccatgaagc gctacgccct ggtgcccggc
      901 accatcgcct ttaccgacgc acatatcgag gtggacatta cctacgccga gtacttcgag
      961 atgagcgttc ggctggcaga agctatgaag cgctatgggc tgaatacaaa ccatcggatc
     1021 gtggtgtgca gcgagaatag cttgcagttc ttcatgcccg tgttgggtgc cctgttcatc
     1081 ggtgtggctg tggccccagc taacgacatc tacaacgagc gcgagctgct gaacagcatg
     1141 ggcatcagcc agcccaccgt cgtattcgtg agcaagaaag ggctgcaaaa gatcctcaac
     1201 gtgcaaaaga agctaccgat catacaaaag atcatcatca tggatagcaa gaccgactac
     1261 cagggcttcc aaagcatgta caccttcgtg acttcccatt tgccacccgg cttcaacgag
     1321 tacgacttcg tgcccgagag cttcgaccgg gacaaaacca tcgccctgat catgaacagt
     1381 agtggcagta ccggattgcc caagggcgta gccctaccgc accgcaccgc ttgtgtccga
     1441 ttcagtcatg cccgcgaccc catcttcggc aaccagatca tccccgacac cgctatcctc
     1501 agcgtggtgc catttcacca cggcttcggc atgttcacca cgctgggcta cttgatctgc
     1561 ggctttcggg tcgtgctcat gtaccgcttc gaggaggagc tattcttgcg cagcttgcaa
     1621 gactataaga ttcaatctgc cctgctggtg cccacactat ttagcttctt cgctaagagc
     1681 actctcatcg acaagtacga cctaagcaac ttgcacgaga tcgccagcgg cggggcgccg
     1741 ctcagcaagg aggtaggtga ggccgtggcc aaacgcttcc acctaccagg catccgccag
     1801 ggctacggcc tgacagaaac aaccagcgcc attctgatca cccccgaagg ggacgacaag
     1861 cctggcgcag taggcaaggt ggtgcccttc ttcgaggcta aggtggtgga cttggacacc
     1921 ggtaagacac tgggtgtgaa ccagcgcggc gagctgtgcg tccgtggccc catgatcatg
     1981 agcggctacg ttaacaaccc cgaggctaca aacgctctca tcgacaagga cggctggctg
     2041 cacagcggcg acatcgccta ctgggacgag gacgagcact tcttcatcgt ggaccggctg
     2101 aagagcctga tcaaatacaa gggctaccag gtagccccag ccgaactgga gagcatcctg
     2161 ctgcaacacc ccaacatctt cgacgccggg gtcgccggcc tgcccgacga cgatgccggc
     2221 gagctgcccg ccgcagtcgt cgtgctggaa cacggtaaaa ccatgaccga gaaggagatc
     2281 gtggactatg tggccagcca ggttacaacc gccaagaagc tgcgcggtgg tgttgtgttc
     2341 gtggacgagg tgcctaaagg actgaccggc aagttggacg cccgcaagat ccgcgagatt
     2401 ctcattaagg ccaagaaggg cggcaagatc gccgtgtaat aattctagag tcggggcggc
     2461 cggccgcttc gagcagacat gataagatac attgatgagt ttggacaaac cacaactaga
     2521 atgcagtgaa aaaaatgctt tatttgtgaa atttgtgatg ctattgcttt atttgtaacc
     2581 attataagct gcaataaaca agttaacaac aacaattgca ttcattttat gtttcaggtt
     2641 cagggggagg tgtgggaggt tttttaaagc aagtaaaacc tctacaaatg tggtaaaatc
     2701 gataaggatc cgtcgaccga tgcccttgag agccttcaac ccagtcagct ccttccggtg
     2761 ggcgcggggc atgactatcg tcgccgcact tatgactgtc ttctttatca tgcaactcgt
     2821 aggacaggtg ccggcagcgc tcttccgctt cctcgctcac tgactcgctg cgctcggtcg
     2881 ttcggctgcg gcgagcggta tcagctcact caaaggcggt aatacggtta tccacagaat
     2941 caggggataa cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta
     3001 aaaaggccgc gttgctggcg tttttccata ggctccgccc ccctgacgag catcacaaaa
     3061 atcgacgctc aagtcagagg tggcgaaacc cgacaggact ataaagatac caggcgtttc
     3121 cccctggaag ctccctcgtg cgctctcctg ttccgaccct gccgcttacc ggatacctgt
     3181 ccgcctttct cccttcggga agcgtggcgc tttctcatag ctcacgctgt aggtatctca
     3241 gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc gttcagcccg
     3301 accgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggtaaga cacgacttat
     3361 cgccactggc agcagccact ggtaacagga ttagcagagc gaggtatgta ggcggtgcta
     3421 cagagttctt gaagtggtgg cctaactacg gctacactag aagaacagta tttggtatct
     3481 gcgctctgct gaagccagtt accttcggaa aaagagttgg tagctcttga tccggcaaac
     3541 aaaccaccgc tggtagcggt ggtttttttg tttgcaagca gcagattacg cgcagaaaaa
     3601 aaggatctca agaagatcct ttgatctttt ctacggggtc tgacgctcag tggaacgaaa
     3661 actcacgtta agggattttg gtcatgagat tatcaaaaag gatcttcacc tagatccttt
     3721 taaattaaaa atgaagtttt aaatcaatct aaagtatata tgagtaaact tggtctgaca
     3781 gcggccgcaa atgctaaacc actgcagtgg ttaccagtgc ttgatcagtg aggcaccgat
     3841 ctcagcgatc tgcctatttc gttcgtccat agtggcctga ctccccgtcg tgtagatcac
     3901 tacgattcgt gagggcttac catcaggccc cagcgcagca atgatgccgc gagagccgcg
     3961 ttcaccggcc cccgatttgt cagcaatgaa ccagccagca gggagggccg agcgaagaag
     4021 tggtcctgct actttgtccg cctccatcca gtctatgagc tgctgtcgtg atgctagagt
     4081 aagaagttcg ccagtgagta gtttccgaag agttgtggcc attgctactg gcatcgtggt
     4141 atcacgctcg tcgttcggta tggcttcgtt caactctggt tcccagcggt caagccgggt
     4201 cacatgatca cccatattat gaagaaatgc agtcagctcc ttagggcctc cgatcgttgt
     4261 cagaagtaag ttggccgcgg tgttgtcgct catggtaatg gcagcactac acaattctct
     4321 taccgtcatg ccatccgtaa gatgcttttc cgtgaccggc gagtactcaa ccaagtcgtt
     4381 ttgtgagtag tgtatacggc gaccaagctg ctcttgcccg gcgtctatac gggacaacac
     4441 cgcgccacat agcagtactt tgaaagtgct catcatcggg aatcgttctt cggggcggaa
     4501 agactcaagg atcttgccgc tattgagatc cagttcgata tagcccactc ttgcacccag
     4561 ttgatcttca gcatctttta ctttcaccag cgtttcgggg tgtgcaaaaa caggcaagca
     4621 aaatgccgca aagaagggaa tgagtgcgac acgaaaatgt tggatgctca tactcgtcct
     4681 ttttcaatat tattgaagca tttatcaggg ttactagtac gtctctcaag gataagtaag
     4741 taatattaag gtacgggagg tattggacag gccgcaataa aatatcttta ttttcattac
     4801 atctgtgtgt tggttttttg tgtgaatcga tagtactaac atacgctctc catcaaaaca
     4861 aaa
//
