LOCUS       sequence-183252-        5629 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  It contains ~1.4Kb of the miR-183/96-182 cluster promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5629)
  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 5629)
  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
FEATURES             Location/Qualifiers
     source          1..5629
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        1389..1420
                     /label=minP
                     /note="minimal TATA-box promoter with low basal activity"
     regulatory      1447..1456
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1453..3105
                     /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    3149..3270
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3160..3179)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     3214..3233
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(3518..3535)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3689..4277)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3769..3788)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4477..5268,5269..5337))
                     /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    5442..5490
                     /note="synthetic polyadenylation signal"
     misc_feature    5504..5595
                     /label=pause site
                     /note="RNA polymerase II transcriptional pause signal from 
                     the human alpha-2 globin gene"
     primer_bind     5544..5563
                     /label=RVprimer3
                     /note="pGL3 vector, forward primer"
ORIGIN
        1 agtaacggcc gccagtgtgc tggaattcgc ccttctcacc cccttctttt gctcgagccc
       61 aagacaaggt gttaacggtc ttcaagtgca agggttaggt ttggtgggag cataggctaa
      121 gcaccaccct cattttacag gggcttctag ggtgccagat ctgcgggggt cgattgggaa
      181 gaactagaag ctaaaaagga gaagggcgga ttaacgtttt attttaatct ggttttgagc
      241 tgccgttgcc gtctggggcg gccgggcggg gcggagctgg gactgccaag ggcagccaaa
      301 gcagcaggga gggagggggc gctccggtgc tcaggatgct ggggaagaag gaagaggcca
      361 ccccaacgct gacagcttgg aggacctagc ccctgccccc cggtcccttc tgaatgctcc
      421 gggtggtgtc agaccaaagg acggttgaga aggatctaat ccgcgtccgc gccccagccc
      481 cggctggccc cgccccctcc cgagggagcc actgtcgcag tggggggagg gggagatggg
      541 aacgggtctg gggggcggct cccctccccc tccctctctt gggacagaca gcctttgttc
      601 ccggcggcgg cggcagcggt ggccagacac ctgtgaggcc gcagaactgg ccgagggtgg
      661 gcgtgtgagg acatggaggc cccttgtccc cattccagcc ctgcgcccct gggtccgggt
      721 gcgccccctg agacaggtca gccttgagca cctttctgcg gggagactgc agagatcgtc
      781 cagtggccct acggtgcgcc ccagctccca acgcacttcc cctcccccct cccagggtcg
      841 gatttccaag cacgccacgc cacacacacc tcggctagct ccgggctcag agggcgctgg
      901 ggtggcaggg cgtccgggcg cacctcggct agcggcaccc tagcccgagg gcaggaggcg
      961 gggatagaag gcgggcgacg ggaccccttg gggccagacc cggccgggtt gggcaagcag
     1021 gcggggctcc cggcgtctcc gcagtcccgg acgggcgtgc cctgcgccgc gcaccaaccg
     1081 gagccacatt caaggtgaat catccccggc ccgccccctg cccctccccg cctcgcggca
     1141 cctgagccaa tggcgctgcg cgaggggcgg gcggggcccg gggcgggggg cggtaaaacc
     1201 ctcctggagt cattctctgt gcactccgtc gagcccccgc gcagccgtgc cggagccgca
     1261 gggcctccgt ccagccgcgc ggtccgggca gcccgcgccc cctgttcttc cggtcctcgg
     1321 gcccaggtac ctgagctcgc tagcctcgag gatatcaaga tctggcctcg gcggccaagc
     1381 ttagacacta gagggtatat aatggaagct cgacttccag cttggcaatc cggtactgtt
     1441 ggtaaagcca ccatggaaga tgccaaaaac attaagaagg gcccagcgcc attctaccca
     1501 ctcgaagacg ggaccgccgg cgagcagctg cacaaagcca tgaagcgcta cgccctggtg
     1561 cccggcacca tcgcctttac cgacgcacat atcgaggtgg acattaccta cgccgagtac
     1621 ttcgagatga gcgttcggct ggcagaagct atgaagcgct atgggctgaa tacaaaccat
     1681 cggatcgtgg tgtgcagcga gaatagcttg cagttcttca tgcccgtgtt gggtgccctg
     1741 ttcatcggtg tggctgtggc cccagctaac gacatctaca acgagcgcga gctgctgaac
     1801 agcatgggca tcagccagcc caccgtcgta ttcgtgagca agaaagggct gcaaaagatc
     1861 ctcaacgtgc aaaagaagct accgatcata caaaagatca tcatcatgga tagcaagacc
     1921 gactaccagg gcttccaaag catgtacacc ttcgtgactt cccatttgcc acccggcttc
     1981 aacgagtacg acttcgtgcc cgagagcttc gaccgggaca aaaccatcgc cctgatcatg
     2041 aacagtagtg gcagtaccgg attgcccaag ggcgtagccc taccgcaccg caccgcttgt
     2101 gtccgattca gtcatgcccg cgaccccatc ttcggcaacc agatcatccc cgacaccgct
     2161 atcctcagcg tggtgccatt tcaccacggc ttcggcatgt tcaccacgct gggctacttg
     2221 atctgcggct ttcgggtcgt gctcatgtac cgcttcgagg aggagctatt cttgcgcagc
     2281 ttgcaagact ataagattca atctgccctg ctggtgccca cactatttag cttcttcgct
     2341 aagagcactc tcatcgacaa gtacgaccta agcaacttgc acgagatcgc cagcggcggg
     2401 gcgccgctca gcaaggaggt aggtgaggcc gtggccaaac gcttccacct accaggcatc
     2461 cgccagggct acggcctgac agaaacaacc agcgccattc tgatcacccc cgaaggggac
     2521 gacaagcctg gcgcagtagg caaggtggtg cccttcttcg aggctaaggt ggtggacttg
     2581 gacaccggta agacactggg tgtgaaccag cgcggcgagc tgtgcgtccg tggccccatg
     2641 atcatgagcg gctacgttaa caaccccgag gctacaaacg ctctcatcga caaggacggc
     2701 tggctgcaca gcggcgacat cgcctactgg gacgaggacg agcacttctt catcgtggac
     2761 cggctgaaga gcctgatcaa atacaagggc taccaggtag ccccagccga actggagagc
     2821 atcctgctgc aacaccccaa catcttcgac gccggggtcg ccggcctgcc cgacgacgat
     2881 gccggcgagc tgcccgccgc agtcgtcgtg ctggaacacg gtaaaaccat gaccgagaag
     2941 gagatcgtgg actatgtggc cagccaggtt acaaccgcca agaagctgcg cggtggtgtt
     3001 gtgttcgtgg acgaggtgcc taaaggactg accggcaagt tggacgcccg caagatccgc
     3061 gagattctca ttaaggccaa gaagggcggc aagatcgccg tgtaataatt ctagagtcgg
     3121 ggcggccggc cgcttcgagc agacatgata agatacattg atgagtttgg acaaaccaca
     3181 actagaatgc agtgaaaaaa atgctttatt tgtgaaattt gtgatgctat tgctttattt
     3241 gtaaccatta taagctgcaa taaacaagtt aacaacaaca attgcattca ttttatgttt
     3301 caggttcagg gggaggtgtg ggaggttttt taaagcaagt aaaacctcta caaatgtggt
     3361 aaaatcgata aggatccgtc gaccgatgcc cttgagagcc ttcaacccag tcagctcctt
     3421 ccggtgggcg cggggcatga ctatcgtcgc cgcacttatg actgtcttct ttatcatgca
     3481 actcgtagga caggtgccgg cagcgctctt ccgcttcctc gctcactgac tcgctgcgct
     3541 cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa ggcggtaata cggttatcca
     3601 cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga
     3661 accgtaaaaa ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc
     3721 acaaaaatcg acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg
     3781 cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat
     3841 acctgtccgc ctttctccct tcgggaagcg tggcgctttc tcatagctca cgctgtaggt
     3901 atctcagttc ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc
     3961 agcccgaccg ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg
     4021 acttatcgcc actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg
     4081 gtgctacaga gttcttgaag tggtggccta actacggcta cactagaaga acagtatttg
     4141 gtatctgcgc tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg
     4201 gcaaacaaac caccgctggt agcggtggtt tttttgtttg caagcagcag attacgcgca
     4261 gaaaaaaagg atctcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga
     4321 acgaaaactc acgttaaggg attttggtca tgagattatc aaaaaggatc ttcacctaga
     4381 tccttttaaa ttaaaaatga agttttaaat caatctaaag tatatatgag taaacttggt
     4441 ctgacagcgg ccgcaaatgc taaaccactg cagtggttac cagtgcttga tcagtgaggc
     4501 accgatctca gcgatctgcc tatttcgttc gtccatagtg gcctgactcc ccgtcgtgta
     4561 gatcactacg attcgtgagg gcttaccatc aggccccagc gcagcaatga tgccgcgaga
     4621 gccgcgttca ccggcccccg atttgtcagc aatgaaccag ccagcaggga gggccgagcg
     4681 aagaagtggt cctgctactt tgtccgcctc catccagtct atgagctgct gtcgtgatgc
     4741 tagagtaaga agttcgccag tgagtagttt ccgaagagtt gtggccattg ctactggcat
     4801 cgtggtatca cgctcgtcgt tcggtatggc ttcgttcaac tctggttccc agcggtcaag
     4861 ccgggtcaca tgatcaccca tattatgaag aaatgcagtc agctccttag ggcctccgat
     4921 cgttgtcaga agtaagttgg ccgcggtgtt gtcgctcatg gtaatggcag cactacacaa
     4981 ttctcttacc gtcatgccat ccgtaagatg cttttccgtg accggcgagt actcaaccaa
     5041 gtcgttttgt gagtagtgta tacggcgacc aagctgctct tgcccggcgt ctatacggga
     5101 caacaccgcg ccacatagca gtactttgaa agtgctcatc atcgggaatc gttcttcggg
     5161 gcggaaagac tcaaggatct tgccgctatt gagatccagt tcgatatagc ccactcttgc
     5221 acccagttga tcttcagcat cttttacttt caccagcgtt tcggggtgtg caaaaacagg
     5281 caagcaaaat gccgcaaaga agggaatgag tgcgacacga aaatgttgga tgctcatact
     5341 cgtccttttt caatattatt gaagcattta tcagggttac tagtacgtct ctcaaggata
     5401 agtaagtaat attaaggtac gggaggtatt ggacaggccg caataaaata tctttatttt
     5461 cattacatct gtgtgttggt tttttgtgtg aatcgatagt actaacatac gctctccatc
     5521 aaaacaaaac gaaacaaaac aaactagcaa aataggctgt ccccagtgca agtgcaggtg
     5581 ccagaacatt tctctggcct aactggccgg taccgagctc ggatccact
//
