LOCUS       sequence-193335-        5628 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 5628)
  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 5628)
  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..5628
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        1388..1419
                     /label=minP
                     /note="minimal TATA-box promoter with low basal activity"
     regulatory      1446..1455
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1452..3104
                     /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    3148..3269
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3159..3178)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     3213..3232
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(3517..3534)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3688..4276)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3768..3787)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4476..5267,5268..5336))
                     /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    5441..5489
                     /note="synthetic polyadenylation signal"
     misc_feature    5503..5594
                     /label=pause site
                     /note="RNA polymerase II transcriptional pause signal from 
                     the human alpha-2 globin gene"
     primer_bind     5543..5562
                     /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 ggggcggctc ccctccccct ccctctcttg ggacagacag cctttgttcc
      601 cggcggcggc ggcagcggtg gccagacacc tgtgaggccg cagaactggc cgagggtggg
      661 cgtgtgagga catggaggcc ccttgtcccc attccagccc tgcgcccctg ggtccgggtg
      721 cgccccctga gacaggtcag ccttgagcac ctttctgcgg ggagactgca gagatcgtcc
      781 agtggcccta cggtgcgccc cagctcccaa cgcacttccc ctcccccctc ccagggtcgg
      841 atttccaagc acgccacgcc acacacacct cggctagctc cgggctcaga gggcgctggg
      901 gtggcagggc gtccgggcgc acctcggcta gcggcaccct agcccgaggg caggaggcgg
      961 ggatagaagg cgggcgacgg gaccccttgg ggccagaccc ggccgggttg ggcaagcagg
     1021 cggggctccc ggcgtctccg cagtcccgga cgggcgtgcc ctgcgccgcg caccaaccgg
     1081 agccacattc aaggtgaatc atccccggcc cgccccctgc ccctccccgc ctcgcggcac
     1141 ctgagccaat ggcgctgcgc gaggggcggg cggggcccgg ggcggggggc ggtaaaaccc
     1201 tcctggagtc attctctgtg cactccgtcg agcccccgcg cagccttgcc ggagccgcag
     1261 ggcctccgtc cagccgcgcg gtccgggcag cccgcgcccc ctgttcttcc ggtcctcggg
     1321 cccaggtacc tgagctcgct agcctcgagg atatcaagat ctggcctcgg cggccaagct
     1381 tagacactag agggtatata atggaagctc gacttccagc ttggcaatcc ggtactgttg
     1441 gtaaagccac catggaagat gccaaaaaca ttaagaaggg cccagcgcca ttctacccac
     1501 tcgaagacgg gaccgccggc gagcagctgc acaaagccat gaagcgctac gccctggtgc
     1561 ccggcaccat cgcctttacc gacgcacata tcgaggtgga cattacctac gccgagtact
     1621 tcgagatgag cgttcggctg gcagaagcta tgaagcgcta tgggctgaat acaaaccatc
     1681 ggatcgtggt gtgcagcgag aatagcttgc agttcttcat gcccgtgttg ggtgccctgt
     1741 tcatcggtgt ggctgtggcc ccagctaacg acatctacaa cgagcgcgag ctgctgaaca
     1801 gcatgggcat cagccagccc accgtcgtat tcgtgagcaa gaaagggctg caaaagatcc
     1861 tcaacgtgca aaagaagcta ccgatcatac aaaagatcat catcatggat agcaagaccg
     1921 actaccaggg cttccaaagc atgtacacct tcgtgacttc ccatttgcca cccggcttca
     1981 acgagtacga cttcgtgccc gagagcttcg accgggacaa aaccatcgcc ctgatcatga
     2041 acagtagtgg cagtaccgga ttgcccaagg gcgtagccct accgcaccgc accgcttgtg
     2101 tccgattcag tcatgcccgc gaccccatct tcggcaacca gatcatcccc gacaccgcta
     2161 tcctcagcgt ggtgccattt caccacggct tcggcatgtt caccacgctg ggctacttga
     2221 tctgcggctt tcgggtcgtg ctcatgtacc gcttcgagga ggagctattc ttgcgcagct
     2281 tgcaagacta taagattcaa tctgccctgc tggtgcccac actatttagc ttcttcgcta
     2341 agagcactct catcgacaag tacgacctaa gcaacttgca cgagatcgcc agcggcgggg
     2401 cgccgctcag caaggaggta ggtgaggccg tggccaaacg cttccaccta ccaggcatcc
     2461 gccagggcta cggcctgaca gaaacaacca gcgccattct gatcaccccc gaaggggacg
     2521 acaagcctgg cgcagtaggc aaggtggtgc ccttcttcga ggctaaggtg gtggacttgg
     2581 acaccggtaa gacactgggt gtgaaccagc gcggcgagct gtgcgtccgt ggccccatga
     2641 tcatgagcgg ctacgttaac aaccccgagg ctacaaacgc tctcatcgac aaggacggct
     2701 ggctgcacag cggcgacatc gcctactggg acgaggacga gcacttcttc atcgtggacc
     2761 ggctgaagag cctgatcaaa tacaagggct accaggtagc cccagccgaa ctggagagca
     2821 tcctgctgca acaccccaac atcttcgacg ccggggtcgc cggcctgccc gacgacgatg
     2881 ccggcgagct gcccgccgca gtcgtcgtgc tggaacacgg taaaaccatg accgagaagg
     2941 agatcgtgga ctatgtggcc agccaggtta caaccgccaa gaagctgcgc ggtggtgttg
     3001 tgttcgtgga cgaggtgcct aaaggactga ccggcaagtt ggacgcccgc aagatccgcg
     3061 agattctcat taaggccaag aagggcggca agatcgccgt gtaataattc tagagtcggg
     3121 gcggccggcc gcttcgagca gacatgataa gatacattga tgagtttgga caaaccacaa
     3181 ctagaatgca gtgaaaaaaa tgctttattt gtgaaatttg tgatgctatt gctttatttg
     3241 taaccattat aagctgcaat aaacaagtta acaacaacaa ttgcattcat tttatgtttc
     3301 aggttcaggg ggaggtgtgg gaggtttttt aaagcaagta aaacctctac aaatgtggta
     3361 aaatcgataa ggatccgtcg accgatgccc ttgagagcct tcaacccagt cagctccttc
     3421 cggtgggcgc ggggcatgac tatcgtcgcc gcacttatga ctgtcttctt tatcatgcaa
     3481 ctcgtaggac aggtgccggc agcgctcttc cgcttcctcg ctcactgact cgctgcgctc
     3541 ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag gcggtaatac ggttatccac
     3601 agaatcaggg gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa
     3661 ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc cgcccccctg acgagcatca
     3721 caaaaatcga cgctcaagtc agaggtggcg aaacccgaca ggactataaa gataccaggc
     3781 gtttccccct ggaagctccc tcgtgcgctc tcctgttccg accctgccgc ttaccggata
     3841 cctgtccgcc tttctccctt cgggaagcgt ggcgctttct catagctcac gctgtaggta
     3901 tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac cccccgttca
     3961 gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag tccaacccgg taagacacga
     4021 cttatcgcca ctggcagcag ccactggtaa caggattagc agagcgaggt atgtaggcgg
     4081 tgctacagag ttcttgaagt ggtggcctaa ctacggctac actagaagaa cagtatttgg
     4141 tatctgcgct ctgctgaagc cagttacctt cggaaaaaga gttggtagct cttgatccgg
     4201 caaacaaacc accgctggta gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag
     4261 aaaaaaagga tctcaagaag atcctttgat cttttctacg gggtctgacg ctcagtggaa
     4321 cgaaaactca cgttaaggga ttttggtcat gagattatca aaaaggatct tcacctagat
     4381 ccttttaaat taaaaatgaa gttttaaatc aatctaaagt atatatgagt aaacttggtc
     4441 tgacagcggc cgcaaatgct aaaccactgc agtggttacc agtgcttgat cagtgaggca
     4501 ccgatctcag cgatctgcct atttcgttcg tccatagtgg cctgactccc cgtcgtgtag
     4561 atcactacga ttcgtgaggg cttaccatca ggccccagcg cagcaatgat gccgcgagag
     4621 ccgcgttcac cggcccccga tttgtcagca atgaaccagc cagcagggag ggccgagcga
     4681 agaagtggtc ctgctacttt gtccgcctcc atccagtcta tgagctgctg tcgtgatgct
     4741 agagtaagaa gttcgccagt gagtagtttc cgaagagttg tggccattgc tactggcatc
     4801 gtggtatcac gctcgtcgtt cggtatggct tcgttcaact ctggttccca gcggtcaagc
     4861 cgggtcacat gatcacccat attatgaaga aatgcagtca gctccttagg gcctccgatc
     4921 gttgtcagaa gtaagttggc cgcggtgttg tcgctcatgg taatggcagc actacacaat
     4981 tctcttaccg tcatgccatc cgtaagatgc ttttccgtga ccggcgagta ctcaaccaag
     5041 tcgttttgtg agtagtgtat acggcgacca agctgctctt gcccggcgtc tatacgggac
     5101 aacaccgcgc cacatagcag tactttgaaa gtgctcatca tcgggaatcg ttcttcgggg
     5161 cggaaagact caaggatctt gccgctattg agatccagtt cgatatagcc cactcttgca
     5221 cccagttgat cttcagcatc ttttactttc accagcgttt cggggtgtgc aaaaacaggc
     5281 aagcaaaatg ccgcaaagaa gggaatgagt gcgacacgaa aatgttggat gctcatactc
     5341 gtcctttttc aatattattg aagcatttat cagggttact agtacgtctc tcaaggataa
     5401 gtaagtaata ttaaggtacg ggaggtattg gacaggccgc aataaaatat ctttattttc
     5461 attacatctg tgtgttggtt ttttgtgtga atcgatagta ctaacatacg ctctccatca
     5521 aaacaaaacg aaacaaaaca aactagcaaa ataggctgtc cccagtgcaa gtgcaggtgc
     5581 cagaacattt ctctggccta actggccggt accgagctcg gatccact
//
