LOCUS       sequence-243846-        5354 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5354)
  AUTHORS   Taganov KD, Boldin MP, Chang KJ, Baltimore D
  TITLE     NF-kappaB-dependent induction of microRNA miR-146, an inhibitor 
            targeted to signaling proteins of innate immune responses.
  JOURNAL   Proc Natl Acad Sci U S A. 2006 Aug 15. 103(33):12481-6.
  PUBMED    16885212
REFERENCE   2  (bases 1 to 5354)
  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"; journalName: "Proc Natl 
            Acad Sci U S A. 2006 Aug 15. 103(33):12481-6."
COMMENT     Sequence Label: A547 (miR-146a promoter)
FEATURES             Location/Qualifiers
     source          1..5354
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     polyA_signal    151..199
                     /note="synthetic polyadenylation signal"
     misc_feature    213..304
                     /label=pause site
                     /note="RNA polymerase II transcriptional pause signal from 
                     the human alpha-2 globin gene"
     primer_bind     253..272
                     /label=RVprimer3
                     /note="pGL3 vector, forward primer"
     regulatory      929..938
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             935..2587
                     /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(1001..1019)
                     /label=LucNrev
                     /note="Firefly luciferase, reverse primer"
     polyA_signal    2628..2749
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2639..2658)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2693..2712
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(2997..3014)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3168..3756)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3248..3267)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3927..4718,4719..4787))
                     /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     4550..4569
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4788..4892)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      4919..5347
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5006..5025)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5216..5237
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
ORIGIN
        1 aatattaacg cttacaattt gccattcgcc attcaggctg cgcaactgtt gggaagggcg
       61 atcggtgcgg gcctcttcgc tattacgcca gcccaagcta ccatgataag taagtaatat
      121 taaggtacgg gaggtacttg gagcggccgc aataaaatat ctttattttc attacatctg
      181 tgtgttggtt ttttgtgtga atcgatagta ctaacatacg ctctccatca aaacaaaacg
      241 aaacaaaaca aactagcaaa ataggctgtc cccagtgcaa gtgcaggtgc cagaacattt
      301 ctctatcgat aggtaccgag ctcttacgcg tgctagcccg ggctcgagat ctttgaaaag
      361 ccaacaggct cattgggcag ccgataaagc tctcgggatt tccccgcggg gctgcggaga
      421 gtacagacag gaagcctggg gacccagcgc ctgaccagaa cttcctcggg ggaggctgca
      481 ggggagcagg cgcatcctgc acagaacgct ctagagcgcg caggccaaag caccaggctg
      541 ctcctgacac gtgctgcaag agggtccccg acccgggggt ccagaccctg cacgcatgat
      601 ggggaaggtg gaggcttccc cctcagctcc gcggagagaa gctgacactg ccaggctgga
      661 accttccatt ccggcccagc ctcttcctcc ctcgctgtgc cgaggaggga tctagaaggg
      721 actttccaga gagggttagc gtgcagggtg tggaaatgga ataaaagcat atgcaaatag
      781 gccttagctg ccttcctcta ccccagcaaa taagagtctc tccagaaaga tgctctttct
      841 ccaagacgct tgaccgctct tcctttcctg gatggcacca gcagggccga ttggagtgga
      901 agcttggcat tccggtactg ttggtaaagc caccatggaa gacgccaaaa acataaagaa
      961 aggcccggcg ccattctatc cgctggaaga tggaaccgct ggagagcaac tgcataaggc
     1021 tatgaagaga tacgccctgg ttcctggaac aattgctttt acagatgcac atatcgaggt
     1081 ggacatcact tacgctgagt acttcgaaat gtccgttcgg ttggcagaag ctatgaaacg
     1141 atatgggctg aatacaaatc acagaatcgt cgtatgcagt gaaaactctc ttcaattctt
     1201 tatgccggtg ttgggcgcgt tatttatcgg agttgcagtt gcgcccgcga acgacattta
     1261 taatgaacgt gaattgctca acagtatggg catttcgcag cctaccgtgg tgttcgtttc
     1321 caaaaagggg ttgcaaaaaa ttttgaacgt gcaaaaaaag ctcccaatca tccaaaaaat
     1381 tattatcatg gattctaaaa cggattacca gggatttcag tcgatgtaca cgttcgtcac
     1441 atctcatcta cctcccggtt ttaatgaata cgattttgtg ccagagtcct tcgataggga
     1501 caagacaatt gcactgatca tgaactcctc tggatctact ggtctgccta aaggtgtcgc
     1561 tctgcctcat agaactgcct gcgtgagatt ctcgcatgcc agagatccta tttttggcaa
     1621 tcaaatcatt ccggatactg cgattttaag tgttgttcca ttccatcacg gttttggaat
     1681 gtttactaca ctcggatatt tgatatgtgg atttcgagtc gtcttaatgt atagatttga
     1741 agaagagctg tttctgagga gccttcagga ttacaagatt caaagtgcgc tgctggtgcc
     1801 aaccctattc tccttcttcg ccaaaagcac tctgattgac aaatacgatt tatctaattt
     1861 acacgaaatt gcttctggtg gcgctcccct ctctaaggaa gtcggggaag cggttgccaa
     1921 gaggttccat ctgccaggta tcaggcaagg atatgggctc actgagacta catcagctat
     1981 tctgattaca cccgaggggg atgataaacc gggcgcggtc ggtaaagttg ttccattttt
     2041 tgaagcgaag gttgtggatc tggataccgg gaaaacgctg ggcgttaatc aaagaggcga
     2101 actgtgtgtg agaggtccta tgattatgtc cggttatgta aacaatccgg aagcgaccaa
     2161 cgccttgatt gacaaggatg gatggctaca ttctggagac atagcttact gggacgaaga
     2221 cgaacacttc ttcatcgttg accgcctgaa gtctctgatt aagtacaaag gctatcaggt
     2281 ggctcccgct gaattggaat ccatcttgct ccaacacccc aacatcttcg acgcaggtgt
     2341 cgcaggtctt cccgacgatg acgccggtga acttcccgcc gccgttgttg ttttggagca
     2401 cggaaagacg atgacggaaa aagagatcgt ggattacgtc gccagtcaag taacaaccgc
     2461 gaaaaagttg cgcggaggag ttgtgtttgt ggacgaagta ccgaaaggtc ttaccggaaa
     2521 actcgacgca agaaaaatca gagagatcct cataaaggcc aagaagggcg gaaagatcgc
     2581 cgtgtaattc tagagtcggg gcggccggcc gcttcgagca gacatgataa gatacattga
     2641 tgagtttgga caaaccacaa ctagaatgca gtgaaaaaaa tgctttattt gtgaaatttg
     2701 tgatgctatt gctttatttg taaccattat aagctgcaat aaacaagtta acaacaacaa
     2761 ttgcattcat tttatgtttc aggttcaggg ggaggtgtgg gaggtttttt aaagcaagta
     2821 aaacctctac aaatgtggta aaatcgataa ggatccgtcg accgatgccc ttgagagcct
     2881 tcaacccagt cagctccttc cggtgggcgc ggggcatgac tatcgtcgcc gcacttatga
     2941 ctgtcttctt tatcatgcaa ctcgtaggac aggtgccggc agcgctcttc cgcttcctcg
     3001 ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag
     3061 gcggtaatac ggttatccac agaatcaggg gataacgcag gaaagaacat gtgagcaaaa
     3121 ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc
     3181 cgcccccctg acgagcatca caaaaatcga cgctcaagtc agaggtggcg aaacccgaca
     3241 ggactataaa gataccaggc gtttccccct ggaagctccc tcgtgcgctc tcctgttccg
     3301 accctgccgc ttaccggata cctgtccgcc tttctccctt cgggaagcgt ggcgctttct
     3361 catagctcac gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt
     3421 gtgcacgaac cccccgttca gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag
     3481 tccaacccgg taagacacga cttatcgcca ctggcagcag ccactggtaa caggattagc
     3541 agagcgaggt atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa ctacggctac
     3601 actagaagaa cagtatttgg tatctgcgct ctgctgaagc cagttacctt cggaaaaaga
     3661 gttggtagct cttgatccgg caaacaaacc accgctggta gcggtggttt ttttgtttgc
     3721 aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag atcctttgat cttttctacg
     3781 gggtctgacg ctcagtggaa cgaaaactca cgttaaggga ttttggtcat gagattatca
     3841 aaaaggatct tcacctagat ccttttaaat taaaaatgaa gttttaaatc aatctaaagt
     3901 atatatgagt aaacttggtc tgacagttac caatgcttaa tcagtgaggc acctatctca
     3961 gcgatctgtc tatttcgttc atccatagtt gcctgactcc ccgtcgtgta gataactacg
     4021 atacgggagg gcttaccatc tggccccagt gctgcaatga taccgcgaga cccacgctca
     4081 ccggctccag atttatcagc aataaaccag ccagccggaa gggccgagcg cagaagtggt
     4141 cctgcaactt tatccgcctc catccagtct attaattgtt gccgggaagc tagagtaagt
     4201 agttcgccag ttaatagttt gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca
     4261 cgctcgtcgt ttggtatggc ttcattcagc tccggttccc aacgatcaag gcgagttaca
     4321 tgatccccca tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat cgttgtcaga
     4381 agtaagttgg ccgcagtgtt atcactcatg gttatggcag cactgcataa ttctcttact
     4441 gtcatgccat ccgtaagatg cttttctgtg actggtgagt actcaaccaa gtcattctga
     4501 gaatagtgta tgcggcgacc gagttgctct tgcccggcgt caatacggga taataccgcg
     4561 ccacatagca gaactttaaa agtgctcatc attggaaaac gttcttcggg gcgaaaactc
     4621 tcaaggatct taccgctgtt gagatccagt tcgatgtaac ccactcgtgc acccaactga
     4681 tcttcagcat cttttacttt caccagcgtt tctgggtgag caaaaacagg aaggcaaaat
     4741 gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa tactcatact cttccttttt
     4801 caatattatt gaagcattta tcagggttat tgtctcatga gcggatacat atttgaatgt
     4861 atttagaaaa ataaacaaat aggggttccg cgcacatttc cccgaaaagt gccacctgac
     4921 gcgccctgta gcggcgcatt aagcgcggcg ggtgtggtgg ttacgcgcag cgtgaccgct
     4981 acacttgcca gcgccctagc gcccgctcct ttcgctttct tcccttcctt tctcgccacg
     5041 ttcgccggct ttccccgtca agctctaaat cgggggctcc ctttagggtt ccgatttagt
     5101 gctttacggc acctcgaccc caaaaaactt gattagggtg atggttcacg tagtgggcca
     5161 tcgccctgat agacggtttt tcgccctttg acgttggagt ccacgttctt taatagtgga
     5221 ctcttgttcc aaactggaac aacactcaac cctatctcgg tctattcttt tgatttataa
     5281 gggattttgc cgatttcggc ctattggtta aaaaatgagc tgatttaaca aaaatttaac
     5341 gcgaatttta acaa
//
