LOCUS       sequence-182857-        6586 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  For dual luciferase report assay.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6586)
  AUTHORS   Zhou W, Pal AS, Hsu AY, Gurol T, Zhu X, Wirbisky-Hershberger SE, 
            Freeman JL, Kasinski AL, Deng Q
  TITLE     MicroRNA-223 Suppresses the Canonical NF-kappaB Pathway in Basal 
            Keratinocytes to Dampen Neutrophilic Inflammation.
  JOURNAL   Cell Rep. 2018 Feb 13;22(7):1810-1823. doi: 
            10.1016/j.celrep.2018.01.058.
  PUBMED    29444433
REFERENCE   2  (bases 1 to 6586)
  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: 
            "10.1016/j.celrep.2018.01"; journalName: "Cell Rep"; date: 
            "2018-02-13- 13"; volume: "22"; issue: "7"; pages: "1810-1823"
COMMENT     Sequence Label: pSI-check2-Cul1b-3'UTR
FEATURES             Location/Qualifiers
     source          1..6586
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     polyA_signal    10..58
                     /note="synthetic polyadenylation signal"
     promoter        67..818
                     /label=HSV TK promoter
                     /note="herpes simplex virus thymidine kinase promoter"
     regulatory      848..857
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             854..2506
                     /codon_start=1
                     /gene="luc+"
                     /product="firefly luciferase"
                     /label=luciferase
                     /note="enhanced luc+ version of the luciferase gene"
                     /db_xref="GI:42718124"
                     /protein_id="AAS38485.1"
                     /translation="MADAKNIKKGPAPFYPLEDGTAGEQLHKAMKRYALVPGTIAFTDA
                     HIEVDITYAEYFEMSVRLAEAMKRYGLNTNHRIVVCSENSLQFFMPVLGALFIGVAVAP
                     ANDIYNERELLNSMGISQPTVVFVSKKGLQKILNVQKKLPIIQKIIIMDSKTDYQGFQS
                     MYTFVTSHLPPGFNEYDFVPESFDRDKTIALIMNSSGSTGLPKGVALPHRTACVRFSHA
                     RDPIFGNQIIPDTAILSVVPFHHGFGMFTTLGYLICGFRVVLMYRFEEELFLRSLQDYK
                     IQSALLVPTLFSFFAKSTLIDKYDLSNLHEIASGGAPLSKEVGEAVAKRFHLPGIRQGY
                     GLTETTSAILITPEGDDKPGAVGKVVPFFEAKVVDLDTGKTLGVNQRGELCVRGPMIMS
                     GYVNNPEATNALIDKDGWLHSGDIAYWDEDEHFFIVDRLKSLIKYKGYQVAPAELESIL
                     LQHPNIFDAGVAGLPDDDAGELPAAVVVLEHGKTMTEKEIVDYVASQVTTAKKLRGGVV
                     FVDEVPKGLTGKLDARKIREILIKAKKGGKIAV"
     RBS             1813..1821
                     /label=Shine-Dalgarno sequence
                     /note="full consensus sequence for ribosome-binding sites 
                     upstream of start codons in E. coli; complementary to a 
                     region in the 3' end of the 16S rRNA (Chen et al., 1994)"
     polyA_signal    2550..2671
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2561..2580)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2615..2634
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     promoter        2804..2908
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2909..2977,2978..3769)
                     /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(3127..3146)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3940..4528
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4429..4448
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        4657..5014
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        4657..4848
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(4680..4700)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4865..5000
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     intron          5084..5216
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     primer_bind     5199..5218
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     promoter        5261..5279
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      5283..5292
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             5289..6224
                     /codon_start=1
                     /product="Renilla luciferase"
                     /label=hRluc
                     /note="human codon-optimized"
                     /translation="MASKVYDPEQRKRMITGPQWWARCKQMNVLDSFINYYDSEKHAEN
                     AVIFLHGNAASSYLWRHVVPHIEPVARCIIPDLIGMGKSGKSGNGSYRLLDHYKYLTAW
                     FELLNLPKKIIFVGHDWGACLAFHYSYEHQDKIKAIVHAESVVDVIESWDEWPDIEEDI
                     ALIKSEEGEKMVLENNFFVETMLPSKIMRKLEPEEFAAYLEPFKEKGEVRRPTLSWPRE
                     IPLVKGGKPDVVQIVRNYNAYLRASDDLPKMFIESDPGFFSNAIVEGAKKFPNTEFVKV
                     KGLHFSQEDAPDEMGKYIKSFVERVLKNEQ"
ORIGIN
        1 gctggccgca ataaaatatc tttattttca ttacatctgt gtgttggttt tttgtgtgag
       61 gatctaaatg agtcttcgga cctcgcgggg gccgcttaag cggtggttag ggtttgtctg
      121 acgcgggggg agggggaagg aacgaaacac tctcattcgg aggcggctcg gggtttggtc
      181 ttggtggcca cgggcacgca gaagagcgcc gcgatcctct taagcacccc cccgccctcc
      241 gtggaggcgg gggtttggtc ggcgggtggt aactggcggg ccgctgactc gggcgggtcg
      301 cgcgccccag agtgtgacct tttcggtctg ctcgcagacc cccgggcggc gccgccgcgg
      361 cggcgacggg ctcgctgggt cctaggctcc atggggaccg tatacgtgga caggctctgg
      421 agcatccgca cgactgcggt gatattaccg gagaccttct gcgggacgag ccgggtcacg
      481 cggctgacgc ggagcgtccg ttgggcgaca aacaccagga cggggcacag gtacactatc
      541 ttgtcacccg gaggcgcgag ggactgcagg agcttcaggg agtggcgcag ctgcttcatc
      601 cccgtggccc gttgctcgcg tttgctggcg gtgtccccgg aagaaatata tttgcatgtc
      661 tttagttcta tgatgacaca aaccccgccc agcgtcttgt cattggcgaa ttcgaacacg
      721 cagatgcagt cggggcggcg cggtcccagg tccacttcgc atattaaggt gacgcgtgtg
      781 gcctcgaaca ccgagcgacc ctgcagcgac ccgcttaaaa gcttggcatt ccggtactgt
      841 tggtaaagcc accatggccg atgctaagaa cattaagaag ggccctgctc ccttctaccc
      901 tctggaggat ggcaccgctg gcgagcagct gcacaaggcc atgaagaggt atgccctggt
      961 gcctggcacc attgccttca ccgatgccca cattgaggtg gacatcacct atgccgagta
     1021 cttcgagatg tctgtgcgcc tggccgaggc catgaagagg tacggcctga acaccaacca
     1081 ccgcatcgtg gtgtgctctg agaactctct gcagttcttc atgccagtgc tgggcgccct
     1141 gttcatcgga gtggccgtgg cccctgctaa cgacatttac aacgagcgcg agctgctgaa
     1201 cagcatgggc atttctcagc ctaccgtggt gttcgtgtct aagaagggcc tgcagaagat
     1261 cctgaacgtg cagaagaagc tgcctatcat ccagaagatc atcatcatgg actctaagac
     1321 cgactaccag ggcttccaga gcatgtacac attcgtgaca tctcatctgc ctcctggctt
     1381 caacgagtac gacttcgtgc cagagtcttt cgacagggac aaaaccattg ccctgatcat
     1441 gaacagctct gggtctaccg gcctgcctaa gggcgtggcc ctgcctcatc gcaccgcctg
     1501 tgtgcgcttc tctcacgccc gcgaccctat tttcggcaac cagatcatcc ccgacaccgc
     1561 tattctgagc gtggtgccat tccaccacgg cttcggcatg ttcaccaccc tgggctacct
     1621 gatttgcggc tttcgggtgg tgctgatgta ccgcttcgag gaggagctgt tcctgcgcag
     1681 cctgcaagac tacaaaattc agtctgccct gctggtgcca accctgttca gcttcttcgc
     1741 taagagcacc ctgatcgaca agtacgacct gtctaacctg cacgagattg cctctggcgg
     1801 cgccccactg tctaaggagg tgggcgaagc cgtggccaag cgctttcatc tgccaggcat
     1861 ccgccagggc tacggcctga ccgagacaac cagcgccatt ctgattaccc cagagggcga
     1921 cgacaagcct ggcgccgtgg gcaaggtggt gccattcttc gaggccaagg tggtggacct
     1981 ggacaccggc aagaccctgg gagtgaacca gcgcggcgag ctgtgtgtgc gcggccctat
     2041 gattatgtcc ggctacgtga ataaccctga ggccacaaac gccctgatcg acaaggacgg
     2101 ctggctgcac tctggcgaca ttgcctactg ggacgaggac gagcacttct tcatcgtgga
     2161 ccgcctgaag tctctgatca agtacaaggg ctaccaggtg gccccagccg agctggagtc
     2221 tatcctgctg cagcacccta acattttcga cgccggagtg gccggcctgc ccgacgacga
     2281 tgccggcgag ctgcctgccg ccgtcgtcgt gctggaacac ggcaagacca tgaccgagaa
     2341 ggagatcgtg gactatgtgg ccagccaggt gacaaccgcc aagaagctgc gcggcggagt
     2401 ggtgttcgtg gacgaggtgc ccaagggcct gaccggcaag ctggacgccc gcaagatccg
     2461 cgagatcctg atcaaggcta agaaaggcgg caagatcgcc gtgtaataat tctagagtcg
     2521 gggcggccgg ccgcttcgag cagacatgat aagatacatt gatgagtttg gacaaaccac
     2581 aactagaatg cagtgaaaaa aatgctttat ttgtgaaatt tgtgatgcta ttgctttatt
     2641 tgtaaccatt ataagctgca ataaacaagt taacaacaac aattgcattc attttatgtt
     2701 tcaggttcag ggggaggtgt gggaggtttt ttaaagcaag taaaacctct acaaatgtgg
     2761 taaaatcgat aaggatccag gtggcacttt tcggggaaat gtgcgcggaa cccctatttg
     2821 tttatttttc taaatacatt caaatatgta tccgctcatg agacaataac cctgataaat
     2881 gcttcaataa tattgaaaaa ggaagagtat gagtattcaa catttccgtg tcgcccttat
     2941 tccctttttt gcggcatttt gccttcctgt ttttgctcac ccagaaacgc tggtgaaagt
     3001 aaaagatgct gaagatcagt tgggtgcacg agtgggttac atcgaactgg atctcaacag
     3061 cggtaagatc cttgagagtt ttcgccccga agaacgtttt ccaatgatga gcacttttaa
     3121 agttctgcta tgtggcgcgg tattatcccg tattgacgcc gggcaagagc aactcggtcg
     3181 ccgcatacac tattctcaga atgacttggt tgagtactca ccagtcacag aaaagcatct
     3241 tacggatggc atgacagtaa gagaattatg cagtgctgcc ataaccatga gtgataacac
     3301 tgcggccaac ttacttctga caacgatcgg aggaccgaag gagctaaccg cttttttgca
     3361 caacatgggg gatcatgtaa ctcgccttga tcgttgggaa ccggagctga atgaagccat
     3421 accaaacgac gagcgtgaca ccacgatgcc tgtagcaatg gcaacaacgt tgcgcaaact
     3481 attaactggc gaactactta ctctagcttc ccggcaacaa ttaatagact ggatggaggc
     3541 ggataaagtt gcaggaccac ttctgcgctc ggcccttccg gctggctggt ttattgctga
     3601 taaatctgga gccggtgagc gtgggtctcg cggtatcatt gcagcactgg ggccagatgg
     3661 taagccctcc cgtatcgtag ttatctacac gacggggagt caggcaacta tggatgaacg
     3721 aaatagacag atcgctgaga taggtgcctc actgattaag cattggtaac tgtcagacca
     3781 agtttactca tatatacttt agattgattt aaaacttcat ttttaattta aaaggatcta
     3841 ggtgaagatc ctttttgata atctcatgac caaaatccct taacgtgagt tttcgttcca
     3901 ctgagcgtca gaccccgtag aaaagatcaa aggatcttct tgagatcctt tttttctgcg
     3961 cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca gcggtggttt gtttgccgga
     4021 tcaagagcta ccaactcttt ttccgaaggt aactggcttc agcagagcgc agataccaaa
     4081 tactgttctt ctagtgtagc cgtagttagg ccaccacttc aagaactctg tagcaccgcc
     4141 tacatacctc gctctgctaa tcctgttacc agtggctgct gccagtggcg ataagtcgtg
     4201 tcttaccggg ttggactcaa gacgatagtt accggataag gcgcagcggt cgggctgaac
     4261 ggggggttcg tgcacacagc ccagcttgga gcgaacgacc tacaccgaac tgagatacct
     4321 acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc
     4381 ggtaagcggc agggtcggaa caggagagcg cacgagggag cttccagggg gaaacgcctg
     4441 gtatctttat agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg
     4501 ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac gcggcctttt tacggttcct
     4561 ggccttttgc tggccttttg ctcacatggc tcgacagatc tgcgcagcac catggcctga
     4621 aataacctct gaaagaggaa cttggttagg taccttctga ggcggaaaga accagctgtg
     4681 gaatgtgtgt cagttagggt gtggaaagtc cccaggctcc ccagcaggca gaagtatgca
     4741 aagcatgcat ctcaattagt cagcaaccag gtgtggaaag tccccaggct ccccagcagg
     4801 cagaagtatg caaagcatgc atctcaatta gtcagcaacc atagtcccgc ccctaactcc
     4861 gcccatcccg cccctaactc cgcccagttc cgcccattct ccgccccatg gctgactaat
     4921 tttttttatt tatgcagagg ccgaggccgc ctcggcctct gagctattcc agaagtagtg
     4981 aggaggcttt tttggaggcc taggcttttg caaaaagctt gattcttctg acacaacagt
     5041 ctcgaactta agctgcagaa gttggtcgtg aggcactggg caggtaagta tcaaggttac
     5101 aagacaggtt taaggagacc aatagaaact gggcttgtcg agacagagaa gactcttgcg
     5161 tttctgatag gcacctattg gtcttactga catccacttt gcctttctct ccacaggtgt
     5221 ccactcccag ttcaattaca gctcttaagg ctagagtact taatacgact cactataggc
     5281 tagccaccat ggcttccaag gtgtacgacc ccgagcaacg caaacgcatg atcactgggc
     5341 ctcagtggtg ggctcgctgc aagcaaatga acgtgctgga ctccttcatc aactactatg
     5401 attccgagaa gcacgccgag aacgccgtga tttttctgca tggtaacgct gcctccagct
     5461 acctgtggag gcacgtcgtg cctcacatcg agcccgtggc tagatgcatc atccctgatc
     5521 tgatcggaat gggtaagtcc ggcaagagcg ggaatggctc atatcgcctc ctggatcact
     5581 acaagtacct caccgcttgg ttcgagctgc tgaaccttcc aaagaaaatc atctttgtgg
     5641 gccacgactg gggggcttgt ctggcctttc actactccta cgagcaccaa gacaagatca
     5701 aggccatcgt ccatgctgag agtgtcgtgg acgtgatcga gtcctgggac gagtggcctg
     5761 acatcgagga ggatatcgcc ctgatcaaga gcgaagaggg cgagaaaatg gtgcttgaga
     5821 ataacttctt cgtcgagacc atgctcccaa gcaagatcat gcggaaactg gagcctgagg
     5881 agttcgctgc ctacctggag ccattcaagg agaagggcga ggttagacgg cctaccctct
     5941 cctggcctcg cgagatccct ctcgttaagg gaggcaagcc cgacgtcgtc cagattgtcc
     6001 gcaactacaa cgcctacctt cgggccagcg acgatctgcc taagatgttc atcgagtccg
     6061 accctgggtt cttttccaac gctattgtcg agggagctaa gaagttccct aacaccgagt
     6121 tcgtgaaggt gaagggcctc cacttcagcc aggaggacgc tccagatgaa atgggtaagt
     6181 acatcaagag cttcgtggag cgcgtgctga agaacgagca gtaattctag gcgatcgctc
     6241 gagaaacaac atcctctgga gtttgccgtc cttcctgaga tgcatctttt tgaggagcgt
     6301 caagagtgga cgcacacata ccgaggactc tcaagacatt gctgagtaag acactggact
     6361 ttttcccttt aaacccagac tgctaaagcg gccagtcttt gtctttatgc ccttaagctg
     6421 ctcaggattg ccatttactt tttaaagatg taaataactg gacattgaac taaactcata
     6481 gtgggaccca gcgtcagggt tgcatgctat aatgcctctc aatataaaag gacaaaaact
     6541 gattaattgt gattccgtct ttgggaaact gacatattga gcggcc
//
