LOCUS       sequence-182888-        6702 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 6702)
  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 6702)
  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-CUL1-3'UTR
FEATURES             Location/Qualifiers
     source          1..6702
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      20..608
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     509..528
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        737..1094
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        737..928
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(760..780)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      945..1080
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     intron          1164..1296
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     primer_bind     1279..1298
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     promoter        1341..1359
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      1363..1372
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1369..2304
                     /codon_start=1
                     /product="Renilla luciferase"
                     /label=hRluc
                     /note="human codon-optimized"
                     /translation="MASKVYDPEQRKRMITGPQWWARCKQMNVLDSFINYYDSEKHAEN
                     AVIFLHGNAASSYLWRHVVPHIEPVARCIIPDLIGMGKSGKSGNGSYRLLDHYKYLTAW
                     FELLNLPKKIIFVGHDWGACLAFHYSYEHQDKIKAIVHAESVVDVIESWDEWPDIEEDI
                     ALIKSEEGEKMVLENNFFVETMLPSKIMRKLEPEEFAAYLEPFKEKGEVRRPTLSWPRE
                     IPLVKGGKPDVVQIVRNYNAYLRASDDLPKMFIESDPGFFSNAIVEGAKKFPNTEFVKV
                     KGLHFSQEDAPDEMGKYIKSFVERVLKNEQ"
     polyA_signal    2792..2840
                     /note="synthetic polyadenylation signal"
     promoter        2849..3600
                     /label=HSV TK promoter
                     /note="herpes simplex virus thymidine kinase promoter"
     regulatory      3630..3639
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             3636..5288
                     /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             4595..4603
                     /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    5332..5453
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5343..5362)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     5397..5416
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     promoter        5586..5690
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5691..5759,5760..6551)
                     /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(5909..5928)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
ORIGIN
        1 aaaagatcaa aggatcttct tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa
       61 caaaaaaacc accgctacca gcggtggttt gtttgccgga tcaagagcta ccaactcttt
      121 ttccgaaggt aactggcttc agcagagcgc agataccaaa tactgttctt ctagtgtagc
      181 cgtagttagg ccaccacttc aagaactctg tagcaccgcc tacatacctc gctctgctaa
      241 tcctgttacc agtggctgct gccagtggcg ataagtcgtg tcttaccggg ttggactcaa
      301 gacgatagtt accggataag gcgcagcggt cgggctgaac ggggggttcg tgcacacagc
      361 ccagcttgga gcgaacgacc tacaccgaac tgagatacct acagcgtgag ctatgagaaa
      421 gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa
      481 caggagagcg cacgagggag cttccagggg gaaacgcctg gtatctttat agtcctgtcg
      541 ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc
      601 tatggaaaaa cgccagcaac gcggcctttt tacggttcct ggccttttgc tggccttttg
      661 ctcacatggc tcgacagatc tgcgcagcac catggcctga aataacctct gaaagaggaa
      721 cttggttagg taccttctga ggcggaaaga accagctgtg gaatgtgtgt cagttagggt
      781 gtggaaagtc cccaggctcc ccagcaggca gaagtatgca aagcatgcat ctcaattagt
      841 cagcaaccag gtgtggaaag tccccaggct ccccagcagg cagaagtatg caaagcatgc
      901 atctcaatta gtcagcaacc atagtcccgc ccctaactcc gcccatcccg cccctaactc
      961 cgcccagttc cgcccattct ccgccccatg gctgactaat tttttttatt tatgcagagg
     1021 ccgaggccgc ctcggcctct gagctattcc agaagtagtg aggaggcttt tttggaggcc
     1081 taggcttttg caaaaagctt gattcttctg acacaacagt ctcgaactta agctgcagaa
     1141 gttggtcgtg aggcactggg caggtaagta tcaaggttac aagacaggtt taaggagacc
     1201 aatagaaact gggcttgtcg agacagagaa gactcttgcg tttctgatag gcacctattg
     1261 gtcttactga catccacttt gcctttctct ccacaggtgt ccactcccag ttcaattaca
     1321 gctcttaagg ctagagtact taatacgact cactataggc tagccaccat ggcttccaag
     1381 gtgtacgacc ccgagcaacg caaacgcatg atcactgggc ctcagtggtg ggctcgctgc
     1441 aagcaaatga acgtgctgga ctccttcatc aactactatg attccgagaa gcacgccgag
     1501 aacgccgtga tttttctgca tggtaacgct gcctccagct acctgtggag gcacgtcgtg
     1561 cctcacatcg agcccgtggc tagatgcatc atccctgatc tgatcggaat gggtaagtcc
     1621 ggcaagagcg ggaatggctc atatcgcctc ctggatcact acaagtacct caccgcttgg
     1681 ttcgagctgc tgaaccttcc aaagaaaatc atctttgtgg gccacgactg gggggcttgt
     1741 ctggcctttc actactccta cgagcaccaa gacaagatca aggccatcgt ccatgctgag
     1801 agtgtcgtgg acgtgatcga gtcctgggac gagtggcctg acatcgagga ggatatcgcc
     1861 ctgatcaaga gcgaagaggg cgagaaaatg gtgcttgaga ataacttctt cgtcgagacc
     1921 atgctcccaa gcaagatcat gcggaaactg gagcctgagg agttcgctgc ctacctggag
     1981 ccattcaagg agaagggcga ggttagacgg cctaccctct cctggcctcg cgagatccct
     2041 ctcgttaagg gaggcaagcc cgacgtcgtc cagattgtcc gcaactacaa cgcctacctt
     2101 cgggccagcg acgatctgcc taagatgttc atcgagtccg accctgggtt cttttccaac
     2161 gctattgtcg agggagctaa gaagttccct aacaccgagt tcgtgaaggt gaagggcctc
     2221 cacttcagcc aggaggacgc tccagatgaa atgggtaagt acatcaagag cttcgtggag
     2281 cgcgtgctga agaacgagca gtaattctag gcgatcgctc gagccctctg gaagggtctg
     2341 actgtgtgac ccgcagcaaa tagttcatgt tggaaagaat gaaaacaact caagttcata
     2401 gcagccagcc tgccgccatt ggacctccct tttaaaaact gagaccaaga ctcccatcag
     2461 ctggtctcgg atttacatcg gaactgctca ggattgatac atttcaagtc tgtaaatacg
     2521 gacaccaacg ccatttaccc taatttaaga acagcgggga ctgaccctcc gtgccgaggg
     2581 ctgcatgcta ccgcactaag tcaatacatg ggctccccga ttcgcagctg tcgtcttggc
     2641 agcacttgtc acgttggcag cactttgaga gcaagtctga gtggacccac atgtaacctg
     2701 ctatgaaaac catttgtata gtgtgtttca ttttttaatg tgtgaaaata aagaaaatta
     2761 aaggatttct gtacaagcgg ccgctggccg caataaaata tctttatttt cattacatct
     2821 gtgtgttggt tttttgtgtg aggatctaaa tgagtcttcg gacctcgcgg gggccgctta
     2881 agcggtggtt agggtttgtc tgacgcgggg ggagggggaa ggaacgaaac actctcattc
     2941 ggaggcggct cggggtttgg tcttggtggc cacgggcacg cagaagagcg ccgcgatcct
     3001 cttaagcacc cccccgccct ccgtggaggc gggggtttgg tcggcgggtg gtaactggcg
     3061 ggccgctgac tcgggcgggt cgcgcgcccc agagtgtgac cttttcggtc tgctcgcaga
     3121 cccccgggcg gcgccgccgc ggcggcgacg ggctcgctgg gtcctaggct ccatggggac
     3181 cgtatacgtg gacaggctct ggagcatccg cacgactgcg gtgatattac cggagacctt
     3241 ctgcgggacg agccgggtca cgcggctgac gcggagcgtc cgttgggcga caaacaccag
     3301 gacggggcac aggtacacta tcttgtcacc cggaggcgcg agggactgca ggagcttcag
     3361 ggagtggcgc agctgcttca tccccgtggc ccgttgctcg cgtttgctgg cggtgtcccc
     3421 ggaagaaata tatttgcatg tctttagttc tatgatgaca caaaccccgc ccagcgtctt
     3481 gtcattggcg aattcgaaca cgcagatgca gtcggggcgg cgcggtccca ggtccacttc
     3541 gcatattaag gtgacgcgtg tggcctcgaa caccgagcga ccctgcagcg acccgcttaa
     3601 aagcttggca ttccggtact gttggtaaag ccaccatggc cgatgctaag aacattaaga
     3661 agggccctgc tcccttctac cctctggagg atggcaccgc tggcgagcag ctgcacaagg
     3721 ccatgaagag gtatgccctg gtgcctggca ccattgcctt caccgatgcc cacattgagg
     3781 tggacatcac ctatgccgag tacttcgaga tgtctgtgcg cctggccgag gccatgaaga
     3841 ggtacggcct gaacaccaac caccgcatcg tggtgtgctc tgagaactct ctgcagttct
     3901 tcatgccagt gctgggcgcc ctgttcatcg gagtggccgt ggcccctgct aacgacattt
     3961 acaacgagcg cgagctgctg aacagcatgg gcatttctca gcctaccgtg gtgttcgtgt
     4021 ctaagaaggg cctgcagaag atcctgaacg tgcagaagaa gctgcctatc atccagaaga
     4081 tcatcatcat ggactctaag accgactacc agggcttcca gagcatgtac acattcgtga
     4141 catctcatct gcctcctggc ttcaacgagt acgacttcgt gccagagtct ttcgacaggg
     4201 acaaaaccat tgccctgatc atgaacagct ctgggtctac cggcctgcct aagggcgtgg
     4261 ccctgcctca tcgcaccgcc tgtgtgcgct tctctcacgc ccgcgaccct attttcggca
     4321 accagatcat ccccgacacc gctattctga gcgtggtgcc attccaccac ggcttcggca
     4381 tgttcaccac cctgggctac ctgatttgcg gctttcgggt ggtgctgatg taccgcttcg
     4441 aggaggagct gttcctgcgc agcctgcaag actacaaaat tcagtctgcc ctgctggtgc
     4501 caaccctgtt cagcttcttc gctaagagca ccctgatcga caagtacgac ctgtctaacc
     4561 tgcacgagat tgcctctggc ggcgccccac tgtctaagga ggtgggcgaa gccgtggcca
     4621 agcgctttca tctgccaggc atccgccagg gctacggcct gaccgagaca accagcgcca
     4681 ttctgattac cccagagggc gacgacaagc ctggcgccgt gggcaaggtg gtgccattct
     4741 tcgaggccaa ggtggtggac ctggacaccg gcaagaccct gggagtgaac cagcgcggcg
     4801 agctgtgtgt gcgcggccct atgattatgt ccggctacgt gaataaccct gaggccacaa
     4861 acgccctgat cgacaaggac ggctggctgc actctggcga cattgcctac tgggacgagg
     4921 acgagcactt cttcatcgtg gaccgcctga agtctctgat caagtacaag ggctaccagg
     4981 tggccccagc cgagctggag tctatcctgc tgcagcaccc taacattttc gacgccggag
     5041 tggccggcct gcccgacgac gatgccggcg agctgcctgc cgccgtcgtc gtgctggaac
     5101 acggcaagac catgaccgag aaggagatcg tggactatgt ggccagccag gtgacaaccg
     5161 ccaagaagct gcgcggcgga gtggtgttcg tggacgaggt gcccaagggc ctgaccggca
     5221 agctggacgc ccgcaagatc cgcgagatcc tgatcaaggc taagaaaggc ggcaagatcg
     5281 ccgtgtaata attctagagt cggggcggcc ggccgcttcg agcagacatg ataagataca
     5341 ttgatgagtt tggacaaacc acaactagaa tgcagtgaaa aaaatgcttt atttgtgaaa
     5401 tttgtgatgc tattgcttta tttgtaacca ttataagctg caataaacaa gttaacaaca
     5461 acaattgcat tcattttatg tttcaggttc agggggaggt gtgggaggtt ttttaaagca
     5521 agtaaaacct ctacaaatgt ggtaaaatcg ataaggatcc aggtggcact tttcggggaa
     5581 atgtgcgcgg aacccctatt tgtttatttt tctaaataca ttcaaatatg tatccgctca
     5641 tgagacaata accctgataa atgcttcaat aatattgaaa aaggaagagt atgagtattc
     5701 aacatttccg tgtcgccctt attccctttt ttgcggcatt ttgccttcct gtttttgctc
     5761 acccagaaac gctggtgaaa gtaaaagatg ctgaagatca gttgggtgca cgagtgggtt
     5821 acatcgaact ggatctcaac agcggtaaga tccttgagag ttttcgcccc gaagaacgtt
     5881 ttccaatgat gagcactttt aaagttctgc tatgtggcgc ggtattatcc cgtattgacg
     5941 ccgggcaaga gcaactcggt cgccgcatac actattctca gaatgacttg gttgagtact
     6001 caccagtcac agaaaagcat cttacggatg gcatgacagt aagagaatta tgcagtgctg
     6061 ccataaccat gagtgataac actgcggcca acttacttct gacaacgatc ggaggaccga
     6121 aggagctaac cgcttttttg cacaacatgg gggatcatgt aactcgcctt gatcgttggg
     6181 aaccggagct gaatgaagcc ataccaaacg acgagcgtga caccacgatg cctgtagcaa
     6241 tggcaacaac gttgcgcaaa ctattaactg gcgaactact tactctagct tcccggcaac
     6301 aattaataga ctggatggag gcggataaag ttgcaggacc acttctgcgc tcggcccttc
     6361 cggctggctg gtttattgct gataaatctg gagccggtga gcgtgggtct cgcggtatca
     6421 ttgcagcact ggggccagat ggtaagccct cccgtatcgt agttatctac acgacgggga
     6481 gtcaggcaac tatggatgaa cgaaatagac agatcgctga gataggtgcc tcactgatta
     6541 agcattggta actgtcagac caagtttact catatatact ttagattgat ttaaaacttc
     6601 atttttaatt taaaaggatc taggtgaaga tcctttttga taatctcatg accaaaatcc
     6661 cttaacgtga gttttcgttc cactgagcgt cagaccccgt ag
//
