LOCUS       sequence-182899-        8143 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 8143)
  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 8143)
  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-TAB2-3'UTR
FEATURES             Location/Qualifiers
     source          1..8143
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      14..602
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     503..522
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        731..1088
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        731..922
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(754..774)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      939..1074
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     intron          1158..1290
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     primer_bind     1273..1292
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     promoter        1335..1353
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      1357..1366
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1363..2298
                     /codon_start=1
                     /product="Renilla luciferase"
                     /label=hRluc
                     /note="human codon-optimized"
                     /translation="MASKVYDPEQRKRMITGPQWWARCKQMNVLDSFINYYDSEKHAEN
                     AVIFLHGNAASSYLWRHVVPHIEPVARCIIPDLIGMGKSGKSGNGSYRLLDHYKYLTAW
                     FELLNLPKKIIFVGHDWGACLAFHYSYEHQDKIKAIVHAESVVDVIESWDEWPDIEEDI
                     ALIKSEEGEKMVLENNFFVETMLPSKIMRKLEPEEFAAYLEPFKEKGEVRRPTLSWPRE
                     IPLVKGGKPDVVQIVRNYNAYLRASDDLPKMFIESDPGFFSNAIVEGAKKFPNTEFVKV
                     KGLHFSQEDAPDEMGKYIKSFVERVLKNEQ"
     polyA_signal    4227..4275
                     /note="synthetic polyadenylation signal"
     promoter        4284..5035
                     /label=HSV TK promoter
                     /note="herpes simplex virus thymidine kinase promoter"
     regulatory      5065..5074
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             5071..6723
                     /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             6030..6038
                     /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    6767..6888
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6778..6797)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     6832..6851
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     promoter        7021..7125
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(7126..7194,7195..7986)
                     /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(7344..7363)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
ORIGIN
        1 tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa
       61 aaccaccgct accagcggtg gtttgtttgc cggatcaaga gctaccaact ctttttccga
      121 aggtaactgg cttcagcaga gcgcagatac caaatactgt tcttctagtg tagccgtagt
      181 taggccacca cttcaagaac tctgtagcac cgcctacata cctcgctctg ctaatcctgt
      241 taccagtggc tgctgccagt ggcgataagt cgtgtcttac cgggttggac tcaagacgat
      301 agttaccgga taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca cagcccagct
      361 tggagcgaac gacctacacc gaactgagat acctacagcg tgagctatga gaaagcgcca
      421 cgcttcccga agggagaaag gcggacaggt atccggtaag cggcagggtc ggaacaggag
      481 agcgcacgag ggagcttcca gggggaaacg cctggtatct ttatagtcct gtcgggtttc
      541 gccacctctg acttgagcgt cgatttttgt gatgctcgtc aggggggcgg agcctatgga
      601 aaaacgccag caacgcggcc tttttacggt tcctggcctt ttgctggcct tttgctcaca
      661 tggctcgaca gatctgcgca gcaccatggc ctgaaataac ctctgaaaga ggaacttggt
      721 taggtacctt ctgaggcgga aagaaccagc tgtggaatgt gtgtcagtta gggtgtggaa
      781 agtccccagg ctccccagca ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa
      841 ccaggtgtgg aaagtcccca ggctccccag caggcagaag tatgcaaagc atgcatctca
      901 attagtcagc aaccatagtc ccgcccctaa ctccgcccat cccgccccta actccgccca
      961 gttccgccca ttctccgccc catggctgac taattttttt tatttatgca gaggccgagg
     1021 ccgcctcggc ctctgagcta ttccagaagt agtgaggagg cttttttgga ggcctaggct
     1081 tttgcaaaaa gcttgattct tctgacacaa cagtctcgaa cttaagctgc agaagttggt
     1141 cgtgaggcac tgggcaggta agtatcaagg ttacaagaca ggtttaagga gaccaataga
     1201 aactgggctt gtcgagacag agaagactct tgcgtttctg ataggcacct attggtctta
     1261 ctgacatcca ctttgccttt ctctccacag gtgtccactc ccagttcaat tacagctctt
     1321 aaggctagag tacttaatac gactcactat aggctagcca ccatggcttc caaggtgtac
     1381 gaccccgagc aacgcaaacg catgatcact gggcctcagt ggtgggctcg ctgcaagcaa
     1441 atgaacgtgc tggactcctt catcaactac tatgattccg agaagcacgc cgagaacgcc
     1501 gtgatttttc tgcatggtaa cgctgcctcc agctacctgt ggaggcacgt cgtgcctcac
     1561 atcgagcccg tggctagatg catcatccct gatctgatcg gaatgggtaa gtccggcaag
     1621 agcgggaatg gctcatatcg cctcctggat cactacaagt acctcaccgc ttggttcgag
     1681 ctgctgaacc ttccaaagaa aatcatcttt gtgggccacg actggggggc ttgtctggcc
     1741 tttcactact cctacgagca ccaagacaag atcaaggcca tcgtccatgc tgagagtgtc
     1801 gtggacgtga tcgagtcctg ggacgagtgg cctgacatcg aggaggatat cgccctgatc
     1861 aagagcgaag agggcgagaa aatggtgctt gagaataact tcttcgtcga gaccatgctc
     1921 ccaagcaaga tcatgcggaa actggagcct gaggagttcg ctgcctacct ggagccattc
     1981 aaggagaagg gcgaggttag acggcctacc ctctcctggc ctcgcgagat ccctctcgtt
     2041 aagggaggca agcccgacgt cgtccagatt gtccgcaact acaacgccta ccttcgggcc
     2101 agcgacgatc tgcctaagat gttcatcgag tccgaccctg ggttcttttc caacgctatt
     2161 gtcgagggag ctaagaagtt ccctaacacc gagttcgtga aggtgaaggg cctccacttc
     2221 agccaggagg acgctccaga tgaaatgggt aagtacatca agagcttcgt ggagcgcgtg
     2281 ctgaagaacg agcagtaatt ctaggcgatc gctcgaggcc aaatggccct gtatcttctc
     2341 taaaaccaca tctaaagttc aagaaactag tctgtcatcg ggaaaaagtt tcactgctac
     2401 ataggatttt gtcaaattga aggtgtgaca agatggtgtt ctgctaatgt taaatgtcag
     2461 cccacagagc taataatacc tcagtataat gtcatgagca gttgaaattc atcacatgaa
     2521 aagtaatctg ctgaaagact tggttgccca ctgcctaact gtgtacagtg ttaccagtgt
     2581 cccattatgg ataattctca atatgttaac acctaggtgt tcccaatacc tttttcccct
     2641 catgtcacta ctgaattttg acaggaggaa ggaatagaat gatagcttgt tttatttgta
     2701 aagctttcag tgaaacacta catacacgaa gaaaaggaac aaggtttaac tatttaagaa
     2761 ccatttgctg ccgcatagtg ccattggata gggaagaact tcagaaatct gtggtactct
     2821 tggccttgtc tttgtcttcc ctgaacgtgt ctccactctg tgaagccagc atctaggggc
     2881 taaagatgca aaggaaagca gcatgcattg tctgtacaaa tgtgcagcga aatacccgaa
     2941 agcttttcct actgtacaga tctctcgagt ctgctttaag tgatttcttt tcttcttgat
     3001 tattttctta tatttctata tgtatagtgt aatagccttt tgttaactaa ttttcttttt
     3061 tccttttagt aattaagcac gatcatgtcc ctttttaagc cttacctgag aggaacaatg
     3121 ccttaaaata aaaaagcatt aatgagatga aagtatgcac agaataactt tcctctactt
     3181 attctgtact ttgccctcat gagttccaat gttgtgtgaa gacaggcaga tgctgcacag
     3241 tgaattgcag atgatattac agaagtgatg tctgtaggtc acattaaata ctgacttgag
     3301 cagtgggtga cacaacacag tgtttgtctt ccacagggaa gcttaaaaca aaagatattt
     3361 ttaacccact gacagaacaa caaggttaag cttcatctgc ttggtgtccc acagaacttg
     3421 cacaagcagt tgttattggg aaagtacagt ctcaaaacca gcaacagcag cagtacctac
     3481 agcccttttt ttggagagaa gtttaaatgc tttactgttg gggcagtcca tttctaatcc
     3541 tgacttggtg acagtatcat gtgtatttat aaaacaaggc tagccatatt taggacaact
     3601 gaagaaaagc tggaaaaaaa aacaagcaaa cttgaacact gaagcaacct caagcatctc
     3661 tttattttga tgatatattt ttgtaaggaa aatattcaga tgatcaggaa tgtatataac
     3721 tgaaatcaag aaaaagaaca gtatgcattt aaaaagacag aattatgaaa ttatatgagt
     3781 gcttagaatg gggctaaggg aagtgctgaa atagagcaaa ggatggaaga taatatagac
     3841 taccacccac tgtaaatgtt tgcaagtggc tgtgttttaa atgggattat tacagttgat
     3901 ctctatgaat gtcagagccc taactttcag gctttgcatt ttgtatatgg gaagaaatat
     3961 gacaatccta ggtaattaaa ccatagaccc aaagccctta cgtttgatgc aatttatttt
     4021 taaaataggc cttgtttttc agcttcatct gcagttctat gtgaagattg ataaatcagt
     4081 ttttacttgt tttattaata aaacgtaatt tggatatctt gagttgatgg ttttgtgatt
     4141 tagctgggta aactatcttt gtaacagata agttatttat aaaaattaaa aaacttatat
     4201 tctaatgtgg agcggccgct ggccgcaata aaatatcttt attttcatta catctgtgtg
     4261 ttggtttttt gtgtgaggat ctaaatgagt cttcggacct cgcgggggcc gcttaagcgg
     4321 tggttagggt ttgtctgacg cggggggagg gggaaggaac gaaacactct cattcggagg
     4381 cggctcgggg tttggtcttg gtggccacgg gcacgcagaa gagcgccgcg atcctcttaa
     4441 gcaccccccc gccctccgtg gaggcggggg tttggtcggc gggtggtaac tggcgggccg
     4501 ctgactcggg cgggtcgcgc gccccagagt gtgacctttt cggtctgctc gcagaccccc
     4561 gggcggcgcc gccgcggcgg cgacgggctc gctgggtcct aggctccatg gggaccgtat
     4621 acgtggacag gctctggagc atccgcacga ctgcggtgat attaccggag accttctgcg
     4681 ggacgagccg ggtcacgcgg ctgacgcgga gcgtccgttg ggcgacaaac accaggacgg
     4741 ggcacaggta cactatcttg tcacccggag gcgcgaggga ctgcaggagc ttcagggagt
     4801 ggcgcagctg cttcatcccc gtggcccgtt gctcgcgttt gctggcggtg tccccggaag
     4861 aaatatattt gcatgtcttt agttctatga tgacacaaac cccgcccagc gtcttgtcat
     4921 tggcgaattc gaacacgcag atgcagtcgg ggcggcgcgg tcccaggtcc acttcgcata
     4981 ttaaggtgac gcgtgtggcc tcgaacaccg agcgaccctg cagcgacccg cttaaaagct
     5041 tggcattccg gtactgttgg taaagccacc atggccgatg ctaagaacat taagaagggc
     5101 cctgctccct tctaccctct ggaggatggc accgctggcg agcagctgca caaggccatg
     5161 aagaggtatg ccctggtgcc tggcaccatt gccttcaccg atgcccacat tgaggtggac
     5221 atcacctatg ccgagtactt cgagatgtct gtgcgcctgg ccgaggccat gaagaggtac
     5281 ggcctgaaca ccaaccaccg catcgtggtg tgctctgaga actctctgca gttcttcatg
     5341 ccagtgctgg gcgccctgtt catcggagtg gccgtggccc ctgctaacga catttacaac
     5401 gagcgcgagc tgctgaacag catgggcatt tctcagccta ccgtggtgtt cgtgtctaag
     5461 aagggcctgc agaagatcct gaacgtgcag aagaagctgc ctatcatcca gaagatcatc
     5521 atcatggact ctaagaccga ctaccagggc ttccagagca tgtacacatt cgtgacatct
     5581 catctgcctc ctggcttcaa cgagtacgac ttcgtgccag agtctttcga cagggacaaa
     5641 accattgccc tgatcatgaa cagctctggg tctaccggcc tgcctaaggg cgtggccctg
     5701 cctcatcgca ccgcctgtgt gcgcttctct cacgcccgcg accctatttt cggcaaccag
     5761 atcatccccg acaccgctat tctgagcgtg gtgccattcc accacggctt cggcatgttc
     5821 accaccctgg gctacctgat ttgcggcttt cgggtggtgc tgatgtaccg cttcgaggag
     5881 gagctgttcc tgcgcagcct gcaagactac aaaattcagt ctgccctgct ggtgccaacc
     5941 ctgttcagct tcttcgctaa gagcaccctg atcgacaagt acgacctgtc taacctgcac
     6001 gagattgcct ctggcggcgc cccactgtct aaggaggtgg gcgaagccgt ggccaagcgc
     6061 tttcatctgc caggcatccg ccagggctac ggcctgaccg agacaaccag cgccattctg
     6121 attaccccag agggcgacga caagcctggc gccgtgggca aggtggtgcc attcttcgag
     6181 gccaaggtgg tggacctgga caccggcaag accctgggag tgaaccagcg cggcgagctg
     6241 tgtgtgcgcg gccctatgat tatgtccggc tacgtgaata accctgaggc cacaaacgcc
     6301 ctgatcgaca aggacggctg gctgcactct ggcgacattg cctactggga cgaggacgag
     6361 cacttcttca tcgtggaccg cctgaagtct ctgatcaagt acaagggcta ccaggtggcc
     6421 ccagccgagc tggagtctat cctgctgcag caccctaaca ttttcgacgc cggagtggcc
     6481 ggcctgcccg acgacgatgc cggcgagctg cctgccgccg tcgtcgtgct ggaacacggc
     6541 aagaccatga ccgagaagga gatcgtggac tatgtggcca gccaggtgac aaccgccaag
     6601 aagctgcgcg gcggagtggt gttcgtggac gaggtgccca agggcctgac cggcaagctg
     6661 gacgcccgca agatccgcga gatcctgatc aaggctaaga aaggcggcaa gatcgccgtg
     6721 taataattct agagtcgggg cggccggccg cttcgagcag acatgataag atacattgat
     6781 gagtttggac aaaccacaac tagaatgcag tgaaaaaaat gctttatttg tgaaatttgt
     6841 gatgctattg ctttatttgt aaccattata agctgcaata aacaagttaa caacaacaat
     6901 tgcattcatt ttatgtttca ggttcagggg gaggtgtggg aggtttttta aagcaagtaa
     6961 aacctctaca aatgtggtaa aatcgataag gatccaggtg gcacttttcg gggaaatgtg
     7021 cgcggaaccc ctatttgttt atttttctaa atacattcaa atatgtatcc gctcatgaga
     7081 caataaccct gataaatgct tcaataatat tgaaaaagga agagtatgag tattcaacat
     7141 ttccgtgtcg cccttattcc cttttttgcg gcattttgcc ttcctgtttt tgctcaccca
     7201 gaaacgctgg tgaaagtaaa agatgctgaa gatcagttgg gtgcacgagt gggttacatc
     7261 gaactggatc tcaacagcgg taagatcctt gagagttttc gccccgaaga acgttttcca
     7321 atgatgagca cttttaaagt tctgctatgt ggcgcggtat tatcccgtat tgacgccggg
     7381 caagagcaac tcggtcgccg catacactat tctcagaatg acttggttga gtactcacca
     7441 gtcacagaaa agcatcttac ggatggcatg acagtaagag aattatgcag tgctgccata
     7501 accatgagtg ataacactgc ggccaactta cttctgacaa cgatcggagg accgaaggag
     7561 ctaaccgctt ttttgcacaa catgggggat catgtaactc gccttgatcg ttgggaaccg
     7621 gagctgaatg aagccatacc aaacgacgag cgtgacacca cgatgcctgt agcaatggca
     7681 acaacgttgc gcaaactatt aactggcgaa ctacttactc tagcttcccg gcaacaatta
     7741 atagactgga tggaggcgga taaagttgca ggaccacttc tgcgctcggc ccttccggct
     7801 ggctggttta ttgctgataa atctggagcc ggtgagcgtg ggtctcgcgg tatcattgca
     7861 gcactggggc cagatggtaa gccctcccgt atcgtagtta tctacacgac ggggagtcag
     7921 gcaactatgg atgaacgaaa tagacagatc gctgagatag gtgcctcact gattaagcat
     7981 tggtaactgt cagaccaagt ttactcatat atactttaga ttgatttaaa acttcatttt
     8041 taatttaaaa ggatctaggt gaagatcctt tttgataatc tcatgaccaa aatcccttaa
     8101 cgtgagtttt cgttccactg agcgtcagac cccgtagaaa aga
//
