LOCUS       sequence-182870-        7636 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 7636)
  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 7636)
  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-Tab1-3'UTR
FEATURES             Location/Qualifiers
     source          1..7636
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      17..605
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     506..525
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        734..1091
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        734..925
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(757..777)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      942..1077
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     intron          1161..1293
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     primer_bind     1276..1295
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     promoter        1338..1356
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      1360..1369
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1366..2301
                     /codon_start=1
                     /product="Renilla luciferase"
                     /label=hRluc
                     /note="human codon-optimized"
                     /translation="MASKVYDPEQRKRMITGPQWWARCKQMNVLDSFINYYDSEKHAEN
                     AVIFLHGNAASSYLWRHVVPHIEPVARCIIPDLIGMGKSGKSGNGSYRLLDHYKYLTAW
                     FELLNLPKKIIFVGHDWGACLAFHYSYEHQDKIKAIVHAESVVDVIESWDEWPDIEEDI
                     ALIKSEEGEKMVLENNFFVETMLPSKIMRKLEPEEFAAYLEPFKEKGEVRRPTLSWPRE
                     IPLVKGGKPDVVQIVRNYNAYLRASDDLPKMFIESDPGFFSNAIVEGAKKFPNTEFVKV
                     KGLHFSQEDAPDEMGKYIKSFVERVLKNEQ"
     polyA_signal    3723..3771
                     /note="synthetic polyadenylation signal"
     promoter        3780..4531
                     /label=HSV TK promoter
                     /note="herpes simplex virus thymidine kinase promoter"
     regulatory      4561..4570
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             4567..6219
                     /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             5526..5534
                     /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    6263..6384
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6274..6293)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     6328..6347
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     promoter        6517..6621
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6622..6690,6691..7482)
                     /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(6840..6859)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
ORIGIN
        1 agatcaaagg atcttcttga gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa
       61 aaaaaccacc gctaccagcg gtggtttgtt tgccggatca agagctacca actctttttc
      121 cgaaggtaac tggcttcagc agagcgcaga taccaaatac tgttcttcta gtgtagccgt
      181 agttaggcca ccacttcaag aactctgtag caccgcctac atacctcgct ctgctaatcc
      241 tgttaccagt ggctgctgcc agtggcgata agtcgtgtct taccgggttg gactcaagac
      301 gatagttacc ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc acacagccca
      361 gcttggagcg aacgacctac accgaactga gatacctaca gcgtgagcta tgagaaagcg
      421 ccacgcttcc cgaagggaga aaggcggaca ggtatccggt aagcggcagg gtcggaacag
      481 gagagcgcac gagggagctt ccagggggaa acgcctggta tctttatagt cctgtcgggt
      541 ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg cggagcctat
      601 ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc cttttgctgg ccttttgctc
      661 acatggctcg acagatctgc gcagcaccat ggcctgaaat aacctctgaa agaggaactt
      721 ggttaggtac cttctgaggc ggaaagaacc agctgtggaa tgtgtgtcag ttagggtgtg
      781 gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag catgcatctc aattagtcag
      841 caaccaggtg tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa agcatgcatc
      901 tcaattagtc agcaaccata gtcccgcccc taactccgcc catcccgccc ctaactccgc
      961 ccagttccgc ccattctccg ccccatggct gactaatttt ttttatttat gcagaggccg
     1021 aggccgcctc ggcctctgag ctattccaga agtagtgagg aggctttttt ggaggcctag
     1081 gcttttgcaa aaagcttgat tcttctgaca caacagtctc gaacttaagc tgcagaagtt
     1141 ggtcgtgagg cactgggcag gtaagtatca aggttacaag acaggtttaa ggagaccaat
     1201 agaaactggg cttgtcgaga cagagaagac tcttgcgttt ctgataggca cctattggtc
     1261 ttactgacat ccactttgcc tttctctcca caggtgtcca ctcccagttc aattacagct
     1321 cttaaggcta gagtacttaa tacgactcac tataggctag ccaccatggc ttccaaggtg
     1381 tacgaccccg agcaacgcaa acgcatgatc actgggcctc agtggtgggc tcgctgcaag
     1441 caaatgaacg tgctggactc cttcatcaac tactatgatt ccgagaagca cgccgagaac
     1501 gccgtgattt ttctgcatgg taacgctgcc tccagctacc tgtggaggca cgtcgtgcct
     1561 cacatcgagc ccgtggctag atgcatcatc cctgatctga tcggaatggg taagtccggc
     1621 aagagcggga atggctcata tcgcctcctg gatcactaca agtacctcac cgcttggttc
     1681 gagctgctga accttccaaa gaaaatcatc tttgtgggcc acgactgggg ggcttgtctg
     1741 gcctttcact actcctacga gcaccaagac aagatcaagg ccatcgtcca tgctgagagt
     1801 gtcgtggacg tgatcgagtc ctgggacgag tggcctgaca tcgaggagga tatcgccctg
     1861 atcaagagcg aagagggcga gaaaatggtg cttgagaata acttcttcgt cgagaccatg
     1921 ctcccaagca agatcatgcg gaaactggag cctgaggagt tcgctgccta cctggagcca
     1981 ttcaaggaga agggcgaggt tagacggcct accctctcct ggcctcgcga gatccctctc
     2041 gttaagggag gcaagcccga cgtcgtccag attgtccgca actacaacgc ctaccttcgg
     2101 gccagcgacg atctgcctaa gatgttcatc gagtccgacc ctgggttctt ttccaacgct
     2161 attgtcgagg gagctaagaa gttccctaac accgagttcg tgaaggtgaa gggcctccac
     2221 ttcagccagg aggacgctcc agatgaaatg ggtaagtaca tcaagagctt cgtggagcgc
     2281 gtgctgaaga acgagcagta attctaggcg atcgctcgag catctgtgag cacagtccgt
     2341 cgtctctgac tgtgtggtgg atggaggagc tggactgact ctataaaagg gaagagactg
     2401 tgttcactca cggtgacgtc tgctcattac tcttcagttt taccccttca cgttctctga
     2461 gcttctgtct gtcctgcttg tgtgacgtct gcttcatgta aagtcaccga gagcagttct
     2521 gcagtcgtct gtctgtctgt ctgtctgttt ctgtgtgtta ttggagtttt agtgccttcc
     2581 tataataaac atagaagctt gattaaaaat ctacaactgt gtgaatacag ctggatgcac
     2641 acttaagcgt ttgggtcttc tgctgcatga aaatatacta tagtaactca tcgtaaatac
     2701 tagtgtttta accacactgt agtgaagtat ttgatttaat ctgttgtgct gattctacag
     2761 ttgccgtggt aacaacatct agaaattact ttctgttgtg gttattctac agttgctaag
     2821 gtaacacaac ttcaaatctg ttgtgctgat tctacagttg ctatggtaac agcatttata
     2881 gagaatactc tgttgagctg attctatagt tgctatggta acacagcacc aaatcgtttg
     2941 tggttattat agttgccatg gtaacattca ttttcttttc agcttagtcc atttattcat
     3001 caaggttcgc cacagtggaa tgaaccgcca agttatccag caaatgtttt acgcagtgga
     3061 tgcccttcca gctgcaaccc atcattggga aacacccata cattctcatt cacgctcata
     3121 cactacagac aatttaaatt acccatttcc cctataacca gagaaaaccg gagaacctga
     3181 aggaaactca cgccaacatg gggagaacat gcaaactcca cacagaaaca ccaactgacc
     3241 cagccgggac tcaagccaac aaccttcttg ctgtgaggcg agcgcgctac ccactaagcc
     3301 actgagtcgc ggccatggta acgacatcta tagaaattac tctctgttgt gctgattcta
     3361 cagttgctat ggtaatacag cagcaaatct gttgtggtga ttctacagtt gctatggtaa
     3421 caacatttat agaaattgtt gttctgattg tatagttgct atatatggta atacagcagc
     3481 aaatcttttg tggtgaatct acagttgtca tggtaacaac atcgagaaat tactctattg
     3541 tgctgattct atagtttcta tggcaataca gcaaaactgt gttgtggtaa ttctattgtt
     3601 gctacggtaa cacaacatct attgtattat aaacaaatga gccaatactg tagttttcta
     3661 gaaatgtagt gtatattata ccacaataca ccagtttact gcagtacgcg gccgctggcc
     3721 gcaataaaat atctttattt tcattacatc tgtgtgttgg ttttttgtgt gaggatctaa
     3781 atgagtcttc ggacctcgcg ggggccgctt aagcggtggt tagggtttgt ctgacgcggg
     3841 gggaggggga aggaacgaaa cactctcatt cggaggcggc tcggggtttg gtcttggtgg
     3901 ccacgggcac gcagaagagc gccgcgatcc tcttaagcac ccccccgccc tccgtggagg
     3961 cgggggtttg gtcggcgggt ggtaactggc gggccgctga ctcgggcggg tcgcgcgccc
     4021 cagagtgtga ccttttcggt ctgctcgcag acccccgggc ggcgccgccg cggcggcgac
     4081 gggctcgctg ggtcctaggc tccatgggga ccgtatacgt ggacaggctc tggagcatcc
     4141 gcacgactgc ggtgatatta ccggagacct tctgcgggac gagccgggtc acgcggctga
     4201 cgcggagcgt ccgttgggcg acaaacacca ggacggggca caggtacact atcttgtcac
     4261 ccggaggcgc gagggactgc aggagcttca gggagtggcg cagctgcttc atccccgtgg
     4321 cccgttgctc gcgtttgctg gcggtgtccc cggaagaaat atatttgcat gtctttagtt
     4381 ctatgatgac acaaaccccg cccagcgtct tgtcattggc gaattcgaac acgcagatgc
     4441 agtcggggcg gcgcggtccc aggtccactt cgcatattaa ggtgacgcgt gtggcctcga
     4501 acaccgagcg accctgcagc gacccgctta aaagcttggc attccggtac tgttggtaaa
     4561 gccaccatgg ccgatgctaa gaacattaag aagggccctg ctcccttcta ccctctggag
     4621 gatggcaccg ctggcgagca gctgcacaag gccatgaaga ggtatgccct ggtgcctggc
     4681 accattgcct tcaccgatgc ccacattgag gtggacatca cctatgccga gtacttcgag
     4741 atgtctgtgc gcctggccga ggccatgaag aggtacggcc tgaacaccaa ccaccgcatc
     4801 gtggtgtgct ctgagaactc tctgcagttc ttcatgccag tgctgggcgc cctgttcatc
     4861 ggagtggccg tggcccctgc taacgacatt tacaacgagc gcgagctgct gaacagcatg
     4921 ggcatttctc agcctaccgt ggtgttcgtg tctaagaagg gcctgcagaa gatcctgaac
     4981 gtgcagaaga agctgcctat catccagaag atcatcatca tggactctaa gaccgactac
     5041 cagggcttcc agagcatgta cacattcgtg acatctcatc tgcctcctgg cttcaacgag
     5101 tacgacttcg tgccagagtc tttcgacagg gacaaaacca ttgccctgat catgaacagc
     5161 tctgggtcta ccggcctgcc taagggcgtg gccctgcctc atcgcaccgc ctgtgtgcgc
     5221 ttctctcacg cccgcgaccc tattttcggc aaccagatca tccccgacac cgctattctg
     5281 agcgtggtgc cattccacca cggcttcggc atgttcacca ccctgggcta cctgatttgc
     5341 ggctttcggg tggtgctgat gtaccgcttc gaggaggagc tgttcctgcg cagcctgcaa
     5401 gactacaaaa ttcagtctgc cctgctggtg ccaaccctgt tcagcttctt cgctaagagc
     5461 accctgatcg acaagtacga cctgtctaac ctgcacgaga ttgcctctgg cggcgcccca
     5521 ctgtctaagg aggtgggcga agccgtggcc aagcgctttc atctgccagg catccgccag
     5581 ggctacggcc tgaccgagac aaccagcgcc attctgatta ccccagaggg cgacgacaag
     5641 cctggcgccg tgggcaaggt ggtgccattc ttcgaggcca aggtggtgga cctggacacc
     5701 ggcaagaccc tgggagtgaa ccagcgcggc gagctgtgtg tgcgcggccc tatgattatg
     5761 tccggctacg tgaataaccc tgaggccaca aacgccctga tcgacaagga cggctggctg
     5821 cactctggcg acattgccta ctgggacgag gacgagcact tcttcatcgt ggaccgcctg
     5881 aagtctctga tcaagtacaa gggctaccag gtggccccag ccgagctgga gtctatcctg
     5941 ctgcagcacc ctaacatttt cgacgccgga gtggccggcc tgcccgacga cgatgccggc
     6001 gagctgcctg ccgccgtcgt cgtgctggaa cacggcaaga ccatgaccga gaaggagatc
     6061 gtggactatg tggccagcca ggtgacaacc gccaagaagc tgcgcggcgg agtggtgttc
     6121 gtggacgagg tgcccaaggg cctgaccggc aagctggacg cccgcaagat ccgcgagatc
     6181 ctgatcaagg ctaagaaagg cggcaagatc gccgtgtaat aattctagag tcggggcggc
     6241 cggccgcttc gagcagacat gataagatac attgatgagt ttggacaaac cacaactaga
     6301 atgcagtgaa aaaaatgctt tatttgtgaa atttgtgatg ctattgcttt atttgtaacc
     6361 attataagct gcaataaaca agttaacaac aacaattgca ttcattttat gtttcaggtt
     6421 cagggggagg tgtgggaggt tttttaaagc aagtaaaacc tctacaaatg tggtaaaatc
     6481 gataaggatc caggtggcac ttttcgggga aatgtgcgcg gaacccctat ttgtttattt
     6541 ttctaaatac attcaaatat gtatccgctc atgagacaat aaccctgata aatgcttcaa
     6601 taatattgaa aaaggaagag tatgagtatt caacatttcc gtgtcgccct tattcccttt
     6661 tttgcggcat tttgccttcc tgtttttgct cacccagaaa cgctggtgaa agtaaaagat
     6721 gctgaagatc agttgggtgc acgagtgggt tacatcgaac tggatctcaa cagcggtaag
     6781 atccttgaga gttttcgccc cgaagaacgt tttccaatga tgagcacttt taaagttctg
     6841 ctatgtggcg cggtattatc ccgtattgac gccgggcaag agcaactcgg tcgccgcata
     6901 cactattctc agaatgactt ggttgagtac tcaccagtca cagaaaagca tcttacggat
     6961 ggcatgacag taagagaatt atgcagtgct gccataacca tgagtgataa cactgcggcc
     7021 aacttacttc tgacaacgat cggaggaccg aaggagctaa ccgctttttt gcacaacatg
     7081 ggggatcatg taactcgcct tgatcgttgg gaaccggagc tgaatgaagc cataccaaac
     7141 gacgagcgtg acaccacgat gcctgtagca atggcaacaa cgttgcgcaa actattaact
     7201 ggcgaactac ttactctagc ttcccggcaa caattaatag actggatgga ggcggataaa
     7261 gttgcaggac cacttctgcg ctcggccctt ccggctggct ggtttattgc tgataaatct
     7321 ggagccggtg agcgtgggtc tcgcggtatc attgcagcac tggggccaga tggtaagccc
     7381 tcccgtatcg tagttatcta cacgacgggg agtcaggcaa ctatggatga acgaaataga
     7441 cagatcgctg agataggtgc ctcactgatt aagcattggt aactgtcaga ccaagtttac
     7501 tcatatatac tttagattga tttaaaactt catttttaat ttaaaaggat ctaggtgaag
     7561 atcctttttg ataatctcat gaccaaaatc ccttaacgtg agttttcgtt ccactgagcg
     7621 tcagaccccg tagaaa
//
