LOCUS       40924_1404        5863 bp DNA     circular SYN 13-MAY-2021
DEFINITION  Reporter construct for NF‐kappa-B activation. Contains two copies of
            immunoglobulin kappa light chain motif (5′‐GGGGACTTTCC‐3′) upstream 
            of the a minimal promoter driving luciferase expression.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5863)
  AUTHORS   Pomerantz JL, Denny EM, Baltimore D
  TITLE     CARD11 mediates factor-specific activation of NF-kappaB by the T 
            cell receptor complex.
  JOURNAL   EMBO J. 2002 Oct 1. 21(19):5184-94.
  PUBMED    12356734
REFERENCE   2  (bases 1 to 5863)
  TITLE     Direct Submission
REFERENCE   3  (bases 1 to 5863)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "EMBO J. 
            2002 Oct 1. 21(19):5184-94."
COMMENT     SGRef: number: 2; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..5863
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        complement(33..51)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(72..88)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    complement(96..112)
                     /label=lac operator
                     /note="The lac repressor binds to the lac operator to
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(120..150)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    complement(165..186)
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     primer_bind     complement(303..320)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(474..1062)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(1236..2093)
                     /codon_start=1
                     /label=AmpR
                     /note="beta-lactamase"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     promoter        complement(2094..2198)
                     /label=AmpR promoter
     rep_origin      complement(2224..2679)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     primer_bind     2821..2837
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        2844..2862
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     polyA_signal    2872..3006
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     CDS             3210..4859
                     /codon_start=1
                     /label=luciferase
                     /note="firefly luciferase"
                     /translation="MEDAKNIKKGPAPFYPLEDGTAGEQLHKAMKRYALVPGTIAFTDA
                     HIEVNITYAEYFEMSVRLAEAMKRYGLNTNHRIVVCSENSLQFFMPVLGALFIGVAVAP
                     ANDIYNERELLNSMNISQPTVVFVSKKGLQKILNVQKKLPIIQKIIIMDSKTDYQGFQS
                     MYTFVTSHLPPGFNEYDFVPESFDRDKTIALIMNSSGSTGLPKGVALPHRTACVRFSHA
                     RDPIFGNQIIPDTAILSVVPFHHGFGMFTTLGYLICGFRVVLMYRFEEELFLRSLQDYK
                     IQSALLVPTLFSFFAKSTLIDKYDLSNLHEIASGGAPLSKEVGEAVAKRFHLPGIRQGY
                     GLTETTSAILITPEGDDKPGAVGKVVPFFEAKVVDLDTGKTLGVNQRGELCVRGPMIMS
                     GYVNNPEATNALIDKDGWLHSGDIAYWDEDEHFFIVDRLKSLIKYKGYQVAPAELESIL
                     LQHPNIFDAGVAGLPDDDAGELPAAVVVLEHGKTMTEKEIVDYVASQVTTAKKLRGGVV
                     FVDEVPKGLTGKLDARKIREILIKAKKGGKSKL"
     intron          5086..5151
                     /label=small t intron
                     /note="SV40 (simian virus 40) small t antigen intron"
     CDS             5281..5301
                     /codon_start=1
                     /label=SV40 NLS
                     /note="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /translation="PKKKRKV"
     polyA_signal    5726..5860
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
ORIGIN
        1 ctcgaggggg ggcccggtac cagcttttgt tccctttagt gagggttaat ttcgagcttg
       61 gcgtaatcat ggtcatagct gtttcctgtg tgaaattgtt atccgctcac aattccacac
      121 aacatacgag ccggaagcat aaagtgtaaa gcctggggtg cctaatgagt gagctaactc
      181 acattaattg cgttgcgctc actgcccgct ttccagtcgg gaaacctgtc gtgccagctg
      241 cattaatgaa tcggccaacg cgcggggaga ggcggtttgc gtattgggcg ctcttccgct
      301 tcctcgctca ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt atcagctcac
      361 tcaaaggcgg taatacggtt atccacagaa tcaggggata acgcaggaaa gaacatgtga
      421 gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc gtttttccat
      481 aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag gtggcgaaac
      541 ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt gcgctctcct
      601 gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg aagcgtggcg
      661 ctttctcata gctcacgctg taggtatctc agttcggtgt aggtcgttcg ctccaagctg
      721 ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg taactatcgt
      781 cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac tggtaacagg
      841 attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg gcctaactac
      901 ggctacacta gaagaacagt atttggtatc tgcgctctgc tgaagccagt taccttcgga
      961 aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg tggttttttt
     1021 gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt
     1081 tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt ggtcatgaga
     1141 ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa aatgaagttt taaatcaatc
     1201 taaagtatat atgagtaaac ttggtctgac agttaccaat gcttaatcag tgaggcacct
     1261 atctcagcga tctgtctatt tcgttcatcc atagttgcct gactccccgt cgtgtagata
     1321 actacgatac gggagggctt accatctggc cccagtgctg caatgatacc gcgagaccca
     1381 cgctcaccgg ctccagattt atcagcaata aaccagccag ccggaagggc cgagcgcaga
     1441 agtggtcctg caactttatc cgcctccatc cagtctatta attgttgccg ggaagctaga
     1501 gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg ccattgctac aggcatcgtg
     1561 gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg gttcccaacg atcaaggcga
     1621 gttacatgat cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt
     1681 gtcagaagta agttggccgc agtgttatca ctcatggtta tggcagcact gcataattct
     1741 cttactgtca tgccatccgt aagatgcttt tctgtgactg gtgagtactc aaccaagtca
     1801 ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat acgggataat
     1861 accgcgccac atagcagaac tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga
     1921 aaactctcaa ggatcttacc gctgttgaga tccagttcga tgtaacccac tcgtgcaccc
     1981 aactgatctt cagcatcttt tactttcacc agcgtttctg ggtgagcaaa aacaggaagg
     2041 caaaatgccg caaaaaaggg aataagggcg acacggaaat gttgaatact catactcttc
     2101 ctttttcaat attattgaag catttatcag ggttattgtc tcatgagcgg atacatattt
     2161 gaatgtattt agaaaaataa acaaataggg gttccgcgca catttccccg aaaagtgcca
     2221 cctaaattgt aagcgttaat attttgttaa aattcgcgtt aaatttttgt taaatcagct
     2281 cattttttaa ccaataggcc gaaatcggca aaatccctta taaatcaaaa gaatagaccg
     2341 agatagggtt gagtgttgtt ccagtttgga acaagagtcc actattaaag aacgtggact
     2401 ccaacgtcaa agggcgaaaa accgtctatc agggcgatgg cccactacgt gaaccatcac
     2461 cctaatcaag ttttttgggg tcgaggtgcc gtaaagcact aaatcggaac cctaaaggga
     2521 gcccccgatt tagagcttga cggggaaagc cggcgaacgt ggcgagaaag gaagggaaga
     2581 aagcgaaagg agcgggcgct agggcgctgg caagtgtagc ggtcacgctg cgcgtaacca
     2641 ccacacccgc cgcgcttaat gcgccgctac agggcgcgtc ccattcgcca ttcaggctgc
     2701 gcaactgttg ggaagggcga tcggtgcggg cctcttcgct attacgccag ctggcgaaag
     2761 ggggatgtgc tgcaaggcga ttaagttggg taacgccagg gttttcccag tcacgacgtt
     2821 gtaaaacgac ggccagtgaa ttgtaatacg actcactata gggcgaattg gaacttgttt
     2881 attgcagctt ataatggtta caaataaagc aatagcatca caaatttcac aaataaagca
     2941 tttttttcac tgcattctag ttgtggtttg tccaaactca tcaatgtatc ttatcatgtc
     3001 tggatcgatc cccccaccgc ggtggcggcc cacagagggg actttccgag agatctacag
     3061 aggggacttt ccgagagcga gcttgggctg caggtcgaga tccgctgaat agagagagga
     3121 ccatctcata taaataggcc atacccatgg agaaaggaca ttctaactgc aacctttcgc
     3181 aagcttggca ttccggtact gttggtaaaa tggaagacgc caaaaacata aagaaaggcc
     3241 cggcgccatt ctatcctcta gaggatggaa ccgctggaga gcaactgcat aaggctatga
     3301 agagatacgc cctggttcct ggaacaattg cttttacaga tgcacatatc gaggtgaaca
     3361 tcacgtacgc ggaatacttc gaaatgtccg ttcggttggc agaagctatg aaacgatatg
     3421 ggctgaatac aaatcacaga atcgtcgtat gcagtgaaaa ctctcttcaa ttctttatgc
     3481 cggtgttggg cgcgttattt atcggagttg cagttgcgcc cgcgaacgac atttataatg
     3541 aacgtgaatt gctcaacagt atgaacattt cgcagcctac cgtagtgttt gtttccaaaa
     3601 aggggttgca aaaaattttg aacgtgcaaa aaaaattacc aataatccag aaaattatta
     3661 tcatggattc taaaacggat taccagggat ttcagtcgat gtacacgttc gtcacatctc
     3721 atctacctcc cggttttaat gaatacgatt ttgtaccaga gtcctttgat cgtgacaaaa
     3781 caattgcact gataatgaat tcctctggat ctactgggtt acctaagggt gtggcccttc
     3841 cgcatagaac tgcctgcgtc agattctcgc atgccagaga tcctattttt ggcaatcaaa
     3901 tcattccgga tactgcgatt ttaagtgttg ttccattcca tcacggtttt ggaatgttta
     3961 ctacactcgg atatttgata tgtggatttc gagtcgtctt aatgtataga tttgaagaag
     4021 agctgttttt acgatccctt caggattaca aaattcaaag tgcgttgcta gtaccaaccc
     4081 tattttcatt cttcgccaaa agcactctga ttgacaaata cgatttatct aatttacacg
     4141 aaattgcttc tgggggcgca cctctttcga aagaagtcgg ggaagcggtt gcaaaacgct
     4201 tccatcttcc agggatacga caaggatatg ggctcactga gactacatca gctattctga
     4261 ttacacccga gggggatgat aaaccgggcg cggtcggtaa agttgttcca ttttttgaag
     4321 cgaaggttgt ggatctggat accgggaaaa cgctgggcgt taatcagaga ggcgaattat
     4381 gtgtcagagg acctatgatt atgtccggtt atgtaaacaa tccggaagcg accaacgcct
     4441 tgattgacaa ggatggatgg ctacattctg gagacatagc ttactgggac gaagacgaac
     4501 acttcttcat agttgaccgc ttgaagtctt taattaaata caaaggatat caggtggccc
     4561 ccgctgaatt ggaatcgata ttgttacaac accccaacat cttcgacgcg ggcgtggcag
     4621 gtcttcccga cgatgacgcc ggtgaacttc ccgccgccgt tgttgttttg gagcacggaa
     4681 agacgatgac ggaaaaagag atcgtggatt acgtcgccag tcaagtaaca accgcgaaaa
     4741 agttgcgcgg aggagttgtg tttgtggacg aagtaccgaa aggtcttacc ggaaaactcg
     4801 acgcaagaaa aatcagagag atcctcataa aggccaagaa gggcggaaag tccaaattgt
     4861 aaaatgtaac tgtattcagc gatgacgaaa ttcttagcta ttgtaatatt atatgcaaat
     4921 tgatgaatgg taattttgta attgtgggtc actgtactat tttaacgaat aataaaatca
     4981 ggtataggta actaaaaagg aattcggggg atctttgtga aggaacctta cttctgtggt
     5041 gtgacataat tggacaaact acctacagag atttaaagct ctaaggtaaa tataaaattt
     5101 ttaagtgtat aatgtgttaa actactgatt ctaattgttt gtgtatttta gattccaacc
     5161 tatggaactg atgaatggga gcagtggtgg aatgccttta atgaggaaaa cctgttttgc
     5221 tcagaagaaa tgccatctag tgatgatgag gctactgctg actctcaaca ttctactcct
     5281 ccaaaaaaga agagaaaggt agaagacccc aaggactttc cttcagaatt gctaagtttt
     5341 ttgagtcatg ctgtgtttag taatagaact cttgcttgct ttgctattta caccacaaag
     5401 gaaaaagctg cactgctata caagaaaatt atggaaaaat attctgtaac ctttataagt
     5461 aggcataaca gttataatca taacatactg ttttttctta ctccacacag gcatagagtg
     5521 tctgctatta ataactatgc tcaaaaattg tgtaccttta gctttttaat ttgtaaaggg
     5581 gttaataagg aatatttgat gtatagtgcc ttgactagag atcataatca gccataccac
     5641 atttgtagag gttttacttg ctttaaaaaa cctcccacac ctccccctga acctgaaaca
     5701 taaaatgaat gcaattgttg ttgttaactt gtttattgca gcttataatg gttacaaata
     5761 aagcaatagc atcacaaatt tcacaaataa agcatttttt tcactgcatt ctagttgtgg
     5821 tttgtccaaa ctcatcaatg tatcttatca tgtctggatc gac
//
