LOCUS       sequence-399622-        5703 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Flourescent reporter for NF-kB activity.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5703)
  AUTHORS   Jutz S, Leitner J, Schmetterer K, Doel-Perez I, Majdic O, 
            Grabmeier-Pfistershammer K, Paster W, Huppa JB, Steinberger P
  TITLE     Assessment of costimulation and coinhibition in a triple parameter T
            cell reporter line: Simultaneous measurement of NF-kappaB, NFAT and 
            AP-1.
  JOURNAL   J Immunol Methods. 2016 Jan 15. pii: S0022-1759(16)30007-2. doi: 
            10.1016/j.jim.2016.01.007.
  PUBMED    26780292
REFERENCE   2  (bases 1 to 5703)
  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"; journalName: "J Immunol 
            Methods. 2016 Jan 15. pii: S0022-1759(16)30007-2. doi: 
            10.1016/j.jim.2016.01.007."
COMMENT     Sequence Label: pSIRV-NF-kB-eCFP
FEATURES             Location/Qualifiers
     source          1..5703
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        87..390
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        391..594
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     545..565
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     misc_feature    833..1190
                     /label=MMLV Psi
                     /note="packaging signal of Moloney murine leukemia virus 
                     (MMLV)"
     CDS             1255..1671
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag 
                     gene lacking the start codon"
     primer_bind     1601..1623
                     /label=pLXSN 5'
                     /note="Murine stem cell virus, forward primer. Also called 
                     MSCV"
     primer_bind     1639..1655
                     /label=pBABE 5'
                     /note="Psi packaging signal, forward primer for pBABE 
                     vectors"
     misc_feature    1681..2055
                     /label=pol region
                     /note="Moloney murine leukemia virus (MMLV) pol region 
                     containing the splice acceptor site"
     primer_bind     1993..2009
                     /label=pBMN 5'
                     /note="MMLV sequence, for inserts in pBMN retroviral 
                     vector"
     CDS             join(2330..2332,2333..2335,2336..3049)
                     /codon_start=1
                     /product="enhanced CFP"
                     /label=ECFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTWGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYISHNVYITADKQKNGIK
                     ANFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(2375..2396)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2636..2655)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2983..3004
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     protein_bind    3538..3554
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /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(join(3562..3568,3569..3586,3587..3592))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3607..3628
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3745..3762)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3916..4504)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3996..4015)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4668..5459,5460..5528))
                     /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     5291..5310
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5529..5633)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 taaagcattt gacttgttct atgccctagt tattaatagt aatcaattac ggggtcatta
       61 gttcatagcc catatatgga gttccgcgtt acataactta cggtaaatgg cccgcctggc
      121 tgaccgccca acgacccccg cccattgacg tcaataatga cgtatgttcc catagtaacg
      181 ccaataggga ctttccattg acgtcaatgg gtggagtatt tacggtaaac tgcccacttg
      241 gcagtacatc aagtgtatca tatgccaagt acgcccccta ttgacgtcaa tgacggtaaa
      301 tggcccgcct ggcattatgc ccagtacatg accttatggg actttcctac ttggcagtac
      361 atctacgtat tagtcatcgc tattaccatg gtgatgcggt tttggcagta catcaatggg
      421 cgtggatagc ggtttgactc acggggattt ccaagtctcc accccattga cgtcaatggg
      481 agtttgtttt ggcaccaaaa tcaacgggac tttccaaaat gtcgtaacaa ctccgcccca
      541 ttgacgcaaa tgggcggtag gcgtgtacgg tgggaggtct atataagcag agctcaataa
      601 aagagcccac aacccctcac tcggcgcgcc agtcctccga ttgactgagt cgcccgggta
      661 cccgtgtatc caataaaccc tcttgcagtt gcatccgact tgtggtctcg ctgttccttg
      721 ggagggtctc ctctgagtga ttgactaccc gtcagcgggg gtctttcatt tgggggctcg
      781 tccgggatcg ggagacccct gcccagggac caccgaccca ccaccgggag gtaagctggc
      841 cagcaactta tctgtgtctg tccgattgtc tagtgtctat gactgatttt atgcgcctgc
      901 gtcggtacta gttagctaac tagctctgta tctggcggac ccgtggtgga actgacgagt
      961 tcggaacacc cggccgcaac cctgggagac gtcccaggga cttcgggggc cgtttttgtg
     1021 gcccgacctg agtccaaaaa tcccgatcgt tttggactct ttggtgcacc ccccttagag
     1081 gagggatatg tggttctggt aggagacgag aacctaaaac agttcccgcc tccgtctgaa
     1141 tttttgcttt cggtttggga ccgaagccgc gccgcgcgtc ttgtctgctg cagcatcgtt
     1201 ctgtgttgtc tctgtctgac tgtgtttctg tatttgtctg aaaatatggg cccgggccag
     1261 actgttacca ctcccttaag tttgacctta ggtcactgga aagatgtcga gcggatcgct
     1321 cacaaccagt cggtagatgt caagaagaga cgttgggtta ccttctgctc tgcagaatgg
     1381 ccaaccttta acgtcggatg gccgcgagac ggcaccttta accgagacct catcacccag
     1441 gttaagatca aggtcttttc acctggcccg catggacacc cagaccaggt cccctacatc
     1501 gtgacctggg aagccttggc ttttgacccc cctccctggg tcaagccctt tgtacaccct
     1561 aagcctccgc ctcctcttcc tccatccgcc ccgtctctcc cccttgaacc tcctcgttcg
     1621 accccgcctc gatcctccct ttatccagcc ctcactcctt ctctaggcgc ccccatatgg
     1681 ccatatgaga tcttatatgg ggcacccccg ccccttgtaa acttccctga ccctgacatg
     1741 acaagagtta ctaacagccc ctctctccaa gctcacttac aggctctcta cttagtccag
     1801 cacgaagtct ggagacctct ggcggcagcc taccaagaac aactggaccg accggtggta
     1861 cctcaccctt accgagtcgg cgacacagtg tgggtccgcc gacaccagac taagaaccta
     1921 gaacctcgct ggaaaggacc ttacacagtc ctgctgacca cccccaccgc cctcaaagta
     1981 gacggcatcg cagcttggat acacgccgcc cacgtgaagg ctgccgaccc cgggggtgga
     2041 ccatcctcta gactgccgga tccggggact ttccgcttgg ggactttccg ctggggactt
     2101 tccgctgggg actttccgct ggggactttc cgaattcagc ttgaataaaa tgaatattag
     2161 aagctgttag aataagagaa aatgacagag gaaaactgaa agggagaact gaaagtggga
     2221 aattcctctg aggcagaaag gaccatccct tataaatagc acaggccatg aaggaagatc
     2281 attctcactg cagcctttga cagcctttgc ctcatcttga agcttcacca tggtgagcaa
     2341 gggcgaggag ctgttcaccg gggtggtgcc catcctggtc gagctggacg gcgacgtaaa
     2401 cggccacaag ttcagcgtgt ccggcgaggg cgagggcgat gccacctacg gcaagctgac
     2461 cctgaagttc atctgcacca ccggcaagct gcccgtgccc tggcccaccc tcgtgaccac
     2521 cctgacctgg ggcgtgcagt gcttcagccg ctaccccgac cacatgaagc agcacgactt
     2581 cttcaagtcc gccatgcccg aaggctacgt ccaggagcgc accatcttct tcaaggacga
     2641 cggcaactac aagacccgcg ccgaggtgaa gttcgagggc gacaccctgg tgaaccgcat
     2701 cgagctgaag ggcatcgact tcaaggagga cggcaacatc ctggggcaca agctggagta
     2761 caactacatc agccacaacg tctatatcac cgccgacaag cagaagaacg gcatcaaggc
     2821 caacttcaag atccgccaca acatcgagga cggcagcgtg cagctcgccg accactacca
     2881 gcagaacacc cccatcggcg acggccccgt gctgctgccc gacaaccact acctgagcac
     2941 ccagtccgcc ctgagcaaag accccaacga gaagcgcgat cacatggtcc tgctggagtt
     3001 cgtgaccgcc gccgggatca ctctcggcat ggacgagctg tacaagtaag cggccgccag
     3061 cacagtggtc gatcgacgat aaaataaaag attttattta gtctccagaa aaagggggga
     3121 atgaaagacc ccacctgtag gtttggcaag ctagcttaag taacgccatt ttgcaaggca
     3181 tggaaaaata cataactgag aatagaaaag ttcagatcaa ggtcaggaac agatggaaca
     3241 gggtcgcgtc ccgcaataaa agagcccaca acccctcact cggggcgcca gtcctccgat
     3301 tgactgagtc gcccgggtac ccgtgtatcc aataaaccct cttgcagttg catccgactt
     3361 gtggtctcgc tgttccttgg gagggtctcc tctgagtgat tgactacccg tcagcggggg
     3421 tctttcacat gcagcatgta tcaaaattaa tttggttttt tttcttaagt atttacatta
     3481 aatggccata gtagttcatt atggacagcg cagaaagagc tggggagaat tgtgaaattg
     3541 ttatccgctc acaattccac acaacatacg agccggaagc ataaagtgta aagcctgggg
     3601 tgcctaatga gtgagctaac tcacattaat tgcgttgcgc tcactgcccg ctttccagtc
     3661 gggaaacctg tcgtgccagc tgcattaatg aatcggccaa cgcgcgggga gaggcggttt
     3721 gcgtattggg cgctcttccg cttcctcgct cactgactcg ctgcgctcgg tcgttcggct
     3781 gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg ttatccacag aatcagggga
     3841 taacgcagga aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc
     3901 cgcgttgctg gcgtttttcc ataggctccg cccccctgac gagcatcaca aaaatcgacg
     3961 ctcaagtcag aggtggcgaa acccgacagg actataaaga taccaggcgt ttccccctgg
     4021 aagctccctc gtgcgctctc ctgttccgac cctgccgctt accggatacc tgtccgcctt
     4081 tctcccttcg ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc tcagttcggt
     4141 gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg
     4201 cgccttatcc ggtaactatc gtcttgagtc caacccggta agacacgact tatcgccact
     4261 ggcagcagcc actggtaaca ggattagcag agcgaggtat gtaggcggtg ctacagagtt
     4321 cttgaagtgg tggcctaact acggctacac tagaagaaca gtatttggta tctgcgctct
     4381 gctgaagcca gttaccttcg gaaaaagagt tggtagctct tgatccggca aacaaaccac
     4441 cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc
     4501 tcaagaagat cctttgatct tttctacggg gtctgacgct cagtggaacg aaaactcacg
     4561 ttaagggatt ttggtcatga gattatcaaa aaggatcttc acctagatcc ttttgcggcc
     4621 ggccgcaaat caatctaaag tatatatgag taaacttggt ctgacagtta ccaatgctta
     4681 atcagtgagg cacctatctc agcgatctgt ctatttcgtt catccatagt tgcctgactc
     4741 cccgtcgtgt agataactac gatacgggag ggcttaccat ctggccccag tgctgcaatg
     4801 ataccgcgag acccacgctc accggctcca gatttatcag caataaacca gccagccgga
     4861 agggccgagc gcagaagtgg tcctgcaact ttatccgcct ccatccagtc tattaattgt
     4921 tgccgggaag ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt
     4981 gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag ctccggttcc
     5041 caacgatcaa ggcgagttac atgatccccc atgttgtgca aaaaagcggt tagctccttc
     5101 ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt tatcactcat ggttatggca
     5161 gcactgcata attctcttac tgtcatgcca tccgtaagat gcttttctgt gactggtgag
     5221 tactcaacca agtcattctg agaatagtgt atgcggcgac cgagttgctc ttgcccggcg
     5281 tcaatacggg ataataccgc gccacatagc agaactttaa aagtgctcat cattggaaaa
     5341 cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa
     5401 cccactcgtg cacccaactg atcttcagca tcttttactt tcaccagcgt ttctgggtga
     5461 gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg gaaatgttga
     5521 atactcatac tcttcctttt tcaatattat tgaagcattt atcagggtta ttgtctcatg
     5581 agcggataca tatttgaatg tatttagaaa aataaacaaa taggggttcc gcgcacattt
     5641 ccccgaaaag tgccaccagc tttgctctta ggagtttcct aatacatccc aaactcaaat
     5701 ata
//
