LOCUS       sequence-227752-        5704 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Fluorescent reporter for NF-kB activity.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5704)
  AUTHORS   Jutz S, Hennig A, Paster W, Asrak O, Dijanovic D, Kellner F, Pickl 
            WF, Huppa JB, Leitner J, Steinberger P
  TITLE     A cellular platform for the evaluation of immune checkpoint 
            molecules.
  JOURNAL   Oncotarget. 2017 May 4;8(39):64892-64906. doi: 
            10.18632/oncotarget.17615. eCollection 2017 Sep 12.
  PUBMED    29029399
REFERENCE   2  (bases 1 to 5704)
  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: 
            "10.18632/oncotarget.17615"; journalName: "Oncotarget"; date: 
            "2017-05-4- 4"; volume: "8"; issue: "39"; pages: "64892-64906"
COMMENT     Sequence Label: pSIRV-NF-kB-eGFP
FEATURES             Location/Qualifiers
     source          1..5704
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        73..376
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        377..580
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     531..551
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     misc_feature    819..1176
                     /label=MMLV Psi
                     /note="packaging signal of Moloney murine leukemia virus 
                     (MMLV)"
     CDS             1241..1657
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag 
                     gene lacking the start codon"
     primer_bind     1587..1609
                     /label=pLXSN 5'
                     /note="Murine stem cell virus, forward primer. Also called 
                     MSCV"
     primer_bind     1625..1641
                     /label=pBABE 5'
                     /note="Psi packaging signal, forward primer for pBABE 
                     vectors"
     misc_feature    1667..2041
                     /label=pol region
                     /note="Moloney murine leukemia virus (MMLV) pol region 
                     containing the splice acceptor site"
     primer_bind     1979..1995
                     /label=pBMN 5'
                     /note="MMLV sequence, for inserts in pBMN retroviral 
                     vector"
     CDS             join(2316..2318,2319..2321,2322..3035)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(2316..2318,2319..2321,2322..3032)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(2361..2382)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2622..2641)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2969..2990
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     protein_bind    3525..3541
                     /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(3549..3555,3556..3573,3574..3579))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3594..3615
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3732..3749)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3903..4491)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3983..4002)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4655..5446,5447..5515))
                     /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     5278..5297
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5516..5620)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 tgttctatgc cctagttatt aatagtaatc aattacgggg tcattagttc atagcccata
       61 tatggagttc cgcgttacat aacttacggt aaatggcccg cctggctgac cgcccaacga
      121 cccccgccca ttgacgtcaa taatgacgta tgttcccata gtaacgccaa tagggacttt
      181 ccattgacgt caatgggtgg agtatttacg gtaaactgcc cacttggcag tacatcaagt
      241 gtatcatatg ccaagtacgc cccctattga cgtcaatgac ggtaaatggc ccgcctggca
      301 ttatgcccag tacatgacct tatgggactt tcctacttgg cagtacatct acgtattagt
      361 catcgctatt accatggtga tgcggttttg gcagtacatc aatgggcgtg gatagcggtt
      421 tgactcacgg ggatttccaa gtctccaccc cattgacgtc aatgggagtt tgttttggca
      481 ccaaaatcaa cgggactttc caaaatgtcg taacaactcc gccccattga cgcaaatggg
      541 cggtaggcgt gtacggtggg aggtctatat aagcagagct caataaaaga gcccacaacc
      601 cctcactcgg cgcgccagtc ctccgattga ctgagtcgcc cgggtacccg tgtatccaat
      661 aaaccctctt gcagttgcat ccgacttgtg gtctcgctgt tccttgggag ggtctcctct
      721 gagtgattga ctacccgtca gcgggggtct ttcatttggg ggctcgtccg ggatcgggag
      781 acccctgccc agggaccacc gacccaccac cgggaggtaa gctggccagc aacttatctg
      841 tgtctgtccg attgtctagt gtctatgact gattttatgc gcctgcgtcg gtactagtta
      901 gctaactagc tctgtatctg gcggacccgt ggtggaactg acgagttcgg aacacccggc
      961 cgcaaccctg ggagacgtcc cagggacttc gggggccgtt tttgtggccc gacctgagtc
     1021 caaaaatccc gatcgttttg gactctttgg tgcacccccc ttagaggagg gatatgtggt
     1081 tctggtagga gacgagaacc taaaacagtt cccgcctccg tctgaatttt tgctttcggt
     1141 ttgggaccga agccgcgccg cgcgtcttgt ctgctgcagc atcgttctgt gttgtctctg
     1201 tctgactgtg tttctgtatt tgtctgaaaa tatgggcccg ggccagactg ttaccactcc
     1261 cttaagtttg accttaggtc actggaaaga tgtcgagcgg atcgctcaca accagtcggt
     1321 agatgtcaag aagagacgtt gggttacctt ctgctctgca gaatggccaa cctttaacgt
     1381 cggatggccg cgagacggca cctttaaccg agacctcatc acccaggtta agatcaaggt
     1441 cttttcacct ggcccgcatg gacacccaga ccaggtcccc tacatcgtga cctgggaagc
     1501 cttggctttt gacccccctc cctgggtcaa gccctttgta caccctaagc ctccgcctcc
     1561 tcttcctcca tccgccccgt ctctccccct tgaacctcct cgttcgaccc cgcctcgatc
     1621 ctccctttat ccagccctca ctccttctct aggcgccccc atatggccat atgagatctt
     1681 atatggggca cccccgcccc ttgtaaactt ccctgaccct gacatgacaa gagttactaa
     1741 cagcccctct ctccaagctc acttacaggc tctctactta gtccagcacg aagtctggag
     1801 acctctggcg gcagcctacc aagaacaact ggaccgaccg gtggtacctc acccttaccg
     1861 agtcggcgac acagtgtggg tccgccgaca ccagactaag aacctagaac ctcgctggaa
     1921 aggaccttac acagtcctgc tgaccacccc caccgccctc aaagtagacg gcatcgcagc
     1981 ttggatacac gccgcccacg tgaaggctgc cgaccccggg ggtggaccat cctctagact
     2041 gccggatccg gggactttcc gcttggggac tttccgctgg ggactttccg ctggggactt
     2101 tccgctgggg actttccgaa ttcagcttga ataaaatgaa tattagaagc tgttagaata
     2161 agagaaaatg acagaggaaa actgaaaggg agaactgaaa gtgggaaatt cctctgaggc
     2221 agaaaggacc atcccttata aatagcacag gccatgaagg aagatcattc tcactgcagc
     2281 ctttgacagc ctttgcctca tcttgaagct tcaccatggt gagcaagggc gaggagctgt
     2341 tcaccggggt ggtgcccatc ctggtcgagc tggacggcga cgtaaacggc cacaagttca
     2401 gcgtgtccgg cgagggcgag ggcgatgcca cctacggcaa gctgaccctg aagttcatct
     2461 gcaccaccgg caagctgccc gtgccctggc ccaccctcgt gaccaccctg acctacggcg
     2521 tgcagtgctt cagccgctac cccgaccaca tgaagcagca cgacttcttc aagtccgcca
     2581 tgcccgaagg ctacgtccag gagcgcacca tcttcttcaa ggacgacggc aactacaaga
     2641 cccgcgccga ggtgaagttc gagggcgaca ccctggtgaa ccgcatcgag ctgaagggca
     2701 tcgacttcaa ggaggacggc aacatcctgg ggcacaagct ggagtacaac tacaacagcc
     2761 acaacgtcta tatcatggcc gacaagcaga agaacggcat caaggtgaac ttcaagatcc
     2821 gccacaacat cgaggacggc agcgtgcagc tcgccgacca ctaccagcag aacaccccca
     2881 tcggcgacgg ccccgtgctg ctgcccgaca accactacct gagcacccag tccgccctga
     2941 gcaaagaccc caacgagaag cgcgatcaca tggtcctgct ggagttcgtg accgccgccg
     3001 ggatcactct cggcatggac gagctgtaca agtaaagcgg ccgccagcac agtggtcgat
     3061 cgacgataaa ataaaagatt ttatttagtc tccagaaaaa ggggggaatg aaagacccca
     3121 cctgtaggtt tggcaagcta gcttaagtaa cgccattttg caaggcatgg aaaaatacat
     3181 aactgagaat agaaaagttc agatcaaggt caggaacaga tggaacaggg tcgcgtcccg
     3241 caataaaaga gcccacaacc cctcactcgg ggcgccagtc ctccgattga ctgagtcgcc
     3301 cgggtacccg tgtatccaat aaaccctctt gcagttgcat ccgacttgtg gtctcgctgt
     3361 tccttgggag ggtctcctct gagtgattga ctacccgtca gcgggggtct ttcacatgca
     3421 gcatgtatca aaattaattt ggtttttttt cttaagtatt tacattaaat ggccatagta
     3481 gttcattatg gacagcgcag aaagagctgg ggagaattgt gaaattgtta tccgctcaca
     3541 attccacaca acatacgagc cggaagcata aagtgtaaag cctggggtgc ctaatgagtg
     3601 agctaactca cattaattgc gttgcgctca ctgcccgctt tccagtcggg aaacctgtcg
     3661 tgccagctgc attaatgaat cggccaacgc gcggggagag gcggtttgcg tattgggcgc
     3721 tcttccgctt cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta
     3781 tcagctcact caaaggcggt aatacggtta tccacagaat caggggataa cgcaggaaag
     3841 aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg
     3901 tttttccata ggctccgccc ccctgacgag catcacaaaa atcgacgctc aagtcagagg
     3961 tggcgaaacc cgacaggact ataaagatac caggcgtttc cccctggaag ctccctcgtg
     4021 cgctctcctg ttccgaccct gccgcttacc ggatacctgt ccgcctttct cccttcggga
     4081 agcgtggcgc tttctcatag ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc
     4141 tccaagctgg gctgtgtgca cgaacccccc gttcagcccg accgctgcgc cttatccggt
     4201 aactatcgtc ttgagtccaa cccggtaaga cacgacttat cgccactggc agcagccact
     4261 ggtaacagga ttagcagagc gaggtatgta ggcggtgcta cagagttctt gaagtggtgg
     4321 cctaactacg gctacactag aagaacagta tttggtatct gcgctctgct gaagccagtt
     4381 accttcggaa aaagagttgg tagctcttga tccggcaaac aaaccaccgc tggtagcggt
     4441 ggtttttttg tttgcaagca gcagattacg cgcagaaaaa aaggatctca agaagatcct
     4501 ttgatctttt ctacggggtc tgacgctcag tggaacgaaa actcacgtta agggattttg
     4561 gtcatgagat tatcaaaaag gatcttcacc tagatccttt tgcggccggc cgcaaatcaa
     4621 tctaaagtat atatgagtaa acttggtctg acagttacca atgcttaatc agtgaggcac
     4681 ctatctcagc gatctgtcta tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga
     4741 taactacgat acgggagggc ttaccatctg gccccagtgc tgcaatgata ccgcgagacc
     4801 cacgctcacc ggctccagat ttatcagcaa taaaccagcc agccggaagg gccgagcgca
     4861 gaagtggtcc tgcaacttta tccgcctcca tccagtctat taattgttgc cgggaagcta
     4921 gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt tgccattgct acaggcatcg
     4981 tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc cggttcccaa cgatcaaggc
     5041 gagttacatg atcccccatg ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg
     5101 ttgtcagaag taagttggcc gcagtgttat cactcatggt tatggcagca ctgcataatt
     5161 ctcttactgt catgccatcc gtaagatgct tttctgtgac tggtgagtac tcaaccaagt
     5221 cattctgaga atagtgtatg cggcgaccga gttgctcttg cccggcgtca atacgggata
     5281 ataccgcgcc acatagcaga actttaaaag tgctcatcat tggaaaacgt tcttcggggc
     5341 gaaaactctc aaggatctta ccgctgttga gatccagttc gatgtaaccc actcgtgcac
     5401 ccaactgatc ttcagcatct tttactttca ccagcgtttc tgggtgagca aaaacaggaa
     5461 ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa atgttgaata ctcatactct
     5521 tcctttttca atattattga agcatttatc agggttattg tctcatgagc ggatacatat
     5581 ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg cacatttccc cgaaaagtgc
     5641 caccagcttt gctcttagga gtttcctaat acatcccaaa ctcaaatata taaagcattt
     5701 gact
//
