LOCUS       sequence-227759-        5690 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Fluorescent reporter for AP-1 activation.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5690)
  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 5690)
  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-AP-1-mCherry
FEATURES             Location/Qualifiers
     source          1..5690
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        37..340
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        341..544
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     495..515
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     misc_feature    783..1140
                     /label=MMLV Psi
                     /note="packaging signal of Moloney murine leukemia virus 
                     (MMLV)"
     CDS             1205..1621
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag 
                     gene lacking the start codon"
     primer_bind     1551..1573
                     /label=pLXSN 5'
                     /note="Murine stem cell virus, forward primer. Also called 
                     MSCV"
     primer_bind     1589..1605
                     /label=pBABE 5'
                     /note="Psi packaging signal, forward primer for pBABE 
                     vectors"
     misc_feature    1631..2005
                     /label=pol region
                     /note="Moloney murine leukemia virus (MMLV) pol region 
                     containing the splice acceptor site"
     primer_bind     1943..1959
                     /label=pBMN 5'
                     /note="MMLV sequence, for inserts in pBMN retroviral 
                     vector"
     CDS             2276..2986
                     /codon_start=1
                     /product="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
                     /label=mCherry
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEG
                     TQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNF
                     EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK
                     GEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERA
                     EGRHSTGGMDELYK"
     primer_bind     complement(2421..2439)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     2689..2708
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     protein_bind    3475..3491
                     /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(3499..3505,3506..3523,3524..3529))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3544..3565
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3682..3699)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3853..4441)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3933..3952)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4605..5396,5397..5465))
                     /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     5228..5247
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5466..5570)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 ggggtcatta gttcatagcc catatatgga gttccgcgtt acataactta cggtaaatgg
       61 cccgcctggc tgaccgccca acgacccccg cccattgacg tcaataatga cgtatgttcc
      121 catagtaacg ccaataggga ctttccattg acgtcaatgg gtggagtatt tacggtaaac
      181 tgcccacttg gcagtacatc aagtgtatca tatgccaagt acgcccccta ttgacgtcaa
      241 tgacggtaaa tggcccgcct ggcattatgc ccagtacatg accttatggg actttcctac
      301 ttggcagtac atctacgtat tagtcatcgc tattaccatg gtgatgcggt tttggcagta
      361 catcaatggg cgtggatagc ggtttgactc acggggattt ccaagtctcc accccattga
      421 cgtcaatggg agtttgtttt ggcaccaaaa tcaacgggac tttccaaaat gtcgtaacaa
      481 ctccgcccca ttgacgcaaa tgggcggtag gcgtgtacgg tgggaggtct atataagcag
      541 agctcaataa aagagcccac aacccctcac tcggcgcgcc agtcctccga ttgactgagt
      601 cgcccgggta cccgtgtatc caataaaccc tcttgcagtt gcatccgact tgtggtctcg
      661 ctgttccttg ggagggtctc ctctgagtga ttgactaccc gtcagcgggg gtctttcatt
      721 tgggggctcg tccgggatcg ggagacccct gcccagggac caccgaccca ccaccgggag
      781 gtaagctggc cagcaactta tctgtgtctg tccgattgtc tagtgtctat gactgatttt
      841 atgcgcctgc gtcggtacta gttagctaac tagctctgta tctggcggac ccgtggtgga
      901 actgacgagt tcggaacacc cggccgcaac cctgggagac gtcccaggga cttcgggggc
      961 cgtttttgtg gcccgacctg agtccaaaaa tcccgatcgt tttggactct ttggtgcacc
     1021 ccccttagag gagggatatg tggttctggt aggagacgag aacctaaaac agttcccgcc
     1081 tccgtctgaa tttttgcttt cggtttggga ccgaagccgc gccgcgcgtc ttgtctgctg
     1141 cagcatcgtt ctgtgttgtc tctgtctgac tgtgtttctg tatttgtctg aaaatatggg
     1201 cccgggccag actgttacca ctcccttaag tttgacctta ggtcactgga aagatgtcga
     1261 gcggatcgct cacaaccagt cggtagatgt caagaagaga cgttgggtta ccttctgctc
     1321 tgcagaatgg ccaaccttta acgtcggatg gccgcgagac ggcaccttta accgagacct
     1381 catcacccag gttaagatca aggtcttttc acctggcccg catggacacc cagaccaggt
     1441 cccctacatc gtgacctggg aagccttggc ttttgacccc cctccctggg tcaagccctt
     1501 tgtacaccct aagcctccgc ctcctcttcc tccatccgcc ccgtctctcc cccttgaacc
     1561 tcctcgttcg accccgcctc gatcctccct ttatccagcc ctcactcctt ctctaggcgc
     1621 ccccatatgg ccatatgaga tcttatatgg ggcacccccg ccccttgtaa acttccctga
     1681 ccctgacatg acaagagtta ctaacagccc ctctctccaa gctcacttac aggctctcta
     1741 cttagtccag cacgaagtct ggagacctct ggcggcagcc taccaagaac aactggaccg
     1801 accggtggta cctcaccctt accgagtcgg cgacacagtg tgggtccgcc gacaccagac
     1861 taagaaccta gaacctcgct ggaaaggacc ttacacagtc ctgctgacca cccccaccgc
     1921 cctcaaagta gacggcatcg cagcttggat acacgccgcc cacgtgaagg ctgccgaccc
     1981 cgggggtgga ccatcctcta gactgccgga tcctgagtca gtgactcagt gagtcagtga
     2041 ctcagtgagt cagtgactca gtgagtcagt gactcaggaa ttcagcttga ataaaatgaa
     2101 tattagaagc tgttagaata agagaaaatg acagaggaaa actgaaaggg agaactgaaa
     2161 gtgggaaatt cctctgaggc agaaaggacc atcccttata aatagcacag gccatgaagg
     2221 aagatcattc tcactgcagc ctttgacagc ctttgcctca tcttgaagct tcaccatggt
     2281 gagcaagggc gaggaggata acatggccat catcaaggag ttcatgcgct tcaaggtgca
     2341 catggagggc tccgtgaacg gccacgagtt cgagatcgag ggcgagggcg agggccgccc
     2401 ctacgagggc acccagaccg ccaagctgaa ggtgaccaag ggtggccccc tgcccttcgc
     2461 ctgggacatc ctgtcccctc agttcatgta cggctccaag gcctacgtga agcaccccgc
     2521 cgacatcccc gactacttga agctgtcctt ccccgagggc ttcaagtggg agcgcgtgat
     2581 gaacttcgag gacggcggcg tggtgaccgt gacccaggac tcctccctgc aggacggcga
     2641 gttcatctac aaggtgaagc tgcgcggcac caacttcccc tccgacggcc ccgtaatgca
     2701 gaagaagacc atgggctggg aggcctcctc cgagcggatg taccccgagg acggcgccct
     2761 gaagggcgag atcaagcaga ggctgaagct gaaggacggc ggccactacg acgctgaggt
     2821 caagaccacc tacaaggcca agaagcccgt gcagctgccc ggcgcctaca acgtcaacat
     2881 caagttggac atcacctccc acaacgagga ctacaccatc gtggaacagt acgaacgcgc
     2941 cgagggccgc cactccaccg gcggcatgga cgagctgtac aagtaggcgg ccgccagcac
     3001 agtggtcgat cgacgataaa ataaaagatt ttatttagtc tccagaaaaa ggggggaatg
     3061 aaagacccca cctgtaggtt tggcaagcta gcttaagtaa cgccattttg caaggcatgg
     3121 aaaaatacat aactgagaat agaaaagttc agatcaaggt caggaacaga tggaacaggg
     3181 tcgcgtcccg caataaaaga gcccacaacc cctcactcgg ggcgccagtc ctccgattga
     3241 ctgagtcgcc cgggtacccg tgtatccaat aaaccctctt gcagttgcat ccgacttgtg
     3301 gtctcgctgt tccttgggag ggtctcctct gagtgattga ctacccgtca gcgggggtct
     3361 ttcacatgca gcatgtatca aaattaattt ggtttttttt cttaagtatt tacattaaat
     3421 ggccatagta gttcattatg gacagcgcag aaagagctgg ggagaattgt gaaattgtta
     3481 tccgctcaca attccacaca acatacgagc cggaagcata aagtgtaaag cctggggtgc
     3541 ctaatgagtg agctaactca cattaattgc gttgcgctca ctgcccgctt tccagtcggg
     3601 aaacctgtcg tgccagctgc attaatgaat cggccaacgc gcggggagag gcggtttgcg
     3661 tattgggcgc tcttccgctt cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg
     3721 gcgagcggta tcagctcact caaaggcggt aatacggtta tccacagaat caggggataa
     3781 cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc
     3841 gttgctggcg tttttccata ggctccgccc ccctgacgag catcacaaaa atcgacgctc
     3901 aagtcagagg tggcgaaacc cgacaggact ataaagatac caggcgtttc cccctggaag
     3961 ctccctcgtg cgctctcctg ttccgaccct gccgcttacc ggatacctgt ccgcctttct
     4021 cccttcggga agcgtggcgc tttctcatag ctcacgctgt aggtatctca gttcggtgta
     4081 ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc gttcagcccg accgctgcgc
     4141 cttatccggt aactatcgtc ttgagtccaa cccggtaaga cacgacttat cgccactggc
     4201 agcagccact ggtaacagga ttagcagagc gaggtatgta ggcggtgcta cagagttctt
     4261 gaagtggtgg cctaactacg gctacactag aagaacagta tttggtatct gcgctctgct
     4321 gaagccagtt accttcggaa aaagagttgg tagctcttga tccggcaaac aaaccaccgc
     4381 tggtagcggt ggtttttttg tttgcaagca gcagattacg cgcagaaaaa aaggatctca
     4441 agaagatcct ttgatctttt ctacggggtc tgacgctcag tggaacgaaa actcacgtta
     4501 agggattttg gtcatgagat tatcaaaaag gatcttcacc tagatccttt tgcggccggc
     4561 cgcaaatcaa tctaaagtat atatgagtaa acttggtctg acagttacca atgcttaatc
     4621 agtgaggcac ctatctcagc gatctgtcta tttcgttcat ccatagttgc ctgactcccc
     4681 gtcgtgtaga taactacgat acgggagggc ttaccatctg gccccagtgc tgcaatgata
     4741 ccgcgagacc cacgctcacc ggctccagat ttatcagcaa taaaccagcc agccggaagg
     4801 gccgagcgca gaagtggtcc tgcaacttta tccgcctcca tccagtctat taattgttgc
     4861 cgggaagcta gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt tgccattgct
     4921 acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc cggttcccaa
     4981 cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa aagcggttag ctccttcggt
     5041 cctccgatcg ttgtcagaag taagttggcc gcagtgttat cactcatggt tatggcagca
     5101 ctgcataatt ctcttactgt catgccatcc gtaagatgct tttctgtgac tggtgagtac
     5161 tcaaccaagt cattctgaga atagtgtatg cggcgaccga gttgctcttg cccggcgtca
     5221 atacgggata ataccgcgcc acatagcaga actttaaaag tgctcatcat tggaaaacgt
     5281 tcttcggggc gaaaactctc aaggatctta ccgctgttga gatccagttc gatgtaaccc
     5341 actcgtgcac ccaactgatc ttcagcatct tttactttca ccagcgtttc tgggtgagca
     5401 aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa atgttgaata
     5461 ctcatactct tcctttttca atattattga agcatttatc agggttattg tctcatgagc
     5521 ggatacatat ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg cacatttccc
     5581 cgaaaagtgc caccagcttt gctcttagga gtttcctaat acatcccaaa ctcaaatata
     5641 taaagcattt gacttgttct atgccctagt tattaatagt aatcaattac
//
