LOCUS       sequence-251289-        5851 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses DsRed.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5851)
  AUTHORS   Wang X, Cato P, Lin HC, Li T, Wan D, Alcocer MJ, Falcone FH
  TITLE     Optimisation and use of humanised RBL NF-AT-GFP and NF-AT-DsRed 
            reporter cell lines suitable for high-throughput scale detection of 
            allergic sensitisation in array format and identification of the 
            ECM-integrin interaction as critical factor.
  JOURNAL   Mol Biotechnol. 2014 Feb;56(2):136-46. doi: 
            10.1007/s12033-013-9689-x.
  PUBMED    23893250
REFERENCE   2  (bases 1 to 5851)
  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"; journalName: 
            "Mol Biotechnol"; date: "2014-02"; volume: "56"; issue: "2"; pages: 
            "136-46"
COMMENT     Sequence Label: pMJA167
FEATURES             Location/Qualifiers
     source          1..5851
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(156..175)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     316..338
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     polyA_signal    469..603
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     polyA_signal    697..831
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     protein_bind    838..867
                     /label=NFAT binding site
                     /bound_moiety="NFAT transcription factor"
                     /note="NFAT binding site from the human interleukin 2 
                     (IL-2) gene (Mattila et al., 1990)"
     protein_bind    868..897
                     /label=NFAT binding site
                     /bound_moiety="NFAT transcription factor"
                     /note="NFAT binding site from the human interleukin 2 
                     (IL-2) gene (Mattila et al., 1990)"
     protein_bind    898..927
                     /label=NFAT binding site
                     /bound_moiety="NFAT transcription factor"
                     /note="NFAT binding site from the human interleukin 2 
                     (IL-2) gene (Mattila et al., 1990)"
     protein_bind    928..957
                     /label=NFAT binding site
                     /bound_moiety="NFAT transcription factor"
                     /note="NFAT binding site from the human interleukin 2 
                     (IL-2) gene (Mattila et al., 1990)"
     CDS             1014..1691
                     /codon_start=1
                     /product="noncytotoxic tetrameric variant of DsRed 
                     fluorescent protein (Strack et al., 2008)"
                     /label=DsRed-Express2
                     /note="mammalian codon-optimized"
                     /translation="MDSTENVIKPFMRFKVHMEGSVNGHEFEIEGEGEGKPYEGTQTAK
                     LQVTKGGPLPFAWDILSPQFQYGSKVYVKHPADIPDYKKLSFPEGFKWERVMNFEDGGV
                     VTVTQDSSLQDGTFIYHVKFIGVNFPSDGPVMQKKTLGWEPSTERLYPRDGVLKGEIHK
                     ALKLKGGGHYLVEFKSIYMAKKPVKLPGYYYVDSKLDITSHNEDYTVVEQYERAEARHH
                     LFQ"
     primer_bind     complement(1194..1214)
                     /label=DsRed1-N
                     /note="DsRed1, reverse primer"
     primer_bind     1412..1431
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     primer_bind     1606..1629
                     /label=DsRed1-C
                     /note="DsRed1, forward primer"
     CDS             1716..1757
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     CDS             1767..1784
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(1807..1824)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    1813..2037
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      2083..2511
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2170..2189)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2380..2401
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(2520..2540)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        2525..2854
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      2705..2840
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     2767..2786
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        2902..2949
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             2968..3366
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
                     VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
                     KAIVKDSDGQPTAVGIRELLPSGYVWEG"
     polyA_signal    3524..3645
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3694..3710)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3694..3710)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3707..3729)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3718..3734
                     /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(3742..3748,3749..3766,3767..3772))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3787..3808
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3925..3942)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4096..4684)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4176..4195)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4855..5646,5647..5715))
                     /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     5478..5497
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5716..5820)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gacggatcgg gagatctgca gaactgtgaa tgcgcaaacc aacccttggc agaacatatc
       61 catcgcgtcc gccatctcca gcagccgcac gcggcgcatc tcggggccga cgcgctgggc
      121 tacgtcttgc tggcgttcgt atggtgcact ctcagtacaa tctgctctga tgccgcatag
      181 ttaagccagt atacactccg ctatcgctac gtgactgggt catggctgcg ccccgacacc
      241 cgccaacacc cgctgacgcg ccctgacggg cttgtctgct cccggcatcc gcttacagac
      301 aagctgtgac cgtctccggg agctgcatgt gtcagaggtt ttcaccgtca tcaccgaaac
      361 gcgcgaggca ggatcagcca tatcacattt gtagaggttt tacttgcttt aaaaaacctc
      421 ccacacctcc ccctgaacct gaaacataaa atgaatgcaa ttgttgttaa cttgtttatt
      481 gcagcttata atggttacaa ataaagcaat agcatcacaa atttcacaaa taaagcattt
      541 ttttcactgc attctagttg tggtttgtcc aaactcatca atgtatctta tcatgtctgg
      601 atcagccata tcacatttgt agaggtttta cttgctttaa aaaacctccc acacctcccc
      661 ctgaacctga aacataaaat gaatgcaatt gttgttaact tgtttattgc agcttataat
      721 ggttacaaat aaagcaatag catcacaaat ttcacaaata aagcattttt ttcactgcat
      781 tctagttgtg gtttgtccaa actcatcaat gtatcttatc atgtctggat ccaagctgga
      841 ggaaaaactg tttcatacag aaggcgtgga ggaaaaactg tttcatacag aaggcgtgga
      901 ggaaaaactg tttcatacag aaggcgtgga ggaaaaactg tttcatacag aaggcgtcgc
      961 ggagactcta gagggtatat aatggatccc cgggtaccga gctcgaattc accatggata
     1021 gcactgagaa cgtcatcaag cccttcatgc gcttcaaggt gcacatggag ggctccgtga
     1081 acggccacga gttcgagatc gagggcgagg gcgagggcaa gccctacgag ggcacccaga
     1141 ccgccaagct gcaggtgacc aagggcggcc ccctgccctt cgcctgggac atcctgtccc
     1201 cccagttcca gtacggctcc aaggtgtacg tgaagcaccc cgccgacatc cccgactaca
     1261 agaagctgtc cttccccgag ggcttcaagt gggagcgcgt gatgaacttc gaggacggcg
     1321 gcgtggtgac cgtgacccag gactcctccc tgcaggacgg caccttcatc taccacgtga
     1381 agttcatcgg cgtgaacttc ccctccgacg gccccgtaat gcagaagaag actctgggct
     1441 gggagccctc caccgagcgc ctgtaccccc gcgacggcgt gctgaagggc gagatccaca
     1501 aggcgctgaa gctgaagggc ggcggccact acctggtgga gttcaagtca atctacatgg
     1561 ccaagaagcc cgtgaagctg cccggctact actacgtgga ctccaagctg gacatcacct
     1621 cccacaacga ggactacacc gtggtggagc agtacgagcg cgccgaggcc cgccaccacc
     1681 tgttccagta gctcgagtct agagggccct tcgaaggtaa gcctatccct aaccctctcc
     1741 tcggtctcga ttctacgcgt accggtcatc atcaccatca ccattgagtt taaacccgct
     1801 gatcagcctc gactgtgcct tctagttgcc agccatctgt tgtttgcccc tcccccgtgc
     1861 cttccttgac cctggaaggt gccactccca ctgtcctttc ctaataaaat gaggaaattg
     1921 catcgcattg tctgagtagg tgtcattcta ttctgggggg tggggtgggg caggacagca
     1981 agggggagga ttgggaagac aatagcaggc atgctgggga tgcggtgggc tctatggctt
     2041 ctgaggcgga aagaaccagc tggggctcta gggggtatcc ccacgcgccc tgtagcggcg
     2101 cattaagcgc ggcgggtgtg gtggttacgc gcagcgtgac cgctacactt gccagcgccc
     2161 tagcgcccgc tcctttcgct ttcttccctt cctttctcgc cacgttcgcc ggctttcccc
     2221 gtcaagctct aaatcggggg ctccctttag ggttccgatt tagtgcttta cggcacctcg
     2281 accccaaaaa acttgattag ggtgatggtt cacgtagtgg gccatcgccc tgatagacgg
     2341 tttttcgccc tttgacgttg gagtccacgt tctttaatag tggactcttg ttccaaactg
     2401 gaacaacact caaccctatc tcggtctatt cttttgattt ataagggatt ttgccgattt
     2461 cggcctattg gttaaaaaat gagctgattt aacaaaaatt taacgcgaat taattctgtg
     2521 gaatgtgtgt cagttagggt gtggaaagtc cccaggctcc ccagcaggca gaagtatgca
     2581 aagcatgcat ctcaattagt cagcaaccag gtgtggaaag tccccaggct ccccagcagg
     2641 cagaagtatg caaagcatgc atctcaatta gtcagcaacc atagtcccgc ccctaactcc
     2701 gcccatcccg cccctaactc cgcccagttc cgcccattct ccgccccatg gctgactaat
     2761 tttttttatt tatgcagagg ccgaggccgc ctctgcctct gagctattcc agaagtagtg
     2821 aggaggcttt tttggaggcc taggcttttg caaaaagctc ccgggagctt gtatatccat
     2881 tttcggatct gatcagcacg tgttgacaat taatcatcgg catagtatat cggcatagta
     2941 taatacgaca aggtgaggaa ctaaaccatg gccaagcctt tgtctcaaga agaatccacc
     3001 ctcattgaaa gagcaacggc tacaatcaac agcatcccca tctctgaaga ctacagcgtc
     3061 gccagcgcag ctctctctag cgacggccgc atcttcactg gtgtcaatgt atatcatttt
     3121 actgggggac cttgtgcaga actcgtggtg ctgggcactg ctgctgctgc ggcagctggc
     3181 aacctgactt gtatcgtcgc gatcggaaat gagaacaggg gcatcttgag cccctgcgga
     3241 cggtgccgac aggtgcttct cgatctgcat cctgggatca aagccatagt gaaggacagt
     3301 gatggacagc cgacggcagt tgggattcgt gaattgctgc cctctggtta tgtgtgggag
     3361 ggctaagcac ttcgtggccg aggagcagga ctgacacgtg ctacgagatt tcgattccac
     3421 cgccgccttc tatgaaaggt tgggcttcgg aatcgttttc cgggacgccg gctggatgat
     3481 cctccagcgc ggggatctca tgctggagtt cttcgcccac cccaacttgt ttattgcagc
     3541 ttataatggt tacaaataaa gcaatagcat cacaaatttc acaaataaag catttttttc
     3601 actgcattct agttgtggtt tgtccaaact catcaatgta tcttatcatg tctgtatacc
     3661 gtcgacctct agctagagct tggcgtaatc atggtcatag ctgtttcctg tgtgaaattg
     3721 ttatccgctc acaattccac acaacatacg agccggaagc ataaagtgta aagcctgggg
     3781 tgcctaatga gtgagctaac tcacattaat tgcgttgcgc tcactgcccg ctttccagtc
     3841 gggaaacctg tcgtgccagc tgcattaatg aatcggccaa cgcgcgggga gaggcggttt
     3901 gcgtattggg cgctcttccg cttcctcgct cactgactcg ctgcgctcgg tcgttcggct
     3961 gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg ttatccacag aatcagggga
     4021 taacgcagga aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc
     4081 cgcgttgctg gcgtttttcc ataggctccg cccccctgac gagcatcaca aaaatcgacg
     4141 ctcaagtcag aggtggcgaa acccgacagg actataaaga taccaggcgt ttccccctgg
     4201 aagctccctc gtgcgctctc ctgttccgac cctgccgctt accggatacc tgtccgcctt
     4261 tctcccttcg ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc tcagttcggt
     4321 gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg
     4381 cgccttatcc ggtaactatc gtcttgagtc caacccggta agacacgact tatcgccact
     4441 ggcagcagcc actggtaaca ggattagcag agcgaggtat gtaggcggtg ctacagagtt
     4501 cttgaagtgg tggcctaact acggctacac tagaagaaca gtatttggta tctgcgctct
     4561 gctgaagcca gttaccttcg gaaaaagagt tggtagctct tgatccggca aacaaaccac
     4621 cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc
     4681 tcaagaagat cctttgatct tttctacggg gtctgacgct cagtggaacg aaaactcacg
     4741 ttaagggatt ttggtcatga gattatcaaa aaggatcttc acctagatcc ttttaaatta
     4801 aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa acttggtctg acagttacca
     4861 atgcttaatc agtgaggcac ctatctcagc gatctgtcta tttcgttcat ccatagttgc
     4921 ctgactcccc gtcgtgtaga taactacgat acgggagggc ttaccatctg gccccagtgc
     4981 tgcaatgata ccgcgagacc cacgctcacc ggctccagat ttatcagcaa taaaccagcc
     5041 agccggaagg gccgagcgca gaagtggtcc tgcaacttta tccgcctcca tccagtctat
     5101 taattgttgc cgggaagcta gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt
     5161 tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc
     5221 cggttcccaa cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa aagcggttag
     5281 ctccttcggt cctccgatcg ttgtcagaag taagttggcc gcagtgttat cactcatggt
     5341 tatggcagca ctgcataatt ctcttactgt catgccatcc gtaagatgct tttctgtgac
     5401 tggtgagtac tcaaccaagt cattctgaga atagtgtatg cggcgaccga gttgctcttg
     5461 cccggcgtca atacgggata ataccgcgcc acatagcaga actttaaaag tgctcatcat
     5521 tggaaaacgt tcttcggggc gaaaactctc aaggatctta ccgctgttga gatccagttc
     5581 gatgtaaccc actcgtgcac ccaactgatc ttcagcatct tttactttca ccagcgtttc
     5641 tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa
     5701 atgttgaata ctcatactct tcctttttca atattattga agcatttatc agggttattg
     5761 tctcatgagc ggatacatat ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg
     5821 cacatttccc cgaaaagtgc cacctgacgt c
//
