LOCUS       sequence-132364-        6531 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of human EBI3 in mammalian cells (V5 tag).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6531)
  AUTHORS   Ramnath D, Tunny K, Hohenhaus DM, Pitts CM, Bergot AS, Hogarth PM, 
            Hamilton JA, Kapetanovic R, Sturm RA, Scholz GM, Sweet MJ
  TITLE     TLR3 drives IRF6-dependent IL-23p19 expression and p19/EBI3 
            heterodimer formation in keratinocytes.
  JOURNAL   Immunol Cell Biol. 2015 Oct;93(9):771-9. doi: 10.1038/icb.2015.77. 
            Epub 2015 Aug 25.
  PUBMED    26303210
REFERENCE   2  (bases 1 to 6531)
  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: 
            "10.1038/icb.2015.77"; journalName: "Immunol Cell Biol"; date: 
            "2015-10"; volume: "93"; issue: "9"; pages: "771-9"
COMMENT     Sequence Label: hEBI3 pEF6-V5 (MJS_002)
FEATURES             Location/Qualifiers
     source          1..6531
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             757..798
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     CDS             808..825
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(848..865)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    854..1078
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      1124..1552
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1211..1230)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1421..1442
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(1561..1581)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        1566..1896
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      1747..1882
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     1809..1828
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        1944..1991
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             2010..2408
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
                     VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
                     KAIVKDSDGQPTAVGIRELLPSGYVWEG"
     polyA_signal    2566..2687
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2603..2622)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2657..2676
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(2736..2752)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2736..2752)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2749..2771)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2760..2776
                     /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(2784..2790,2791..2808,2809..2814))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2829..2850
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2967..2984)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3138..3726)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3218..3237)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3897..4688,4689..4757))
                     /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     4520..4539
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4758..4862)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(4937..4956)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        5246..6424
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation 
                     factor EF-1-alpha"
     intron          5477..6415
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
     primer_bind     6372..6392
                     /label=EF1a-F
                     /note="Human elongation factor-1a promoter, forward primer"
     primer_bind     6441..6460
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        6441..6459
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
ORIGIN
        1 gccatgaccc cgcagcttct cctggccctt gtcctctggg ccagctgccc gccctgcagt
       61 ggaaggaaag ggcccccagc agctctgaca ctgccccggg tgcaatgccg agcctctcgg
      121 tacccgatcg ccgtggattg ctcctggacc ctgccgcctg ctccaaactc caccagcccc
      181 gtgtccttca ttgccacgta caggctcggc atggctgccc ggggccacag ctggccctgc
      241 ctgcagcaga cgccaacgtc caccagctgc accatcacgg atgtccagct gttctccatg
      301 gctccctacg tgctcaatgt caccgccgtc cacccctggg gctccagcag cagcttcgtg
      361 cctttcataa cagagcacat catcaagccc gaccctccag aaggcgtgcg cctaagcccc
      421 ctcgctgagc gccagctaca ggtgcagtgg gagcctcccg ggtcctggcc cttcccagag
      481 atcttctcac tgaagtactg gatccgttac aagcgtcagg gagctgcgcg cttccaccgg
      541 gtggggccca ttgaagccac gtccttcatc ctcagggctg tgcggccccg agccaggtac
      601 tacgtccaag tggcggctca ggacctcaca gactacgggg aactgagtga ctggagtctc
      661 cccgccactg ccacaatgag cctgggcaag aagggcaatt ctgcagatat ccagcacagt
      721 ggcggccgct cgagtctaga gggcccgcgg ttcgaaggta agcctatccc taaccctctc
      781 ctcggtctcg attctacgcg taccggtcat catcaccatc accattgagt ttaaacccgc
      841 tgatcagcct cgactgtgcc ttctagttgc cagccatctg ttgtttgccc ctcccccgtg
      901 ccttccttga ccctggaagg tgccactccc actgtccttt cctaataaaa tgaggaaatt
      961 gcatcgcatt gtctgagtag gtgtcattct attctggggg gtggggtggg gcaggacagc
     1021 aagggggagg attgggaaga caatagcagg catgctgggg atgcggtggg ctctatggct
     1081 tctgaggcgg aaagaaccag ctggggctct agggggtatc cccacgcgcc ctgtagcggc
     1141 gcattaagcg cggcgggtgt ggtggttacg cgcagcgtga ccgctacact tgccagcgcc
     1201 ctagcgcccg ctcctttcgc tttcttccct tcctttctcg ccacgttcgc cggctttccc
     1261 cgtcaagctc taaatcgggg catcccttta gggttccgat ttagtgcttt acggcacctc
     1321 gaccccaaaa aacttgatta gggtgatggt tcacgtagtg ggccatcgcc ctgatagacg
     1381 gtttttcgcc ctttgacgtt ggagtccacg ttctttaata gtggactctt gttccaaact
     1441 ggaacaacac tcaaccctat ctcggtctat tcttttgatt tataagggat tttggggatt
     1501 tcggcctatt ggttaaaaaa tgagctgatt taacaaaaat ttaacgcgaa ttaattctgt
     1561 ggaatgtgtg tcagttaggg tgtggaaagt ccccaggctc cccaggcagg cagaagtatg
     1621 caaagcatgc atctcaatta gtcagcaacc aggtgtggaa agtccccagg ctccccagca
     1681 ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa ccatagtccc gcccctaact
     1741 ccgcccatcc cgcccctaac tccgcccagt tccgcccatt ctccgcccca tggctgacta
     1801 atttttttta tttatgcaga ggccgaggcc gcctctgcct ctgagctatt ccagaagtag
     1861 tgaggaggct tttttggagg cctaggcttt tgcaaaaagc tcccgggagc ttgtatatcc
     1921 attttcggat ctgatcagca cgtgttgaca attaatcatc ggcatagtat atcggcatag
     1981 tataatacga caaggtgagg aactaaacca tggccaagcc tttgtctcaa gaagaatcca
     2041 ccctcattga aagagcaacg gctacaatca acagcatccc catctctgaa gactacagcg
     2101 tcgccagcgc agctctctct agcgacggcc gcatcttcac tggtgtcaat gtatatcatt
     2161 ttactggggg accttgtgca gaactcgtgg tgctgggcac tgctgctgct gcggcagctg
     2221 gcaacctgac ttgtatcgtc gcgatcggaa atgagaacag gggcatcttg agcccctgcg
     2281 gacggtgtcg acaggtgctt ctcgatctgc atcctgggat caaagcgata gtgaaggaca
     2341 gtgatggaca gccgacggca gttgggattc gtgaattgct gccctctggt tatgtgtggg
     2401 agggctaagc acttcgtggc cgaggagcag gactgacacg tgctacgaga tttcgattcc
     2461 accgccgcct tctatgaaag gttgggcttc ggaatcgttt tccgggacgc cggctggatg
     2521 atcctccagc gcggggatct catgctggag ttcttcgccc accccaactt gtttattgca
     2581 gcttataatg gttacaaata aagcaatagc atcacaaatt tcacaaataa agcatttttt
     2641 tcactgcatt ctagttgtgg tttgtccaaa ctcatcaatg tatcttatca tgtctgtata
     2701 ccgtcgacct ctagctagag cttggcgtaa tcatggtcat agctgtttcc tgtgtgaaat
     2761 tgttatccgc tcacaattcc acacaacata cgagccggaa gcataaagtg taaagcctgg
     2821 ggtgcctaat gagtgagcta actcacatta attgcgttgc gctcactgcc cgctttccag
     2881 tcgggaaacc tgtcgtgcca gctgcattaa tgaatcggcc aacgcgcggg gagaggcggt
     2941 ttgcgtattg ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg
     3001 ctgcggcgag cggtatcagc tcactcaaag gcggtaatac ggttatccac agaatcaggg
     3061 gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag
     3121 gccgcgttgc tggcgttttt ccataggctc cgcccccctg acgagcatca caaaaatcga
     3181 cgctcaagtc agaggtggcg aaacccgaca ggactataaa gataccaggc gtttccccct
     3241 ggaagctccc tcgtgcgctc tcctgttccg accctgccgc ttaccggata cctgtccgcc
     3301 tttctccctt cgggaagcgt ggcgctttct caatgctcac gctgtaggta tctcagttcg
     3361 gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac cccccgttca gcccgaccgc
     3421 tgcgccttat ccggtaacta tcgtcttgag tccaacccgg taagacacga cttatcgcca
     3481 ctggcagcag ccactggtaa caggattagc agagcgaggt atgtaggcgg tgctacagag
     3541 ttcttgaagt ggtggcctaa ctacggctac actagaagga cagtatttgg tatctgcgct
     3601 ctgctgaagc cagttacctt cggaaaaaga gttggtagct cttgatccgg caaacaaacc
     3661 accgctggta gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga
     3721 tctcaagaag atcctttgat cttttctacg gggtctgacg ctcagtggaa cgaaaactca
     3781 cgttaaggga ttttggtcat gagattatca aaaaggatct tcacctagat ccttttaaat
     3841 taaaaatgaa gttttaaatc aatctaaagt atatatgagt aaacttggtc tgacagttac
     3901 caatgcttaa tcagtgaggc acctatctca gcgatctgtc tatttcgttc atccatagtt
     3961 gcctgactcc ccgtcgtgta gataactacg atacgggagg gcttaccatc tggccccagt
     4021 gctgcaatga taccgcgaga cccacgctca ccggctccag atttatcagc aataaaccag
     4081 ccagccggaa gggccgagcg cagaagtggt cctgcaactt tatccgcctc catccagtct
     4141 attaattgtt gccgggaagc tagagtaagt agttcgccag ttaatagttt gcgcaacgtt
     4201 gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc
     4261 tccggttccc aacgatcaag gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt
     4321 agctccttcg gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg
     4381 gttatggcag cactgcataa ttctcttact gtcatgccat ccgtaagatg cttttctgtg
     4441 actggtgagt actcaaccaa gtcattctga gaatagtgta tgcggcgacc gagttgctct
     4501 tgcccggcgt caatacggga taataccgcg ccacatagca gaactttaaa agtgctcatc
     4561 attggaaaac gttcttcggg gcgaaaactc tcaaggatct taccgctgtt gagatccagt
     4621 tcgatgtaac ccactcgtgc acccaactga tcttcagcat cttttacttt caccagcgtt
     4681 tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg
     4741 aaatgttgaa tactcatact cttccttttt caatattatt gaagcattta tcagggttat
     4801 tgtctcatga gcggatacat atttgaatgt atttagaaaa ataaacaaat aggggttccg
     4861 cgcacatttc cccgaaaagt gccacctgac gtcgacggat cgggagatct cccgatcccc
     4921 tatggtcgac tctcagtaca atctgctctg atgccgcata gttaagccag tatctgctcc
     4981 ctgcttgtgt gttggaggtc gctgagtagt gcgcgagcaa aatttaagct acaacaaggc
     5041 aaggcttgac cgacaattgc atgaagaatc tgcttagggt taggcgtttt gcgctgcttc
     5101 gcgatgtacg ggccagatat acgcgttgac attgattatt gactaggctt ttgcaaaaag
     5161 ctttgcaaag atggataaag ttttaaacag agaggaatct ttgcagctaa tggaccttct
     5221 aggtcttgaa aggagtgcct cgtgaggctc cggtgcccgt cagtgggcag agcgcacatc
     5281 gcccacagtc cccgagaagt tggggggagg ggtcggcaat tgaaccggtg cctagagaag
     5341 gtggcgcggg gtaaactggg aaagtgatgt cgtgtactgg ctccgccttt ttcccgaggg
     5401 tgggggagaa ccgtatataa gtgcagtagt cgccgtgaac gttctttttc gcaacgggtt
     5461 tgccgccaga acacaggtaa gtgccgtgtg tggttcccgc gggcctggcc tctttacggg
     5521 ttatggccct tgcgtgcctt gaattacttc cacctggctg cagtacgtga ttcttgatcc
     5581 cgagcttcgg gttggaagtg ggtgggagag ttcgaggcct tgcgcttaag gagccccttc
     5641 gcctcgtgct tgagttgagg cctggcctgg gcgctggggc cgccgcgtgc gaatctggtg
     5701 gcaccttcgc gcctgtctcg ctgctttcga taagtctcta gccatttaaa atttttgatg
     5761 acctgctgcg acgctttttt tctggcaaga tagtcttgta aatgcgggcc aagatctgca
     5821 cactggtatt tcggtttttg gggccgcggg cggcgacggg gcccgtgcgt cccagcgcac
     5881 atgttcggcg aggcggggcc tgcgagcgcg gccaccgaga atcggacggg ggtagtctca
     5941 agctggccgg cctgctctgg tgcctggcct cgcgccgccg tgtatcgccc cgccctgggc
     6001 ggcaaggctg gcccggtcgg caccagttgc gtgagcggaa agatggccgc ttcccggccc
     6061 tgctgcaggg agctcaaaat ggaggacgcg gcgctcggga gagcgggcgg gtgagtcacc
     6121 cacacaaagg aaaagggcct ttccgtcctc agccgtcgct tcatgtgact ccacggagta
     6181 ccgggcgccg tccaggcacc tcgattagtt ctcgagcttt tggagtacgt cgtctttagg
     6241 ttggggggag gggttttatg cgatggagtt tccccacact gagtgggtgg agactgaagt
     6301 taggccagct tggcacttga tgtaattctc cttggaattt gccctttttg agtttggatc
     6361 ttggttcatt ctcaagcctc agacagtggt tcaaagtttt tttcttccat ttcaggtgtc
     6421 gtgaggaatt agcttggtac taatacgact cactataggg agacccaagc tggctaggta
     6481 agcttggtac cgagctcgga tccactagtc cagtgtggtg gaattgccct t
//
