LOCUS       sequence-274502-        5427 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Express Prkd2 without kinase domain in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5427)
  AUTHORS   Misawa T, SoRelle JA, Choi JH, Yue T, Wang KW, McAlpine W, Wang J, 
            Liu A, Tabeta K, Turer EE, Evers B, Nair-Gill E, Poddar S, Su L, Ou 
            F, Yu L, Russell J, Ludwig S, Zhan X, Hildebrand S, Li X, Tang M, 
            Murray AR, Moresco EMY, Beutler B
  TITLE     Mutual inhibition between Prkd2 and Bcl6 controls T follicular 
            helper cell differentiation.
  JOURNAL   Sci Immunol. 2020 Jan 24;5(43):eaaz0085. doi: 
            10.1126/sciimmunol.aaz0085.
  PUBMED    31980486
REFERENCE   2  (bases 1 to 5427)
  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: "Sci 
            Immunol."; date: "2020-01-24"; volume: "5(43):eaaz0085. doi"; pages:
            " 10.1126/sciimmunol.aaz0085"
COMMENT     Sequence Label: pCMV-HA_N-human Prkd2 without kinase domain
FEATURES             Location/Qualifiers
     source          1..5427
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        1..304
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        305..508
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     459..479
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     505..529
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     intron          646..742
                     /label=SV40 intron
                     /note="modified SV40 intron with splice donor and acceptor 
                     sites"
     CDS             806..832
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     polyA_signal    2540..2674
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2564..2583)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2618..2637
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(2785..2801)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2785..2801)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2798..2820)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2809..2825
                     /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(2833..2839,2840..2857,2858..2863))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2878..2899
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3016..3033)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3187..3775)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3267..3286)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3946..4737,4738..4806))
                     /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     4569..4588
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4807..4911)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     4979..4997
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(5035..5057)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     5157..5176
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5370..5392
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     5384..5401
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     5385..5401
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
ORIGIN
        1 cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt
       61 gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca
      121 atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc
      181 aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta
      241 catgacctta tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac
      301 catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg actcacgggg
      361 atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg
      421 ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg gtaggcgtgt
      481 acggtgggag gtctatataa gcagagctcg tttagtgaac cgtcagatcg cctggagacg
      541 ccatccacgc tgttttgacc tccatagaag acaccgggac cgatccagcc tccggactct
      601 agaggatccg gtactagagg aactgaaaaa ccagaaagtt aactggtaag tttagtcttt
      661 ttgtctttta tttcaggtcc cggatccggt ggtggtgcaa atcaaagaac tgctcctcag
      721 tggatgttgc ctttacttct aggcctgtac ggaagtgtta cttctgctct aaaagctgcg
      781 gaattgtacc cgcgggccca ccatgtaccc atacgatgtt ccagattacg ctcttatggc
      841 catggaggcc cgaattcggt cgaccgagat ctctcgaatg gccaccgccc cctcttatcc
      901 cgccgggctc cctggctctc ccgggccggg gtctcctccg ccccccggcg gcctagagct
      961 gcagtcgccg ccaccgctac tgccccagat cccggccccg ggttccgggg tctcctttca
     1021 catccagatc gggctgaccc gcgagttcgt gctgttgccc gccgcctccg agctggctca
     1081 tgtgaagcag ctggcctgtt ccatcgtgga ccagaagttc cctgagtgtg gcttctacgg
     1141 cctttacgac aagatcctgc ttttcaaaca tgaccccacg tcggccaacc tcctgcagct
     1201 ggtgcgctcg tccggagaca tccaggaggg cgacctggtg gaggtggtgc tgtcggcctc
     1261 ggccaccttc gaggacttcc agatccgccc gcacgccctc acggtgcact cctatcgggc
     1321 gcctgccttc tgtgatcact gcggggagat gctcttcggc ctagtgcgcc agggcctcaa
     1381 gtgcgatggc tgcgggctga actaccacaa gcgctgtgcc ttcagcatcc ccaacaactg
     1441 tagtggggcc cgcaaacggc gcctgtcatc cacgtctctg gccagtggcc actcggtgcg
     1501 cctcggcacc tccgagtccc tgccctgcac ggctgaagag ctgagccgta gcaccaccga
     1561 actcctgcct cgccgtcccc cgtcatcctc ttcctcctct tctgcctcat cgtatacggg
     1621 ccgccccatt gagctggaca agatgctgct ctccaaggtc aaggtgccgc acaccttcct
     1681 catccacagc tatacacggc ccaccgtttg ccaggcttgc aagaaactcc tcaagggcct
     1741 cttccggcag ggcctgcaat gcaaagactg caagtttaac tgtcacaaac gctgcgccac
     1801 ccgcgtccct aatgactgcc tgggggaggc ccttatcaat ggagatgtgc cgatggagga
     1861 ggccaccgat ttcagcgagg ctgacaagag cgccctcatg gatgagtcag aggactccgg
     1921 tgtcatccct ggctcccact cagagaatgc gctccacgcc agtgaggagg aggaaggcga
     1981 gggaggcaag gcccagagct ccctggggta catcccccta atgagggtgg tgcaatcggt
     2041 gcgacacacg acgcggaaat ccagcaccac gctgcgggag ggttgggtgg ttcattacag
     2101 caacaaggac acgctgagaa agcggcacta ttggcgcctg gactgcaagt gtatcacgct
     2161 cttccagaac aacacgacca acagatacta taaggaaatt ccgctgtcag aaatcctcac
     2221 ggtggagtcc gcccagaact tcagccttgt gccgccgggc accaacccac actgctttga
     2281 gatcgtcact gccaatgcca cctacttcgt gggcgagatg cctggcggga ctccgggtgg
     2341 gccaagtggg cagggggctg aggccgcccg gggctgggag acagccatcc gccaggccct
     2401 gatgcccgtc atccttcagg acgcacccag cgccccaggc cacgcgcccc acagacaagc
     2461 ttctctgagc atctctgtgt ccaacagtca gatccaagag aatgtggaca ttgccactgt
     2521 ctaccagtga ggccgcgggg atccagacat gataagatac attgatgagt ttggacaaac
     2581 cacaactaga atgcagtgaa aaaaatgctt tatttgtgaa atttgtgatg ctattgcttt
     2641 atttgtaacc attataagct gcaataaaca agttaacaac aacaattgca ttcattttat
     2701 gtttcaggtt cagggggagg tgtgggaggt tttttcggat cctctagagt cgatctgcag
     2761 gcatgctagc ttggcgtaat catggtcata gctgtttcct gtgtgaaatt gttatccgct
     2821 cacaattcca cacaacatac gagccggaag cataaagtgt aaagcctggg gtgcctaatg
     2881 agtgagctaa ctcacattaa ttgcgttgcg ctcactgccc gctttccagt cgggaaacct
     2941 gtcgtgccag ctgcattaat gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg
     3001 gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc
     3061 ggtatcagct cactcaaagg cggtaatacg gttatccaca gaatcagggg ataacgcagg
     3121 aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct
     3181 ggcgtttttc cataggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagtca
     3241 gaggtggcga aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct
     3301 cgtgcgctct cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc
     3361 gggaagcgtg gcgctttctc atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt
     3421 tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc
     3481 cggtaactat cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc
     3541 cactggtaac aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg
     3601 gtggcctaac tacggctaca ctagaagaac agtatttggt atctgcgctc tgctgaagcc
     3661 agttaccttc ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag
     3721 cggtggtttt tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga
     3781 tcctttgatc ttttctacgg ggtctgacgc tcagtggaac gaaaactcac gttaagggat
     3841 tttggtcatg agattatcaa aaaggatctt cacctagatc cttttaaatt aaaaatgaag
     3901 ttttaaatca atctaaagta tatatgagta aacttggtct gacagttacc aatgcttaat
     3961 cagtgaggca cctatctcag cgatctgtct atttcgttca tccatagttg cctgactccc
     4021 cgtcgtgtag ataactacga tacgggaggg cttaccatct ggccccagtg ctgcaatgat
     4081 accgcgagac ccacgctcac cggctccaga tttatcagca ataaaccagc cagccggaag
     4141 ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc atccagtcta ttaattgttg
     4201 ccgggaagct agagtaagta gttcgccagt taatagtttg cgcaacgttg ttgccattgc
     4261 tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct tcattcagct ccggttccca
     4321 acgatcaagg cgagttacat gatcccccat gttgtgcaaa aaagcggtta gctccttcgg
     4381 tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg ttatggcagc
     4441 actgcataat tctcttactg tcatgccatc cgtaagatgc ttttctgtga ctggtgagta
     4501 ctcaaccaag tcattctgag aatagtgtat gcggcgaccg agttgctctt gcccggcgtc
     4561 aatacgggat aataccgcgc cacatagcag aactttaaaa gtgctcatca ttggaaaacg
     4621 ttcttcgggg cgaaaactct caaggatctt accgctgttg agatccagtt cgatgtaacc
     4681 cactcgtgca cccaactgat cttcagcatc ttttactttc accagcgttt ctgggtgagc
     4741 aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat
     4801 actcatactc ttcctttttc aatattattg aagcatttat cagggttatt gtctcatgag
     4861 cggatacata tttgaatgta tttagaaaaa taaacaaata ggggttccgc gcacatttcc
     4921 ccgaaaagtg ccacctgacg tctaagaaac cattattatc atgacattaa cctataaaaa
     4981 taggcgtatc acgaggccct ttcgtctcgc gcgtttcggt gatgacggtg aaaacctctg
     5041 acacatgcag ctcccggaga cggtcacagc ttgtctgtaa gcggatgccg ggagcagaca
     5101 agcccgtcag ggcgcgtcag cgggtgttgg cgggtgtcgg ggctggctta actatgcggc
     5161 atcagagcag attgtactga gagtgcacca tatgcggtgt gaaataccgc acagatgcgt
     5221 aaggagaaaa taccgcatca ggcgccattc gccattcagg ctgcgcaact gttgggaagg
     5281 gcgatcggtg cgggcctctt cgctattacg ccagctggcg aaagggggat gtgctgcaag
     5341 gcgattaagt tgggtaacgc cagggttttc ccagtcacga cgttgtaaaa cgacggccag
     5401 tgagttcgag cttgcatgcc tgcaggt
//
