LOCUS       sequence-274503-        6411 bp DNA     circular SYN 26-AUG-2024
DEFINITION  Express Prkd2 in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6411)
  TITLE     Mutual inhibition between Prkd2 and Bcl6 controls T follicular 
            helper cell differentiation
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 6411)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Aug 26, 2024 from SnapGene Server 7.0.3
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: 
            "Unpublished"
FEATURES             Location/Qualifiers
     source          1..6411
                     /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    3524..3658
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3548..3567)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     3602..3621
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(3769..3785)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3769..3785)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3782..3804)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3793..3809
                     /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(3817..3847)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3862..3883
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4000..4017)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4171..4759)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4251..4270)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(4930..5790)
                     /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     5553..5572
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5791..5895)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     5963..5981
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(6019..6041)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     6141..6160
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     6354..6376
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     6368..6385
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     6369..6385
                     /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 ctaccagatc ttccctgacg aagtgctggg ctcagggcag tttggagtgg tctatggagg
     2581 aaaacaccgg aagacaggcc gggacgtggc agttaaggtc attgacaaac tgcgcttccc
     2641 taccaagcag gagagccagc tccggaatga agtggccatt ctgcagagcc tgcggcatcc
     2701 cgggatcgtg aacctggagt gcatgttcga gacgcctgag aaagtgtttg tggtgatgga
     2761 gaagctgcat ggggacatgt tggagatgat cctgtccagt gagaagggcc ggctgcctga
     2821 gcgcctcacc aagttcctca tcacccagat cctggtggct ttgagacacc ttcacttcaa
     2881 gaacattgtc cactgtgact tgaaaccaga aaacgtgttg ctggcatcag cagacccatt
     2941 tcctcaggtg aagctgtgtg actttggctt tgctcgcatc atcggcgaga agtcgttccg
     3001 ccgctcagtg gtgggcacgc cggcctacct ggcacccgag gtgctgctca accagggcta
     3061 caaccgctcg ctggacatgt ggtcagtggg cgtgatcatg tacgtcagcc tcagcggcac
     3121 cttccctttc aacgaggatg aggacatcaa tgaccagatc cagaacgccg ccttcatgta
     3181 cccggccagc ccctggagcc acatctcagc tggagccatt gacctcatca acaacctgct
     3241 gcaggtgaag atgcgcaaac gctacagcgt ggacaaatct ctcagccacc cctggttaca
     3301 ggagtaccag acgtggctgg acctccgaga gctggagggg aagatgggag agcgatacat
     3361 cacgcatgag agtgacgacg cgcgctggga gcagtttgca gcagagcatc cgctgcctgg
     3421 gtctgggctg cccacggaca gggatctcgg tggggcctgt ccaccacagg accacgacat
     3481 gcaggggctg gcggagcgca tcagtgttct ctgaggccgc ggggatccag acatgataag
     3541 atacattgat gagtttggac aaaccacaac tagaatgcag tgaaaaaaat gctttatttg
     3601 tgaaatttgt gatgctattg ctttatttgt aaccattata agctgcaata aacaagttaa
     3661 caacaacaat tgcattcatt ttatgtttca ggttcagggg gaggtgtggg aggttttttc
     3721 ggatcctcta gagtcgatct gcaggcatgc tagcttggcg taatcatggt catagctgtt
     3781 tcctgtgtga aattgttatc cgctcacaat tccacacaac atacgagccg gaagcataaa
     3841 gtgtaaagcc tggggtgcct aatgagtgag ctaactcaca ttaattgcgt tgcgctcact
     3901 gcccgctttc cagtcgggaa acctgtcgtg ccagctgcat taatgaatcg gccaacgcgc
     3961 ggggagaggc ggtttgcgta ttgggcgctc ttccgcttcc tcgctcactg actcgctgcg
     4021 ctcggtcgtt cggctgcggc gagcggtatc agctcactca aaggcggtaa tacggttatc
     4081 cacagaatca ggggataacg caggaaagaa catgtgagca aaaggccagc aaaaggccag
     4141 gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg ctccgccccc ctgacgagca
     4201 tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg acaggactat aaagatacca
     4261 ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg
     4321 atacctgtcc gcctttctcc cttcgggaag cgtggcgctt tctcatagct cacgctgtag
     4381 gtatctcagt tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt
     4441 tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggtaagaca
     4501 cgacttatcg ccactggcag cagccactgg taacaggatt agcagagcga ggtatgtagg
     4561 cggtgctaca gagttcttga agtggtggcc taactacggc tacactagaa gaacagtatt
     4621 tggtatctgc gctctgctga agccagttac cttcggaaaa agagttggta gctcttgatc
     4681 cggcaaacaa accaccgctg gtagcggtgg tttttttgtt tgcaagcagc agattacgcg
     4741 cagaaaaaaa ggatctcaag aagatccttt gatcttttct acggggtctg acgctcagtg
     4801 gaacgaaaac tcacgttaag ggattttggt catgagatta tcaaaaagga tcttcaccta
     4861 gatcctttta aattaaaaat gaagttttaa atcaatctaa agtatatatg agtaaacttg
     4921 gtctgacagt taccaatgct taatcagtga ggcacctatc tcagcgatct gtctatttcg
     4981 ttcatccata gttgcctgac tccccgtcgt gtagataact acgatacggg agggcttacc
     5041 atctggcccc agtgctgcaa tgataccgcg agacccacgc tcaccggctc cagatttatc
     5101 agcaataaac cagccagccg gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc
     5161 ctccatccag tctattaatt gttgccggga agctagagta agtagttcgc cagttaatag
     5221 tttgcgcaac gttgttgcca ttgctacagg catcgtggtg tcacgctcgt cgtttggtat
     5281 ggcttcattc agctccggtt cccaacgatc aaggcgagtt acatgatccc ccatgttgtg
     5341 caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt
     5401 gttatcactc atggttatgg cagcactgca taattctctt actgtcatgc catccgtaag
     5461 atgcttttct gtgactggtg agtactcaac caagtcattc tgagaatagt gtatgcggcg
     5521 accgagttgc tcttgcccgg cgtcaatacg ggataatacc gcgccacata gcagaacttt
     5581 aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct
     5641 gttgagatcc agttcgatgt aacccactcg tgcacccaac tgatcttcag catcttttac
     5701 tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat
     5761 aagggcgaca cggaaatgtt gaatactcat actcttcctt tttcaatatt attgaagcat
     5821 ttatcagggt tattgtctca tgagcggata catatttgaa tgtatttaga aaaataaaca
     5881 aataggggtt ccgcgcacat ttccccgaaa agtgccacct gacgtctaag aaaccattat
     5941 tatcatgaca ttaacctata aaaataggcg tatcacgagg ccctttcgtc tcgcgcgttt
     6001 cggtgatgac ggtgaaaacc tctgacacat gcagctcccg gagacggtca cagcttgtct
     6061 gtaagcggat gccgggagca gacaagcccg tcagggcgcg tcagcgggtg ttggcgggtg
     6121 tcggggctgg cttaactatg cggcatcaga gcagattgta ctgagagtgc accatatgcg
     6181 gtgtgaaata ccgcacagat gcgtaaggag aaaataccgc atcaggcgcc attcgccatt
     6241 caggctgcgc aactgttggg aagggcgatc ggtgcgggcc tcttcgctat tacgccagct
     6301 ggcgaaaggg ggatgtgctg caaggcgatt aagttgggta acgccagggt tttcccagtc
     6361 acgacgttgt aaaacgacgg ccagtgagtt cgagcttgca tgcctgcagg t
//
