LOCUS       40924_20156        6306 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Mammalian expression vector pFLAG-p65-S536A, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6306)
  AUTHORS   De Schamphelaire W, Olbrechts A, Meert J, Verhelst K, Roggeman 
            Fonseca M, Vanhoucke M, Beyaert R.
  TITLE     BCCM/LMBP Plasmid collection
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 6306)
  AUTHORS   De Schamphelaire W.
  TITLE     Direct Submission
  JOURNAL   Submitted (02-FEB-2017) BCCM/LMBP, Universiteit Gent, 
            Technologiepark 927, 9052, BELGIUM
REFERENCE   3  (bases 1 to 6306)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 6306)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: 
            "Unpublished"
COMMENT     SGRef: number: 2; type: "Journal Article"; journalName: "Submitted 
            (02-FEB-2017) BCCM/LMBP, Universiteit Gent, Technologiepark 927, 
            9052, BELGIUM"
COMMENT     SGRef: number: 3; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..6306
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     141..157
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     enhancer        318..697
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        698..901
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
     CDS             935..958
                     /codon_start=1
                     /label=FLAG
                     /note="FLAG(R) epitope tag, followed by an enterokinase
                     cleavage site"
                     /translation="DYKDDDDK"
     CDS             2240..2611
                     /codon_start=1
                     /label=RelA (p65) AD
                     /note="transcriptional activation domain of human RelA,
                     also known as p65 (O'Shea and Perkins, 2008)"
                     /translation="PTQAGEGTLSEALLQLQFDDEDLGALLGNSTDPAVFTDLASVDNS
                     EFQQLLNQGIPVAPHTTEPMLMEYPEAITRLVTGAQRPPDPAPAPLGAPGLPNGLLSGD
                     EDFSAIADMDFSALLSQISS"
     polyA_signal    2647..3269
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     promoter        3298..3627
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     promoter        complement(3666..3684)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(3698..3714)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    complement(3722..3738)
                     /label=lac operator
                     /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(3746..3776)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    complement(3791..3812)
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     rep_origin      complement(4100..4688)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(4862..5719)
                     /codon_start=1
                     /label=AmpR
                     /note="beta-lactamase"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     promoter        complement(5720..5824)
                     /label=AmpR promoter
     rep_origin      5851..6306
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
ORIGIN
        1 ccattcgcca ttcaggctgc gcaactgttg ggaagggcga tcggtgcggg cctcttcgct
       61 attacgccag ctggcgaaag ggggatgtgc tgcaaggcga ttaagttggg taacgccagg
      121 gttttcccag tcacgacgtt gtaaaacgac ggccagtgcc aagctgatct atacattgaa
      181 tcaatattgg caattagcca tattagtcat tggttatata gcataaatca atattggcta
      241 ttggccattg catacgttgt atctatatca taatatgtac atttatattg gctcatgtcc
      301 aatatgaccg ccatgttgac attgattatt gactagttat taatagtaat caattacggg
      361 gtcattagtt catagcccat atatggagtt ccgcgttaca taacttacgg taaatggccc
      421 gcctggctga ccgcccaacg acccccgccc attgacgtca ataatgacgt atgttcccat
      481 agtaacgcca atagggactt tccattgacg tcaatgggtg gagtatttac ggtaaactgc
      541 ccacttggca gtacatcaag tgtatcatat gccaagtccg ccccctattg acgtcaatga
      601 cggtaaatgg cccgcctggc attatgccca gtacatgacc ttacgggact ttcctacttg
      661 gcagtacatc tacgtattag tcatcgctat taccatggtg atgcggtttt ggcagtacac
      721 caatgggcgt ggatagcggt ttgactcacg gggatttcca agtctccacc ccattgacgt
      781 caatgggagt ttgttttggc accaaaatca acgggacttt ccaaaatgtc gtaataaccc
      841 cgccccgttg acgcaaatgg gcggtaggcg tgtacggtgg gaggtctata taagcagagc
      901 tcgtttagtg aaccgtcaga attgatctac catggactac aaagacgatg acgacaagct
      961 tgacgaactg ttccccctca tcttcccggc agagccagcc caggcctctg gcccctatgt
     1021 ggagatcatt gagcagccca agcagcgggg catgcgcttc cgctacaagt gcgaggggcg
     1081 ctccgcgggc agcatcccag gcgagaggag cacagatacc accaagaccc accccaccat
     1141 caagatcaat ggctacacag gaccagggac agtgcgcatc tccctggtca ccaaggaccc
     1201 tcctcaccgg cctcaccccc acgagcttgt aggaaaggac tgccgggatg gcttctatga
     1261 ggctgagctc tgcccggacc gctgcatcca cagtttccag aacctgggaa tccagtgtgt
     1321 gaagaagcgg gacctggagc aggctatcag tcagcgcatc cagaccaaca acaacccctt
     1381 ccaagttcct atagaagagc agcgtgggga ctacgacctg aatgctgtgc ggctctgctt
     1441 ccaggtgaca gtgcgggacc catcaggcag gcccctccgc ctgccgcctg tccttcctca
     1501 tcccatcttt gacaatcgtg cccccaacac tgccgagctc aagatctgcc gagtgaaccg
     1561 aaactctggc agctgcctcg gtggggatga gatcttccta ctgtgtgaca aggtgcagaa
     1621 agaggacatt gaggtgtatt tcacgggacc aggctgggag gcccgaggct ccttttcgca
     1681 agctgatgtg caccgacaag tggccattgt gttccggacc cctccctacg cagaccccag
     1741 cctgcaggct cctgtgcgtg tctccatgca gctgcggcgg ccttccgacc gggagctcag
     1801 tgagcccatg gaattccagt acctgccaga tacagacgat cgtcaccgga ttgaggagaa
     1861 acgtaaaagg acatatgaga ccttcaagag catcatgaag aagagtcctt tcagcggacc
     1921 caccgacccc cggcctccac ctcgacgcat tgctgtgcct tcccgcagct cagcttctgt
     1981 ccccaagcca gcaccccagc cctatccctt tacgtcatcc ctgagcacca tcaactatga
     2041 tgagtttccc accatggtgt ttccttctgg gcagatcagc caggcctcgg ccttggcccc
     2101 ggcccctccc caagtcctgc cccaggctcc agcccctgcc cctgctccag ccatggtatc
     2161 agctctggcc caggccccag cccctgtccc agtcctagcc ccaggccctc ctcaggctgt
     2221 ggccccacct gcccccaagc ccacccaggc tggggaagga acgctgtcag aggccctgct
     2281 gcagctgcag tttgatgatg aagacctggg ggccttgctt ggcaacagca cagacccagc
     2341 tgtgttcaca gacctggcat ccgtcgacaa ctccgagttt cagcagctgc tgaaccaggg
     2401 catacctgtg gccccccaca caactgagcc catgctgatg gagtaccctg aggctataac
     2461 tcgcctagtg acaggggccc agaggccccc cgacccagct cctgctccac tgggggcccc
     2521 ggggctcccc aatggcctcc tttcaggaga tgaagacttc tccgccattg cggacatgga
     2581 cttctcagcc ctgctgagtc agatcagctc ctaagatatc ggtaccagtc gactctagag
     2641 gatcccgggt ggcatccctg tgacccctcc ccagtgcctc tcctggccct ggaagttgcc
     2701 actccagtgc ccaccagcct tgtcctaata aaattaagtt gcatcatttt gtctgactag
     2761 gtgtccttct ataatattat ggggtggagg ggggtggtat ggagcaaggg gcaagttggg
     2821 aagacaacct gtagggcctg cggggtctat tgggaaccaa gctggagtgc agtggcacaa
     2881 tcttggctca ctgcaatctc cgcctcctgg gttcaagcga ttctcctgcc tcagcctccc
     2941 gagttgttgg gattccaggc atgcatgacc aggctcagct aatttttgtt tttttggtag
     3001 agacggggtt tcaccatatt ggccaggctg gtctccaact cctaatctca ggtgatctac
     3061 ccaccttggc ctcccaaatt gctgggatta caggcgtgaa ccactgctcc cttccctgtc
     3121 cttctgattt taaaataact ataccagcag gaggacgtcc agacacagca taggctacct
     3181 ggccatgccc aaccggtggg acatttgagt tgcttgcttg gcactgtcct ctcatgcgtt
     3241 gggtccactc agtagatgcc tgttgaattg ggtacgcggc cagcttggct gtggaatgtg
     3301 tgtcagttag ggtgtggaaa gtccccaggc tccccagcag gcagaagtat gcaaagcatg
     3361 catctcaatt agtcagcaac caggtgtgga aagtccccag gctccccagc aggcagaagt
     3421 atgcaaagca tgcatctcaa ttagtcagca accatagtcc cgcccctaac tccgcccatc
     3481 ccgcccctaa ctccgcccag ttccgcccat tctccgcccc atggctgact aatttttttt
     3541 atttatgcag aggccgaggc cgcctcggcc tctgagctat tccagaagta gtgaggaggc
     3601 ttttttggag gcctaggctt ttgcaaaaag ctcctcgagg aactgaaaaa ccagaaagtt
     3661 aattccctat agtgagtcgt attaaattcg taatcatgtc atagctgttt cctgtgtgaa
     3721 attgttatcc gctcacaatt ccacacaaca tacgagccgg aagcataaag tgtaaagcct
     3781 ggggtgccta atgagtgagc taactcacat taattgcgtt gcgctcactg cccgctttcc
     3841 agtcgggaaa cctgtcgtgc cagctgcatt aatgaatcgg ccaacgcgcg gggagaggcg
     3901 gtttgcgtat tgggcgctct tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc
     3961 ggctgcggcg agcggtatca gctcactcaa aggcggtaat acggttatcc acagaatcag
     4021 gggataacgc aggaaagaac atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa
     4081 aggccgcgtt gctggcgttt ttccataggc tccgcccccc tgacgagcat cacaaaaatc
     4141 gacgctcaag tcagaggtgg cgaaacccga caggactata aagataccag gcgtttcccc
     4201 ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga tacctgtccg
     4261 cctttctccc ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg tatctcagtt
     4321 cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga accccccgtt cagcccgacc
     4381 gctgcgcctt atccggtaac tatcgtcttg agtccaaccc ggtaagacac gacttatcgc
     4441 cactggcagc agccactggt aacaggatta gcagagcgag gtatgtaggc ggtgctacag
     4501 agttcttgaa gtggtggcct aactacggct acactagaag aacagtattt ggtatctgcg
     4561 ctctgctgaa gccagttacc ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa
     4621 ccaccgctgg tagcggtggt ttttttgttt gcaagcagca gattacgcgc agaaaaaaag
     4681 gatctcaaga agatcctttg atcttttcta cggggtctga cgctcagtgg aacgaaaact
     4741 cacgttaagg gattttggtc atgagattat caaaaaggat cttcacctag atccttttaa
     4801 attaaaaatg aagttttaaa tcaatctaaa gtatatatga gtaaacttgg tctgacagtt
     4861 accaatgctt aatcagtgag gcacctatct cagcgatctg tctatttcgt tcatccatag
     4921 ttgcctgact ccccgtcgtg tagataacta cgatacggga gggcttacca tctggcccca
     4981 gtgctgcaat gataccgcga gacccacgct caccggctcc agatttatca gcaataaacc
     5041 agccagccgg aagggccgag cgcagaagtg gtcctgcaac tttatccgcc tccatccagt
     5101 ctattaattg ttgccgggaa gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg
     5161 ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg gcttcattca
     5221 gctccggttc ccaacgatca aggcgagtta catgatcccc catgttgtgc aaaaaagcgg
     5281 ttagctcctt cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg ttatcactca
     5341 tggttatggc agcactgcat aattctctta ctgtcatgcc atccgtaaga tgcttttctg
     5401 tgactggtga gtactcaacc aagtcattct gagaatagtg tatgcggcga ccgagttgct
     5461 cttgcccggc gtcaatacgg gataataccg cgccacatag cagaacttta aaagtgctca
     5521 tcattggaaa acgttcttcg gggcgaaaac tctcaaggat cttaccgctg ttgagatcca
     5581 gttcgatgta acccactcgt gcacccaact gatcttcagc atcttttact ttcaccagcg
     5641 tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata agggcgacac
     5701 ggaaatgttg aatactcata ctcttccttt ttcaatatta ttgaagcatt tatcagggtt
     5761 attgtctcat gagcggatac atatttgaat gtatttagaa aaataaacaa ataggggttc
     5821 cgcgcacatt tccccgaaaa gtgccacctg acgcgccctg tagcggcgca ttaagcgcgg
     5881 cgggtgtggt ggttacgcgc agcgtgaccg ctacacttgc cagcgcccta gcgcccgctc
     5941 ctttcgcttt cttcccttcc tttctcgcca cgttcgccgg ctttccccgt caagctctaa
     6001 atcgggggct ccctttaggg ttccgattta gtgctttacg gcacctcgac cccaaaaaac
     6061 ttgattaggg tgatggttca cgtagtgggc catcgccctg atagacggtt tttcgccctt
     6121 tgacgttgga gtccacgttc tttaatagtg gactcttgtt ccaaactgga acaacactca
     6181 accctatctc ggtctattct tttgatttat aagggatttt gccgatttcg gcctattggt
     6241 taaaaaatga gctgatttaa caaaaattta acgcgaattt taacaaaata ttaacgctta
     6301 caattt
//
