LOCUS       66_Zdk3-VPR             5354 bp    DNA     circular UNA 28-NOV-2024
DEFINITION  Generated by Golden Gate/Type IIS Cloning (CMV-bb, VPR).
ACCESSION   urn.local...4jx-fwjre5t
VERSION     urn.local...4jx-fwjre5t
KEYWORDS    .
SOURCE      
  ORGANISM  .
FEATURES             Location/Qualifiers
     CDS             7..24
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /transl_table=1
                     /translation="HHHHHH"
                     /standard_name="6xHis"
     polyA_signal    53..277
                     /note="bovine growth hormone polyadenylation signal"
                     /standard_name="bGH poly(A) signal"
     protein_bind    372..388
                     /bound_moiety="<i>lac</i> repressor encoded by
                     <i>lacI</i>"
                     /note="The <i>lac</i> repressor binds to the <i>lac</i>
                     operator to inhibit transcription in <i>E. coli</i>. This
                     inhibition can be relieved by adding lactose or
                     isopropyl-β-D-thiogalactopyranoside (IPTG)."
                     /standard_name="lac operator"
     promoter        join(complement(421..426),complement(403..420),
                     complement(396..402))
                     /note="promoter for the <i>E. coli lac</i> operon"
                     /standard_name="lac promoter"
     protein_bind    441..462
                     /bound_moiety="<i>E. coli</i> catabolite activator
                     protein"
                     /note="CAP binding activates transcription in the
                     presenceof cAMP."
                     /standard_name="CAP binding site"
     rep_origin      complement(750..1338)
     CDS             join(complement(2301..2369),complement(1509..2300))
                     /codon_start=1
                     /gene="<i>bla</i>"
                     /note="confers resistance to ampicillin, carbenicillin,
                     and related antibiotics"
                     /product="β-lactamase"
                     /transl_table=1
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGY
                     IELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVE
                     YSPVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRL
                     DRWEPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPL
                     LRSALPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIA
                     EIGASLIKHW*"
                     /standard_name="AmpR"
     promoter        complement(2370..2474)
                     /gene="<i>bla</i>"
                     /standard_name="AmpR promoter"
     enhancer        2744..3123
                     /note="human cytomegalovirus immediate early enhancer"
                     /standard_name="CMV enhancer"
     promoter        3124..3327
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
                     /standard_name="CMV promoter"
     promoter        3369..3387
                     /note="promoter for bacteriophage T7 RNA polymerase"
                     /standard_name="T7 promoter"
     CDS             3434..3454
                     /product="nuclear localization signal of SV40 large T
                     antigen"
                     /codon_start=1
                     /translation="PKKKRKV"
                     /transl_table=1
                     /standard_name="SV40 NLS"
     CDS             3488..3661
                     /translation="VLVARQEIYWLPNLNWEQKFAFISSLTNDPSQSANLLAEAKKLN
                     GAQP"
                     /transl_table=1
                     /standard_name="Zdk3"
     CDS             3716..3865
                     /translation="DALDDFDLDMLGSDALDDFDLDMLGSDALDDFDLDMLGSDALDD
                     FDLDML"
                     /codon_start=1
                     /transl_table=1
                     /product="tetrameric repeat of the minimal activation
                     domain of herpes simplex virus VP16 (<a
                     href=""http://www.ncbi.nlm.nih.gov/pubmed/9843940""
                     title=""http://www.ncbi.nlm.nih.gov/pubmed/9843940"">Beerl
                     i et al., 1998</a>)"
                     /standard_name="VP64"
     CDS             4334..4690
                     /translation="PTQAGEGTLSEALLQLQFDDEDLGALLGNSTDPAVFTDLASVDN
                     SEFQQLLNQGIPVAPHTTEPMLMEYPEAITRLVTGAQRPPDPAPAPLGAPGLPNGLLS
                     GDEDFSSIADMDFSALL"
                     /codon_start=1
                     /transl_table=1
                     /product="transcriptional activation domain of human
                     RelA,also known as p65
                     (<ahref=""http://www.ncbi.nlm.nih.gov/pubmed/18631125""tit
                     le=""http://www.ncbi.nlm.nih.gov/pubmed/18631125"">O'Shea
                     and Perkins, 2008</a>)"
                     /standard_name="RelA (p65) AD"
     CDS             4709..5278
                     /codon_start=1
                     /transl_table=1
                     /product="transcriptional activation domain from the
                     humanherpesvirus 4 (Epstein-Barr virus) replication
                     andtranscription activator Rta/BRLF1
                     (ahref=""http://www.ncbi.nlm.nih.gov/pubmed/1323708""title
                     =""http://www.ncbi.nlm.nih.gov/pubmed/1323708""Hardwick et
                     al.,
                     1992/a;ahref=""http://www.ncbi.nlm.nih.gov/pubmed/25730490
                     ""title=""http://www.ncbi.nlm.nih.gov/pubmed/25730490""Cha
                     vez et al., 2015/a)"
                     /standard_name="Rta AD"
     CDS             5300..5320
                     /product="nuclear localization signal of SV40 large T
                     antigen"
                     /codon_start=1
                     /translation="PKKKRKV"
                     /transl_table=1
                     /standard_name="SV40 NLS"
ORIGIN      
        1 accggtcatc atcaccatca ccattgagtt taaacccgct gatcagcctc gactgtgcct
       61 tctagttgcc agccatctgt tgtttgcccc tcccccgtgc cttccttgac cctggaaggt
      121 gccactccca ctgtcctttc ctaataaaat gaggaaattg catcgcattg tctgagtagg
      181 tgtcattcta ttctgggggg tggggtgggg caggacagca agggggagga ttgggaagac
      241 aatagcaggc atgctgggga tgcggtgggc tctatggctt ctgaggcgga aagaaccagc
      301 tggggctcga taccgtcgac ctctagctag agcttggcgt aatcatggtc atagctgttt
      361 cctgtgtgaa attgttatcc gctcacaatt ccacacaaca tacgagccgg aagcataaag
      421 tgtaaagcct agggtgccta atgagtgagc taactcacat taattgcgtt gcgctcactg
      481 cccgctttcc agtcgggaaa cctgtcgtgc cagctgcatt aatgaatcgg ccaacgcgcg
      541 gggagaggcg gtttgcgtat tgggcgctct tccgcttcct cgctcactga ctcgctgcgc
      601 tcggtcgttc ggctgcggcg agcggtatca gctcactcaa aggcggtaat acggttatcc
      661 acagaatcag gggataacgc aggaaagaac atgtgagcaa aaggccagca aaaggccagg
      721 aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc tccgcccccc tgacgagcat
      781 cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga caggactata aagataccag
      841 gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga
      901 tacctgtccg cctttctccc ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg
      961 tatctcagtt cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga accccccgtt
     1021 cagcccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc ggtaagacac
     1081 gacttatcgc cactggcagc agccactggt aacaggatta gcagagcgag gtatgtaggc
     1141 ggtgctacag agttcttgaa gtggtggcct aactacggct acactagaag aacagtattt
     1201 ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa gagttggtag ctcttgatcc
     1261 ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt gcaagcagca gattacgcgc
     1321 agaaaaaaag gatctcaaga agatcctttg atcttttcta cggggtctga cactcagtgg
     1381 aacgaaaact cacgttaagg gattttggtc atgagattat caaaaaggat cttcacctag
     1441 atccttttaa attaaaaatg aagttttaaa tcaatctaaa gtatatatga gtaaacttgg
     1501 tctgacagtt accaatgctt aatcagtgag gcacctatct cagcgatctg tctatttcgt
     1561 tcatccatag ttgcctgact ccccgtcgtg tagataacta cgatacggga gggcttacca
     1621 tctggcccca gtgctgcaat gataccgcga gacccacgct caccggctcc agatttatca
     1681 gcaataaacc agccagccgg aagggccgag cgcagaagtg gtcctgcaac tttatccgcc
     1741 tccatccagt ctattaattg ttgccgggaa gctagagtaa gtagttcgcc agttaatagt
     1801 ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg
     1861 gcttcattca gctccggttc ccaacgatca aggcgagtta catgatcccc catgttgtgc
     1921 aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg
     1981 ttatcactca tggttatggc agcactgcat aattctctta ctgtcatgcc atccgtaaga
     2041 tgcttttctg tgactggtga gtactcaacc aagtcattct gagaatagtg tatgcggcga
     2101 ccgagttgct cttgcccggc gtcaatacgg gataataccg cgccacatag cagaacttta
     2161 aaagtgctca tcattggaaa acgttcttcg gggcgaaaac tctcaaggat cttaccgctg
     2221 ttgagatcca gttcgatgta acccactcgt gcacccaact gatcttcagc atcttttact
     2281 ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata
     2341 agggcgacac ggaaatgttg aatactcata ctcttccttt ttcaatatta ttgaagcatt
     2401 tatcagggtt attgtctcat gagcggatac atatttgaat gtatttagaa aaataaacaa
     2461 ataggggttc cgcgcacatt tccccgaaaa gtgccacctg acgtcgacgg atcgggagat
     2521 cgatctcccg atcccctagg gtcgactctc agtacaatct gctctgatgc cgcatagtta
     2581 agccagtatc tgctccctgc ttgtgtgttg gaggtcgctg agtagtgcgc gagcaaaatt
     2641 taagctacaa caaggcaagg cttgaccgac aattgcatga agaatctgct tagggttagg
     2701 cgttttgcgc tgcttcgcga tgtacgggcc agatatacgc gttgacattg attattgact
     2761 agttattaat agtaatcaat tacggggtca ttagttcata gcccatatat tgagttccgc
     2821 gttacataac ttacggtaaa tggcccgcct ggctgaccgc ccaacgaccc ccgcccattg
     2881 acgtcaataa tgacgtatgt tcccatagta acgccaatag ggactttcca ttgacgtcaa
     2941 tgggtggagt atttacggta aactgcccac ttggcagtac atcaagtgta tcatatgcca
     3001 agtacgcccc ctattgacgt caatgacggt aaatggcccg cctggcatta tgcccagtac
     3061 atgaccttat gggactttcc tacttggcag tacatctacg tattagtcat cgctattacc
     3121 atggtgatgc ggttttggca gtacatcaat gggcgtggat agcggtttga ctcacgggga
     3181 tttccaagtc tccaccccat tgacgtcaat gggagtttgt tttggcacca aaatcaacgg
     3241 gactttccaa aatgtcgtaa caactccgcc ccattgacgc aaatgggcgg taggcgtgta
     3301 cggtgggagg tctatataag cagagctggt ttagtgaacc gtcagatccg ctagagatcc
     3361 gcggccgcta atacgactca ctatagggag agccgccacc gtgtcgtgac gtacggccac
     3421 catgggtagc ggacctaaga aaaagaggaa ggtggcggcc gctggaggtt ctggaagtgg
     3481 atccatggtg gataacaaat tcaataaaga agtgctggtg gcccggcagg aaatctattg
     3541 gctgccaaac ctgaattggg agcagaagtt tgccttcatc agtagcctga ctaatgatcc
     3601 atctcagagc gccaatctgc tggccgaagc caaaaaactg aacggtgccc agcccccaaa
     3661 aggaggaggt ggaagcggag gcggcggaag tgcggccgct ggatccggac gggctgacgc
     3721 attggacgat tttgatctgg atatgctggg aagtgacgcc ctcgatgatt ttgaccttga
     3781 catgcttggt tcggatgccc ttgatgactt tgacctcgac atgctcggca gtgacgccct
     3841 tgatgatttc gacctggaca tgctgattaa cactagtggc ggctctggcg gcggctccgg
     3901 aggctctagc cagtacctgc ccgacaccga cgacaggcac cggatcgagg aaaagcggaa
     3961 gcggacctac gagacattca agagcatcat gaagaagtcc cccttcagcg gccccaccga
     4021 ccctagacct ccacctagaa gaatcgccgt gcccagcaga tccagcgcca gcgtgccaaa
     4081 acctgccccc cagccttacc ccttcaccag cagcctgagc accatcaact acgacgagtt
     4141 ccctaccatg gtgttcccca gcggccagat ctctcaggcc tctgctctgg ctccagcccc
     4201 tcctcaggtg ctgcctcagg ctcctgctcc tgcaccagct ccagccatgg tgtctgcact
     4261 ggctcaggca ccagcacccg tgcctgtgct ggctcctgga cctccacagg ctgtggctcc
     4321 accagcccct aaacctacac aggccggcga gggcacactg tctgaagctc tgctgcagct
     4381 gcagttcgac gacgaggatc tgggagccct gctgggaaac agcaccgatc ctgccgtgtt
     4441 caccgacctg gccagcgtgg acaacagcga gttccagcag ctgctgaacc agggcatccc
     4501 tgtggcccct cacaccaccg agcccatgct gatggaatac cccgaggcca tcacccggct
     4561 cgtgacaggc gctcagaggc ctcctgatcc agctcctgcc cctctgggag caccaggcct
     4621 gcctaatgga ctgctgtctg gcgacgagga cttcagctct atcgccgata tggatttctc
     4681 agccttgctg ggctctggca gcggcagccg ggattccagg gaagggatgt ttttgccgaa
     4741 gcctgaggcc ggctccgcta ttagtgacgt gtttgagggc cgcgaggtgt gccagccaaa
     4801 acgaatccgg ccatttcatc ctccaggaag tccatgggcc aaccgcccac tccccgccag
     4861 cctcgcacca acaccaaccg gtccagtaca tgagccagtc gggtcactga ccccggcacc
     4921 agtccctcag ccactggacc cagcgcccgc agtgactccc gaggccagtc acctgttgga
     4981 ggaccccgat gaagagacga gccaggctgt caaagccctt cgggagatgg ccgatactgt
     5041 gattccccag aaggaagagg ctgcaatctg tggccaaatg gacctttccc atccgccccc
     5101 aaggggccat ctggatgagc tgacaaccac acttgagtcc atgaccgagg atctgaacct
     5161 ggactcaccc ctgaccccgg aattgaacga gattctggat accttcctga acgacgagtg
     5221 cctcttgcat gccatgcata tcagcacagg actgtccatc ttcgacacat ctctgtttgg
     5281 ttctggaagt ggatccagcc ccaagaagaa gagaaaggtg gaggccagct aaagcgaagt
     5341 caatcccaga attc
//
