LOCUS       V005190                10074 bp    DNA     circular SYN 13-MAY-2021
DEFINITION  Exported.
ACCESSION   V005190
VERSION     V005190
KEYWORDS    pHRdSV40-scFv-GCN4-sfGFP-VP64-GB1-NLS
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
            .
REFERENCE   1  (bases 1 to 10074)
  AUTHORS   Tanenbaum ME, Gilbert LA, Qi LS, Weissman JS, Vale RD
  TITLE     A Protein-Tagging System for Signal Amplification in Gene Expression
            and Fluorescence Imaging.
  JOURNAL   Cell. 2014 Oct 8. pii: S0092-8674(14)01227-6. doi:
            10.1016/j.cell.2014.09.039.
   PUBMED   25307933
REFERENCE   2  (bases 1 to 10074)
  TITLE     Direct Submission
REFERENCE   3  (bases 1 to 10074)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Cell. 2014
            Oct 8. pii: S0092-8674(14)01227-6. doi: 10.1016/j.cell.2014.09.039."
            SGRef: number: 2; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..10074
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        complement(119..137)
                     /label="SP6 promoter"
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(427..446)
                     /label="pBRrevBam"
                     /note="pBR322 vectors, tet region, downstream of BamHI,
                     reverse primer"
     primer_bind     780..802
                     /label="pGEX 3'"
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(840..858)
                     /label="pBRforEco"
                     /note="pBR322 vectors, upsteam of EcoRI site, forward
                     primer"
     promoter        926..1030
                     /label="AmpR promoter"
     CDS             1031..1888
                     /label="AmpR"
                     /note="beta-lactamase"
     rep_origin      2062..2650
                     /label="ori"
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
     primer_bind     2804..2821
                     /label="L4440"
                     /note="L4440 vector, forward primer"
     promoter        2896..3225
                     /label="SV40 promoter"
                     /note="SV40 enhancer and early promoter"
     intron          4359..4424
                     /label="small t intron"
                     /note="SV40 (simian virus 40) small t antigen intron"
     CDS             4554..4574
                     /label="SV40 NLS"
                     /note="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
     polyA_signal    4999..5133
                     /label="SV40 poly(A) signal"
                     /note="SV40 polyadenylation signal"
     LTR             5302..5935
                     /label="3' LTR"
                     /note="3' long terminal repeat (LTR) from HIV-1"
     misc_feature    5982..6107
                     /label="HIV-1 Psi"
                     /note="packaging signal of human immunodeficiency virus
                     type 1"
     misc_feature    6599..6832
                     /label="RRE"
                     /note="The Rev response element (RRE) of HIV-1 allows for
                     Rev-dependent mRNA export from the nucleus to the
                     cytoplasm."
     CDS             7017..7061
                     /label="gp41 peptide"
                     /note="antigenic peptide corresponding to amino acids 655
                     to 669 of the HIV envelope protein gp41 (Lutje Hulsik et
                     al., 2013)"
     CDS             7210..7251
                     /note="Protein Tat from Human immunodeficiency virus type 1
                     group M subtype B (isolate WMJ22). Accession#: P12509"
                     /label="Protein Tat"
     misc_feature    7346..7463
                     /label="cPPT/CTS"
                     /note="central polypurine tract and central termination
                     sequence of HIV-1"
     promoter        7526..7736
                     /label="SV40 promoter"
                     /note="SV40 early promoter"
     rep_origin      7595..7730
                     /label="SV40 ori"
                     /note="SV40 origin of replication"
     regulatory      7805..7814
                     /label="Kozak sequence"
                     /note="vertebrate consensus sequence for strong initiation
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             8552..8578
                     /label="HA"
                     /note="HA (human influenza hemagglutinin) epitope tag"
     CDS             8669..9379
                     /label="superfolder GFP"
                     /note="GFP variant that folds robustly even when fused to
                     poorly folded proteins (Nager et al., 2011)"
     CDS             9413..9562
                     /codon_start=1
                     /product="tetrameric repeat of the minimal activation
                     domain of herpes simplex virus VP16 (Beerli et al., 1998)"
                     /label="VP64"
                     /translation="DALDDFDLDMLGSDALDDFDLDMLGSDALDDFDLDMLGSDALDDF
                     DLDML"
     CDS             9593..9754
                     /codon_start=1
                     /product="B1 domain of Streptococcal protein G   "
                     /label="GB1"
                     /note="effective as a solubilizing fusion partner (Cheng
                     and Patel, 2004)"
                     /translation="YKLILNGKTLKGETTTEAVDAATAEKVFKQYANDNGVDGEWTYDD
                     ATKTFTVTE"
     CDS             9782..9832
                     /codon_start=1
                     /product="nuclear localization signal from the human T-cell
                     leukemia virus type 1 (HTLV-1) Rex protein"
                     /label="Rex NLS"
                     /translation="PKTRRRPRRSQRKRPPT"
ORIGIN
        1 tgagtgcttc aagtagtgtg tgcccgtctg ttgtgtgact ctggtaacta gagatccctc
       61 agaccctttt agtcagtgtg gaaaatctct agcagcatct agaattaatt ccgtgtattc
      121 tatagtgtca cctaaatcgt atgtgtatga tacataaggt tatgtattaa ttgtagccgc
      181 gttctaacga caatatgtac aagcctaatt gtgtagcatc tggcttactg aagcagaccc
      241 tatcatctct ctcgtaaact gccgtcagag tcggtttggt tggacgaacc ttctgagttt
      301 ctggtaacgc cgtcccgcac ccggaaatgg tcagcgaacc aatcagcagg gtcatcgcta
      361 gccagatcct ctacgccgga cgcatcgtgg ccggcatcac cggcgccaca ggtgcggttg
      421 ctggcgccta tatcgccgac atcaccgatg gggaagatcg ggctcgccac ttcgggctca
      481 tgagcgcttg tttcggcgtg ggtatggtgg caggccccgt ggccggggga ctgttgggcg
      541 ccatctcctt gcatgcacca ttccttgcgg cggcggtgct caacggcctc aacctactac
      601 tgggctgctt cctaatgcag gagtcgcata agggagagcg tcgaatggtg cactctcagt
      661 acaatctagc tctgatgccg catagttaag ccagccccga cacccgccaa cacccgctga
      721 cgcgccctga cgggcttgtc tgctcccggc atccgcttac agacaagctg tgaccgtctc
      781 cgggagctgc atgtgtcaga ggttttcacc gtcatcaccg aaacgcgcga gacgaaaggg
      841 cctcgtgata cgcctatttt tataggttaa tgtcatgata ataatggttt cttagacgtc
      901 aggtggcact tttcggggaa atgtgcgcgg aacccctatt tgtttatttt tctaaataca
      961 ttcaaatatg tatccgctca tgagacaata accctgataa atgcttcaat aatattgaaa
     1021 aaggaagagt atgagtattc aacatttccg tgtcgccctt attccctttt ttgcggcatt
     1081 ttgccttcct gtttttgctc acccagaaac gctggtgaaa gtaaaagatg ctgaagatca
     1141 gttgggtgca cgagtgggtt acatcgaact ggatctcaac agcggtaaga tccttgagag
     1201 ttttcgcccc gaagaacgtt ttccaatgat gagcactttt aaagttctgc tatgtggcgc
     1261 ggtattatcc cgtattgacg ccgggcaaga gcaactcggt cgccgcatac actattctca
     1321 gaatgacttg gttgagtact caccagtcac agaaaagcat cttacggatg gcatgacagt
     1381 aagagaatta tgcagtgctg ccataaccat gagtgataac actgcggcca acttacttct
     1441 gacaacgatc ggaggaccga aggagctaac cgcttttttg cacaacatgg gggatcatgt
     1501 aactcgcctt gatcgttggg aaccggagct gaatgaagcc ataccaaacg acgagcgtga
     1561 caccacgatg cctgtagcaa tggcaacaac gttgcgcaaa ctattaactg gcgaactact
     1621 tactctagct tcccggcaac aattaataga ctggatggag gcggataaag ttgcaggacc
     1681 acttctgcgc tcggcccttc cggctggctg gtttattgct gataaatctg gagccggtga
     1741 gcgtgggtct cgcggtatca ttgcagcact ggggccagat ggtaagccct cccgtatcgt
     1801 agttatctac acgacgggga gtcaggcaac tatggatgaa cgaaatagac agatcgctga
     1861 gataggtgcc tcactgatta agcattggta actgtcagac caagtttact catatatact
     1921 ttagattgat ttaaaacttc atttttaatt taaaaggatc taggtgaaga tcctttttga
     1981 taatctcatg accaaaatcc cttaacgtga gttttcgttc cactgagcgt cagaccccgt
     2041 agaaaagatc aaaggatctt cttgagatcc tttttttctg cgcgtaatct gctgcttgca
     2101 aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg gatcaagagc taccaactct
     2161 ttttccgaag gtaactggct tcagcagagc gcagatacca aatactgtcc ttctagtgta
     2221 gccgtagtta ggccaccact tcaagaactc tgtagcaccg cctacatacc tcgctctgct
     2281 aatcctgtta ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg ggttggactc
     2341 aagacgatag ttaccggata aggcgcagcg gtcgggctga acggggggtt cgtgcacaca
     2401 gcccagcttg gagcgaacga cctacaccga actgagatac ctacagcgtg agctatgaga
     2461 aagcgccacg cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg
     2521 aacaggagag cgcacgaggg agcttccagg gggaaacgcc tggtatcttt atagtcctgt
     2581 cgggtttcgc cacctctgac ttgagcgtcg atttttgtga tgctcgtcag gggggcggag
     2641 cctatggaaa aacgccagca acgcggcctt tttacggttc ctggcctttt gctggccttt
     2701 tgctcacatg ttctttcctg cgttatcccc tgattctgtg gataaccgta ttaccgcctt
     2761 tgagtgagct gataccgctc gccgcagccg aacgaccgag cgcagcgagt cagtgagcga
     2821 ggaagcggaa gagcgcccaa tacgcaaacc gcctctcccc gcgcgttggc cgattcatta
     2881 atgcagctgt ggaatgtgtg tcagttaggg tgtggaaagt ccccaggctc cccagcaggc
     2941 agaagtatgc aaagcatgca tctcaattag tcagcaacca ggtgtggaaa gtccccaggc
     3001 tccccagcag gcagaagtat gcaaagcatg catctcaatt agtcagcaac catagtcccg
     3061 cccctaactc cgcccatccc gcccctaact ccgcccagtt ccgcccattc tccgccccat
     3121 ggctgactaa ttttttttat ttatgcagag gccgaggccg cctcggcctc tgagctattc
     3181 cagaagtagt gaggaggctt ttttggaggc ctaggctttt gcaaaaagct tggacacaag
     3241 acaggcttgc gagatatgtt tgagaatacc actttatccc gcgtcaggga gaggcagtgc
     3301 gtaaaaagac gcggactcat gtgaaatact ggtttttagt gcgccagatc tctataatct
     3361 cgcgcaacct attttcccct cgaacacttt ttaagccgta gataaacagg ctgggacact
     3421 tcacatgagc gaaaaataca tcgtcacctg ggacatgttg cagatccatg cacgtaaact
     3481 cgcaagccga ctgatgcctt ctgaacaatg gaaaggcatt attgccgtaa gccgtggcgg
     3541 tctgtaccgg gtgcgttact ggcgcgtgaa ctgggtattc gtcatgtcga taccgtttgt
     3601 atttccagct acgatcacga caaccagcgc gagcttaaag tgctgaaacg cgcagaaggc
     3661 gatggcgaag gcttcatcgt tattgatgac ctggtggata ccggtggtac tgcggttgcg
     3721 attcgtgaaa tgtatccaaa agcgcacttt gtcaccatct tcgcaaaacc ggctggtcgt
     3781 ccgctggttg atgactatgt tgttgatatc ccgcaagata cctggattga acagccgtgg
     3841 gatatgggcg tcgtattcgt cccgccaatc tccggtcgct aatcttttca acgcctggca
     3901 ctgccgggcg ttgttctttt taacttcagg cgggttacaa tagtttccag taagtattct
     3961 ggaggctgca tccatgacac aggcaaacct gagcgaaacc ctgttcaaac cccgctttaa
     4021 acatcctgaa acctcgacgc tagtccgccg ctttaatcac ggcgcacaac cgcctgtgca
     4081 gtcggccctt gatggtaaaa ccatccctca ctggtatcgc atgattaacc gtctgatgtg
     4141 gatctggcgc ggcattgacc cacgcgaaat cctcgacgtc caggcacgta ttgtgatgag
     4201 cgatgccgaa cgtaccgacg atgatttata cgatacggtg attggctacc gtggcggcaa
     4261 ctggatttat gagtgggccc cggatctttg tgaaggaacc ttacttctgt ggtgtgacat
     4321 aattggacaa actacctaca gagatttaaa gctctaaggt aaatataaaa tttttaagtg
     4381 tataatgtgt taaactactg attctaattg tttgtgtatt ttagattcca acctatggaa
     4441 ctgatgaatg ggagcagtgg tggaatgcct ttaatgagga aaacctgttt tgctcagaag
     4501 aaatgccatc tagtgatgat gaggctactg ctgactctca acattctact cctccaaaaa
     4561 agaagagaaa ggtagaagac cccaaggact ttccttcaga attgctaagt tttttgagtc
     4621 atgctgtgtt tagtaataga actcttgctt gctttgctat ttacaccaca aaggaaaaag
     4681 ctgcactgct atacaagaaa attatggaaa aatattctgt aacctttata agtaggcata
     4741 acagttataa tcataacata ctgttttttc ttactccaca caggcataga gtgtctgcta
     4801 ttaataacta tgctcaaaaa ttgtgtacct ttagcttttt aatttgtaaa ggggttaata
     4861 aggaatattt gatgtatagt gccttgacta gagatcataa tcagccatac cacatttgta
     4921 gaggttttac ttgctttaaa aaacctccca cacctccccc tgaacctgaa acataaaatg
     4981 aatgcaattg ttgttgttaa cttgtttatt gcagcttata atggttacaa ataaagcaat
     5041 agcatcacaa atttcacaaa taaagcattt ttttcactgc attctagttg tggtttgtcc
     5101 aaactcatca atgtatctta tcatgtctgg atcaactgga taactcaagc taaccaaaat
     5161 catcccaaac ttcccacccc ataccctatt accactgcca attacctagt ggtttcattt
     5221 actctaaacc tgtgattcct ctgaattatt ttcattttaa agaaattgta tttgttaaat
     5281 atgtactaca aacttagtag ttggaagggc taattcactc ccaaagaaga caagatatcc
     5341 ttgatctgtg gatctaccac acacaaggct acttccctga ttagcagaac tacacaccag
     5401 ggccaggggt cagatatcca ctgacctttg gatggtgcta caagctagta ccagttgagc
     5461 cagataaggt agaagaggcc aataaaggag agaacaccag cttgttacac cctgtgagcc
     5521 tgcatgggat ggatgacccg gagagagaag tgttagagtg gaggtttgac agccgcctag
     5581 catttcatca cgtggcccga gagctgcatc cggagtactt caagaactgc tgatatcgag
     5641 cttgctacaa gggactttcc gctggggact ttccagggag gcgtggcctg ggcgggactg
     5701 gggagtggcg agccctcaga tcctgcatat aagcagctgc tttttgcctg tactgggtct
     5761 ctctggttag accagatctg agcctgggag ctctctggct aactagggaa cccactgctt
     5821 aagcctcaat aaagcttgcc ttgagtgctt caagtagtgt gtgcccgtct gttgtgtgac
     5881 tctggtaact agagatccct cagacccttt tagtcagtgt ggaaaatctc tagcagtggc
     5941 gcccgaacag ggacttgaaa gcgaaaggga aaccagagga gctctctcga cgcaggactc
     6001 ggcttgctga agcgcgcacg gcaagaggcg aggggcggcg actggtgagt acgccaaaaa
     6061 ttttgactag cggaggctag aaggagagag atgggtgcga gagcgtcagt attaagcggg
     6121 ggagaattag atcgcgatgg gaaaaaattc ggttaaggcc agggggaaag aaaaaatata
     6181 aattaaaaca tatagtatgg gcaagcaggg agctagaacg attcgcagtt aatcctggcc
     6241 tgttagaaac atcagaaggc tgtagacaaa tactgggaca gctacaacca tcccttcaga
     6301 caggatcaga agaacttaga tcattatata atacagtagc aaccctctat tgtgtgcatc
     6361 aaaggataga gataaaagac accaaggaag ctttagacaa gatagaggaa gagcaaaaca
     6421 aaagtaagac caccgcacag caagcggccg gtgatcttca gacctggacg atatatatga
     6481 gggacaattg gagaagtgaa ttatataaat ataaagtagt aaaaattgaa ccattaggag
     6541 tagcacccac caaggcaaag agaagagtgg tgcagagaga aaaaagagca gtgggaatag
     6601 gagctttgtt ccttgggttc ttgggagcag caggaagcac tatgggcgca gcgtcaatga
     6661 cgctgacggt acaggccaga caattattgt ctggtatagt gcagcagcag aacaatttgc
     6721 tgagggctat tgaggcgcaa cagcatctgt tgcaactcac agtctggggc atcaagcagc
     6781 tccaggcaag aatcctggct gtggaaagat acctaaagga tcaacagctc ctggggattt
     6841 ggggttgctc tggaaaactc atttgcacca ctgctgtgcc ttggaatgct agttggagta
     6901 ataaatctct ggaacagatt tggaatcaca cgacctggat ggagtgggac agagaaatta
     6961 acaattacac aagcttaata cactccttaa ttgaagaatc gcaaaaccag caagaaaaga
     7021 atgaacaaga attattggaa ttagataaat gggcaagttt gtggaattgg tttaacataa
     7081 caaattggct gtggtatata aaattattca taatgatagt aggaggcttg gtaggtttaa
     7141 gaatagtttt tgctgtactt tctatagtga atagagttag gcagggatat tcaccattat
     7201 cgtttcagac ccacctccca accccgaggg gacccgacag gcccgaagga atagaagaag
     7261 aaggtggaga gagagacaga gacagatcca ttcgattagt gaacggatct cgacggtatc
     7321 gccaaatggc agtattcatc cacaatttta aaagaaaagg ggggattggg gggtacagtg
     7381 caggggaaag aatagtagac ataatagcaa cagacataca aactaaagaa ttacaaaaac
     7441 aaattacaaa aattcaaaat tttcgggttt attacaggga cagcagagat ccagtttgga
     7501 tcgataagct tgatatcgaa ttcaggcagg cagaagtatg caaagcatgc atctcaatta
     7561 gtcagcaacc atagtcccgc ccctaactcc gcccatcccg cccctaactc cgcccagttc
     7621 cgcccattct ccgccccatg gctgactaat tttttttatt tatgcagagg ccgaggccgc
     7681 ctctgcctct gagctattcc agaagtagtg aggaggcttt tttggaggcc taggctacgc
     7741 gccagtcctc cgacagactg agtcgcccgg gggggatctg gagctctcga gaattctcac
     7801 gcgtgccacc atgggccccg acatcgtgat gacccagagc cccagcagcc tgagcgccag
     7861 cgtgggcgac cgcgtgacca tcacctgccg cagcagcacc ggcgccgtga ccaccagcaa
     7921 ctacgccagc tgggtgcagg agaagcccgg caagctgttc aagggcctga tcggcggcac
     7981 caacaaccgc gcccccggcg tgcccagccg cttcagcggc agcctgatcg gcgacaaggc
     8041 caccctgacc atcagcagcc tgcagcccga ggacttcgcc acctacttct gcgccctgtg
     8101 gtacagcaac cactgggtgt tcggccaggg caccaaggtg gagctgaagc gcggcggcgg
     8161 cggcagcggc ggcggcggca gcggcggcgg cggcagcagc ggcggcggca gcgaggtgaa
     8221 gctgctggag agcggcggcg gcctggtgca gcccggcggc agcctgaagc tgagctgcgc
     8281 cgtgagcggc ttcagcctga ccgactacgg cgtgaactgg gtgcgccagg cccccggccg
     8341 cggcctggag tggatcggcg tgatctgggg cgacggcatc accgactaca acagcgccct
     8401 gaaggaccgc ttcatcatca gcaaggacaa cggcaagaac accgtgtacc tgcagatgag
     8461 caaggtgcgc agcgacgaca ccgccctgta ctactgcgtg accggcctgt tcgactactg
     8521 gggccagggc accctggtga ccgtgagcag ctacccatac gatgttccag attacgctgg
     8581 tggaggcgga ggttctgggg gaggaggtag tggcggtggt ggttcaggag gcggcggaag
     8641 cttggatcca ggtggaggtg gaagcggtag caaaggagaa gaacttttca ctggagttgt
     8701 cccaattctt gttgaattag atggtgatgt taatgggcac aaattttctg tccgtggaga
     8761 gggtgaaggt gatgctacaa acggaaaact cacccttaaa tttatttgca ctactggaaa
     8821 actacctgtt ccgtggccaa cacttgtcac tactctgacc tatggtgttc aatgcttttc
     8881 ccgttatccg gatcacatga aacggcatga ctttttcaag agtgccatgc ccgaaggtta
     8941 tgtacaggaa cgcactatat ctttcaaaga tgacgggacc tacaagacgc gtgctgaagt
     9001 caagtttgaa ggtgataccc ttgttaatcg tatcgagtta aagggtattg attttaaaga
     9061 agatggaaac attcttggac acaaactcga gtacaacttt aactcacaca atgtatacat
     9121 cacggcagac aaacaaaaga atggaatcaa agctaacttc aaaattcgcc acaacgttga
     9181 agatggttcc gttcaactag cagaccatta tcaacaaaat actccaattg gcgatggccc
     9241 tgtcctttta ccagacaacc attacctgtc gacacaatct gtcctttcga aagatcccaa
     9301 cgaaaagcgt gaccacatgg tccttcttga gtttgtaact gctgctggga ttacacatgg
     9361 catggatgag ctctacaaag gtggaggtcg gaccggtggc ggtggcggag gggatgcttt
     9421 agacgatttt gacttagata tgcttggttc agacgcgtta gacgacttcg acctagacat
     9481 gttaggctca gatgcattgg acgacttcga tttagatatg ttgggctccg atgccctaga
     9541 tgactttgat ctagatatgc taggtagtgg aggaggatct cggaccgaag agtacaagct
     9601 tatcctgaac ggtaaaaccc tgaaaggtga aaccaccacc gaagctgttg acgctgctac
     9661 cgcggaaaaa gttttcaaac agtacgctaa cgacaacggt gttgacggtg aatggaccta
     9721 cgacgacgct accaaaacct tcacggtaac cgaaggtggt ggtagcggtg gtggtactag
     9781 tccaaaaaca aggaggagac cgcgaagatc acaacggaaa aggccgccta cgccatggcc
     9841 gtaagcggcc gcgactctag agtcgaccct ttaagaccaa tgacttacaa ggcagctgta
     9901 gatcttagcc actttttaaa agaaaagggg ggactggaag ggctaattca ctcccaacga
     9961 agacaagatc tgctttttgc ttgtactggg tctctctggt tagaccagat ctgagcctgg
    10021 gagctctctg gctaactagg gaacccactg cttaagcctc aataaagctt gcct
//
