LOCUS       sequence-196760-        3198 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  Transcription activator (transactivation domain of viral VP16 
            protein) that is constitutively recruited tetO arrays.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 3198)
  AUTHORS   Rademacher A, Erdel F, Trojanowski J, Schumacher S, Rippe K
  TITLE     Real-time observation of light-controlled transcription in living 
            cells.
  JOURNAL   J Cell Sci. 2017 Dec 15;130(24):4213-4224. doi: 10.1242/jcs.205534. 
            Epub 2017 Nov 9.
  PUBMED    29122982
REFERENCE   2  (bases 1 to 3198)
  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"; doi: 
            "10.1242/jcs.205534"; journalName: "J Cell Sci"; date: "2017-12-15- 
            15"; volume: "130"; issue: "24"; pages: "4213-4224"
COMMENT     Sequence Label: pEGFP-TetR-NLS-VP16
FEATURES             Location/Qualifiers
     source          1..3198
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     primer_bind     225..244
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     complement(452..472)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        457..786
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      637..772
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     regulatory      825..834
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(831..833,834..836,837..1547)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(831..833,834..836,837..1547)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(876..897)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1137..1156)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1484..1505
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     CDS             1554..2177
                     /codon_start=1
                     /gene="tetR from transposon Tn10"
                     /product="tetracycline repressor TetR"
                     /label=TetR
                     /note="TetR binds to the tetracycline operator tetO to 
                     inhibit transcription. This inhibition can be relieved by 
                     adding tetracycline or doxycycline."
                     /translation="MVSRLDKSKVINSALELLNEVGIEGLTTRKLAQKLGVEQPTLYWH
                     VKNKRALLDALAIEMLDRHHTHFCPLEGESWQDFLRNNAKSFRCALLSHRDGAKVHLGT
                     RPTEKQYETLENQLAFLCQQGFSLENALYALSAVGHFTLGCVLEDQEHQVAKEERETPT
                     TDSMPPLLRQAIELFDHQGAEPAFLFGLELIICGLEKQLKCESGS"
     primer_bind     complement(1630..1649)
                     /label=Tet-R
                     /note="Tetracycline resistance gene, reverse primer"
     CDS             2199..2219
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     polyA_signal    2602..2723
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2639..2658)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2693..2712
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(2730..3184)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2867..2888)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3078..3097
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
ORIGIN
        1 accgggttgg actcaagacg atagttaccg gataaggcgc agcggtcggg ctgaacgggg
       61 ggttcgtgca cacagcccag cttggagcga acgacctaca ccgaactgag atacctacag
      121 cgtgagctat gagaaagcgc cacgcttccc gaagggagaa aggcggacag gtatccggta
      181 agcggcaggg tcggaacagg agagcgcacg agggagcttc cagggggaaa cgcctggtat
      241 ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt gtgatgctcg
      301 tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc
      361 ttttgctggc cttttgctca catgttcttt cctgcgttat cccctgattc tgtggataac
      421 cgtattaccg ccatgcatta gttattactg tggaatgtgt gtcagttagg gtgtggaaag
      481 tccccaggct ccccagcagg cagaagtatg caaagcatgc atctcaatta gtcagcaacc
      541 aggtgtggaa agtccccagg ctccccagca ggcagaagta tgcaaagcat gcatctcaat
      601 tagtcagcaa ccatagtccc gcccctaact ccgcccatcc cgcccctaac tccgcccagt
      661 tccgcccatt ctccgcccca tggctgacta atttttttta tttatgcaga ggccgaggcc
      721 gcctcggcct ctgagctatt ccagaagtag tgaggaggct tttttggagg cctaggcttt
      781 tgcaaaaagc tagcgctacc ggggccgctc tagcgctacc ggtcgccacc atggtgagca
      841 agggcgagga gctgttcacc ggggtggtgc ccatcctggt cgagctggac ggcgacgtaa
      901 acggccacaa gttcagcgtg tccggcgagg gcgagggcga tgccacctac ggcaagctga
      961 ccctgaagtt catctgcacc accggcaagc tgcccgtgcc ctggcccacc ctcgtgacca
     1021 ccctgaccta cggcgtgcag tgcttcagcc gctaccccga ccacatgaag cagcacgact
     1081 tcttcaagtc cgccatgccc gaaggctacg tccaggagcg caccatcttc ttcaaggacg
     1141 acggcaacta caagacccgc gccgaggtga agttcgaggg cgacaccctg gtgaaccgca
     1201 tcgagctgaa gggcatcgac ttcaaggagg acggcaacat cctggggcac aagctggagt
     1261 acaactacaa cagccacaac gtctatatca tggccgacaa gcagaagaac ggcatcaagg
     1321 tgaacttcaa gatccgccac aacatcgagg acggcagcgt gcagctcgcc gaccactacc
     1381 agcagaacac ccccatcggc gacggccccg tgctgctgcc cgacaaccac tacctgagca
     1441 cccagtccgc cctgagcaaa gaccccaacg agaagcgcga tcacatggtc ctgctggagt
     1501 tcgtgaccgc cgccgggatc actctcggca tggacgagct gtacaagtcc ggaatggtgt
     1561 ctagattaga taaaagtaaa gtgattaaca gcgcattaga gctgcttaat gaggtcggaa
     1621 tcgaaggttt aacaacccgt aaactcgccc agaagctagg tgtagagcag cctacattgt
     1681 attggcatgt aaaaaataag cgggctttgc tcgacgcctt agccattgag atgttagata
     1741 ggcaccatac tcacttttgc cctttagaag gggaaagctg gcaagatttt ttacgtaata
     1801 acgctaaaag ttttagatgt gctttactaa gtcatcgcga tggagcaaaa gtacatttag
     1861 gtacacggcc tacagaaaaa cagtatgaaa ctctcgaaaa tcaattagcc tttttatgcc
     1921 aacaaggttt ttcactagag aatgcattat atgcactcag cgctgtgggg cattttactt
     1981 taggttgcgt attggaagat caagagcatc aagtcgctaa agaagaaagg gaaacaccta
     2041 ctactgatag tatgccgcca ttattacgac aagctatcga attatttgat caccaaggtg
     2101 cagagccagc cttcttattc ggccttgaat tgatcatatg cggattagaa aaacaactta
     2161 aatgtgaaag tgggtctgac cgtcgactgg gtacccctcc taagaagaag aggaaggttg
     2221 cccccaccat ggataaaaag aagagaaagg cccccccgac cgatgtcagc ctgggggacg
     2281 agctccactt agacggcgag gacgtggcga tggcgcatgc cgacgcgcta gacgatttcg
     2341 atctggacat gttgggggac ggggattccc cgggtccggg atttaccccc cacgactccg
     2401 ccccctacgg cgctctggat atggccgact tcgagtttga gcagatgttt accgatgccc
     2461 ctctagacgg ggggatccac cggatctaga taactgatca taatcagcca taccacattt
     2521 gtagaggttt tacttgcttt aaaaaacctc ccacacctcc ccctgaacct gaaacataaa
     2581 atgaatgcaa ttgttgttgt taacttgttt attgcagctt ataatggtta caaataaagc
     2641 aatagcatca caaatttcac aaataaagca tttttttcac tgcattctag ttgtggtttg
     2701 tccaaactca tcaatgtatc ttaaggcgta aattgtaagc gttaatattt tgttaaaatt
     2761 cgcgttaaat ttttgttaaa tcagctcatt ttttaaccaa taggccgaaa tcggcaaaat
     2821 cccttataaa tcaaaagaat agaccgagat agggttgagt gttgttccag tttggaacaa
     2881 gagtccacta ttaaagaacg tggactccaa cgtcaaaggg cgaaaaaccg tctatcaggg
     2941 cgatggccca ctacgtgaac catcacccta atcaagtttt ttggggtcga ggtgccgtaa
     3001 gcactaaatc ggaaccctaa agggagcccc cgatttagag cttgacgggg aaagccggcg
     3061 aacgtggcga gaaaggaagg gaagaaagcg aaaggagcgg gcgctagggc gctggcaagt
     3121 gtagcggtca cgctgcgcgt aaccaccaca cccgccgcgc ttaatgcgcc gctacagggc
     3181 gcgtcaggtg gcactttt
//
