LOCUS       sequence-364164-        5791 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  PCP tandem dimer binding to PP7 RNA stem loops coupled to VP16 
            activation domain; labeled with GFP.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5791)
  AUTHORS   Trojanowski J, Frank L, Rademacher A, Mucke N, Grigaitis P, Rippe K
  TITLE     Transcription activation is enhanced by multivalent interactions 
            independent of phase separation.
  JOURNAL   Mol Cell. 2022 May 4. pii: S1097-2765(22)00328-8. doi: 
            10.1016/j.molcel.2022.04.017.
  PUBMED    35537448
REFERENCE   2  (bases 1 to 5791)
  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"; journalName: "Mol Cell. 
            2022 May 4. pii: S1097-2765(22)00328-8. doi: 
            10.1016/j.molcel.2022.04.017."
COMMENT     Sequence Label: NLS-tdPCP-GFP-VP16
FEATURES             Location/Qualifiers
     source          1..5791
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        68..371
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        372..575
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     526..546
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     CDS             644..664
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             join(1433..1435,1436..1438,1439..2149)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(1433..1435,1436..1438,1439..2149)
                     /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(1478..1499)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1739..1758)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2086..2107
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     CDS             2183..2203
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     polyA_signal    2586..2707
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2623..2642)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2677..2696
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(2714..3169)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2851..2872)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3063..3082
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3196..3300
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3302..3659
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3302..3493
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3325..3345)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3510..3645
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3694..4488
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     CDS             3694..4488
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(3748..3767)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4358..4377
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(4676..4695)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    4720..4767
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5096..5684
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5585..5604
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 catgcattag ttattaatag taatcaatta cggggtcatt agttcatagc ccatatatgg
       61 agttccgcgt tacataactt acggtaaatg gcccgcctgg ctgaccgccc aacgaccccc
      121 gcccattgac gtcaataatg acgtatgttc ccatagtaac gccaataggg actttccatt
      181 gacgtcaatg ggtggagtat ttacggtaaa ctgcccactt ggcagtacat caagtgtatc
      241 atatgccaag tacgccccct attgacgtca atgacggtaa atggcccgcc tggcattatg
      301 cccagtacat gaccttatgg gactttccta cttggcagta catctacgta ttagtcatcg
      361 ctattaccat ggtgatgcgg ttttggcagt acatcaatgg gcgtggatag cggtttgaca
      421 cacggggatt tccaagtctc caccccattg acgtcaatgg gagtttgttt tggcaccaaa
      481 atcaacggga ctttccaaaa tgtcgtaaca actccgcccc attgacgcaa atgggcggta
      541 ggcgtgtacg gtgggaggtc ggggggggca gagctggttt agtgaaccgt cagatccgct
      601 agcgctaccg gactcagatc tcgagctcaa gcttctcatg ggcccaaaaa agaaaagaaa
      661 agttgggtca catatgctag cctccaaaac catcgttctt tcggtcggcg aggctactcg
      721 cactctgact gagatccagt ccaccgcaga ccgtcagatc ttcgaagaga aggtcgggcc
      781 tctggtgggt cggctgcgcc tcacggcttc gctccgtcaa aacggagcca agaccgcgta
      841 tcgcgtcaac ctaaaactgg atcaggcgga cgtcgttgat tccggacttc cgaaagtgcg
      901 ctacactcag gtatggtcgc acgacgtgac aatcgttgcg aatagcaccg aggcctcgcg
      961 caaatcgttg tacgatttga ccaagtccct cgtcgcgacc tcgcaggtcg aagatcttgt
     1021 cgtcaacctt gtgccgctgg gccgtgcgga tccgctagcc tccaaaacca tcgttctttc
     1081 ggtcggcgag gctactcgca ctctgactga gatccagtcc accgcagacc gtcagatctt
     1141 cgaagagaag gtcgggcctc tggtgggtcg gctgcgcctc acggcttcgc tccgtcaaaa
     1201 cggagccaag accgcgtatc gcgtcaacct aaaactggat caggcggacg tcgttgattc
     1261 cggacttccg aaagtgcgct acactcaggt atggtcgcac gacgtgacaa tcgttgcgaa
     1321 tagcaccgag gcctcgcgca aatcgttgta cgatttgacc aagtccctcg tcgcgacctc
     1381 gcaggtcgaa gatcttgtcg tcaaccttgt gccgctgggc cgtccaccgg tcatggtgag
     1441 caagggcgag gagctgttca ccggggtggt gcccatcctg gtcgagctgg acggcgacgt
     1501 aaacggccac aagttcagcg tgtccggcga gggcgagggc gatgccacct acggcaagct
     1561 gaccctgaag ttcatctgca ccaccggcaa gctgcccgtg ccctggccca ccctcgtgac
     1621 caccctgacc tacggcgtgc agtgcttcag ccgctacccc gaccacatga agcagcacga
     1681 cttcttcaag tccgccatgc ccgaaggcta cgtccaggag cgcaccatct tcttcaagga
     1741 cgacggcaac tacaagaccc gcgccgaggt gaagttcgag ggcgacaccc tggtgaaccg
     1801 catcgagctg aagggcatcg acttcaagga ggacggcaac atcctggggc acaagctgga
     1861 gtacaactac aacagccaca acgtctatat catggccgac aagcagaaga acggcatcaa
     1921 ggtgaacttc aagatccgcc acaacatcga ggacggcagc gtgcagctcg ccgaccacta
     1981 ccagcagaac acccccatcg gcgacggccc cgtgctgctg cccgacaacc actacctgag
     2041 cacccagtcc gccctgagca aagaccccaa cgagaagcgc gatcacatgg tcctgctgga
     2101 gttcgtgacc gccgccggga tcactctcgg catggacgag ctgtacaaga gcggccgcag
     2161 ggggcgtcga ctgggtaccc ctcctaagaa gaagaggaag gttgccccca ccatggataa
     2221 aaagaagaga aaggcccccc cgaccgatgt cagcctgggg gacgagctcc acttagacgg
     2281 cgaggacgtg gcgatggcgc atgccgacgc gctagacgat ttcgatctgg acatgttggg
     2341 ggacggggat tccccgggtc cgggatttac cccccacgac tccgccccct acggcgctct
     2401 ggatatggcc gacttcgagt ttgagcagat gtttaccgat gcccctctag acggggggat
     2461 ccaccggatc tagcggctag atcataatca gccataccac atttgtagag gttttacttg
     2521 ctttaaaaaa cctcccacac ctccccctga acctgaaaca taaaatgaat gcaattgttg
     2581 ttgttaactt gtttattgca gcttataatg gttacaaata aagcaatagc atcacaaatt
     2641 tcacaaataa agcatttttt tcactgcatt ctagttgtgg tttgtccaaa ctcatcaatg
     2701 tatcttaagg cgtaaattgt aagcgttaat attttgttaa aattcgcgtt aaatttttgt
     2761 taaatcagct cattttttaa ccaataggcc gaaatcggca aaatccctta taaatcaaaa
     2821 gaatagaccg agatagggtt gagtgttgtt ccagtttgga acaagagtcc actattaaag
     2881 aacgtggact ccaacgtcaa agggcgaaaa accgtctatc agggcgatgg cccactacgt
     2941 gaaccatcac cctaatcaag ttttttgggg tcgaggtgcc gtaaagcact aaatcggaac
     3001 cctaaaggga gcccccgatt tagagcttga cggggaaagc cggcgaacgt ggcgagaaag
     3061 gaagggaaga aagcgaaagg agcgggcgct agggcgctgg caagtgtagc ggtcacgctg
     3121 cgcgtaacca ccacacccgc cgcgcttaat gcgccgctac agggcgcgtc aggtggcact
     3181 tttcggggaa atgtgcgcgg aacccctatt tgtttatttt tctaaataca ttcaaatatg
     3241 tatccgctca tgagacaata accctgataa atgcttcaat aatattgaaa aaggaagagt
     3301 cctgaggcgg aaagaaccag ctgtggaatg tgtgtcagtt agggtgtgga aagtccccag
     3361 gctccccagc aggcagaagt atgcaaagca tgcatctcaa ttagtcagca accaggtgtg
     3421 gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag catgcatctc aattagtcag
     3481 caaccatagt cccgccccta actccgccca tcccgcccct aactccgccc agttccgccc
     3541 attctccgcc ccatggctga ctaatttttt ttatttatgc agaggccgag gccgcctcgg
     3601 cctctgagct attccagaag tagtgaggag gcttttttgg aggcctaggc ttttgcaaag
     3661 atcgatcaag agacaggatg aggatcgttt cgcatgattg aacaagatgg attgcacgca
     3721 ggttctccgg ccgcttgggt ggagaggcta ttcggctatg actgggcaca acagacaatc
     3781 ggctgctctg atgccgccgt gttccggctg tcagcgcagg ggcgcccggt tctttttgtc
     3841 aagaccgacc tgtccggtgc cctgaatgaa ctgcaagacg aggcagcgcg gctatcgtgg
     3901 ctggccacga cgggcgttcc ttgcgcagct gtgctcgacg ttgtcactga agcgggaagg
     3961 gactggctgc tattgggcga agtgccgggg caggatctcc tgtcatctca ccttgctcct
     4021 gccgagaaag tatccatcat ggctgatgca atgcggcggc tgcatacgct tgatccggct
     4081 acctgcccat tcgaccacca agcgaaacat cgcatcgagc gagcacgtac tcggatggaa
     4141 gccggtcttg tcgatcagga tgatctggac gaagagcatc aggggctcgc gccagccgaa
     4201 ctgttcgcca ggctcaaggc gagcatgccc gacggcgagg atctcgtcgt gacccatggc
     4261 gatgcctgct tgccgaatat catggtggaa aatggccgct tttctggatt catcgactgt
     4321 ggccggctgg gtgtggcgga ccgctatcag gacatagcgt tggctacccg tgatattgct
     4381 gaagagcttg gcggcgaatg ggctgaccgc ttcctcgtgc tttacggtat cgccgctccc
     4441 gattcgcagc gcatcgcctt ctatcgcctt cttgacgagt tcttctgagc gggactctgg
     4501 ggttcgaaat gaccgaccaa gcgacgccca acctgccatc acgagatttc gattccaccg
     4561 ccgccttcta tgaaaggttg ggcttcggaa tcgttttccg ggacgccggc tggatgatcc
     4621 tccagcgcgg ggatctcatg ctggagttct tcgcccaccc tagggggagg ctaactgaaa
     4681 cacggaagga gacaataccg gaaggaaccc gcgctatgac ggcaataaaa agacagaata
     4741 aaacgcacgg tgttgggtcg tttgttcata aacgcggggt tcggtcccag ggctggcact
     4801 ctgtcgatac cccaccgaga ccccattggg gccaatacgc ccgcgtttct tccttttccc
     4861 caccccaccc cccaagttcg ggtgaaggcc cagggctcgc agccaacgtc ggggcggcag
     4921 gccctgccat agcctcaggt tactcatata tactttagat tgatttaaaa cttcattttt
     4981 aatttaaaag gatctaggtg aagatccttt ttgataatct catgaccaaa atcccttaac
     5041 gtgagttttc gttccactga gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag
     5101 atcctttttt tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg
     5161 tggtttgttt gccggatcaa gagctaccaa ctctttttcc gaaggtaact ggcttcagca
     5221 gagcgcagat accaaatact gttcttctag tgtagccgta gttaggccac cacttcaaga
     5281 actctgtagc accgcctaca tacctcgctc tgctaatcct gttaccagtg gctgctgcca
     5341 gtggcgataa gtcgtgtctt accgggttgg actcaagacg atagttaccg gataaggcgc
     5401 agcggtcggg ctgaacgggg ggttcgtgca cacagcccag cttggagcga acgacctaca
     5461 ccgaactgag atacctacag cgtgagctat gagaaagcgc cacgcttccc gaagggagaa
     5521 aggcggacag gtatccggta agcggcaggg tcggaacagg agagcgcacg agggagcttc
     5581 cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc
     5641 gtcgattttt gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg
     5701 cctttttacg gttcctggcc ttttgctggc cttttgctca catgttcttt cctgcgttat
     5761 cccctgattc tgtggataac cgtattaccg c
//
