LOCUS       sequence-79487-e        7485 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Artificial transcription factor with VP64.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7485)
  AUTHORS   Senecal A, Munsky B, Proux F, Ly N, Braye FE, Zimmer C, Mueller F, 
            Darzacq X
  TITLE     Transcription Factors Modulate c-Fos Transcriptional Bursts.
  JOURNAL   Cell Rep. 2014 Jun 25. pii: S2211-1247(14)00447-1. doi: 
            10.1016/j.celrep.2014.05.053.
  PUBMED    24981864
REFERENCE   2  (bases 1 to 7485)
  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: "Cell Rep. 
            2014 Jun 25. pii: S2211-1247(14)00447-1. doi: 
            10.1016/j.celrep.2014.05.053."
COMMENT     Sequence Label: pAF1-VP64
FEATURES             Location/Qualifiers
     source          1..7485
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(44..63)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        235..614
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        615..818
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     769..789
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     promoter        863..881
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      917..926
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             complement(1679..2086)
                     /codon_start=1
                     /gene="lacZ fragment"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /label=lacZ-alpha
                     /translation="MTMITPSAQLTLTKGNKSWRTSRGGPVPNSPYSESYYARSLAVVL
                     QRRDWENPGVTQLNRLAAHPPFASWRNSEEARTDRPSQQLRSLNGEWDAPCSGARLQGG
                     TRQCIARCMHCLHCGHAATFCNLHKLACVQL"
     primer_bind     1936..1958
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     1950..1967
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     1951..1967
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        1977..1995
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2043..2063)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(2043..2061)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(2082..2098)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    2106..2122
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /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(join(2130..2136,2137..2154,2155..2160))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2175..2196
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     CDS             2889..3038
                     /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"
     primer_bind     complement(3079..3096)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3085..3309
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      3355..3783
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3442..3461)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3652..3673
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3792..3812)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3797..4126
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3977..4112
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     4039..4058
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             4193..4987
                     /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
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(4247..4266)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4857..4876
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    5161..5282
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5198..5217)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5252..5271
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5331..5347)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    5355..5371
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /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(join(5379..5385,5386..5403,5404..5409))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5424..5445
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5562..5579)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5733..6318)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5813..5832)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6489..7280,7281..7349))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     7112..7131
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7350..7454)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gacggatcgg gagatctccc gatcccctat ggtgcactct cagtacaatc tgctctgatg
       61 ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg
      121 cgagcaaaat ttaagctaca acaaggcaag gcttgaccga caattgcatg aagaatctgc
      181 ttagggttag gcgttttgcg ctgcttcgcg atgtacgggc cagatatacg cgttgacatt
      241 gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata
      301 tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc
      361 cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc
      421 attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt
      481 atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt
      541 atgcccagta catgacctta tgggactttc ctacttggca gtacatctac gtattagtca
      601 tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg
      661 actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc
      721 aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg
      781 gtaggcgtgt acggtgggag gtctatataa gcagagctct ctggctaact agagaaccca
      841 ctgcttactg gcttatcgaa attaatacga ctcactatag ggagacccaa gctggctagc
      901 gtttaaactt aagcttgcca ccatggaccc cattcgtccg cgcaggccaa gtcctgcccg
      961 cgagcttctg cccggacccc aaccggatag ggttcagccg actgcagatc gtggggtgtc
     1021 tgcgcctgct ggcagccctc tggatggctt gcccgctcgg cggacggtgt cccggacccg
     1081 gctgccatct ccccctgcgc cctcacctgc gttctcggcg ggcagcttca gcgatctgct
     1141 ccgtccgttc gatccgtcgc ttcttgatac atcgcttctt gattcgatgc ctgccgtcgg
     1201 cacgccgcat acagcggctg ccccagcaga gtgggatgag gcgcaatcgg ctctgcgtgc
     1261 agccgatgac ccgccaccca ccgtgcgtgt cgctgtcact gccgcgcggc cgccgcgcgc
     1321 caagccggcc ccgcgacggc gtgctgcgca accctccgac gcttcgccgg ccgcgcaggt
     1381 ggatctacgc acgctcggct acagtcagca gcagcaagag aagatcaaac cgaaggtgcg
     1441 ttcgacagtg gcgcagcacc acgaggcact ggtgggccat gggtttacac acgcgcacat
     1501 cgttgcgctc agccaacacc cggcagcgtt agggaccgtc gctgtcacgt atcagcacat
     1561 aatcacggcg ttgccagagg cgacacacga agacatcgtt ggcgtcggca aacagtggtc
     1621 cggcgcacgc gccctgtagg ccttgctcac ggatgcgggg gagttgagag gtccgccgtt
     1681 acagttggac acaggccaac ttgtgaagat tgcaaaacgt ggcggcgtga ccgcaatgga
     1741 ggcagtgcat gcatcgcgca atgcactgac gggtgccccc ctggagacgg gcgccgctac
     1801 agggcgcgtc ccattcgcca ttcaggctgc gcaactgttg ggaagggcga tcggtgcggg
     1861 cctcttcgct attacgccag ctggcgaaag ggggatgtgc tgcaaggcga ttaagttggg
     1921 taacgccagg gttttcccag tcacgacgtt gtaaaacgac ggccagtgag cgcgcgtaat
     1981 acgactcact atagggcgaa ttgggtaccg ggccccccct cgaggtcctc cagcttttgt
     2041 tccctttagt gagggttaat tgcgcgcttg gcgtaatcat ggtcatagct gtttcctgtg
     2101 tgaaattgtt atccgctcac aattccacac aacatacgag ccggaagcat aaagtgtaaa
     2161 gcctggggtg cctaatgagt gagctaactc acattaattg cgttgcgctc actgcccgct
     2221 ttccaccggt cgtctccaac gaccacctcg tcgccttggc ctgcctcggc ggacgtcctg
     2281 ccatggatgc agtgaaaaag ggattgccgc acgcgccgga attgatcaga agagtcaatc
     2341 gccgtattgg cgaacgcacg tcccatcgcg ttgccgacta cgcgcaagtg gttcgcgtgc
     2401 tggagttttt ccagtgccac tcccacccag cgtacgcatt tgatgaggcc atgacgcagt
     2461 tcgggatgag caggaacggg ttggtacagc tctttcgcag agtgggcgtc accgaactcg
     2521 aagcccgcgg tggaacgctc cccccagcct cgcagcgttg ggaccgtatc ctccaggcat
     2581 cagggatgaa aagggccaaa ccgtccccta cttcagctca aacaccggat caggcgtctt
     2641 tgcatgcatt cgccgattcg ctggagcgtg accttgatgc gcccagccca atgcacgagg
     2701 gagatcagac gcgggcaagc agccgtaaac ggtcccgatc ggatcgtgct gtcaccggcc
     2761 cctccgcaca gcaggctgtc gaggtgcgcg ttcccgaaca gcgcgatgcg ctgcatttgc
     2821 ccctcagctg gagggtaaaa cgcccgcgta ccaggatctg gggcggcctc ccggatcccg
     2881 gacgggctga cgcattggac gattttgatc tggatatgct gggaagtgac gccctcgatg
     2941 attttgacct tgacatgctt ggttcggatg cccttgatga ctttgacctc gacatgctcg
     3001 gcagtgacgc ccttgatgat ttcgacctgg acatgctgat taactaatct agagggcccg
     3061 tttaaacccg ctgatcagcc tcgactgtgc cttctagttg ccagccatct gttgtttgcc
     3121 cctcccccgt gccttccttg accctggaag gtgccactcc cactgtcctt tcctaataaa
     3181 atgaggaaat tgcatcgcat tgtctgagta ggtgtcattc tattctgggg ggtggggtgg
     3241 ggcaggacag caagggggag gattgggaag acaatagcag gcatgctggg gatgcggtgg
     3301 gctctatggc ttctgaggcg gaaagaacca gctggggctc tagggggtat ccccacgcgc
     3361 cctgtagcgg cgcattaagc gcggcgggtg tggtggttac gcgcagcgtg accgctacac
     3421 ttgccagcgc cctagcgccc gctcctttcg ctttcttccc ttcctttctc gccacgttcg
     3481 ccggctttcc ccgtcaagct ctaaatcggg ggctcccttt agggttccga tttagtgctt
     3541 tacggcacct cgaccccaaa aaacttgatt agggtgatgg ttcacgtagt gggccatcgc
     3601 cctgatagac ggtttttcgc cctttgacgt tggagtccac gttctttaat agtggactct
     3661 tgttccaaac tggaacaaca ctcaacccta tctcggtcta ttcttttgat ttataaggga
     3721 ttttgccgat ttcggcctat tggttaaaaa atgagctgat ttaacaaaaa tttaacgcga
     3781 attaattctg tggaatgtgt gtcagttagg gtgtggaaag tccccaggct ccccagcagg
     3841 cagaagtatg caaagcatgc atctcaatta gtcagcaacc aggtgtggaa agtccccagg
     3901 ctccccagca ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa ccatagtccc
     3961 gcccctaact ccgcccatcc cgcccctaac tccgcccagt tccgcccatt ctccgcccca
     4021 tggctgacta atttttttta tttatgcaga ggccgaggcc gcctctgcct ctgagctatt
     4081 ccagaagtag tgaggaggct tttttggagg cctaggcttt tgcaaaaagc tcccgggagc
     4141 ttgtatatcc attttcggat ctgatcaaga gacaggatga ggatcgtttc gcatgattga
     4201 acaagatgga ttgcacgcag gttctccggc cgcttgggtg gagaggctat tcggctatga
     4261 ctgggcacaa cagacaatcg gctgctctga tgccgccgtg ttccggctgt cagcgcaggg
     4321 gcgcccggtt ctttttgtca agaccgacct gtccggtgcc ctgaatgaac tgcaggacga
     4381 ggcagcgcgg ctatcgtggc tggccacgac gggcgttcct tgcgcagctg tgctcgacgt
     4441 tgtcactgaa gcgggaaggg actggctgct attgggcgaa gtgccggggc aggatctcct
     4501 gtcatctcac cttgctcctg ccgagaaagt atccatcatg gctgatgcaa tgcggcggct
     4561 gcatacgctt gatccggcta cctgcccatt cgaccaccaa gcgaaacatc gcatcgagcg
     4621 agcacgtact cggatggaag ccggtcttgt cgatcaggat gatctggacg aagagcatca
     4681 ggggctcgcg ccagccgaac tgttcgccag gctcaaggcg cgcatgcccg acggcgagga
     4741 tctcgtcgtg acccatggcg atgcctgctt gccgaatatc atggtggaaa atggccgctt
     4801 ttctggattc atcgactgtg gccggctggg tgtggcggac cgctatcagg acatagcgtt
     4861 ggctacccgt gatattgctg aagagcttgg cggcgaatgg gctgaccgct tcctcgtgct
     4921 ttacggtatc gccgctcccg attcgcagcg catcgccttc tatcgccttc ttgacgagtt
     4981 cttctgagcg ggactctggg gttcgaaatg accgaccaag cgacgcccaa cctgccatca
     5041 cgagatttcg attccaccgc cgccttctat gaaaggttgg gcttcggaat cgttttccgg
     5101 gacgccggct ggatgatcct ccagcgcggg gatctcatgc tggagttctt cgcccacccc
     5161 aacttgttta ttgcagctta taatggttac aaataaagca atagcatcac aaatttcaca
     5221 aataaagcat ttttttcact gcattctagt tgtggtttgt ccaaactcat caatgtatct
     5281 tatcatgtct gtataccgtc gacctctagc tagagcttgg cgtaatcatg gtcatagctg
     5341 tttcctgtgt gaaattgtta tccgctcaca attccacaca acatacgagc cggaagcata
     5401 aagtgtaaag cctggggtgc ctaatgagtg agctaactca cattaattgc gttgcgctca
     5461 ctgcccgctt tccagtcggg aaacctgtcg tgccagctgc attaatgaat cggccaacgc
     5521 gcggggagag gcggtttgcg tattgggcgc tcttccgctt cctcgctcac tgactcgctg
     5581 cgctcggtcg ttcggctgcg gcgagcggta tcagctcact caaaggcggt aatacggtta
     5641 tccacagaat caggggataa cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc
     5701 aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc ccctgacgag
     5761 catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact ataaagatac
     5821 caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct gccgcttacc
     5881 ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcatag ctcacgctgt
     5941 aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc
     6001 gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggtaaga
     6061 cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc gaggtatgta
     6121 ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag aagaacagta
     6181 tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg tagctcttga
     6241 tccggcaaac aaaccaccgc tggtagcggt ttttttgttt gcaagcagca gattacgcgc
     6301 agaaaaaaag gatctcaaga agatcctttg atcttttcta cggggtctga cgctcagtgg
     6361 aacgaaaact cacgttaagg gattttggtc atgagattat caaaaaggat cttcacctag
     6421 atccttttaa attaaaaatg aagttttaaa tcaatctaaa gtatatatga gtaaacttgg
     6481 tctgacagtt accaatgctt aatcagtgag gcacctatct cagcgatctg tctatttcgt
     6541 tcatccatag ttgcctgact ccccgtcgtg tagataacta cgatacggga gggcttacca
     6601 tctggcccca gtgctgcaat gataccgcga gacccacgct caccggctcc agatttatca
     6661 gcaataaacc agccagccgg aagggccgag cgcagaagtg gtcctgcaac tttatccgcc
     6721 tccatccagt ctattaattg ttgccgggaa gctagagtaa gtagttcgcc agttaatagt
     6781 ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg
     6841 gcttcattca gctccggttc ccaacgatca aggcgagtta catgatcccc catgttgtgc
     6901 aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg
     6961 ttatcactca tggttatggc agcactgcat aattctctta ctgtcatgcc atccgtaaga
     7021 tgcttttctg tgactggtga gtactcaacc aagtcattct gagaatagtg tatgcggcga
     7081 ccgagttgct cttgcccggc gtcaatacgg gataataccg cgccacatag cagaacttta
     7141 aaagtgctca tcattggaaa acgttcttcg gggcgaaaac tctcaaggat cttaccgctg
     7201 ttgagatcca gttcgatgta acccactcgt gcacccaact gatcttcagc atcttttact
     7261 ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata
     7321 agggcgacac ggaaatgttg aatactcata ctcttccttt ttcaatatta ttgaagcatt
     7381 tatcagggtt attgtctcat gagcggatac atatttgaat gtatttagaa aaataaacaa
     7441 ataggggttc cgcgcacatt tccccgaaaa gtgccacctg acgtc
//
