LOCUS       sequence-79486-e        7705 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Artificial transcription factor with VP16.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7705)
  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 7705)
  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-VP16
FEATURES             Location/Qualifiers
     source          1..7705
                     /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             3030..3263
                     /codon_start=1
                     /gene="UL48"
                     /product="transcriptional activation domain of herpes 
                     simplex virus protein VP16 (Triezenberg et al., 1988; 
                     Cousens et al., 1989)"
                     /label=VP16 AD
                     /translation="APPTDVSLGDELHLDGEDVAMAHADALDDFDLDMLGDGDSPGPGF
                     TPHDSAPYGALDMADFEFEQMFTDALGIDEYGG"
     primer_bind     complement(3299..3316)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3305..3529
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      3575..4003
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3662..3681)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3872..3893
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4012..4032)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        4017..4346
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      4197..4332
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     4259..4278
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             4413..5207
                     /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(4467..4486)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5077..5096
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    5381..5502
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5418..5437)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5472..5491
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5551..5567)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    5575..5591
                     /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(5599..5605,5606..5623,5624..5629))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5644..5665
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5782..5799)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5953..6538)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6033..6052)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6709..7500,7501..7569))
                     /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     7332..7351
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7570..7674)
                     /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 gccctggagg 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 ccggatccac
     2881 cgtacagccg cgcgcgtacg aaaaacaatt acgggtctac catcgagggc ctgctcgatc
     2941 tcccggacga cgacgccccc gaagaggcgg ggctggcggc tccgcgcctg tcctttctcc
     3001 ccgcgggaca cacgcgcaga ctgtcgacgg cccccccgac cgatgtcagc ctgggggacg
     3061 agctccactt agacggcgag gacgtggcga tggcgcatgc cgacgcgcta gacgatttcg
     3121 atctggacat gttgggggac ggggattccc cgggtccggg atttaccccc cacgactccg
     3181 ccccctacgg cgctctggat atggccgact tcgagtttga gcagatgttt accgatgccc
     3241 ttggaattga cgagtacggt gggtaggtct agagggcccg tttaaacccg ctgatcagcc
     3301 tcgactgtgc cttctagttg ccagccatct gttgtttgcc cctcccccgt gccttccttg
     3361 accctggaag gtgccactcc cactgtcctt tcctaataaa atgaggaaat tgcatcgcat
     3421 tgtctgagta ggtgtcattc tattctgggg ggtggggtgg ggcaggacag caagggggag
     3481 gattgggaag acaatagcag gcatgctggg gatgcggtgg gctctatggc ttctgaggcg
     3541 gaaagaacca gctggggctc tagggggtat ccccacgcgc cctgtagcgg cgcattaagc
     3601 gcggcgggtg tggtggttac gcgcagcgtg accgctacac ttgccagcgc cctagcgccc
     3661 gctcctttcg ctttcttccc ttcctttctc gccacgttcg ccggctttcc ccgtcaagct
     3721 ctaaatcggg ggctcccttt agggttccga tttagtgctt tacggcacct cgaccccaaa
     3781 aaacttgatt agggtgatgg ttcacgtagt gggccatcgc cctgatagac ggtttttcgc
     3841 cctttgacgt tggagtccac gttctttaat agtggactct tgttccaaac tggaacaaca
     3901 ctcaacccta tctcggtcta ttcttttgat ttataaggga ttttgccgat ttcggcctat
     3961 tggttaaaaa atgagctgat ttaacaaaaa tttaacgcga attaattctg tggaatgtgt
     4021 gtcagttagg gtgtggaaag tccccaggct ccccagcagg cagaagtatg caaagcatgc
     4081 atctcaatta gtcagcaacc aggtgtggaa agtccccagg ctccccagca ggcagaagta
     4141 tgcaaagcat gcatctcaat tagtcagcaa ccatagtccc gcccctaact ccgcccatcc
     4201 cgcccctaac tccgcccagt tccgcccatt ctccgcccca tggctgacta atttttttta
     4261 tttatgcaga ggccgaggcc gcctctgcct ctgagctatt ccagaagtag tgaggaggct
     4321 tttttggagg cctaggcttt tgcaaaaagc tcccgggagc ttgtatatcc attttcggat
     4381 ctgatcaaga gacaggatga ggatcgtttc gcatgattga acaagatgga ttgcacgcag
     4441 gttctccggc cgcttgggtg gagaggctat tcggctatga ctgggcacaa cagacaatcg
     4501 gctgctctga tgccgccgtg ttccggctgt cagcgcaggg gcgcccggtt ctttttgtca
     4561 agaccgacct gtccggtgcc ctgaatgaac tgcaggacga ggcagcgcgg ctatcgtggc
     4621 tggccacgac gggcgttcct tgcgcagctg tgctcgacgt tgtcactgaa gcgggaaggg
     4681 actggctgct attgggcgaa gtgccggggc aggatctcct gtcatctcac cttgctcctg
     4741 ccgagaaagt atccatcatg gctgatgcaa tgcggcggct gcatacgctt gatccggcta
     4801 cctgcccatt cgaccaccaa gcgaaacatc gcatcgagcg agcacgtact cggatggaag
     4861 ccggtcttgt cgatcaggat gatctggacg aagagcatca ggggctcgcg ccagccgaac
     4921 tgttcgccag gctcaaggcg cgcatgcccg acggcgagga tctcgtcgtg acccatggcg
     4981 atgcctgctt gccgaatatc atggtggaaa atggccgctt ttctggattc atcgactgtg
     5041 gccggctggg tgtggcggac cgctatcagg acatagcgtt ggctacccgt gatattgctg
     5101 aagagcttgg cggcgaatgg gctgaccgct tcctcgtgct ttacggtatc gccgctcccg
     5161 attcgcagcg catcgccttc tatcgccttc ttgacgagtt cttctgagcg ggactctggg
     5221 gttcgaaatg accgaccaag cgacgcccaa cctgccatca cgagatttcg attccaccgc
     5281 cgccttctat gaaaggttgg gcttcggaat cgttttccgg gacgccggct ggatgatcct
     5341 ccagcgcggg gatctcatgc tggagttctt cgcccacccc aacttgttta ttgcagctta
     5401 taatggttac aaataaagca atagcatcac aaatttcaca aataaagcat ttttttcact
     5461 gcattctagt tgtggtttgt ccaaactcat caatgtatct tatcatgtct gtataccgtc
     5521 gacctctagc tagagcttgg cgtaatcatg gtcatagctg tttcctgtgt gaaattgtta
     5581 tccgctcaca attccacaca acatacgagc cggaagcata aagtgtaaag cctggggtgc
     5641 ctaatgagtg agctaactca cattaattgc gttgcgctca ctgcccgctt tccagtcggg
     5701 aaacctgtcg tgccagctgc attaatgaat cggccaacgc gcggggagag gcggtttgcg
     5761 tattgggcgc tcttccgctt cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg
     5821 gcgagcggta tcagctcact caaaggcggt aatacggtta tccacagaat caggggataa
     5881 cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc
     5941 gttgctggcg tttttccata ggctccgccc ccctgacgag catcacaaaa atcgacgctc
     6001 aagtcagagg tggcgaaacc cgacaggact ataaagatac caggcgtttc cccctggaag
     6061 ctccctcgtg cgctctcctg ttccgaccct gccgcttacc ggatacctgt ccgcctttct
     6121 cccttcggga agcgtggcgc tttctcatag ctcacgctgt aggtatctca gttcggtgta
     6181 ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc gttcagcccg accgctgcgc
     6241 cttatccggt aactatcgtc ttgagtccaa cccggtaaga cacgacttat cgccactggc
     6301 agcagccact ggtaacagga ttagcagagc gaggtatgta ggcggtgcta cagagttctt
     6361 gaagtggtgg cctaactacg gctacactag aagaacagta tttggtatct gcgctctgct
     6421 gaagccagtt accttcggaa aaagagttgg tagctcttga tccggcaaac aaaccaccgc
     6481 tggtagcggt ttttttgttt gcaagcagca gattacgcgc agaaaaaaag gatctcaaga
     6541 agatcctttg atcttttcta cggggtctga cgctcagtgg aacgaaaact cacgttaagg
     6601 gattttggtc atgagattat caaaaaggat cttcacctag atccttttaa attaaaaatg
     6661 aagttttaaa tcaatctaaa gtatatatga gtaaacttgg tctgacagtt accaatgctt
     6721 aatcagtgag gcacctatct cagcgatctg tctatttcgt tcatccatag ttgcctgact
     6781 ccccgtcgtg tagataacta cgatacggga gggcttacca tctggcccca gtgctgcaat
     6841 gataccgcga gacccacgct caccggctcc agatttatca gcaataaacc agccagccgg
     6901 aagggccgag cgcagaagtg gtcctgcaac tttatccgcc tccatccagt ctattaattg
     6961 ttgccgggaa gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat
     7021 tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg gcttcattca gctccggttc
     7081 ccaacgatca aggcgagtta catgatcccc catgttgtgc aaaaaagcgg ttagctcctt
     7141 cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg ttatcactca tggttatggc
     7201 agcactgcat aattctctta ctgtcatgcc atccgtaaga tgcttttctg tgactggtga
     7261 gtactcaacc aagtcattct gagaatagtg tatgcggcga ccgagttgct cttgcccggc
     7321 gtcaatacgg gataataccg cgccacatag cagaacttta aaagtgctca tcattggaaa
     7381 acgttcttcg gggcgaaaac tctcaaggat cttaccgctg ttgagatcca gttcgatgta
     7441 acccactcgt gcacccaact gatcttcagc atcttttact ttcaccagcg tttctgggtg
     7501 agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata agggcgacac ggaaatgttg
     7561 aatactcata ctcttccttt ttcaatatta ttgaagcatt tatcagggtt attgtctcat
     7621 gagcggatac atatttgaat gtatttagaa aaataaacaa ataggggttc cgcgcacatt
     7681 tccccgaaaa gtgccacctg acgtc
//
