LOCUS       sequence-79489-e        8675 bp    DNA     circular SYN 15-APR-2025
DEFINITION  Artificial transcription factor with VP64 targeting c-Fos (TALE A).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8675)
  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 8675)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 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-SeqFosA
FEATURES             Location/Qualifiers
     source          1..8675
                     /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"
     primer_bind     863..882
                     /label=T7
                     /note="T7 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             4079..4228
                     /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(4269..4286)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    4275..4499
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      4545..4973
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4632..4651)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4842..4863
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4982..5002)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        4987..5316
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      5167..5302
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     5229..5248
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             5383..6177
                     /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(5437..5456)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     6047..6066
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    6351..6472
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6388..6407)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     6442..6461
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(6521..6537)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6521..6537)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(6534..6556)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    6545..6561
                     /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(6569..6575,6576..6593,6594..6599))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6614..6635
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(6752..6769)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6923..7508)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(7003..7022)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7679..8470,8471..8539))
                     /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     8302..8321
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(8540..8644)
                     /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 ctgaacctga ccccggacca
     1801 agtggtggct atcgccagcc acgatggcgg caagcaagcg ctcgaaacgg tgcagcggct
     1861 gttgccggtg ctgtgccagg accatggcct gactccggac caagtggtgg ctatcgccag
     1921 ccacgatggc ggcaagcaag cgctcgaaac ggtgcagcgg ctgttgccgg tgctgtgcca
     1981 ggaccatggc ctgaccccgg accaagtggt ggctatcgcc agcaacaatg gcggcaagca
     2041 agcgctcgaa acggtgcagc ggctgttgcc ggtgctgtgc caggaccatg gcctgactcc
     2101 ggaccaagtg gtggctatcg ccagccacga tggcggcaag caagcgctcg aaacggtgca
     2161 gcggctgttg ccggtgctgt gccaggacca tggcctgacc ccggaccaag tggtggctat
     2221 cgccagcaac attggcggca agcaagcgct cgaaacggtg cagcggctgt tgccggtgct
     2281 gtgccaggac catggcctga ccccggacca agtggtggct atcgccagca acggtggcgg
     2341 caagcaagcg ctcgaaacgg tgcagcggct gttgccggtg ctgtgccagg accatggcct
     2401 gaccccggac caagtggtgg ctatcgccag caacggtggc ggcaagcaag cgctcgaaac
     2461 ggtgcagcgg ctgttgccgg tgctgtgcca ggaccatggc ctgaccccgg accaagtggt
     2521 ggctatcgcc agcaacaatg gcggcaagca agcgctcgaa acggtgcagc ggctgttgcc
     2581 ggtgctgtgc caggaccatg gcctgacccc ggaccaagtg gtggctatcg ccagcaacat
     2641 tggcggcaag caagcgctcg aaacggtgca gcggctgttg ccggtgctgt gccaggacca
     2701 tggcctgacc ccggaccaag tggtggctat cgccagcaac attggcggca agcaagcgct
     2761 cgaaacggtg cagcggctgt tgccggtgct gtgccaggac catggcctga ccccggacca
     2821 agtggtggct atcgccagcc acgatggcgg caagcaagcg ctcgaaacgg tgcagcggct
     2881 gttgccggtg ctgtgccagg accatggcct gactccggac caagtggtgg ctatcgccag
     2941 ccacgatggc ggcaagcaag cgctcgaaac ggtgcagcgg ctgttgccgg tgctgtgcca
     3001 ggaccatggc ctgaccccgg accaagtggt ggctatcgcc agcaacattg gcggcaagca
     3061 agcgctcgaa acggtgcagc ggctgttgcc ggtgctgtgc caggaccatg gcctgacccc
     3121 ggaccaagtg gtggctatcg ccagcaacaa tggcggcaag caagcgctcg aaacggtgca
     3181 gcggctgttg ccggtgctgt gccaggacca tggcctgacc ccggaccaag tggtggctat
     3241 cgccagcaac aatggcggca agcaagcgct cgaaacggtg cagcggctgt tgccggtgct
     3301 gtgccaggac catggcctga ccccggacca agtggtggct atcgccagca acggtggcgg
     3361 caagcaagcg ctcgaaagca ttgtggccca gctgagccgg cctgatccgg cgttggccgc
     3421 gttgaccaac gaccacctcg tcgccttggc ctgcctcggc ggacgtcctg ccatggatgc
     3481 agtgaaaaag ggattgccgc acgcgccgga attgatcaga agagtcaatc gccgtattgg
     3541 cgaacgcacg tcccatcgcg ttgccgacta cgcgcaagtg gttcgcgtgc tggagttttt
     3601 ccagtgccac tcccacccag cgtacgcatt tgatgaggcc atgacgcagt tcgggatgag
     3661 caggaacggg ttggtacagc tctttcgcag agtgggcgtc accgaactcg aagcccgcgg
     3721 tggaacgctc cccccagcct cgcagcgttg ggaccgtatc ctccaggcat cagggatgaa
     3781 aagggccaaa ccgtccccta cttcagctca aacaccggat caggcgtctt tgcatgcatt
     3841 cgccgattcg ctggagcgtg accttgatgc gcccagccca atgcacgagg gagatcagac
     3901 gcgggcaagc agccgtaaac ggtcccgatc ggatcgtgct gtcaccggcc cctccgcaca
     3961 gcaggctgtc gaggtgcgcg ttcccgaaca gcgcgatgcg ctgcatttgc ccctcagctg
     4021 gagggtaaaa cgcccgcgta ccaggatctg gggcggcctc ccggatcccg gacgggctga
     4081 cgcattggac gattttgatc tggatatgct gggaagtgac gccctcgatg attttgacct
     4141 tgacatgctt ggttcggatg cccttgatga ctttgacctc gacatgctcg gcagtgacgc
     4201 ccttgatgat ttcgacctgg acatgctgat taactaatct agagggcccg tttaaacccg
     4261 ctgatcagcc tcgactgtgc cttctagttg ccagccatct gttgtttgcc cctcccccgt
     4321 gccttccttg accctggaag gtgccactcc cactgtcctt tcctaataaa atgaggaaat
     4381 tgcatcgcat tgtctgagta ggtgtcattc tattctgggg ggtggggtgg ggcaggacag
     4441 caagggggag gattgggaag acaatagcag gcatgctggg gatgcggtgg gctctatggc
     4501 ttctgaggcg gaaagaacca gctggggctc tagggggtat ccccacgcgc cctgtagcgg
     4561 cgcattaagc gcggcgggtg tggtggttac gcgcagcgtg accgctacac ttgccagcgc
     4621 cctagcgccc gctcctttcg ctttcttccc ttcctttctc gccacgttcg ccggctttcc
     4681 ccgtcaagct ctaaatcggg ggctcccttt agggttccga tttagtgctt tacggcacct
     4741 cgaccccaaa aaacttgatt agggtgatgg ttcacgtagt gggccatcgc cctgatagac
     4801 ggtttttcgc cctttgacgt tggagtccac gttctttaat agtggactct tgttccaaac
     4861 tggaacaaca ctcaacccta tctcggtcta ttcttttgat ttataaggga ttttgccgat
     4921 ttcggcctat tggttaaaaa atgagctgat ttaacaaaaa tttaacgcga attaattctg
     4981 tggaatgtgt gtcagttagg gtgtggaaag tccccaggct ccccagcagg cagaagtatg
     5041 caaagcatgc atctcaatta gtcagcaacc aggtgtggaa agtccccagg ctccccagca
     5101 ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa ccatagtccc gcccctaact
     5161 ccgcccatcc cgcccctaac tccgcccagt tccgcccatt ctccgcccca tggctgacta
     5221 atttttttta tttatgcaga ggccgaggcc gcctctgcct ctgagctatt ccagaagtag
     5281 tgaggaggct tttttggagg cctaggcttt tgcaaaaagc tcccgggagc ttgtatatcc
     5341 attttcggat ctgatcaaga gacaggatga ggatcgtttc gcatgattga acaagatgga
     5401 ttgcacgcag gttctccggc cgcttgggtg gagaggctat tcggctatga ctgggcacaa
     5461 cagacaatcg gctgctctga tgccgccgtg ttccggctgt cagcgcaggg gcgcccggtt
     5521 ctttttgtca agaccgacct gtccggtgcc ctgaatgaac tgcaggacga ggcagcgcgg
     5581 ctatcgtggc tggccacgac gggcgttcct tgcgcagctg tgctcgacgt tgtcactgaa
     5641 gcgggaaggg actggctgct attgggcgaa gtgccggggc aggatctcct gtcatctcac
     5701 cttgctcctg ccgagaaagt atccatcatg gctgatgcaa tgcggcggct gcatacgctt
     5761 gatccggcta cctgcccatt cgaccaccaa gcgaaacatc gcatcgagcg agcacgtact
     5821 cggatggaag ccggtcttgt cgatcaggat gatctggacg aagagcatca ggggctcgcg
     5881 ccagccgaac tgttcgccag gctcaaggcg cgcatgcccg acggcgagga tctcgtcgtg
     5941 acccatggcg atgcctgctt gccgaatatc atggtggaaa atggccgctt ttctggattc
     6001 atcgactgtg gccggctggg tgtggcggac cgctatcagg acatagcgtt ggctacccgt
     6061 gatattgctg aagagcttgg cggcgaatgg gctgaccgct tcctcgtgct ttacggtatc
     6121 gccgctcccg attcgcagcg catcgccttc tatcgccttc ttgacgagtt cttctgagcg
     6181 ggactctggg gttcgaaatg accgaccaag cgacgcccaa cctgccatca cgagatttcg
     6241 attccaccgc cgccttctat gaaaggttgg gcttcggaat cgttttccgg gacgccggct
     6301 ggatgatcct ccagcgcggg gatctcatgc tggagttctt cgcccacccc aacttgttta
     6361 ttgcagctta taatggttac aaataaagca atagcatcac aaatttcaca aataaagcat
     6421 ttttttcact gcattctagt tgtggtttgt ccaaactcat caatgtatct tatcatgtct
     6481 gtataccgtc gacctctagc tagagcttgg cgtaatcatg gtcatagctg tttcctgtgt
     6541 gaaattgtta tccgctcaca attccacaca acatacgagc cggaagcata aagtgtaaag
     6601 cctggggtgc ctaatgagtg agctaactca cattaattgc gttgcgctca ctgcccgctt
     6661 tccagtcggg aaacctgtcg tgccagctgc attaatgaat cggccaacgc gcggggagag
     6721 gcggtttgcg tattgggcgc tcttccgctt cctcgctcac tgactcgctg cgctcggtcg
     6781 ttcggctgcg gcgagcggta tcagctcact caaaggcggt aatacggtta tccacagaat
     6841 caggggataa cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta
     6901 aaaaggccgc gttgctggcg tttttccata ggctccgccc ccctgacgag catcacaaaa
     6961 atcgacgctc aagtcagagg tggcgaaacc cgacaggact ataaagatac caggcgtttc
     7021 cccctggaag ctccctcgtg cgctctcctg ttccgaccct gccgcttacc ggatacctgt
     7081 ccgcctttct cccttcggga agcgtggcgc tttctcatag ctcacgctgt aggtatctca
     7141 gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc gttcagcccg
     7201 accgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggtaaga cacgacttat
     7261 cgccactggc agcagccact ggtaacagga ttagcagagc gaggtatgta ggcggtgcta
     7321 cagagttctt gaagtggtgg cctaactacg gctacactag aagaacagta tttggtatct
     7381 gcgctctgct gaagccagtt accttcggaa aaagagttgg tagctcttga tccggcaaac
     7441 aaaccaccgc tggtagcggt ttttttgttt gcaagcagca gattacgcgc agaaaaaaag
     7501 gatctcaaga agatcctttg atcttttcta cggggtctga cgctcagtgg aacgaaaact
     7561 cacgttaagg gattttggtc atgagattat caaaaaggat cttcacctag atccttttaa
     7621 attaaaaatg aagttttaaa tcaatctaaa gtatatatga gtaaacttgg tctgacagtt
     7681 accaatgctt aatcagtgag gcacctatct cagcgatctg tctatttcgt tcatccatag
     7741 ttgcctgact ccccgtcgtg tagataacta cgatacggga gggcttacca tctggcccca
     7801 gtgctgcaat gataccgcga gacccacgct caccggctcc agatttatca gcaataaacc
     7861 agccagccgg aagggccgag cgcagaagtg gtcctgcaac tttatccgcc tccatccagt
     7921 ctattaattg ttgccgggaa gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg
     7981 ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg gcttcattca
     8041 gctccggttc ccaacgatca aggcgagtta catgatcccc catgttgtgc aaaaaagcgg
     8101 ttagctcctt cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg ttatcactca
     8161 tggttatggc agcactgcat aattctctta ctgtcatgcc atccgtaaga tgcttttctg
     8221 tgactggtga gtactcaacc aagtcattct gagaatagtg tatgcggcga ccgagttgct
     8281 cttgcccggc gtcaatacgg gataataccg cgccacatag cagaacttta aaagtgctca
     8341 tcattggaaa acgttcttcg gggcgaaaac tctcaaggat cttaccgctg ttgagatcca
     8401 gttcgatgta acccactcgt gcacccaact gatcttcagc atcttttact ttcaccagcg
     8461 tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata agggcgacac
     8521 ggaaatgttg aatactcata ctcttccttt ttcaatatta ttgaagcatt tatcagggtt
     8581 attgtctcat gagcggatac atatttgaat gtatttagaa aaataaacaa ataggggttc
     8641 cgcgcacatt tccccgaaaa gtgccacctg acgtc
//
