LOCUS       sequence-350965-        7042 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Transcription Factor chassis for activation domains. mCherry, Zif268
            DNA binding domain, estrogen response domain.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7042)
  AUTHORS   Staller MV, Holehouse AS, Swain-Lenz D, Das RK, Pappu RV, Cohen BA
  TITLE     A High-Throughput Mutational Scan of an Intrinsically Disordered 
            Acidic Transcriptional Activation Domain.
  JOURNAL   Cell Syst. 2018 Mar 1. pii: S2405-4712(18)30052-8. doi: 
            10.1016/j.cels.2018.01.015.
  PUBMED    29525204
REFERENCE   2  (bases 1 to 7042)
  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 Syst. 
            2018 Mar 1. pii: S2405-4712(18)30052-8. doi: 
            10.1016/j.cels.2018.01.015."
COMMENT     Sequence Label: pMVS142_pACT1_mCherry_Zif268_EBD_MCS_KAN
FEATURES             Location/Qualifiers
     source          1..7042
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        53..71
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     53..70
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(1251..1269)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     1519..1538
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     terminator      3026..3213
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae 
                     alcohol dehydrogenase 1 (ADH1) gene"
     primer_bind     complement(3081..3105)
                     /label=pHybLex-R
                     /note="ADH terminator, reverse primer"
     gene            3288..4644
                     /label=kanMX
                     /note="yeast selectable marker conferring kanamycin 
                     resistance (Wach et al., 1994)"
     promoter        3288..3631
                     /label=TEF promoter
                     /note="Ashbya gossypii TEF promoter"
     CDS             3632..4441
                     /codon_start=1
                     /gene="aph(3')-Ia"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin"
                     /translation="MGKEKTHVSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYGKP
                     DAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGKTA
                     FQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDASD
                     FDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGIAD
                     RYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     complement(3699..3718)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     terminator      4447..4644
                     /label=TEF terminator
                     /note="Ashbya gossypii TEF terminator"
     primer_bind     complement(4748..4767)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(4749..4767)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(4854..4871)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5025..5613)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5105..5124)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5784..6575,6576..6644))
                     /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     6407..6426
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6645..6749)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     6817..6835
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(6873..6895)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     6995..7014
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 cgttagaacg cggctacaat taatacataa ccttatgtat catacacata cgatttaggt
       61 gacactatag aacgcggccg ccagctgaag cttcgtacgc tgcaggtcga cgtggctgca
      121 agagattgat cggtatgcaa cctcaacagt gttgaagcta ttggtaggta acaagtgtga
      181 tttaaaggac aagcgtgtcg tggaatatga cgtggcaaag gaatttgcgg acgcgaataa
      241 gatgccgttc ttagagacta gtgctttgga ctccaccaac gtcgaggatg cgtttttaac
      301 catggctaga caaatcaagg aaagtatgtc ccaacaaaac ctgaacgaaa ccactcagaa
      361 gaaggaagac aaagggaacg tcaacctgaa gggacagagt ttaaccaaca ccggtggggg
      421 ctgctgttga acaagcgcgc ctctaccttg cagacccata taatataata actaaataag
      481 taaataagac acacgcgaga acatatatac acaattacag taacaataac aagaggacag
      541 atactaccaa aatgtgtggg gaagcgggta agctgccaca gcaattaatg cacaacattt
      601 aacctacatt cttccttatc ggatcctcaa aacccttaaa aacatatgcc tcaccctaac
      661 atattttcca attaaccctc aatatttctc tgtcacccgg cctctatttt ccattttctt
      721 ctttacccgc cacgcgtttt tttctttcaa atttttttct tccttcttct ttttcttcca
      781 cgtcctcttg cataaataaa taaaccgttt tgaaaccaaa ctcgcctctc tctctccttt
      841 ttgaaatatt tttgggtttg tttgatcctt tccttcccaa tctctcttgt ttaatatata
      901 ttcatttata tcacgctctc tttttatctt cctttttttc ctctctcttg tattcttcct
      961 tcccctttct actcaaacca agaagaaaaa gaaaaggtca atctttgtta aagaatagga
     1021 tcttctacta catcagcttt tagatttttc acgcttactg cttttttctt cccaagatcg
     1081 aaaatttact gaattaacaa tgggtaccgt gagcaagggc gaggaggata acatggccat
     1141 catcaaggag ttcatgagat tcaaggtgca catggagggc tccgtgaacg gccacgagtt
     1201 cgagatcgag ggcgagggcg agggcagacc ctacgagggc acccagaccg ccaagctgaa
     1261 ggtgaccaag ggtggccccc tgcccttcgc ctgggacatc ctgtcccctc agttcatgta
     1321 cggctccaag gcctacgtga agcaccccgc cgacatcccc gactacttga agctgtcctt
     1381 ccccgagggc ttcaagtggg agagagtgat gaacttcgag gacggcggcg tggtgaccgt
     1441 gacccaggac tcctccctgc aggacggcga gttcatctac aaggtgaagt tgagaggcac
     1501 caacttcccc tccgacggcc ccgtaatgca gaagaagacc atgggctggg aggcctcctc
     1561 cgagagaatg taccccgagg acggcgccct gaagggcgag atcaagcaga ggctgaagct
     1621 gaaggacggc ggccactacg acgctgaggt caagaccacc tacaaggcca agaagcccgt
     1681 gcagctgccc ggcgcctaca acgtcaacat caagttggac atcacctccc acaacgagga
     1741 ctacaccatt gtggaacaat atgaaagagc tgaaggtaga cattctactg gtggtatgga
     1801 tgaattgtac aaaggtaccc gcccatatgc ttgccctgtc gagtcctgcg atcgccgctt
     1861 ttctcgctcg gatgagctta cccgccatat ccgcatccat accggtcaga agcccttcca
     1921 gtgtcgaatc tgcatgcgta acttcagtcg tagtgaccac cttaccaccc acatccgcac
     1981 ccacacaggc gagaagcctt ttgcctgtga catttgtggg aggaagtttg ccaggagtga
     2041 tgaacgcaag aggcatacca aaatccatac aggtcctgca gctgcgtcaa ctctagagga
     2101 tccatctgct ggagacatga gagctgccaa cctttggcca agcccgctca tgatcaaacg
     2161 ctctaagaag aacagcctgg ccttgtccct gacggccgac cagatggtca gtgccttgtt
     2221 ggatgctgag ccccccatac tctattccga gtatgatcct accagaccct tcagtgaagc
     2281 ttcgatgatg ggcttactga ccaacctggc agacagggag ctggttcaca tgatcaactg
     2341 ggcgaagagg gtgccaggct ttgtggattt gaccctccat gatcaggtcc accttctaga
     2401 atgtgcctgg ctagagatcc tgatgattgg tctcgtctgg cgctccatgg agcacccagt
     2461 gaagctactg tttgctccta acttgctctt ggacaggaac cagggaaaat gtgtagaggg
     2521 catggtggag atcttcgaca tgctgctggc tacatcatct cggttccgca tgatgaatct
     2581 gcagggagag gagtttgtgt gcctcaaatc tattattttg cttaattctg gagtgtacac
     2641 atttctgtcc agcaccctga agtctctgga agagaaggac catatccacc gagtcctgga
     2701 caagatcaca gacactttga tccacctgat ggccaaggca ggcctgaccc tgcagcagca
     2761 gcaccagcgg ctggcccagc tcctcctcat cctctcccac atcaggcaca tgagtaacaa
     2821 aggcatggag catctgtaca gcatgaagtg caagaacgtg gtgcccctct atgacctgct
     2881 gctggagatg ctggacgccc accgcctaca tgcgcccact agccgtggag gggcatccgt
     2941 ggaggagacg gaccaaagcc acttggccac tgcgggctct acttcatcgg ctagcggtta
     3001 attaaggcgc gccacttcta aataagcgaa tttcttatga tttatgattt ttattattaa
     3061 ataagttata aaaaaaataa gtgtatacaa attttaaagt gactcttagg ttttaaaacg
     3121 aaaattctta ttcttgagta actctttcct gtaggtcagg ttgctttctc aggtatagta
     3181 tgaggtcgct cttattgacc acacctctac cggcagatcc gctagggata acagggtaat
     3241 atagatctgt ttagcttgcc tcgtccccgc cgggtcaccc ggccagcgac atggaggccc
     3301 agaataccct ccttgacagt cttgacgtgc gcagctcagg ggcatgatgt gactgtcgcc
     3361 cgtacattta gcccatacat ccccatgtat aatcatttgc atccatacat tttgatggcc
     3421 gcacggcgcg aagcaaaaat tacggctcct cgctgcagac ctgcgagcag ggaaacgctc
     3481 ccctcacaga cgcgttgaat tgtccccacg ccgcgcccct gtagagaaat ataaaaggtt
     3541 aggatttgcc actgaggttc ttctttcata tacttccttt taaaatcttg ctaggataca
     3601 gttctcacat cacatccgaa cataaacaac catgggtaag gaaaagactc acgtttcgag
     3661 gccgcgatta aattccaaca tggatgctga tttatatggg tataaatggg ctcgcgataa
     3721 tgtcgggcaa tcaggtgcga caatctatcg attgtatggg aagcccgatg cgccagagtt
     3781 gtttctgaaa catggcaaag gtagcgttgc caatgatgtt acagatgaga tggtcagact
     3841 aaactggctg acggaattta tgcctcttcc gaccatcaag cattttatcc gtactcctga
     3901 tgatgcatgg ttactcacca ctgcgatccc cggcaaaaca gcattccagg tattagaaga
     3961 atatcctgat tcaggtgaaa atattgttga tgcgctggca gtgttcctgc gccggttgca
     4021 ttcgattcct gtttgtaatt gtccttttaa cagcgatcgc gtatttcgtc tcgctcaggc
     4081 gcaatcacga atgaataacg gtttggttga tgcgagtgat tttgatgacg agcgtaatgg
     4141 ctggcctgtt gaacaagtct ggaaagaaat gcataagctt ttgccattct caccggattc
     4201 agtcgtcact catggtgatt tctcacttga taaccttatt tttgacgagg ggaaattaat
     4261 aggttgtatt gatgttggac gagtcggaat cgcagaccga taccaggatc ttgccatcct
     4321 atggaactgc ctcggtgagt tttctccttc attacagaaa cggctttttc aaaaatatgg
     4381 tattgataat cctgatatga ataaattgca gtttcatttg atgctcgatg agtttttcta
     4441 atcagtactg acaataaaaa gattcttgtt ttcaagaact tgtcatttgt atagtttttt
     4501 tatattgtag ttgttctatt ttaatcaaat gttagcgtga tttatatttt ttttcgcctc
     4561 gacatcatct gcccagatgc gaagttaagt gcgcagaaag taatatcatg cgtcaatcgt
     4621 atgtgaatgc tggtcgctat actgctgtcg attcgatact aacgccgcca tccagtttaa
     4681 acgagctcga attcatcgat gatatcagat ccactagtgg cctatgcggc cgcggatctg
     4741 ccggtctccc tatagtgagt cgtattaatt tcgataagcc aggttaacct gcattaatga
     4801 atcggccaac gcgcggggag aggcggtttg cgtattgggc gctcttccgc ttcctcgctc
     4861 actgactcgc tgcgctcggt cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg
     4921 gtaatacggt tatccacaga atcaggggat aacgcaggaa agaacatgtg agcaaaaggc
     4981 cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca taggctccgc
     5041 ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga
     5101 ctataaagat accaggcgtt tccccctgga agctccctcg tgcgctctcc tgttccgacc
     5161 ctgccgctta ccggatacct gtccgccttt ctcccttcgg gaagcgtggc gctttctcat
     5221 agctcacgct gtaggtatct cagttcggtg taggtcgttc gctccaagct gggctgtgtg
     5281 cacgaacccc ccgttcagcc cgaccgctgc gccttatccg gtaactatcg tcttgagtcc
     5341 aacccggtaa gacacgactt atcgccactg gcagcagcca ctggtaacag gattagcaga
     5401 gcgaggtatg taggcggtgc tacagagttc ttgaagtggt ggcctaacta cggctacact
     5461 agaagaacag tatttggtat ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt
     5521 ggtagctctt gatccggcaa acaaaccacc gctggtagcg gtggtttttt tgtttgcaag
     5581 cagcagatta cgcgcagaaa aaaaggatct caagaagatc ctttgatctt ttctacgggg
     5641 tctgacgctc agtggaacga aaactcacgt taagggattt tggtcatgag attatcaaaa
     5701 aggatcttca cctagatcct tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata
     5761 tatgagtaaa cttggtctga cagttaccaa tgcttaatca gtgaggcacc tatctcagcg
     5821 atctgtctat ttcgttcatc catagttgcc tgactccccg tcgtgtagat aactacgata
     5881 cgggagggct taccatctgg ccccagtgct gcaatgatac cgcgagaccc acgctcaccg
     5941 gctccagatt tatcagcaat aaaccagcca gccggaaggg ccgagcgcag aagtggtcct
     6001 gcaactttat ccgcctccat ccagtctatt aattgttgcc gggaagctag agtaagtagt
     6061 tcgccagtta atagtttgcg caacgttgtt gccattgcta caggcatcgt ggtgtcacgc
     6121 tcgtcgtttg gtatggcttc attcagctcc ggttcccaac gatcaaggcg agttacatga
     6181 tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt tgtcagaagt
     6241 aagttggccg cagtgttatc actcatggtt atggcagcac tgcataattc tcttactgtc
     6301 atgccatccg taagatgctt ttctgtgact ggtgagtact caaccaagtc attctgagaa
     6361 tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca
     6421 catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg aaaactctca
     6481 aggatcttac cgctgttgag atccagttcg atgtaaccca ctcgtgcacc caactgatct
     6541 tcagcatctt ttactttcac cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc
     6601 gcaaaaaagg gaataagggc gacacggaaa tgttgaatac tcatactctt cctttttcaa
     6661 tattattgaa gcatttatca gggttattgt ctcatgagcg gatacatatt tgaatgtatt
     6721 tagaaaaata aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc acctgacgtc
     6781 taagaaacca ttattatcat gacattaacc tataaaaata ggcgtatcac gaggcccttt
     6841 cgtctcgcgc gtttcggtga tgacggtgaa aacctctgac acatgcagct cccggagacg
     6901 gtcacagctt gtctgtaagc ggatgccggg agcagacaag cccgtcaggg cgcgtcagcg
     6961 ggtgttggcg ggtgtcgggg ctggcttaac tatgcggcat cagagcagat tgtactgaga
     7021 gtgcaccata tggacatatt gt
//
