LOCUS       sequence-227013-        7332 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Vector to express tAD candidates from a library cloned from 
            fragmented transcription factor coding sequences fused to the Gal4 
            DNA binding domain..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7332)
  AUTHORS   Arnold CD, Nemcko F, Woodfin AR, Wienerroither S, Vlasova A, 
            Schleiffer A, Pagani M, Rath M, Stark A
  TITLE     A high-throughput method to identify trans-activation domains within
            transcription factor sequences.
  JOURNAL   EMBO J. 2018 Aug 15;37(16). pii: embj.201798896. doi: 
            10.15252/embj.201798896. Epub 2018 Jul 13.
  PUBMED    30006452
REFERENCE   2  (bases 1 to 7332)
  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: "EMBO J."; 
            date: "2018-08-15"; volume: "37(16). pii: embj.201798896. doi"; 
            pages: " 10.15252/embj.201798896. Epub 2018 Jul 13"
COMMENT     Sequence Label: ptAD-seq-ubi63E-Gal4-DBD
FEATURES             Location/Qualifiers
     source          1..7332
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     polyA_signal    1..49
                     /note="synthetic polyadenylation signal"
     misc_feature    63..154
                     /label=pause site
                     /note="RNA polymerase II transcriptional pause signal from 
                     the human alpha-2 globin gene"
     primer_bind     103..122
                     /label=RVprimer3
                     /note="pGL3 vector, forward primer"
     regulatory      2374..2383
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     intron          2389..2455
                     /label=Mhc intron
                     /note="modified Drosophila myosin heavy chain intron 16 
                     (Pfeiffer et al., 2010)"
     CDS             2467..2907
                     /codon_start=1
                     /gene="Saccharomyces cerevisiae GAL4 (truncated)"
                     /product="DNA binding domain of the GAL4 transcriptional 
                     activator"
                     /label=GAL4 DNA binding domain
                     /translation="MKLLSSIEQACDICRLKKLKCSKEKPKCAKCLKNNWECRYSPKTK
                     RSPLTRAHLTEVESRLERLEQLFLLIFPREDLDMILKMDSLQDIKALLTGLFVQDNVNK
                     DAVTDRLASVETDMPLTLRQHRISATSSSEESSNKGQRQLTVS"
     promoter        join(2972..2977,2978..2995,2996..3002)
                     /label=lac UV5 promoter
                     /note="E. coli lac promoter with an ""up"" mutation"
     CDS             3056..3715
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(3122..3141)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     CDS             4057..4362
                     /codon_start=1
                     /gene="ccdB"
                     /product="CcdB, a bacterial toxin that poisons DNA gyrase"
                     /label=ccdB
                     /note="Plasmids containing the ccdB gene cannot be 
                     propagated in standard E. coli strains."
                     /translation="MQFKVYTYKRESRYRLFVDVQSDIIDTPGRRMVIPLASARLLSDK
                     VSRELYPVVHIGDESWRMMTTDMASVPVSVIGEEVADLSHRENDIKNAINLMFWGI"
     primer_bind     4241..4260
                     /label=ccdB-fwd
                     /note="ccdB gene, forward primer"
     polyA_signal    4459..4580
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4470..4489)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     4524..4543
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(4825..4842)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4996..5584)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5076..5095)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5755..6546,6547..6615))
                     /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     6378..6397
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6616..6720)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      6747..7202
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6834..6853)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     7044..7065
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
ORIGIN
        1 aataaaatat ctttattttc attacatctg tgtgttggtt ttttgtgtga atcgatagta
       61 ctaacatacg ctctccatca aaacaaaacg aaacaaaaca aactagcaaa ataggctgtc
      121 cccagtgcaa gtgcaggtgc cagaacattt ctctatcgat aggtacctgt cgccggaacg
      181 cagcgacaga gattccaatg tgtccgtatc tttcaggctt ttgcccttca gttccagacg
      241 aagcgactgg cgattcgcgt gtggggtctg cttcagggtc ttgtgaatta gggcgcgcag
      301 atcgccgatg ggcgtggcgc cggagggcac cttcaccttg ccgtacggct tgctgttctt
      361 cgcgttcaaa atctccagct ccattttgct ttcggtgcgc ttgcaatcag tactgtccaa
      421 aatcgaaaat cgccgaaccg tagtgtgacc gtgcggggct ctgcgaaaat aaactttttt
      481 aggtatatgg ccacacacgg ggaaagcaca gtggattata tgttttaata ttataatatg
      541 caggttttca ttacttatcc agatgtaagc ccacttaaag cgatttaaca attatttgcc
      601 gaaagagtat aaacaaattt cacttaaaaa tggattaaga aaagcttgtg taagattatg
      661 cgcagcgttg ccagatagct ccatttaaaa cacttcaaaa acaataagtt ttgaaaatat
      721 atacataaat agcagtcgtt gccgcaacgc tcaacacatc acacttttaa aacacccttt
      781 acctacacag aattactttt taaatttcca gtcaagctgc gagtttcaaa attatagccg
      841 gtagagaaga cagtgctatt tcaaaagcaa actaacaagg gtcttaaatt ccaaaacacc
      901 aatcctaaca agccttggac ttttgtaagt ttagatcaaa ggtggcattg cattcaatgt
      961 catggtaaga agtaggtcgt ctaggtagaa atcctcattc agccggtcaa gtcagtacga
     1021 gaaaggtctc aatttgaaat tgtcttaaaa atattttatt gttttgtact gtggtgagtt
     1081 taaacgaaaa acacaaaaaa aaagtgatac acagaaatca taaaaaattt taatacaagg
     1141 tattcgtacg tatcaaaaac atttcggcac aatttttttt ctctgtacta aagtgttacg
     1201 aacactacgg tattttttag tgattttcaa cggacaccga aggtatataa acagcgttcg
     1261 cgaacggtcg ccttcaaaac caattgacat ttgcagcagc aagtacaagt agaaagtaaa
     1321 gcgcaatcag cgaaaaattt atacttaatt gttggtgatt aaagtacaat taaaagaaca
     1381 ttctcgaaag tcacaagaaa cgtaagtttt taactcgctg ttaccaatta gtaataagag
     1441 caacaagacg ttgagtaatt tcaagaaaaa ctgcatttca aggtctttgt tcggccattt
     1501 tttttttatt caacgctcta cgtaattaca aaataagaaa ttggcagcca cgcatcttgt
     1561 tttcccaatg aattggcatc aaaacgcaaa caaatctata aataaaactt gcgtgttgat
     1621 tttcgccaag atttattggc aaattgtgaa attcgcagtg acgcatttga aaattcgaga
     1681 aatcacgaac gcactcgagc atttgtgtgc atgttattag ttagttgttt agttaattga
     1741 agtattttac caacgaaatc cacttatttt tagctgaaat agagtaggtt gcttaaacaa
     1801 agccacgtct gaaaatttct tattgcttgt agttgtgacg tcaccatata cacacaaaat
     1861 aatgtgtatg catgcatttc agctgtgtat atatacatgc acacactcgc aacacgaaaa
     1921 cgatgacgaa gcaacggaac aaaggtttct caactaccct ttgttccctg tttcttcgct
     1981 ttcctttgtt ccaatattcg tagagggtta ataggggttt ctcaacaaag ttggcgtcga
     2041 taaataagtt tcccattttt attccccagc caggaagtta gtttcaatag ttttgtaatt
     2101 tcaacgaaac tcatttgatt tcgtactaat tttccacatc tctattttct gcccgcagaa
     2161 taatccaaac tgcagagatc tgcgatcgag ctcgcccggg gatcgagcgc agcggtataa
     2221 aagggcgcgg ggtggctgag agcatcagtt gtgaatgaat gttcgagccg agcagacgtg
     2281 ccgctgcctt cgttaatatc ctttgaataa gccaactttg aatcacaaga cgcataccaa
     2341 accctgcagg ggcattccgg tactgttggt aaagccacca tggaaaaggt aggttcaacc
     2401 actgatgcct aggcacaccg aaacgactaa ccctaattct tatcctttac ttcaggccac
     2461 ttcaaaatga agctactgtc ttctatcgaa caagcatgcg atatttgccg acttaaaaag
     2521 ctcaagtgct ccaaagaaaa accgaagtgc gccaagtgtc tgaagaacaa ctgggagtgt
     2581 cgctactctc ccaaaaccaa aaggtctccg ctgactaggg cacatctgac agaagtggaa
     2641 tcaaggctag aaagactgga acagctattt ctactgattt ttcctcgaga agaccttgac
     2701 atgattttga aaatggattc tttacaggat ataaaagcat tgttaacagg attatttgta
     2761 caagataatg tgaataaaga tgccgtcaca gatagattgg cttcagtgga gactgatatg
     2821 cctctaacat tgagacagca tagaataagt gcgacatcat catcggaaga gagtagtaac
     2881 aaaggtcaaa gacagttgac tgtatcgtcg ggtggtggtg gtagtttgag catgcaccgg
     2941 tgatatcgcg gccgcattag gcaccccagg ctttacactt tatgcttccg gctcgtataa
     3001 tgtgtggatt ttgagttagg atccgtcgag attttcagga gctaaggaag ctaaaatgga
     3061 gaaaaaaatc actggatata ccaccgttga tatatcccaa tggcatcgta aagaacattt
     3121 tgaggcattt cagtcagttg ctcaatgtac ctataaccag accgttcagc tggatattac
     3181 ggccttttta aagaccgtaa agaaaaataa gcacaagttt tatccggcct ttattcacat
     3241 tcttgcccgc ctgatgaatg ctcatccgga attccgtatg gcaatgaaag acggtgagct
     3301 ggtgatatgg gatagtgttc acccttgtta caccgttttc catgagcaaa ctgaaacgtt
     3361 ttcatcgctc tggagtgaat accacgacga tttccggcag tttctacaca tatattcgca
     3421 agatgtggcg tgttacggtg aaaacctggc ctatttccct aaagggttta ttgagaatat
     3481 gtttttcgtc tcagccaatc cctgggtgag tttcaccagt tttgatttaa acgtggccaa
     3541 tatggacaac ttcttcgccc ccgttttcac catgggcaaa tattatacgc aaggcgacaa
     3601 ggtgctgatg ccgctggcga ttcaggttca tcatgccgtt tgtgatggct tccatgtcgg
     3661 cagaatgctt aatgaattac aacagtactg cgatgagtgg cagggcgggg cgtaaacgcg
     3721 tggatccggc ttactaaaag ccagataaca gtatgcgtat ttgcgcgctg atttttgcgg
     3781 tataagaata tatactgata tgtatacccg aagtatgtca aaaagaggta tgctatgaag
     3841 cagcgtatta cagtgacagt tgacagcgac agctatcagt tgctcaaggc atatatgatg
     3901 tcaatatctc cggtctggta agcacaacca tgcagaatga agcccgtcgt ctgcgtgccg
     3961 aacgctggaa agcggaaaat caggaaggga tggctgaggt cgcccggttt attgaaatga
     4021 acggctcttt tgctgacgag aacaggggct ggtgaaatgc agtttaaggt ttacacctat
     4081 aaaagagaga gccgttatcg tctgtttgtg gatgtacaga gtgatattat tgacacgccc
     4141 gggcgacgga tggtgatccc cctggccagt gcacgtctgc tgtcagataa agtctcccgt
     4201 gaactttacc cggtggtgca tatcggggat gaaagctggc gcatgatgac caccgatatg
     4261 gccagtgtgc cggtctccgt tatcggggaa gaagtggctg atctcagcca ccgcgaaaat
     4321 gacatcaaaa acgccattaa cctgatgttc tggggaatat aaatgtcagg ctcccttata
     4381 cacagccagt ctgcaggata tcgtcgacgt agatagatcg tgacgtaagc tagggccggc
     4441 cgcttcgagc agacatgata agatacattg atgagtttgg acaaaccaca actagaatgc
     4501 agtgaaaaaa atgctttatt tgtgaaattt gtgatgctat tgctttattt gtaaccatta
     4561 taagctgcaa taaacaagtt aacaacaaca attgcattca ttttatgttt caggttcagg
     4621 gggaggtgtg ggaggttttt taaagcaagt aaaacctcta caaatgtggt aaaatcgata
     4681 aggatccgac cgatgccctt gagagccttc aacccagtca gctccttccg gtgggcgcgg
     4741 ggcatgacta tcgtcgccgc acttatgact gtcttcttta tcatgcaact cgtaggacag
     4801 gtgccggcag cgctcttccg cttcctcgct cactgactcg ctgcgctcgg tcgttcggct
     4861 gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg ttatccacag aatcagggga
     4921 taacgcagga aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc
     4981 cgcgttgctg gcgtttttcc ataggctccg cccccctgac gagcatcaca aaaatcgacg
     5041 ctcaagtcag aggtggcgaa acccgacagg actataaaga taccaggcgt ttccccctgg
     5101 aagctccctc gtgcgctctc ctgttccgac cctgccgctt accggatacc tgtccgcctt
     5161 tctcccttcg ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc tcagttcggt
     5221 gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg
     5281 cgccttatcc ggtaactatc gtcttgagtc caacccggta agacacgact tatcgccact
     5341 ggcagcagcc actggtaaca ggattagcag agcgaggtat gtaggcggtg ctacagagtt
     5401 cttgaagtgg tggcctaact acggctacac tagaagaaca gtatttggta tctgcgctct
     5461 gctgaagcca gttaccttcg gaaaaagagt tggtagctct tgatccggca aacaaaccac
     5521 cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc
     5581 tcaagaagat cctttgatct tttctacggg gtctgacgct cagtggaacg aaaactcacg
     5641 ttaagggatt ttggtcatga gattatcaaa aaggatcttc acctagatcc ttttaaatta
     5701 aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa acttggtctg acagttacca
     5761 atgcttaatc agtgaggcac ctatctcagc gatctgtcta tttcgttcat ccatagttgc
     5821 ctgactcccc gtcgtgtaga taactacgat acgggagggc ttaccatctg gccccagtgc
     5881 tgcaatgata ccgcgagacc cacgctcacc ggctccagat ttatcagcaa taaaccagcc
     5941 agccggaagg gccgagcgca gaagtggtcc tgcaacttta tccgcctcca tccagtctat
     6001 taattgttgc cgggaagcta gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt
     6061 tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc
     6121 cggttcccaa cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa aagcggttag
     6181 ctccttcggt cctccgatcg ttgtcagaag taagttggcc gcagtgttat cactcatggt
     6241 tatggcagca ctgcataatt ctcttactgt catgccatcc gtaagatgct tttctgtgac
     6301 tggtgagtac tcaaccaagt cattctgaga atagtgtatg cggcgaccga gttgctcttg
     6361 cccggcgtca atacgggata ataccgcgcc acatagcaga actttaaaag tgctcatcat
     6421 tggaaaacgt tcttcggggc gaaaactctc aaggatctta ccgctgttga gatccagttc
     6481 gatgtaaccc actcgtgcac ccaactgatc ttcagcatct tttactttca ccagcgtttc
     6541 tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa
     6601 atgttgaata ctcatactct tcctttttca atattattga agcatttatc agggttattg
     6661 tctcatgagc ggatacatat ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg
     6721 cacatttccc cgaaaagtgc cacctgacgc gccctgtagc ggcgcattaa gcgcggcggg
     6781 tgtggtggtt acgcgcagcg tgaccgctac acttgccagc gccctagcgc ccgctccttt
     6841 cgctttcttc ccttcctttc tcgccacgtt cgccggcttt ccccgtcaag ctctaaatcg
     6901 ggggctccct ttagggttcc gatttagtgc tttacggcac ctcgacccca aaaaacttga
     6961 ttagggtgat ggttcacgta gtgggccatc gccctgatag acggtttttc gccctttgac
     7021 gttggagtcc acgttcttta atagtggact cttgttccaa actggaacaa cactcaaccc
     7081 tatctcggtc tattcttttg atttataagg gattttgccg atttcggcct attggttaaa
     7141 aaatgagctg atttaacaaa aatttaacgc gaattttaac aaaatattaa cgcttacaat
     7201 ttgccattcg ccattcaggc tgcgcaactg ttgggaaggg cgatcggtgc gggcctcttc
     7261 gctattacgc cagcccaagc taccatgata agtaagtaat attaaggtac gggaggtact
     7321 tggagcggcc gc
//
