LOCUS       sequence-254778-        4575 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Vector to measure the activity of CP candidates in response to a 
            tethered (4xUAS) GAL4-DBD-COF by determining the abundance of 
            transcripts originating from each candidate in human cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4575)
  AUTHORS   Haberle V, Arnold CD, Pagani M, Rath M, Schernhuber K, Stark A
  TITLE     Transcriptional cofactors display specificity for distinct types of 
            core promoters.
  JOURNAL   Nature. 2019 Jun;570(7759):122-126. doi: 10.1038/s41586-019-1210-7. 
            Epub 2019 May 15.
  PUBMED    31092928
REFERENCE   2  (bases 1 to 4575)
  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"; doi: 
            "10.1038/s41586-019-1210-7"; journalName: "Nature"; date: "2019-06";
            volume: "570"; issue: "7759"; pages: "122-126"
COMMENT     Sequence Label: pSTAP-seq_human-4xUAS
FEATURES             Location/Qualifiers
     source          1..4575
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    complement(104..128)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     promoter        join(180..185,186..203,204..210)
                     /label=lac UV5 promoter
                     /note="E. coli lac promoter with an ""up"" mutation"
     CDS             264..923
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(330..349)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     CDS             1265..1570
                     /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     1449..1468
                     /label=ccdB-fwd
                     /note="ccdB gene, forward primer"
     regulatory      1658..1667
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     intron          1670..1802
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     primer_bind     complement(1846..1874)
                     /label=GFP-R
                     /note="GFP, reverse primer. Does NOT anneal to EGFP"
     polyA_signal    2041..2162
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2052..2071)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2106..2125
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(2426..2443)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2597..3185)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2677..2696)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3385..4176,4177..4245))
                     /codon_start=1
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     polyA_signal    4350..4398
                     /note="synthetic polyadenylation signal"
     misc_feature    4412..4503
                     /label=pause site
                     /note="RNA polymerase II transcriptional pause signal from 
                     the human alpha-2 globin gene"
     primer_bind     4452..4471
                     /label=RVprimer3
                     /note="pGL3 vector, forward primer"
     protein_bind    4550..4574
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
ORIGIN
        1 ccgggagtac tgtcctccga gcggagtact gtcctccgag cgggaagctt gcatgcctgc
       61 aggtcggagt actgtcctcc gagcggagta ctgtcctccg aggacccagc tttcttgtac
      121 aaagtggtag atcttagagc atgcaccggt gatatccggc cgcattaggc accccaggct
      181 ttacacttta tgcttccggc tcgtataatg tgtggatttt gagttaggat ccgtcgagat
      241 tttcaggagc taaggaagct aaaatggaga aaaaaatcac tggatatacc accgttgata
      301 tatcccaatg gcatcgtaaa gaacattttg aggcatttca gtcagttgct caatgtacct
      361 ataaccagac cgttcagctg gatattacgg cctttttaaa gaccgtaaag aaaaataagc
      421 acaagtttta tccggccttt attcacattc ttgcccgcct gatgaatgct catccggaat
      481 tccgtatggc aatgaaagac ggtgagctgg tgatatggga tagtgttcac ccttgttaca
      541 ccgttttcca tgagcaaact gaaacgtttt catcgctctg gagtgaatac cacgacgatt
      601 tccggcagtt tctacacata tattcgcaag atgtggcgtg ttacggtgaa aacctggcct
      661 atttccctaa agggtttatt gagaatatgt ttttcgtctc agccaatccc tgggtgagtt
      721 tcaccagttt tgatttaaac gtggccaata tggacaactt cttcgccccc gttttcacca
      781 tgggcaaata ttatacgcaa ggcgacaagg tgctgatgcc gctggcgatt caggttcatc
      841 atgccgtttg tgatggcttc catgtcggca gaatgcttaa tgaattacaa cagtactgcg
      901 atgagtggca gggcggggcg taaacgcgtg gatccggctt actaaaagcc agataacagt
      961 atgcgtattt gcgcgctgat ttttgcggta taagaatata tactgatatg tatacccgaa
     1021 gtatgtcaaa aagaggtatg ctatgaagca gcgtattaca gtgacagttg acagcgacag
     1081 ctatcagttg ctcaaggcat atatgatgtc aatatctccg gtctggtaag cacaaccatg
     1141 cagaatgaag cccgtcgtct gcgtgccgaa cgctggaaag cggaaaatca ggaagggatg
     1201 gctgaggtcg cccggtttat tgaaatgaac ggctcttttg ctgacgagaa caggggctgg
     1261 tgaaatgcag tttaaggttt acacctataa aagagagagc cgttatcgtc tgtttgtgga
     1321 tgtacagagt gatattattg acacgcccgg gcgacggatg gtgatccccc tggccagtgc
     1381 acgtctgctg tcagataaag tctcccgtga actttacccg gtggtgcata tcggggatga
     1441 aagctggcgc atgatgacca ccgatatggc cagtgtgccg gtctccgtta tcggggaaga
     1501 agtggctgat ctcagccacc gcgaaaatga catcaaaaac gccattaacc tgatgttctg
     1561 gggaatataa atgtcaggct cccttataca cagccagtct gcaggatatc gtcgacgaat
     1621 tcggcccctg caggggcatt ccggtactgt tggtaaagcc accatggaag taagtatcaa
     1681 ggttacaaga caggtttaag gagaccaata gaaactgggc ttgtcgagac agagaagact
     1741 cttgcgtttc tgataggcac ctattggtct tactgacatc cactttgcct ttctctccac
     1801 agccacttaa gcatggctag caaaggagaa gaactcttca ctggagttgt cccaattctt
     1861 gttgaattag atggtgatgt taacggccac aagttctctg tcagtggaga gggtgaaggt
     1921 gatgcaacat acggaaaact taccctgaag ttcatctgca ctactggcaa actgcctgtt
     1981 ccctggccaa cactagtctg atctagagtc ggggcggccg gccgcttcga gcagacatga
     2041 taagatacat tgatgagttt ggacaaacca caactagaat gcagtgaaaa aaatgcttta
     2101 tttgtgaaat ttgtgatgct attgctttat ttgtaaccat tataagctgc aataaacaag
     2161 ttaacaacaa caattgcatt cattttatgt ttcaggttca gggggaggtg tgggaggttt
     2221 tttaaagcaa gtaaaacctc tacaaatgtg gtaaaatcga taaggatccg accgatgccc
     2281 tcaagctttg agaggggccc ccttcaaccc agtcagctcc gacgtcttgg cgcccccgcg
     2341 gggcatgact atcgtcgtcg cacttatgac tgtcttcttt atcatgcaac tcgtaggaca
     2401 ggtgccggca gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc
     2461 tgcggcgagc ggtatcagct cactcaaagg cggtaatacg gttatccaca gaatcagggg
     2521 ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg
     2581 ccgcgttgct ggcgtttttc cataggctcc gcccccctga cgagcatcac aaaaatcgac
     2641 gctcaagtca gaggtggcga aacccgacag gactataaag ataccaggcg tttccccctg
     2701 gaagctccct cgtgcgctct cctgttccga ccctgccgct taccggatac ctgtccgcct
     2761 ttctcccttc gggaagcgtg gcgctttctc atagctcacg ctgtaggtat ctcagttcgg
     2821 tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct
     2881 gcgccttatc cggtaactat cgtcttgagt ccaacccggt aagacacgac ttatcgccac
     2941 tggcagcagc cactggtaac aggattagca gagcgaggta tgtaggcggt gctacagagt
     3001 tcttgaagtg gtggcctaac tacggctaca ctagaagaac agtatttggt atctgcgctc
     3061 tgctgaagcc agttaccttc ggaaaaagag ttggtagctc ttgatccggc aaacaaacca
     3121 ccgctggtag cggtggtttt tttgtttgca agcagcagat tacgcgcaga aaaaaaggat
     3181 ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc tcagtggaac gaaaactcac
     3241 gttaagggat tttggtcatg agattatcaa aaaggatctt cacctagatc cttttaaatt
     3301 aaaaatgaag ttttaaatca atctaaagta tatatgagta aacttggtct gacagcggcc
     3361 gcaaatgcta aaccactgca gtggttacca gtgcttgatc agtgaggcac cgatctcagc
     3421 gatctgccta tttcgttcgt ccatagtggc ctgactcccc gtcgtgtaga tcactacgat
     3481 tcgtgagggc ttaccatcag gccccagcgc agcaatgatg ccgcgagagc cgcgttcacc
     3541 ggcccccgat ttgtcagcaa tgaaccagcc agcagggagg gccgagcgaa gaagtggtcc
     3601 tgctactttg tccgcctcca tccagtctat gagctgctgt cgtgatgcta gagtaagaag
     3661 ttcgccagtg agtagtttcc gaagagttgt ggccattgct actggcatcg tggtatcacg
     3721 ctcgtcgttc ggtatggctt cgttcaactc tggttcccag cggtcaagcc gggtcacatg
     3781 atcacccata ttatgaagaa atgcagtcag ctccttaggg cctccgatcg ttgtcagaag
     3841 taagttggcc gcggtgttgt cgctcatggt aatggcagca ctacacaatt ctcttaccgt
     3901 catgccatcc gtaagatgct tttccgtgac cggcgagtac tcaaccaagt cgttttgtga
     3961 gtagtgtata cggcgaccaa gctgctcttg cccggcgtct atacgggaca acaccgcgcc
     4021 acatagcagt actttgaaag tgctcatcat cgggaatcgt tcttcggggc ggaaagactc
     4081 aaggatcttg ccgctattga gatccagttc gatatagccc actcttgcac ccagttgatc
     4141 ttcagcatct tttactttca ccagcgtttc ggggtgtgca aaaacaggca agcaaaatgc
     4201 cgcaaagaag ggaatgagtg cgacacgaaa atgttggatg ctcatactcg tcctttttca
     4261 atattattga agcatttatc agggttacta gtacgtctct caaggataag taagtaatat
     4321 taaggtacgg gaggtattgg acaggccgca ataaaatatc tttattttca ttacatctgt
     4381 gtgttggttt tttgtgtgaa tcgatagtac taacatacgc tctccatcaa aacaaaacga
     4441 aacaaaacaa actagcaaaa taggctgtcc ccagtgcaag tgcaggtgcc agaacatttc
     4501 tctggcctaa ctggccggta cctgagctcg ctagcctcga ggatatcaaa caagtttgta
     4561 caaaaaagca ggctt
//
