LOCUS       sequence-240528-        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    47..71
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     protein_bind    complement(176..200)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     promoter        join(252..257,258..275,276..282)
                     /label=lac UV5 promoter
                     /note="E. coli lac promoter with an ""up"" mutation"
     CDS             336..995
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(402..421)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     CDS             1337..1642
                     /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     1521..1540
                     /label=ccdB-fwd
                     /note="ccdB gene, forward primer"
     regulatory      1730..1739
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     intron          1742..1874
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     primer_bind     complement(1918..1946)
                     /label=GFP-R
                     /note="GFP, reverse primer. Does NOT anneal to EGFP"
     polyA_signal    2113..2234
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2124..2143)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2178..2197
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(2498..2515)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2669..3257)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2749..2768)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3457..4248,4249..4317))
                     /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    4422..4470
                     /note="synthetic polyadenylation signal"
     misc_feature    4484..4575
                     /label=pause site
                     /note="RNA polymerase II transcriptional pause signal from 
                     the human alpha-2 globin gene"
     primer_bind     4524..4543
                     /label=RVprimer3
                     /note="pGL3 vector, forward primer"
ORIGIN
        1 ggcctaactg gccggtacct gagctcgcta gcctcgagga tatcaaacaa gtttgtacaa
       61 aaaagcaggc ttccgggagt actgtcctcc gagcggagta ctgtcctccg agcgggaagc
      121 ttgcatgcct gcaggtcgga gtactgtcct ccgagcggag tactgtcctc cgaggaccca
      181 gctttcttgt acaaagtggt agatcttaga gcatgcaccg gtgatatccg gccgcattag
      241 gcaccccagg ctttacactt tatgcttccg gctcgtataa tgtgtggatt ttgagttagg
      301 atccgtcgag attttcagga gctaaggaag ctaaaatgga gaaaaaaatc actggatata
      361 ccaccgttga tatatcccaa tggcatcgta aagaacattt tgaggcattt cagtcagttg
      421 ctcaatgtac ctataaccag accgttcagc tggatattac ggccttttta aagaccgtaa
      481 agaaaaataa gcacaagttt tatccggcct ttattcacat tcttgcccgc ctgatgaatg
      541 ctcatccgga attccgtatg gcaatgaaag acggtgagct ggtgatatgg gatagtgttc
      601 acccttgtta caccgttttc catgagcaaa ctgaaacgtt ttcatcgctc tggagtgaat
      661 accacgacga tttccggcag tttctacaca tatattcgca agatgtggcg tgttacggtg
      721 aaaacctggc ctatttccct aaagggttta ttgagaatat gtttttcgtc tcagccaatc
      781 cctgggtgag tttcaccagt tttgatttaa acgtggccaa tatggacaac ttcttcgccc
      841 ccgttttcac catgggcaaa tattatacgc aaggcgacaa ggtgctgatg ccgctggcga
      901 ttcaggttca tcatgccgtt tgtgatggct tccatgtcgg cagaatgctt aatgaattac
      961 aacagtactg cgatgagtgg cagggcgggg cgtaaacgcg tggatccggc ttactaaaag
     1021 ccagataaca gtatgcgtat ttgcgcgctg atttttgcgg tataagaata tatactgata
     1081 tgtatacccg aagtatgtca aaaagaggta tgctatgaag cagcgtatta cagtgacagt
     1141 tgacagcgac agctatcagt tgctcaaggc atatatgatg tcaatatctc cggtctggta
     1201 agcacaacca tgcagaatga agcccgtcgt ctgcgtgccg aacgctggaa agcggaaaat
     1261 caggaaggga tggctgaggt cgcccggttt attgaaatga acggctcttt tgctgacgag
     1321 aacaggggct ggtgaaatgc agtttaaggt ttacacctat aaaagagaga gccgttatcg
     1381 tctgtttgtg gatgtacaga gtgatattat tgacacgccc gggcgacgga tggtgatccc
     1441 cctggccagt gcacgtctgc tgtcagataa agtctcccgt gaactttacc cggtggtgca
     1501 tatcggggat gaaagctggc gcatgatgac caccgatatg gccagtgtgc cggtctccgt
     1561 tatcggggaa gaagtggctg atctcagcca ccgcgaaaat gacatcaaaa acgccattaa
     1621 cctgatgttc tggggaatat aaatgtcagg ctcccttata cacagccagt ctgcaggata
     1681 tcgtcgacga attcggcccc tgcaggggca ttccggtact gttggtaaag ccaccatgga
     1741 agtaagtatc aaggttacaa gacaggttta aggagaccaa tagaaactgg gcttgtcgag
     1801 acagagaaga ctcttgcgtt tctgataggc acctattggt cttactgaca tccactttgc
     1861 ctttctctcc acagccactt aagcatggct agcaaaggag aagaactctt cactggagtt
     1921 gtcccaattc ttgttgaatt agatggtgat gttaacggcc acaagttctc tgtcagtgga
     1981 gagggtgaag gtgatgcaac atacggaaaa cttaccctga agttcatctg cactactggc
     2041 aaactgcctg ttccctggcc aacactagtc tgatctagag tcggggcggc cggccgcttc
     2101 gagcagacat gataagatac attgatgagt ttggacaaac cacaactaga atgcagtgaa
     2161 aaaaatgctt tatttgtgaa atttgtgatg ctattgcttt atttgtaacc attataagct
     2221 gcaataaaca agttaacaac aacaattgca ttcattttat gtttcaggtt cagggggagg
     2281 tgtgggaggt tttttaaagc aagtaaaacc tctacaaatg tggtaaaatc gataaggatc
     2341 cgaccgatgc cctcaagctt tgagaggggc cctgagagcc ttcaacccag tcagctcctt
     2401 ccggtgggcg cggggcatga ctatcgtcgc cgcacttatg actgtcttct ttatcatgca
     2461 actcgtagga caggtgccgg cagcgctctt ccgcttcctc gctcactgac tcgctgcgct
     2521 cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa ggcggtaata cggttatcca
     2581 cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga
     2641 accgtaaaaa ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc
     2701 acaaaaatcg acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg
     2761 cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat
     2821 acctgtccgc ctttctccct tcgggaagcg tggcgctttc tcatagctca cgctgtaggt
     2881 atctcagttc ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc
     2941 agcccgaccg ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg
     3001 acttatcgcc actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg
     3061 gtgctacaga gttcttgaag tggtggccta actacggcta cactagaaga acagtatttg
     3121 gtatctgcgc tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg
     3181 gcaaacaaac caccgctggt agcggtggtt tttttgtttg caagcagcag attacgcgca
     3241 gaaaaaaagg atctcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga
     3301 acgaaaactc acgttaaggg attttggtca tgagattatc aaaaaggatc ttcacctaga
     3361 tccttttaaa ttaaaaatga agttttaaat caatctaaag tatatatgag taaacttggt
     3421 ctgacagcgg ccgcaaatgc taaaccactg cagtggttac cagtgcttga tcagtgaggc
     3481 accgatctca gcgatctgcc tatttcgttc gtccatagtg gcctgactcc ccgtcgtgta
     3541 gatcactacg attcgtgagg gcttaccatc aggccccagc gcagcaatga tgccgcgaga
     3601 gccgcgttca ccggcccccg atttgtcagc aatgaaccag ccagcaggga gggccgagcg
     3661 aagaagtggt cctgctactt tgtccgcctc catccagtct atgagctgct gtcgtgatgc
     3721 tagagtaaga agttcgccag tgagtagttt ccgaagagtt gtggccattg ctactggcat
     3781 cgtggtatca cgctcgtcgt tcggtatggc ttcgttcaac tctggttccc agcggtcaag
     3841 ccgggtcaca tgatcaccca tattatgaag aaatgcagtc agctccttag ggcctccgat
     3901 cgttgtcaga agtaagttgg ccgcggtgtt gtcgctcatg gtaatggcag cactacacaa
     3961 ttctcttacc gtcatgccat ccgtaagatg cttttccgtg accggcgagt actcaaccaa
     4021 gtcgttttgt gagtagtgta tacggcgacc aagctgctct tgcccggcgt ctatacggga
     4081 caacaccgcg ccacatagca gtactttgaa agtgctcatc atcgggaatc gttcttcggg
     4141 gcggaaagac tcaaggatct tgccgctatt gagatccagt tcgatatagc ccactcttgc
     4201 acccagttga tcttcagcat cttttacttt caccagcgtt tcggggtgtg caaaaacagg
     4261 caagcaaaat gccgcaaaga agggaatgag tgcgacacga aaatgttgga tgctcatact
     4321 cgtccttttt caatattatt gaagcattta tcagggttac tagtacgtct ctcaaggata
     4381 agtaagtaat attaaggtac gggaggtatt ggacaggccg caataaaata tctttatttt
     4441 cattacatct gtgtgttggt tttttgtgtg aatcgatagt actaacatac gctctccatc
     4501 aaaacaaaac gaaacaaaac aaactagcaa aataggctgt ccccagtgca agtgcaggtg
     4561 ccagaacatt tctct
//
