LOCUS       sequence-247994-        5097 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 fly cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5097)
  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 5097)
  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_fly-4xUAS
FEATURES             Location/Qualifiers
     source          1..5097
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     74..95
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     polyA_signal    363..411
                     /note="synthetic polyadenylation signal"
     misc_feature    425..516
                     /label=pause site
                     /note="RNA polymerase II transcriptional pause signal from 
                     the human alpha-2 globin gene"
     primer_bind     465..484
                     /label=RVprimer3
                     /note="pGL3 vector, forward primer"
     protein_bind    557..581
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     protein_bind    complement(686..710)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     promoter        join(768..773,774..791,792..798)
                     /label=lac UV5 promoter
                     /note="E. coli lac promoter with an ""up"" mutation"
     CDS             852..1511
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(918..937)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     CDS             1853..2158
                     /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     2037..2056
                     /label=ccdB-fwd
                     /note="ccdB gene, forward primer"
     regulatory      2246..2255
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     intron          2261..2327
                     /label=Mhc intron
                     /note="modified Drosophila myosin heavy chain intron 16 
                     (Pfeiffer et al., 2010)"
     primer_bind     complement(2372..2400)
                     /label=GFP-R
                     /note="GFP, reverse primer. Does NOT anneal to EGFP"
     polyA_signal    2567..2688
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2578..2597)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2632..2651
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(2952..2969)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3123..3711)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3203..3222)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3882..4673,4674..4742))
                     /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     4505..4524
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4743..4847)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      join(4874..5097,1..232)
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4961..4980)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
ORIGIN
        1 ggttcacgta gtgggccatc gccctgatag acggtttttc gccctttgac gttggagtcc
       61 acgttcttta atagtggact cttgttccaa actggaacaa cactcaaccc tatctcggtc
      121 tattcttttg atttataagg gattttgccg atttcggcct attggttaaa aaatgagctg
      181 atttaacaaa aatttaacgc gaattttaac aaaatattaa cgcttacaat ttgccattcg
      241 ccattcaggc tgcgcaactg ttgggaaggg cgatcggtgc gggcctcttc gctattacgc
      301 cagcccaagc taccatgata agtaagtaat attaaggtac gggaggtact tggagcggcc
      361 gcaataaaat atctttattt tcattacatc tgtgtgttgg ttttttgtgt gaatcgatag
      421 tactaacata cgctctccat caaaacaaaa cgaaacaaaa caaactagca aaataggctg
      481 tccccagtgc aagtgcaggt gccagaacat ttctctatcg ataggtacct gagctcgcta
      541 gcctcgagga tatcaaacaa gtttgtacaa aaaagcaggc ttccgggagt actgtcctcc
      601 gagcggagta ctgtcctccg agcgggaagc ttgcatgcct gcaggtcgga gtactgtcct
      661 ccgagcggag tactgtcctc cgaggaccca gctttcttgt acaaagtggt atatcaagat
      721 cttagagcat gcaccggtga tatccggccg cattaggcac cccaggcttt acactttatg
      781 cttccggctc gtataatgtg tggattttga gttaggatcc gtcgagattt tcaggagcta
      841 aggaagctaa aatggagaaa aaaatcactg gatataccac cgttgatata tcccaatggc
      901 atcgtaaaga acattttgag gcatttcagt cagttgctca atgtacctat aaccagaccg
      961 ttcagctgga tattacggcc tttttaaaga ccgtaaagaa aaataagcac aagttttatc
     1021 cggcctttat tcacattctt gcccgcctga tgaatgctca tccggaattc cgtatggcaa
     1081 tgaaagacgg tgagctggtg atatgggata gtgttcaccc ttgttacacc gttttccatg
     1141 agcaaactga aacgttttca tcgctctgga gtgaatacca cgacgatttc cggcagtttc
     1201 tacacatata ttcgcaagat gtggcgtgtt acggtgaaaa cctggcctat ttccctaaag
     1261 ggtttattga gaatatgttt ttcgtctcag ccaatccctg ggtgagtttc accagttttg
     1321 atttaaacgt ggccaatatg gacaacttct tcgcccccgt tttcaccatg ggcaaatatt
     1381 atacgcaagg cgacaaggtg ctgatgccgc tggcgattca ggttcatcat gccgtttgtg
     1441 atggcttcca tgtcggcaga atgcttaatg aattacaaca gtactgcgat gagtggcagg
     1501 gcggggcgta aacgcgtgga tccggcttac taaaagccag ataacagtat gcgtatttgc
     1561 gcgctgattt ttgcggtata agaatatata ctgatatgta tacccgaagt atgtcaaaaa
     1621 gaggtatgct atgaagcagc gtattacagt gacagttgac agcgacagct atcagttgct
     1681 caaggcatat atgatgtcaa tatctccggt ctggtaagca caaccatgca gaatgaagcc
     1741 cgtcgtctgc gtgccgaacg ctggaaagcg gaaaatcagg aagggatggc tgaggtcgcc
     1801 cggtttattg aaatgaacgg ctcttttgct gacgagaaca ggggctggtg aaatgcagtt
     1861 taaggtttac acctataaaa gagagagccg ttatcgtctg tttgtggatg tacagagtga
     1921 tattattgac acgcccgggc gacggatggt gatccccctg gccagtgcac gtctgctgtc
     1981 agataaagtc tcccgtgaac tttacccggt ggtgcatatc ggggatgaaa gctggcgcat
     2041 gatgaccacc gatatggcca gtgtgccggt ctccgttatc ggggaagaag tggctgatct
     2101 cagccaccgc gaaaatgaca tcaaaaacgc cattaacctg atgttctggg gaatataaat
     2161 gtcaggctcc cttatacaca gccagtctgc aggatatcgt cgacgaattc ggcccctgca
     2221 ggggcattcc ggtactgttg gtaaagccac catggaaaag gtaggttcaa ccactgatgc
     2281 ctaggcacac cgaaacgact aaccctaatt cttatccttt acttcaggcc acttaagcat
     2341 ggctagcaaa ggagaagaac tcttcactgg agttgtccca attcttgttg aattagatgg
     2401 tgatgttaac ggccacaagt tctctgtcag tggagagggt gaaggtgatg caacatacgg
     2461 aaaacttacc ctgaagttca tctgcactac tggcaaactg cctgttccct ggccaacact
     2521 agtctgatct agagtcgggg cggccggccg cttcgagcag acatgataag atacattgat
     2581 gagtttggac aaaccacaac tagaatgcag tgaaaaaaat gctttatttg tgaaatttgt
     2641 gatgctattg ctttatttgt aaccattata agctgcaata aacaagttaa caacaacaat
     2701 tgcattcatt ttatgtttca ggttcagggg gaggtgtggg aggtttttta aagcaagtaa
     2761 aacctctaca aatgtggtaa aatcgataag gatccgaccg atgccctcaa gctttgagag
     2821 gggccccctt caacccagtc agctccgacg tcttggcgcc cccgcggggc atgactatcg
     2881 tcgccgcact tatgactgtc ttctttatca tgcaactcgt aggacaggtg ccggcagcgc
     2941 tcttccgctt cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta
     3001 tcagctcact caaaggcggt aatacggtta tccacagaat caggggataa cgcaggaaag
     3061 aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg
     3121 tttttccata ggctccgccc ccctgacgag catcacaaaa atcgacgctc aagtcagagg
     3181 tggcgaaacc cgacaggact ataaagatac caggcgtttc cccctggaag ctccctcgtg
     3241 cgctctcctg ttccgaccct gccgcttacc ggatacctgt ccgcctttct cccttcggga
     3301 agcgtggcgc tttctcatag ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc
     3361 tccaagctgg gctgtgtgca cgaacccccc gttcagcccg accgctgcgc cttatccggt
     3421 aactatcgtc ttgagtccaa cccggtaaga cacgacttat cgccactggc agcagccact
     3481 ggtaacagga ttagcagagc gaggtatgta ggcggtgcta cagagttctt gaagtggtgg
     3541 cctaactacg gctacactag aagaacagta tttggtatct gcgctctgct gaagccagtt
     3601 accttcggaa aaagagttgg tagctcttga tccggcaaac aaaccaccgc tggtagcggt
     3661 ggtttttttg tttgcaagca gcagattacg cgcagaaaaa aaggatctca agaagatcct
     3721 ttgatctttt ctacggggtc tgacgctcag tggaacgaaa actcacgtta agggattttg
     3781 gtcatgagat tatcaaaaag gatcttcacc tagatccttt taaattaaaa atgaagtttt
     3841 aaatcaatct aaagtatata tgagtaaact tggtctgaca gttaccaatg cttaatcagt
     3901 gaggcaccta tctcagcgat ctgtctattt cgttcatcca tagttgcctg actccccgtc
     3961 gtgtagataa ctacgatacg ggagggctta ccatctggcc ccagtgctgc aatgataccg
     4021 cgagacccac gctcaccggc tccagattta tcagcaataa accagccagc cggaagggcc
     4081 gagcgcagaa gtggtcctgc aactttatcc gcctccatcc agtctattaa ttgttgccgg
     4141 gaagctagag taagtagttc gccagttaat agtttgcgca acgttgttgc cattgctaca
     4201 ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat tcagctccgg ttcccaacga
     4261 tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag cggttagctc cttcggtcct
     4321 ccgatcgttg tcagaagtaa gttggccgca gtgttatcac tcatggttat ggcagcactg
     4381 cataattctc ttactgtcat gccatccgta agatgctttt ctgtgactgg tgagtactca
     4441 accaagtcat tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata
     4501 cgggataata ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg aaaacgttct
     4561 tcggggcgaa aactctcaag gatcttaccg ctgttgagat ccagttcgat gtaacccact
     4621 cgtgcaccca actgatcttc agcatctttt actttcacca gcgtttctgg gtgagcaaaa
     4681 acaggaaggc aaaatgccgc aaaaaaggga ataagggcga cacggaaatg ttgaatactc
     4741 atactcttcc tttttcaata ttattgaagc atttatcagg gttattgtct catgagcgga
     4801 tacatatttg aatgtattta gaaaaataaa caaatagggg ttccgcgcac atttccccga
     4861 aaagtgccac ctgacgcgcc ctgtagcggc gcattaagcg cggcgggtgt ggtggttacg
     4921 cgcagcgtga ccgctacact tgccagcgcc ctagcgcccg ctcctttcgc tttcttccct
     4981 tcctttctcg ccacgttcgc cggctttccc cgtcaagctc taaatcgggg gctcccttta
     5041 gggttccgat ttagtgcttt acggcacctc gaccccaaaa aacttgatta gggtgat
//
