LOCUS       sequence-240527-        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"
     rep_origin      1..456
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(88..107)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     298..319
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     polyA_signal    587..635
                     /note="synthetic polyadenylation signal"
     misc_feature    649..740
                     /label=pause site
                     /note="RNA polymerase II transcriptional pause signal from 
                     the human alpha-2 globin gene"
     primer_bind     689..708
                     /label=RVprimer3
                     /note="pGL3 vector, forward primer"
     protein_bind    781..805
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     protein_bind    complement(910..934)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     promoter        join(992..997,998..1015,1016..1022)
                     /label=lac UV5 promoter
                     /note="E. coli lac promoter with an ""up"" mutation"
     CDS             1076..1735
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(1142..1161)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     CDS             2077..2382
                     /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     2261..2280
                     /label=ccdB-fwd
                     /note="ccdB gene, forward primer"
     regulatory      2470..2479
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     intron          2485..2551
                     /label=Mhc intron
                     /note="modified Drosophila myosin heavy chain intron 16 
                     (Pfeiffer et al., 2010)"
     primer_bind     complement(2596..2624)
                     /label=GFP-R
                     /note="GFP, reverse primer. Does NOT anneal to EGFP"
     polyA_signal    2791..2912
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2802..2821)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2856..2875
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(3176..3193)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3347..3935)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3427..3446)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4106..4897,4898..4966))
                     /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     4729..4748
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4967..5071)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 acgcgccctg tagcggcgca ttaagcgcgg cgggtgtggt ggttacgcgc agcgtgaccg
       61 ctacacttgc cagcgcccta gcgcccgctc ctttcgcttt cttcccttcc tttctcgcca
      121 cgttcgccgg ctttccccgt caagctctaa atcgggggct ccctttaggg ttccgattta
      181 gtgctttacg gcacctcgac cccaaaaaac ttgattaggg tgatggttca cgtagtgggc
      241 catcgccctg atagacggtt tttcgccctt tgacgttgga gtccacgttc tttaatagtg
      301 gactcttgtt ccaaactgga acaacactca accctatctc ggtctattct tttgatttat
      361 aagggatttt gccgatttcg gcctattggt taaaaaatga gctgatttaa caaaaattta
      421 acgcgaattt taacaaaata ttaacgctta caatttgcca ttcgccattc aggctgcgca
      481 actgttggga agggcgatcg gtgcgggcct cttcgctatt acgccagccc aagctaccat
      541 gataagtaag taatattaag gtacgggagg tacttggagc ggccgcaata aaatatcttt
      601 attttcatta catctgtgtg ttggtttttt gtgtgaatcg atagtactaa catacgctct
      661 ccatcaaaac aaaacgaaac aaaacaaact agcaaaatag gctgtcccca gtgcaagtgc
      721 aggtgccaga acatttctct atcgataggt acctgagctc gctagcctcg aggatatcaa
      781 acaagtttgt acaaaaaagc aggcttccgg gagtactgtc ctccgagcgg agtactgtcc
      841 tccgagcggg aagcttgcat gcctgcaggt cggagtactg tcctccgagc ggagtactgt
      901 cctccgagga cccagctttc ttgtacaaag tggtatatca agatcttaga gcatgcaccg
      961 gtgatatccg gccgcattag gcaccccagg ctttacactt tatgcttccg gctcgtataa
     1021 tgtgtggatt ttgagttagg atccgtcgag attttcagga gctaaggaag ctaaaatgga
     1081 gaaaaaaatc actggatata ccaccgttga tatatcccaa tggcatcgta aagaacattt
     1141 tgaggcattt cagtcagttg ctcaatgtac ctataaccag accgttcagc tggatattac
     1201 ggccttttta aagaccgtaa agaaaaataa gcacaagttt tatccggcct ttattcacat
     1261 tcttgcccgc ctgatgaatg ctcatccgga attccgtatg gcaatgaaag acggtgagct
     1321 ggtgatatgg gatagtgttc acccttgtta caccgttttc catgagcaaa ctgaaacgtt
     1381 ttcatcgctc tggagtgaat accacgacga tttccggcag tttctacaca tatattcgca
     1441 agatgtggcg tgttacggtg aaaacctggc ctatttccct aaagggttta ttgagaatat
     1501 gtttttcgtc tcagccaatc cctgggtgag tttcaccagt tttgatttaa acgtggccaa
     1561 tatggacaac ttcttcgccc ccgttttcac catgggcaaa tattatacgc aaggcgacaa
     1621 ggtgctgatg ccgctggcga ttcaggttca tcatgccgtt tgtgatggct tccatgtcgg
     1681 cagaatgctt aatgaattac aacagtactg cgatgagtgg cagggcgggg cgtaaacgcg
     1741 tggatccggc ttactaaaag ccagataaca gtatgcgtat ttgcgcgctg atttttgcgg
     1801 tataagaata tatactgata tgtatacccg aagtatgtca aaaagaggta tgctatgaag
     1861 cagcgtatta cagtgacagt tgacagcgac agctatcagt tgctcaaggc atatatgatg
     1921 tcaatatctc cggtctggta agcacaacca tgcagaatga agcccgtcgt ctgcgtgccg
     1981 aacgctggaa agcggaaaat caggaaggga tggctgaggt cgcccggttt attgaaatga
     2041 acggctcttt tgctgacgag aacaggggct ggtgaaatgc agtttaaggt ttacacctat
     2101 aaaagagaga gccgttatcg tctgtttgtg gatgtacaga gtgatattat tgacacgccc
     2161 gggcgacgga tggtgatccc cctggccagt gcacgtctgc tgtcagataa agtctcccgt
     2221 gaactttacc cggtggtgca tatcggggat gaaagctggc gcatgatgac caccgatatg
     2281 gccagtgtgc cggtctccgt tatcggggaa gaagtggctg atctcagcca ccgcgaaaat
     2341 gacatcaaaa acgccattaa cctgatgttc tggggaatat aaatgtcagg ctcccttata
     2401 cacagccagt ctgcaggata tcgtcgacga attcggcccc tgcaggggca ttccggtact
     2461 gttggtaaag ccaccatgga aaaggtaggt tcaaccactg atgcctaggc acaccgaaac
     2521 gactaaccct aattcttatc ctttacttca ggccacttaa gcatggctag caaaggagaa
     2581 gaactcttca ctggagttgt cccaattctt gttgaattag atggtgatgt taacggccac
     2641 aagttctctg tcagtggaga gggtgaaggt gatgcaacat acggaaaact taccctgaag
     2701 ttcatctgca ctactggcaa actgcctgtt ccctggccaa cactagtctg atctagagtc
     2761 ggggcggccg gccgcttcga gcagacatga taagatacat tgatgagttt ggacaaacca
     2821 caactagaat gcagtgaaaa aaatgcttta tttgtgaaat ttgtgatgct attgctttat
     2881 ttgtaaccat tataagctgc aataaacaag ttaacaacaa caattgcatt cattttatgt
     2941 ttcaggttca gggggaggtg tgggaggttt tttaaagcaa gtaaaacctc tacaaatgtg
     3001 gtaaaatcga taaggatccg accgatgccc tcaagctttg agaggggccc ccttcaaccc
     3061 agtcagctcc gacgtcttgg cgcccccgcg gggcatgact atcgtcgccg cacttatgac
     3121 tgtcttcttt atcatgcaac tcgtaggaca ggtgccggca gcgctcttcc gcttcctcgc
     3181 tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg
     3241 cggtaatacg gttatccaca gaatcagggg ataacgcagg aaagaacatg tgagcaaaag
     3301 gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc
     3361 gcccccctga cgagcatcac aaaaatcgac gctcaagtca gaggtggcga aacccgacag
     3421 gactataaag ataccaggcg tttccccctg gaagctccct cgtgcgctct cctgttccga
     3481 ccctgccgct taccggatac ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc
     3541 atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg
     3601 tgcacgaacc ccccgttcag cccgaccgct gcgccttatc cggtaactat cgtcttgagt
     3661 ccaacccggt aagacacgac ttatcgccac tggcagcagc cactggtaac aggattagca
     3721 gagcgaggta tgtaggcggt gctacagagt tcttgaagtg gtggcctaac tacggctaca
     3781 ctagaagaac agtatttggt atctgcgctc tgctgaagcc agttaccttc ggaaaaagag
     3841 ttggtagctc ttgatccggc aaacaaacca ccgctggtag cggtggtttt tttgtttgca
     3901 agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg
     3961 ggtctgacgc tcagtggaac gaaaactcac gttaagggat tttggtcatg agattatcaa
     4021 aaaggatctt cacctagatc cttttaaatt aaaaatgaag ttttaaatca atctaaagta
     4081 tatatgagta aacttggtct gacagttacc aatgcttaat cagtgaggca cctatctcag
     4141 cgatctgtct atttcgttca tccatagttg cctgactccc cgtcgtgtag ataactacga
     4201 tacgggaggg cttaccatct ggccccagtg ctgcaatgat accgcgagac ccacgctcac
     4261 cggctccaga tttatcagca ataaaccagc cagccggaag ggccgagcgc agaagtggtc
     4321 ctgcaacttt atccgcctcc atccagtcta ttaattgttg ccgggaagct agagtaagta
     4381 gttcgccagt taatagtttg cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac
     4441 gctcgtcgtt tggtatggct tcattcagct ccggttccca acgatcaagg cgagttacat
     4501 gatcccccat gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc gttgtcagaa
     4561 gtaagttggc cgcagtgtta tcactcatgg ttatggcagc actgcataat tctcttactg
     4621 tcatgccatc cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag tcattctgag
     4681 aatagtgtat gcggcgaccg agttgctctt gcccggcgtc aatacgggat aataccgcgc
     4741 cacatagcag aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct
     4801 caaggatctt accgctgttg agatccagtt cgatgtaacc cactcgtgca cccaactgat
     4861 cttcagcatc ttttactttc accagcgttt ctgggtgagc aaaaacagga aggcaaaatg
     4921 ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat actcatactc ttcctttttc
     4981 aatattattg aagcatttat cagggttatt gtctcatgag cggatacata tttgaatgta
     5041 tttagaaaaa taaacaaata ggggttccgc gcacatttcc ccgaaaagtg ccacctg
//
