LOCUS       sequence-239300-        6414 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Gateway-compatible vector to express a fusion protein containing the
            GAL4-DBD (N-terminally) followed by the protein of interest under 
            the control of a D.pseudoobscura enhancer and CP.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6414)
  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 6414)
  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: pAGW-dpse-GAL4-DBD
FEATURES             Location/Qualifiers
     source          1..6414
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             1433..1474
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     CDS             1484..1924
                     /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"
     protein_bind    1946..2070
                     /gene="mutant version of attR"
                     /label=attR1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     promoter        join(2095..2100,2101..2118,2119..2125)
                     /label=lac UV5 promoter
                     /note="E. coli lac promoter with an ""up"" mutation"
     CDS             2179..2838
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(2245..2264)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     CDS             3180..3485
                     /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
                     VPRELYPVVHIGDESWRMMTTDMASVPVSVIGEEVADLSHRENDIKNAINLMFWGI"
     primer_bind     3364..3383
                     /label=ccdB-fwd
                     /note="ccdB gene, forward primer"
     protein_bind    complement(3526..3650)
                     /gene="mutant version of attR"
                     /label=attR2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     intron          3742..3807
                     /label=small t intron
                     /note="SV40 (simian virus 40) small t antigen intron"
     CDS             3937..3957
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     polyA_signal    4382..4516
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4419..4438)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4473..4492
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     CDS             join(4613..4681,4682..5473)
                     /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     complement(4831..4850)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5644..6232
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6133..6152
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6386..6403
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 gcgcccaata cgcaaaccgc ctctccccgc gcgttggccg attcattaat gcaggcaact
       61 cgtgaaaggt aggcggatca gcggccgcag catgcaattc tatattctaa aaacacaaat
      121 gatacttcta aaaaaaaatc atgaatggca tcaactctga atcaaatctt tgcagatgca
      181 cctacttctc atttccactg tcacatcatt tttccagatc tatctgagga tgccgcaccc
      241 tgccccacct gtggctgggc attcgacctt caggggccat tgtcccttgt aatgcccacc
      301 cactcctatt agtactctta taaatctttt cttatagatt attgttcatt taaagtatgt
      361 tcttcccaaa acaagagctt catggttcat ggccgctaca cccggaagca actgcagccg
      421 ttcacacgct gacagcgaac tgacacgaat actcataatc atttcaatta gacgtgttgc
      481 aattagcctt gtcgacccgc caccactgca ggttcattcc gtacgagtat gttgcacaat
      541 tacgtaaatt agcggcgagg aattcaatat gcttgggcag gcaaagaggg agtcgagcga
      601 gagtccctgt gtgcgcatta tctttcaggg ccccgagcgc ttgtaattgc caatgcttaa
      661 cacgattgag tgaatcatcg tgaatcactg aatcattcat tgtgtgactc gcggaatgcg
      721 atatcaattc catgcgagct tctggatgtc aaaacactcc acggaagacc agattctacg
      781 aaaagctggt tcttataatg cgaaccatta atttagtaga tcactctaca tatacatact
      841 atctccgact gattcttggt gcgaactatt tgttttttgc tgacagatcc actgaaaagg
      901 taaagaataa tgatatattt ccgcttccgt tcaaggtctt ggatcagaga tacttcatcg
      961 ttgtcccatg tttttttaga tttttgctta tttttggttg gttttttatt gtcacatttt
     1021 ttcgcacgtt ttatttgatt tatgtctgtg acggctgctg accgccctgc tcctcctatt
     1081 atttctttta tccctgatac atggctgttg tgtggcgcaa atttccttat ctcaggcctt
     1141 gtcggactgg ccatatttta tcgcatgctt tgcctttctt ttctgctttg caggcaaatt
     1201 gaaaaggcaa ttcccatcag acgtctggct cgagatctac tgcatcgatc agtgcggtcg
     1261 actggcgttg taattttttt atatagcggg cttttgcgca ctccgtgcat catttagtaa
     1321 cgaagcttaa accgaaccgc atcgctctga agctctaaag cgcgcgcgct ctccttcccc
     1381 gcggaaccat cgagaaccaa catcgacaag aactacaaca acagcgagaa tggggaagcc
     1441 aatccctaat ccccttctgg gactcgactc taccggcggc tctatgaagc tactgtcttc
     1501 tatcgaacaa gcatgcgata tttgccgact taaaaagctc aagtgctcca aagaaaaacc
     1561 gaagtgcgcc aagtgtctga agaacaactg ggagtgtcgc tactctccca aaaccaaaag
     1621 gtctccgctg actagggcac atctgacaga agtggaatca aggctagaaa gactggaaca
     1681 gctatttcta ctgatttttc ctcgagaaga ccttgacatg attttgaaaa tggattcttt
     1741 acaggatata aaagcattgt taacaggatt atttgtacaa gataatgtga ataaagatgc
     1801 cgtcacagat agattggctt cagtggagac tgatatgcct ctaacattga gacagcatag
     1861 aataagtgcg acatcatcat cggaagagag tagtaacaaa ggtcaaagac agttgactgt
     1921 atcgtcgggc ggcggccacc ggtatacaag tttgtacaaa aaagctgaac gagaaacgta
     1981 aaatgatata aatatcaata tattaaatta gattttgcat aaaaaacaga ctacataata
     2041 ctgtaaaaca caacatatcc agtcactatg gcggccgcat taggcacccc aggctttaca
     2101 ctttatgctt ccggctcgta taatgtgtgg attttgagtt aggatccgtc gagattttca
     2161 ggagctaagg aagctaaaat ggagaaaaaa atcactggat ataccaccgt tgatatatcc
     2221 caatggcatc gtaaagaaca ttttgaggca tttcagtcag ttgctcaatg tacctataac
     2281 cagaccgttc agctggatat tacggccttt ttaaagaccg taaagaaaaa taagcacaag
     2341 ttttatccgg cctttattca cattcttgcc cgcctgatga atgctcatcc ggaattccgt
     2401 atggcaatga aagacggtga gctggtgata tgggatagtg ttcacccttg ttacaccgtt
     2461 ttccatgagc aaactgaaac gttttcatcg ctctggagtg aataccacga cgatttccgg
     2521 cagtttctac acatatattc gcaagatgtg gcgtgttacg gtgaaaacct ggcctatttc
     2581 cctaaagggt ttattgagaa tatgtttttc gtctcagcca atccctgggt gagtttcacc
     2641 agttttgatt taaacgtggc caatatggac aacttcttcg cccccgtttt caccatgggc
     2701 aaatattata cgcaaggcga caaggtgctg atgccgctgg cgattcaggt tcatcatgcc
     2761 gtttgtgatg gcttccatgt cggcagaatg cttaatgaat tacaacagta ctgcgatgag
     2821 tggcagggcg gggcgtaatc tagaggatcc ggcttactaa aagccagata acagtatgcg
     2881 tatttgcgcg ctgatttttg cggtataaga atatatactg atatgtatac ccgaagtatg
     2941 tcaaaaagag gtatgctatg aagcagcgta ttacagtgac agttgacagc gacagctatc
     3001 agttgctcaa ggcatatatg atgtcaatat ctccggtctg gtaagcacaa ccatgcagaa
     3061 tgaagcccgt cgtctgcgtg ccgaacgctg gaaagcggaa aatcaggaag ggatggctga
     3121 ggtcgcccgg tttattgaaa tgaacggctc ttttgctgac gagaacaggg gctggtgaaa
     3181 tgcagtttaa ggtttacacc tataaaagag agagccgtta tcgtctgttt gtggatgtac
     3241 agagtgatat tattgacacg cccgggcgac ggatggtgat ccccctggcc agtgcacgtc
     3301 tgctgtcaga taaagtcccc cgtgaacttt acccggtggt gcatatcggg gatgaaagct
     3361 ggcgcatgat gaccaccgat atggccagtg tgccggtctc cgttatcggg gaagaagtgg
     3421 ctgatctcag ccaccgcgaa aatgacatca aaaacgccat taacctgatg ttctggggaa
     3481 tataaatgtc aggctccctt atacacagcc agtctgcagg tcgaccatag tgactggata
     3541 tgttgtgttt tacagtatta tgtagtctgt tttttatgca aaatctaatt taatatattg
     3601 atatttatat cattttacgt ttctcgttca gctttcttgt acaaagtggt gacgtaagct
     3661 agcaggatct ttgtgaagga accttacttc tgtggtgtga cataattgga caaactacct
     3721 acagagattt aaagctctaa ggtaaatata aaatttttaa gtgtataatg tgttaaacta
     3781 ctgattctaa ttgtttgtgt attttagatt ccaacctatg gaactgatga atgggagcag
     3841 tggtggaatg cctttaatga ggaaaacctg ttttgctcag aagaaatgcc atctagtgat
     3901 gatgaggcta ctgctgactc tcaacattct actcctccaa aaaagaagag aaaggtagaa
     3961 gaccccaagg actttccttc agaattgcta agttttttga gtcatgctgt gtttagtaat
     4021 agaactcttg cttgctttgc tatttacacc acaaaggaaa aagctgcact gctatacaag
     4081 aaaattatgg aaaaatattc tgtaaccttt ataagtaggc ataacagtta taatcataac
     4141 atactgtttt ttcttactcc acacaggcat agagtgtctg ctattaataa ctatgctcaa
     4201 aaattgtgta cctttagctt tttaatttgt aaaggggtta ataaggaata tttgatgtat
     4261 agtgccttga ctagagatca taatcagcca taccacattt gtagaggttt tacttgcttt
     4321 aaaaaacctc ccacacctcc ccctgaacct gaaacataaa atgaatgcaa ttgttgttgt
     4381 taacttgttt attgcagctt ataatggtta caaataaagc aatagcatca caaatttcac
     4441 aaataaagca tttttttcac tgcattctag ttgtggtttg tccaaactca tcaatgtatc
     4501 ttatcatgtc tggatcccgt ttaaactacg cgtaattcaa acagggttct ggcgtcgttc
     4561 tcgtactgtt ttccccaggc cagtgcttta gcgttattga aaaaggaaga gtatgagtat
     4621 tcaacatttc cgtgtcgccc ttattccctt ttttgcggca ttttgccttc ctgtttttgc
     4681 tcacccagaa acgctggtga aagtaaaaga tgctgaagat cagttgggtg cacgagtggg
     4741 ttacatcgaa ctggatctca acagcggtaa gatccttgag agttttcgcc ccgaagaacg
     4801 ttttccaatg atgagcactt ttaaagttct gctatgtggc gcggtattat cccgtattga
     4861 cgccgggcaa gagcaactcg gtcgccgcat acactattct cagaatgact tggttgagta
     4921 ctcaccagtc acagaaaagc atcttacgga tggcatgaca gtaagagaat tatgcagtgc
     4981 tgccataacc atgagtgata acactgcggc caacttactt ctgacaacga tcggaggacc
     5041 gaaggagcta accgcttttt tgcacaacat gggggatcat gtaactcgcc ttgatcgttg
     5101 ggaaccggag ctgaatgaag ccataccaaa cgacgagcgt gacaccacga tgcctgtagc
     5161 aatggcaaca acgttgcgca aactattaac tggcgaacta cttactctag cttcccggca
     5221 acaattaata gactggatgg aggcggataa agttgcagga ccacttctgc gctcggccct
     5281 tccggctggc tggtttattg ctgataaatc tggagccggt gagcgtgggt ctcgcggtat
     5341 cattgcagca ctggggccag atggtaagcc ctcccgtatc gtagttatct acacgacggg
     5401 gagtcaggca actatggatg aacgaaatag acagatcgct gagataggtg cctcactgat
     5461 taagcattgg taactgtcag accaagttta ctcatatata ctttagattg atttaaaact
     5521 tcatttttaa tttaaaagga tctaggtgaa gatccttttt gataatctca tgaccaaaat
     5581 cccttaacgt gagttttcgt tccactgagc gtcagacccc gtagaaaaga tcaaaggatc
     5641 ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct
     5701 accagcggtg gtttgtttgc cggatcaaga gctaccaact ctttttccga aggtaactgg
     5761 cttcagcaga gcgcagatac caaatactgt ccttctagtg tagccgtagt taggccacca
     5821 cttcaagaac tctgtagcac cgcctacata cctcgctctg ctaatcctgt taccagtggc
     5881 tgctgccagt ggcgataagt cgtgtcttac cgggttggac tcaagacgat agttaccgga
     5941 taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca cagcccagct tggagcgaac
     6001 gacctacacc gaactgagat acctacagcg tgagctatga gaaagcgcca cgcttcccga
     6061 agggagaaag gcggacaggt atccggtaag cggcagggtc ggaacaggag agcgcacgag
     6121 ggagcttcca gggggaaacg cctggtatct ttatagtcct gtcgggtttc gccacctctg
     6181 acttgagcgt cgatttttgt gatgctcgtc aggggggcgg agcctatgga aaaacgccag
     6241 caacgcggcc tttttacggt tcctggcctt ttgctggcct tttgctcaca tgttctttcc
     6301 tgcgttatcc cctgattctg tggataaccg tattaccgcc tttgagtgag ctgataccgc
     6361 tcgccgcagc cgaacgaccg agcgcagcga gtcagtgagc gaggaagcgg aaga
//
