LOCUS       sequence-248016-        6413 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 6413)
  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 6413)
  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..6413
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             152..193
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     CDS             203..643
                     /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    665..789
                     /gene="mutant version of attR"
                     /label=attR1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     promoter        join(814..819,820..837,838..844)
                     /label=lac UV5 promoter
                     /note="E. coli lac promoter with an ""up"" mutation"
     CDS             898..1578
                     /codon_start=1
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQAGRNLE
                     DPAY"
     primer_bind     complement(964..983)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     CDS             1898..2203
                     /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     2082..2101
                     /label=ccdB-fwd
                     /note="ccdB gene, forward primer"
     protein_bind    complement(2244..2368)
                     /gene="mutant version of attR"
                     /label=attR2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     intron          2460..2525
                     /label=small t intron
                     /note="SV40 (simian virus 40) small t antigen intron"
     CDS             2655..2675
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     polyA_signal    3100..3234
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3137..3156)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3191..3210
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     CDS             join(3331..3399,3400..4191)
                     /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(3549..3568)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4362..4950
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4851..4870
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5104..5121
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 tatagcgggc ttttgcgcac tccgtgcatc atttagtaac gaagcttaaa ccgaaccgca
       61 tcgctctgaa gctctaaagc gcgcgcgctc tccttccccg cggaaccatc gagaaccaac
      121 atcgacaaga actacaacaa cagcgagaat ggggaagcca atccctaatc cccttctggg
      181 actcgactct accggcggct ctatgaagct actgtcttct atcgaacaag catgcgatat
      241 ttgccgactt aaaaagctca agtgctccaa agaaaaaccg aagtgcgcca agtgtctgaa
      301 gaacaactgg gagtgtcgct actctcccaa aaccaaaagg tctccgctga ctagggcaca
      361 tctgacagaa gtggaatcaa ggctagaaag actggaacag ctatttctac tgatttttcc
      421 tcgagaagac cttgacatga ttttgaaaat ggattcttta caggatataa aagcattgtt
      481 aacaggatta tttgtacaag ataatgtgaa taaagatgcc gtcacagata gattggcttc
      541 agtggagact gatatgcctc taacattgag acagcataga ataagtgcga catcatcatc
      601 ggaagagagt agtaacaaag gtcaaagaca gttgactgta tcgtcgggcg gcggccaccg
      661 gtatacaagt ttgtacaaaa aagctgaacg agaaacgtaa aatgatataa atatcaatat
      721 attaaattag attttgcata aaaaacagac tacataatac tgtaaaacac aacatatcca
      781 gtcactatgg cggccgcatt aggcacccca ggctttacac tttatgcttc cggctcgtat
      841 aatgtgtgga ttttgagtta ggatccgtcg agattttcag gagctaagga agctaaaatg
      901 gagaaaaaaa tcactggata taccaccgtt gatatatccc aatggcatcg taaagaacat
      961 tttgaggcat ttcagtcagt tgctcaatgt acctataacc agaccgttca gctggatatt
     1021 acggcctttt taaagaccgt aaagaaaaat aagcacaagt tttatccggc ctttattcac
     1081 attcttgccc gcctgatgaa tgctcatccg gaattccgta tggcaatgaa agacggtgag
     1141 ctggtgatat gggatagtgt tcacccttgt tacaccgttt tccatgagca aactgaaacg
     1201 ttttcatcgc tctggagtga ataccacgac gatttccggc agtttctaca catatattcg
     1261 caagatgtgg cgtgttacgg tgaaaacctg gcctatttcc ctaaagggtt tattgagaat
     1321 atgtttttcg tctcagccaa tccctgggtg agtttcacca gttttgattt aaacgtggcc
     1381 aatatggaca acttcttcgc ccccgttttc accatgggca aatattatac gcaaggcgac
     1441 aaggtgctga tgccgctggc gattcaggtt catcatgccg tttgtgatgg cttccatgtc
     1501 ggcagaatgc ttaatgaatt acaacagtac tgcgatgagt ggcaggcggg gcgtaatcta
     1561 gaggatccgg cttactaaaa gccagataac agtatgcgta tttgcgcgct gatttttgcg
     1621 gtataagaat atatactgat atgtataccc gaagtatgtc aaaaagaggt atgctatgaa
     1681 gcagcgtatt acagtgacag ttgacagcga cagctatcag ttgctcaagg catatatgat
     1741 gtcaatatct ccggtctggt aagcacaacc atgcagaatg aagcccgtcg tctgcgtgcc
     1801 gaacgctgga aagcggaaaa tcaggaaggg atggctgagg tcgcccggtt tattgaaatg
     1861 aacggctctt ttgctgacga gaacaggggc tggtgaaatg cagtttaagg tttacaccta
     1921 taaaagagag agccgttatc gtctgtttgt ggatgtacag agtgatatta ttgacacgcc
     1981 cgggcgacgg atggtgatcc ccctggccag tgcacgtctg ctgtcagata aagtcccccg
     2041 tgaactttac ccggtggtgc atatcgggga tgaaagctgg cgcatgatga ccaccgatat
     2101 ggccagtgtg ccggtctccg ttatcgggga agaagtggct gatctcagcc accgcgaaaa
     2161 tgacatcaaa aacgccatta acctgatgtt ctggggaata taaatgtcag gctcccttat
     2221 acacagccag tctgcaggtc gaccatagtg actggatatg ttgtgtttta cagtattatg
     2281 tagtctgttt tttatgcaaa atctaattta atatattgat atttatatca ttttacgttt
     2341 ctcgttcagc tttcttgtac aaagtggtga cgtaagctag caggatcttt gtgaaggaac
     2401 cttacttctg tggtgtgaca taattggaca aactacctac agagatttaa agctctaagg
     2461 taaatataaa atttttaagt gtataatgtg ttaaactact gattctaatt gtttgtgtat
     2521 tttagattcc aacctatgga actgatgaat gggagcagtg gtggaatgcc tttaatgagg
     2581 aaaacctgtt ttgctcagaa gaaatgccat ctagtgatga tgaggctact gctgactctc
     2641 aacattctac tcctccaaaa aagaagagaa aggtagaaga ccccaaggac tttccttcag
     2701 aattgctaag ttttttgagt catgctgtgt ttagtaatag aactcttgct tgctttgcta
     2761 tttacaccac aaaggaaaaa gctgcactgc tatacaagaa aattatggaa aaatattctg
     2821 taacctttat aagtaggcat aacagttata atcataacat actgtttttt cttactccac
     2881 acaggcatag agtgtctgct attaataact atgctcaaaa attgtgtacc tttagctttt
     2941 taatttgtaa aggggttaat aaggaatatt tgatgtatag tgccttgact agagatcata
     3001 atcagccata ccacatttgt agaggtttta cttgctttaa aaaacctccc acacctcccc
     3061 ctgaacctga aacataaaat gaatgcaatt gttgttgtta acttgtttat tgcagcttat
     3121 aatggttaca aataaagcaa tagcatcaca aatttcacaa ataaagcatt tttttcactg
     3181 cattctagtt gtggtttgtc caaactcatc aatgtatctt atcatgtctg gatcccgttt
     3241 aaactacgcg taattcaaac agggttctgg cgtcgttctc gtactgtttt ccccaggcca
     3301 gtgctttagc gttattgaaa aaggaagagt atgagtattc aacatttccg tgtcgccctt
     3361 attccctttt ttgcggcatt ttgccttcct gtttttgctc acccagaaac gctggtgaaa
     3421 gtaaaagatg ctgaagatca gttgggtgca cgagtgggtt acatcgaact ggatctcaac
     3481 agcggtaaga tccttgagag ttttcgcccc gaagaacgtt ttccaatgat gagcactttt
     3541 aaagttctgc tatgtggcgc ggtattatcc cgtattgacg ccgggcaaga gcaactcggt
     3601 cgccgcatac actattctca gaatgacttg gttgagtact caccagtcac agaaaagcat
     3661 cttacggatg gcatgacagt aagagaatta tgcagtgctg ccataaccat gagtgataac
     3721 actgcggcca acttacttct gacaacgatc ggaggaccga aggagctaac cgcttttttg
     3781 cacaacatgg gggatcatgt aactcgcctt gatcgttggg aaccggagct gaatgaagcc
     3841 ataccaaacg acgagcgtga caccacgatg cctgtagcaa tggcaacaac gttgcgcaaa
     3901 ctattaactg gcgaactact tactctagct tcccggcaac aattaataga ctggatggag
     3961 gcggataaag ttgcaggacc acttctgcgc tcggcccttc cggctggctg gtttattgct
     4021 gataaatctg gagccggtga gcgtgggtct cgcggtatca ttgcagcact ggggccagat
     4081 ggtaagccct cccgtatcgt agttatctac acgacgggga gtcaggcaac tatggatgaa
     4141 cgaaatagac agatcgctga gataggtgcc tcactgatta agcattggta actgtcagac
     4201 caagtttact catatatact ttagattgat ttaaaacttc atttttaatt taaaaggatc
     4261 taggtgaaga tcctttttga taatctcatg accaaaatcc cttaacgtga gttttcgttc
     4321 cactgagcgt cagaccccgt agaaaagatc aaaggatctt cttgagatcc tttttttctg
     4381 cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg
     4441 gatcaagagc taccaactct ttttccgaag gtaactggct tcagcagagc gcagatacca
     4501 aatactgttc ttctagtgta gccgtagtta ggccaccact tcaagaactc tgtagcaccg
     4561 cctacatacc tcgctctgct aatcctgtta ccagtggctg ctgccagtgg cgataagtcg
     4621 tgtcttaccg ggttggactc aagacgatag ttaccggata aggcgcagcg gtcgggctga
     4681 acggggggtt cgtgcacaca gcccagcttg gagcgaacga cctacaccga actgagatac
     4741 ctacagcgtg agctatgaga aagcgccacg cttcccgaag ggagaaaggc ggacaggtat
     4801 ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg agcttccagg gggaaacgcc
     4861 tggtatcttt atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg atttttgtga
     4921 tgctcgtcag gggggcggag cctatggaaa aacgccagca acgcggcctt tttacggttc
     4981 ctggcctttt gctggccttt tgctcacatg ttctttcctg cgttatcccc tgattctgtg
     5041 gataaccgta ttaccgcctt tgagtgagct gataccgctc gccgcagccg aacgaccgag
     5101 cgcagcgagt cagtgagcga ggaagcggaa gagcgcccaa tacgcaaacc gcctctcccc
     5161 gcgcgttggc cgattcatta atgcaggcaa ctcgtgaaag gtaggcggat cagcggccgc
     5221 agcatgcaat tctatattct aaaaacacaa atgatacttc taaaaaaaaa tcatgaatgg
     5281 catcaactct gaatcaaatc tttgcagatg cacctacttc tcatttccac tgtcacatca
     5341 tttttccaga tctatctgag gatgccgcac cctgccccac ctgtggctgg gcattcgacc
     5401 ttcaggggcc attgtccctt gtaatgccca cccactccta ttagtactct tataaatctt
     5461 ttcttataga ttattgttca tttaaagtat gttcttccca aaacaagagc ttcatggttc
     5521 atggccgcta cacccggaag caactgcagc cgttcacacg ctgacagcga actgacacga
     5581 atactcataa tcatttcaat tagacgtgtt gcaattagcc ttgtcgaccc gccaccactg
     5641 caggttcatt ccgtacgagt atgttgcaca attacgtaaa ttagcggcga ggaattcaat
     5701 atgcttgggc aggcaaagag ggagtcgagc gagagtccct gtgtgcgcat tatctttcag
     5761 ggccccgagc gcttgtaatt gccaatgctt aacacgattg agtgaatcat cgtgaatcac
     5821 tgaatcattc attgtgtgac tcgcggaatg cgatatcaat tccatgcgag cttctggatg
     5881 tcaaaacact ccacggaaga ccagattcta cgaaaagctg gttcttataa tgcgaaccat
     5941 taatttagta gatcactcta catatacata ctatctccga ctgattcttg gtgcgaacta
     6001 tttgtttttt gctgacagat ccactgaaaa ggtaaagaat aatgatatat ttccgcttcc
     6061 gttcaaggtc ttggatcaga gatacttcat cgttgtccca tgttttttta gatttttgct
     6121 tatttttggt tggtttttta ttgtcacatt ttttcgcacg ttttatttga tttatgtctg
     6181 tgacggctgc tgaccgccct gctcctccta ttatttcttt tatccctgat acatggctgt
     6241 tgtgtggcgc aaatttcctt atctcaggcc ttgtcggact ggccatattt tatcgcatgc
     6301 tttgcctttc ttttctgctt tgcaggcaaa ttgaaaaggc aattcccatc agacgtctgg
     6361 ctcgagatct actgcatcga tcagtgcggt cgactggcgt tgtaattttt tta
//
