LOCUS       sequence-335554-        3194 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  The resulting plasmid was used to generate a Gal4x5-E4-Gless 210bp 
            construct for ampilifying the immobilized template for assays. 
            Originally came from Michael Carey, was submitted with his 
            permission.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3194)
  AUTHORS   He Y, Sato S, Tomomori-Sato C, Chen S, Goode ZH, Conaway JW, Conaway
            RC
  TITLE     Elongin functions as a loading factor for Mediator at 
            ATF6alpha-regulated ER stress response genes.
  JOURNAL   Proc Natl Acad Sci U S A. 2021 Sep 28;118(39):e2108751118. doi: 
            10.1073/pnas.2108751118.
  PUBMED    34544872
REFERENCE   2  (bases 1 to 3194)
  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"; journalName: "Proc Natl 
            Acad Sci U S A."; date: "2021-09-28"; volume: "118(39):e2108751118. 
            doi"; pages: " 10.1073/pnas.2108751118"
COMMENT     Sequence Label: pGal4x5-E4T-G210
FEATURES             Location/Qualifiers
     source          1..3194
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        130..148
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     130..147
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     protein_bind    174..268
                     /label=5X UAS
                     /bound_moiety="GAL4"
                     /note="five tandem copies of the ""ScaI site"" 17-mer 
                     CGGAGTACTGTCCTCCG, an upstream activating sequence (UAS) 
                     that efficiently binds yeast Gal4 (Webster et al., 1988; 
                     Pfeiffer et al., 2010)"
     primer_bind     complement(545..564)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(546..564)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(645..662)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(816..1404)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(896..915)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(1575..2366,2367..2435))
                     /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     2198..2217
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(2436..2540)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     2608..2626
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(2664..2686)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     2786..2805
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3015..3034
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
ORIGIN
        1 gacggcagtt tacgagagag atgatagggt ctgcttcagt aagccagatg ctacacaatt
       61 aggcttgtac atattgtcgt tagaacgcgg ctacaattaa tacataacct tatgtatcat
      121 acacatacga tttaggtgac actatagaat acaagcttgc atgcctgcag gtcctcggag
      181 gacagtactc cgctcggagg acagtactcc gctcggagga cagtactccg ctcggaggac
      241 agtactccgc tcggaggaca gtactccgac tctagaggat ccccagtcct atatatactc
      301 gctctgcact tggccctttt ttacacccat acccttcctc catctatacc accctactct
      361 cctttcctca ttattcctcc tattatcttc tcctcttctc tccttcttct atatttccca
      421 aatctatcat cattcactct catcccctct tccttcactc ccattctatt ctactccttt
      481 ccctttccat atcccctcca ccccccttcc tcccctcttt caatcttatc ccgaattccg
      541 gtctccctat agtgagtcgt attaatttcg ataagccagc tgcattaatg aatcggccaa
      601 cgcgcgggga gaggcggttt gcgtattggg cgctcttccg cttcctcgct cactgactcg
      661 ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg
      721 ttatccacag aatcagggga taacgcagga aagaacatgt gagcaaaagg ccagcaaaag
      781 gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg cccccctgac
      841 gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg actataaaga
      901 taccaggcgt ttccccctgg aagctccctc gtgcgctctc ctgttccgac cctgccgctt
      961 accggatacc tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca tagctcacgc
     1021 tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc
     1081 cccgttcagc ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc caacccggta
     1141 agacacgact tatcgccact ggcagcagcc actggtaaca ggattagcag agcgaggtat
     1201 gtaggcggtg ctacagagtt cttgaagtgg tggcctaact acggctacac tagaagaaca
     1261 gtatttggta tctgcgctct gctgaagcca gttaccttcg gaaaaagagt tggtagctct
     1321 tgatccggca aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt
     1381 acgcgcagaa aaaaaggatc tcaagaagat cctttgatct tttctacggg gtctgacgct
     1441 cagtggaacg aaaactcacg ttaagggatt ttggtcatga gattatcaaa aaggatcttc
     1501 acctagatcc ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa
     1561 acttggtctg acagttacca atgcttaatc agtgaggcac ctatctcagc gatctgtcta
     1621 tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga taactacgat acgggagggc
     1681 ttaccatctg gccccagtgc tgcaatgata ccgcgagacc cacgctcacc ggctccagat
     1741 ttatcagcaa taaaccagcc agccggaagg gccgagcgca gaagtggtcc tgcaacttta
     1801 tccgcctcca tccagtctat taattgttgc cgggaagcta gagtaagtag ttcgccagtt
     1861 aatagtttgc gcaacgttgt tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt
     1921 ggtatggctt cattcagctc cggttcccaa cgatcaaggc gagttacatg atcccccatg
     1981 ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg ttgtcagaag taagttggcc
     2041 gcagtgttat cactcatggt tatggcagca ctgcataatt ctcttactgt catgccatcc
     2101 gtaagatgct tttctgtgac tggtgagtac tcaaccaagt cattctgaga atagtgtatg
     2161 cggcgaccga gttgctcttg cccggcgtca atacgggata ataccgcgcc acatagcaga
     2221 actttaaaag tgctcatcat tggaaaacgt tcttcggggc gaaaactctc aaggatctta
     2281 ccgctgttga gatccagttc gatgtaaccc actcgtgcac ccaactgatc ttcagcatct
     2341 tttactttca ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag
     2401 ggaataaggg cgacacggaa atgttgaata ctcatactct tcctttttca atattattga
     2461 agcatttatc agggttattg tctcatgagc ggatacatat ttgaatgtat ttagaaaaat
     2521 aaacaaatag gggttccgcg cacatttccc cgaaaagtgc cacctgacgt ctaagaaacc
     2581 attattatca tgacattaac ctataaaaat aggcgtatca cgaggccctt tcgtctcgcg
     2641 cgtttcggtg atgacggtga aaacctctga cacatgcagc tcccggagac ggtcacagct
     2701 tgtctgtaag cggatgccgg gagcagacaa gcccgtcagg gcgcgtcagc gggtgttggc
     2761 gggtgtcggg gctggcttaa ctatgcggca tcagagcaga ttgtactgag agtgcaccat
     2821 tcgacgctct cccttatgcg actcctgcat taggaagcag cccagtagta ggttgaggcc
     2881 gttgagcacc gccgccgcaa ggaatggtgc aaggagatgg cgcccaacag tcccccggcc
     2941 acggggcctg ccaccatacc cacgccgaaa caagcgctca tgagcccgaa gtggcgagcc
     3001 cgatcttccc catcggtgat gtcggcgata taggcgccag caaccgcacc tgtggcgccg
     3061 gtgatgccgg ccacgatgcg tccggcgtag aggatctggc tagcgatgac cctgctgatt
     3121 ggttcgctga ccatttccgg gtgcgggacg gcgttaccag aaactcagaa ggttcgtcca
     3181 accaaaccga ctct
//
