LOCUS       sequence-450933-        6232 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  For overexpression of Gal4-DBD EGR1 WT.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6232)
  AUTHORS   Naderi J, Magalhaes AP, Kibar G, Stik G, Zhang Y, Mackowiak SD, 
            Wieler HM, Rossi F, Buschow R, Christou-Kent M, Alcoverro-Bertran M,
            Graf T, Vingron M, Hnisz D
  TITLE     An activity-specificity trade-off encoded in human transcription 
            factors.
  JOURNAL   Nat Cell Biol. 2024 Aug;26(8):1309-1321. doi: 
            10.1038/s41556-024-01411-0. Epub 2024 Jul 5.
  PUBMED    38969762
REFERENCE   2  (bases 1 to 6232)
  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/s41556-024-01411-0"; journalName: "Nat Cell Biol"; date: 
            "2024-08"; volume: "26"; issue: "8"; pages: "1309-1321"
COMMENT     Sequence Label: pGAL4-DBD-EGR1-WT
FEATURES             Location/Qualifiers
     source          1..6232
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      57..645
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     546..565
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     799..816
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    933..954
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(969..974,975..992,993..999)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1007..1023
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     primer_bind     1012..1034
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     1031..1047
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     1031..1047
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     polyA_signal    1096..1217
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1107..1126)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     1161..1180
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     CDS             complement(1391..2185)
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(1502..1521)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     2112..2131
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     promoter        complement(2252..2581)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      2266..2401
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(2320..2339)
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     primer_bind     2566..2586
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      complement(2595..3023)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2705..2726)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2917..2936
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     polyA_signal    3069..3293
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     3282..3299
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     promoter        3319..3337
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     3319..3336
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     CDS             complement(3738..4178)
                     /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"
     primer_bind     complement(4240..4259)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(4241..4259)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     promoter        complement(4304..4507)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(4333..4353)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        4508..4887
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     primer_bind     5059..5078
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        5153..5257
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5258..5326,5327..6118)
                     /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(5476..5495)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
ORIGIN
        1 cgtgagtttt cgttccactg agcgtcagac cccgtagaaa agatcaaagg atcttcttga
       61 gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg
      121 gtggtttgtt tgccggatca agagctacca actctttttc cgaaggtaac tggcttcagc
      181 agagcgcaga taccaaatac tgttcttcta gtgtagccgt agttaggcca ccacttcaag
      241 aactctgtag caccgcctac atacctcgct ctgctaatcc tgttaccagt ggctgctgcc
      301 agtggcgata agtcgtgtct taccgggttg gactcaagac gatagttacc ggataaggcg
      361 cagcggtcgg gctgaacggg gggttcgtgc acacagccca gcttggagcg aacgacctac
      421 accgaactga gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga
      481 aaggcggaca ggtatccggt aagcggcagg gtcggaacag gagagcgcac gagggagctt
      541 ccagggggaa acgcctggta tctttatagt cctgtcgggt ttcgccacct ctgacttgag
      601 cgtcgatttt tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg
      661 gcctttttac ggttcctggc cttttgctgg ccttttgctc acatgttctt tcctgcgtta
      721 tcccctgatt ctgtggataa ccgtattacc gcctttgagt gagctgatac cgctcgccgc
      781 agccgaacga ccgagcgcag cgagtcagtg agcgaggaag cggaagagcg cccaatacgc
      841 aaaccgcctc tccccgcgcg ttggccgatt cattaatgca gctggcacga caggtttccc
      901 gactggaaag cgggcagtga gcgcaacgca attaatgtga gttagctcac tcattaggca
      961 ccccaggctt tacactttat gcttccggct cgtatgttgt gtggaattgt gagcggataa
     1021 caatttcaca caggaaacag ctatgaccat gattacgcca agctctagct agaggtcgac
     1081 ggtatacaga catgataaga tacattgatg agtttggaca aaccacaact agaatgcagt
     1141 gaaaaaaatg ctttatttgt gaaatttgtg atgctattgc tttatttgta accattataa
     1201 gctgcaataa acaagttggg gtgggcgaag aactccagca tgagatcccc gcgctggagg
     1261 atcatccagc cggcgtcccg gaaaacgatt ccgaagccca acctttcata gaaggcggcg
     1321 gtggaatcga aatctcgtga tggcaggttg ggcgtcgctt ggtcggtcat ttcgaacccc
     1381 agagtcccgc tcagaagaac tcgtcaagaa ggcgatagaa ggcgatgcgc tgcgaatcgg
     1441 gagcggcgat accgtaaagc acgaggaagc ggtcagccca ttcgccgcca agctcttcag
     1501 caatatcacg ggtagccaac gctatgtcct gatagcggtc cgccacaccc agccggccac
     1561 agtcgatgaa tccagaaaag cggccatttt ccaccatgat attcggcaag caggcatcgc
     1621 catgggtcac gacgagatcc tcgccgtcgg gcatgcgcgc cttgagcctg gcgaacagtt
     1681 cggctggcgc gagcccctga tgctcttcgt ccagatcatc ctgatcgaca agaccggctt
     1741 ccatccgagt acgtgctcgc tcgatgcgat gtttcgcttg gtggtcgaat gggcaggtag
     1801 ccggatcaag cgtatgcagc cgccgcattg catcagccat gatggatact ttctcggcag
     1861 gagcaaggtg agatgacagg agatcctgcc ccggcacttc gcccaatagc agccagtccc
     1921 ttcccgcttc agtgacaacg tcgagcacag ctgcgcaagg aacgcccgtc gtggccagcc
     1981 acgatagccg cgctgcctcg tcctgcagtt cattcagggc accggacagg tcggtcttga
     2041 caaaaagaac cgggcgcccc tgcgctgaca gccggaacac ggcggcatca gagcagccga
     2101 ttgtctgttg tgcccagtca tagccgaata gcctctccac ccaagcggcc ggagaacctg
     2161 cgtgcaatcc atcttgttca atcatgcgaa acgatcctca tcctgtctct tgatcagatc
     2221 cgaaaatgga tatacaagct cccgggagct ttttgcaaaa gcctaggcct ccaaaaaagc
     2281 ctcctcacta cttctggaat agctcagagg cagaggcggc ctcggcctct gcataaataa
     2341 aaaaaattag tcagccatgg ggcggagaat gggcggaact gggcggagtt aggggcggga
     2401 tgggcggagt taggggcggg actatggttg ctgactaatt gagatgcatg ctttgcatac
     2461 ttctgcctgc tggggagcct ggggactttc cacacctggt tgctgactaa ttgagatgca
     2521 tgctttgcat acttctgcct gctggggagc ctggggactt tccacaccct aactgacaca
     2581 cattccacag aattaattcg cgttaaattt ttgttaaatc agctcatttt ttaaccaata
     2641 ggccgaaatc ggcaaaatcc cttataaatc aaaagaatag accgagatag ggttgagtgt
     2701 tgttccagtt tggaacaaga gtccactatt aaagaacgtg gactccaacg tcaaagggcg
     2761 aaaaaccgtc tatcagggcg atggcccact acgtgaacca tcaccctaat caagtttttt
     2821 ggggtcgagg tgccgtaaag cactaaatcg gaaccctaaa gggagccccc gatttagagc
     2881 ttgacgggga aagccggcga acgtggcgag aaaggaaggg aagaaagcga aaggagcggg
     2941 cgctagggcg ctggcaagtg tagcggtcac gctgcgcgta accaccacac ccgccgcgct
     3001 taatgcgccg ctacagggcg cgtggggata ccccctagag ccccagctgg ttctttccgc
     3061 ctcagaagcc atagagccca ccgcatcccc agcatgcctg ctattgtctt cccaatcctc
     3121 ccccttgctg tcctgcccca ccccaccccc cagaatagaa tgacacctac tcagacaatg
     3181 cgatgcaatt tcctcatttt attaggaaag gacagtggga gtggcacctt ccagggtcaa
     3241 ggaaggcacg ggggaggggc aaacaacaga tggctggcaa ctagaaggca cagtcgaggc
     3301 tgatcagcga gctctagcat ttaggtgaca ctatagaata gggccctcta gactcagcaa
     3361 atttcaattg tcctgggaga aaaggttgct gtcatgtccg aaagccctgt ggaggcgctg
     3421 aaggagttgg tgacagctga ggaagggaag ctgctgacct gggccgggaa agcaggggga
     3481 acagaggagt acgtggtggc caccgacggg gaggggaagc cactgtgcac aggggatggg
     3541 taggtcgagg agccgggaga ggagaaggag gtgggcacag gggatgggta tgaggtggtg
     3601 gccggggatg gataagaggt agtaaccggg gacgggtaag aggtagcaac cggggacggg
     3661 taggaagaga gagaggaggt ggccgaagag gccacaacac ttttgtctgc tttcttgtcc
     3721 ttctgccgca atatcgccga tacagtcaac tgtctttgac ctttgttact actctcttcc
     3781 gatgatgatg tcgcacttat tctatgctgt ctcaatgtta gaggcatatc agtctccact
     3841 gaagccaatc tatctgtgac ggcatcttta ttcacattat cttgtacaaa taatcctgtt
     3901 aacaatgctt ttatatcctg taaagaatcc attttcaaaa tcatgtcaag gtcttctcga
     3961 ggaaaaatca gtagaaatag ctgttccagt ctttctagcc ttgattccac ttctgtcaga
     4021 tgtgccctag tcagcggaga ccttttggtt ttgggagagt agcgacactc ccagttgttc
     4081 ttcagacact tggcgcactt cggtttttct ttggagcact tgagcttttt aagtcggcaa
     4141 atatcgcatg cttgttcgat agaagacagt agcttcatga attccagcac actggcggcc
     4201 gttactagtg gatccgagct cggtaccaag cttgggtctc cctatagtga gtcgtattaa
     4261 tttcgataag ccagtaagca gtgggttctc tagttagcca gagagctctg cttatataga
     4321 cctcccaccg tacacgccta ccgcccattt gcgtcaatgg ggcggagttg ttacgacatt
     4381 ttggaaagtc ccgttgattt tggtgccaaa acaaactccc attgacgtca atggggtgga
     4441 gacttggaaa tccccgtgag tcaaaccgct atccacgccc attgatgtac tgccaaaacc
     4501 gcatcaccat ggtaatagcg atgactaata cgtagatgta ctgccaagta ggaaagtccc
     4561 ataaggtcat gtactgggca taatgccagg cgggccattt accgtcattg acgtcaatag
     4621 ggggcgtact tggcatatga tacacttgat gtactgccaa gtgggcagtt taccgtaaat
     4681 actccaccca ttgacgtcaa tggaaagtcc ctattggcgt tactatggga acatacgtca
     4741 ttattgacgt caatgggcgg gggtcgttgg gcggtcagcc aggcgggcca tttaccgtaa
     4801 gttatgtaac gcggaactcc atatatgggc tatgaactaa tgaccccgta attgattact
     4861 attaataact agtcaataat caatgtcaac gcgtatatct ggcccgtaca tcgcgaagca
     4921 gcgcaaaacg cctaacccta agcagattct tcatgcaatt gtcggtcaag ccttgccttg
     4981 ttgtagctta aattttgctc gcgcactact cagcgacctc caacacacaa gcagggagca
     5041 gatactggct taactatgcg gcatcagagc agattgtact gagagtgcac cataggggat
     5101 cgggagatct cccgatccgt cgacgtcagg tggcactttt cggggaaatg tgcgcggaac
     5161 ccctatttgt ttatttttct aaatacattc aaatatgtat ccgctcatga gacaataacc
     5221 ctgataaatg cttcaataat attgaaaaag gaagagtatg agtattcaac atttccgtgt
     5281 cgcccttatt cccttttttg cggcattttg ccttcctgtt tttgctcacc cagaaacgct
     5341 ggtgaaagta aaagatgctg aagatcagtt gggtgcacga gtgggttaca tcgaactgga
     5401 tctcaacagc ggtaagatcc ttgagagttt tcgccccgaa gaacgttttc caatgatgag
     5461 cacttttaaa gttctgctat gtggcgcggt attatcccgt attgacgccg ggcaagagca
     5521 actcggtcgc cgcatacact attctcagaa tgacttggtt gagtactcac cagtcacaga
     5581 aaagcatctt acggatggca tgacagtaag agaattatgc agtgctgcca taaccatgag
     5641 tgataacact gcggccaact tacttctgac aacgatcgga ggaccgaagg agctaaccgc
     5701 ttttttgcac aacatggggg atcatgtaac tcgccttgat cgttgggaac cggagctgaa
     5761 tgaagccata ccaaacgacg agcgtgacac cacgatgcct gtagcaatgg caacaacgtt
     5821 gcgcaaacta ttaactggcg aactacttac tctagcttcc cggcaacaat taatagactg
     5881 gatggaggcg gataaagttg caggaccact tctgcgctcg gcccttccgg ctggctggtt
     5941 tattgctgat aaatctggag ccggtgagcg tgggtctcgc ggtatcattg cagcactggg
     6001 gccagatggt aagccctccc gtatcgtagt tatctacacg acgggaagtc aggcaactat
     6061 ggatgaacga aatagacaga tcgctgagat aggtgcctca ctgattaagc attggtaact
     6121 gtcagaccaa gtttactcat atatacttta gattgattta aaacttcatt tttaatttaa
     6181 aaggatctag gtgaagatcc tttttgataa tctcatgacc aaaatccctt aa
//
