LOCUS       sequence-443028-        6154 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  For overexpression of Gal4-DBD D6Y AroPERFECT.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6154)
  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 6154)
  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-D6Y_AroPERFECT
FEATURES             Location/Qualifiers
     source          1..6154
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     69..86
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     promoter        106..124
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     106..123
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     CDS             complement(447..887)
                     /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(949..968)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(950..968)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     promoter        complement(1013..1216)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(1042..1062)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        1217..1596
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     primer_bind     1768..1787
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        1862..1966
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1967..2035,2036..2827)
                     /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(2185..2204)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2998..3586
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3487..3506
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3740..3757
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    3874..3895
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(3910..3915,3916..3933,3934..3940)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3948..3964
                     /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     3953..3975
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     3972..3988
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     3972..3988
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     polyA_signal    4037..4158
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4048..4067)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     4102..4121
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     CDS             complement(4332..5126)
                     /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(4443..4462)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     5053..5072
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     promoter        complement(5193..5522)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      5207..5342
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(5261..5280)
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     primer_bind     5507..5527
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      complement(5536..5964)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5646..5667)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     5858..5877
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     polyA_signal    join(6010..6154,1..80)
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
ORIGIN
        1 agtgggagtg gcaccttcca gggtcaagga aggcacgggg gaggggcaaa caacagatgg
       61 ctggcaacta gaaggcacag tcgaggctga tcagcgagct ctagcattta ggtgacacta
      121 tagaataggg ccctctagac tcaatacgat ccatcagagc tgccataccc ggagtcactg
      181 ccggagtatg agccatcacc ggaagaatag ctagaatccc cgccgctgta cccgctatct
      241 cctgaagaat aggatgagtc accagagctg tacccggaat ctgagccgct gtatgaacca
      301 tcgcttgatc catagccact gtctgagccg gagtaggagc cgtcaccgga agaataggat
      361 gaatcgccac cactgtagcc gctatctcca gaggaataag atgagtcacc gctactatat
      421 ccagatgaag aaccactcat gatcgccgat acagtcaact gtctttgacc tttgttacta
      481 ctctcttccg atgatgatgt cgcacttatt ctatgctgtc tcaatgttag aggcatatca
      541 gtctccactg aagccaatct atctgtgacg gcatctttat tcacattatc ttgtacaaat
      601 aatcctgtta acaatgcttt tatatcctgt aaagaatcca ttttcaaaat catgtcaagg
      661 tcttctcgag gaaaaatcag tagaaatagc tgttccagtc tttctagcct tgattccact
      721 tctgtcagat gtgccctagt cagcggagac cttttggttt tgggagagta gcgacactcc
      781 cagttgttct tcagacactt ggcgcacttc ggtttttctt tggagcactt gagcttttta
      841 agtcggcaaa tatcgcatgc ttgttcgata gaagacagta gcttcatgaa ttccagcaca
      901 ctggcggccg ttactagtgg atccgagctc ggtaccaagc ttgggtctcc ctatagtgag
      961 tcgtattaat ttcgataagc cagtaagcag tgggttctct agttagccag agagctctgc
     1021 ttatatagac ctcccaccgt acacgcctac cgcccatttg cgtcaatggg gcggagttgt
     1081 tacgacattt tggaaagtcc cgttgatttt ggtgccaaaa caaactccca ttgacgtcaa
     1141 tggggtggag acttggaaat ccccgtgagt caaaccgcta tccacgccca ttgatgtact
     1201 gccaaaaccg catcaccatg gtaatagcga tgactaatac gtagatgtac tgccaagtag
     1261 gaaagtccca taaggtcatg tactgggcat aatgccaggc gggccattta ccgtcattga
     1321 cgtcaatagg gggcgtactt ggcatatgat acacttgatg tactgccaag tgggcagttt
     1381 accgtaaata ctccacccat tgacgtcaat ggaaagtccc tattggcgtt actatgggaa
     1441 catacgtcat tattgacgtc aatgggcggg ggtcgttggg cggtcagcca ggcgggccat
     1501 ttaccgtaag ttatgtaacg cggaactcca tatatgggct atgaactaat gaccccgtaa
     1561 ttgattacta ttaataacta gtcaataatc aatgtcaacg cgtatatctg gcccgtacat
     1621 cgcgaagcag cgcaaaacgc ctaaccctaa gcagattctt catgcaattg tcggtcaagc
     1681 cttgccttgt tgtagcttaa attttgctcg cgcactactc agcgacctcc aacacacaag
     1741 cagggagcag atactggctt aactatgcgg catcagagca gattgtactg agagtgcacc
     1801 ataggggatc gggagatctc ccgatccgtc gacgtcaggt ggcacttttc ggggaaatgt
     1861 gcgcggaacc cctatttgtt tatttttcta aatacattca aatatgtatc cgctcatgag
     1921 acaataaccc tgataaatgc ttcaataata ttgaaaaagg aagagtatga gtattcaaca
     1981 tttccgtgtc gcccttattc ccttttttgc ggcattttgc cttcctgttt ttgctcaccc
     2041 agaaacgctg gtgaaagtaa aagatgctga agatcagttg ggtgcacgag tgggttacat
     2101 cgaactggat ctcaacagcg gtaagatcct tgagagtttt cgccccgaag aacgttttcc
     2161 aatgatgagc acttttaaag ttctgctatg tggcgcggta ttatcccgta ttgacgccgg
     2221 gcaagagcaa ctcggtcgcc gcatacacta ttctcagaat gacttggttg agtactcacc
     2281 agtcacagaa aagcatctta cggatggcat gacagtaaga gaattatgca gtgctgccat
     2341 aaccatgagt gataacactg cggccaactt acttctgaca acgatcggag gaccgaagga
     2401 gctaaccgct tttttgcaca acatggggga tcatgtaact cgccttgatc gttgggaacc
     2461 ggagctgaat gaagccatac caaacgacga gcgtgacacc acgatgcctg tagcaatggc
     2521 aacaacgttg cgcaaactat taactggcga actacttact ctagcttccc ggcaacaatt
     2581 aatagactgg atggaggcgg ataaagttgc aggaccactt ctgcgctcgg cccttccggc
     2641 tggctggttt attgctgata aatctggagc cggtgagcgt gggtctcgcg gtatcattgc
     2701 agcactgggg ccagatggta agccctcccg tatcgtagtt atctacacga cgggaagtca
     2761 ggcaactatg gatgaacgaa atagacagat cgctgagata ggtgcctcac tgattaagca
     2821 ttggtaactg tcagaccaag tttactcata tatactttag attgatttaa aacttcattt
     2881 ttaatttaaa aggatctagg tgaagatcct ttttgataat ctcatgacca aaatccctta
     2941 acgtgagttt tcgttccact gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg
     3001 agatcctttt tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac cgctaccagc
     3061 ggtggtttgt ttgccggatc aagagctacc aactcttttt ccgaaggtaa ctggcttcag
     3121 cagagcgcag ataccaaata ctgttcttct agtgtagccg tagttaggcc accacttcaa
     3181 gaactctgta gcaccgccta catacctcgc tctgctaatc ctgttaccag tggctgctgc
     3241 cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac cggataaggc
     3301 gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc agcttggagc gaacgaccta
     3361 caccgaactg agatacctac agcgtgagct atgagaaagc gccacgcttc ccgaagggag
     3421 aaaggcggac aggtatccgg taagcggcag ggtcggaaca ggagagcgca cgagggagct
     3481 tccaggggga aacgcctggt atctttatag tcctgtcggg tttcgccacc tctgacttga
     3541 gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc
     3601 ggccttttta cggttcctgg ccttttgctg gccttttgct cacatgttct ttcctgcgtt
     3661 atcccctgat tctgtggata accgtattac cgcctttgag tgagctgata ccgctcgccg
     3721 cagccgaacg accgagcgca gcgagtcagt gagcgaggaa gcggaagagc gcccaatacg
     3781 caaaccgcct ctccccgcgc gttggccgat tcattaatgc agctggcacg acaggtttcc
     3841 cgactggaaa gcgggcagtg agcgcaacgc aattaatgtg agttagctca ctcattaggc
     3901 accccaggct ttacacttta tgcttccggc tcgtatgttg tgtggaattg tgagcggata
     3961 acaatttcac acaggaaaca gctatgacca tgattacgcc aagctctagc tagaggtcga
     4021 cggtatacag acatgataag atacattgat gagtttggac aaaccacaac tagaatgcag
     4081 tgaaaaaaat gctttatttg tgaaatttgt gatgctattg ctttatttgt aaccattata
     4141 agctgcaata aacaagttgg ggtgggcgaa gaactccagc atgagatccc cgcgctggag
     4201 gatcatccag ccggcgtccc ggaaaacgat tccgaagccc aacctttcat agaaggcggc
     4261 ggtggaatcg aaatctcgtg atggcaggtt gggcgtcgct tggtcggtca tttcgaaccc
     4321 cagagtcccg ctcagaagaa ctcgtcaaga aggcgataga aggcgatgcg ctgcgaatcg
     4381 ggagcggcga taccgtaaag cacgaggaag cggtcagccc attcgccgcc aagctcttca
     4441 gcaatatcac gggtagccaa cgctatgtcc tgatagcggt ccgccacacc cagccggcca
     4501 cagtcgatga atccagaaaa gcggccattt tccaccatga tattcggcaa gcaggcatcg
     4561 ccatgggtca cgacgagatc ctcgccgtcg ggcatgcgcg ccttgagcct ggcgaacagt
     4621 tcggctggcg cgagcccctg atgctcttcg tccagatcat cctgatcgac aagaccggct
     4681 tccatccgag tacgtgctcg ctcgatgcga tgtttcgctt ggtggtcgaa tgggcaggta
     4741 gccggatcaa gcgtatgcag ccgccgcatt gcatcagcca tgatggatac tttctcggca
     4801 ggagcaaggt gagatgacag gagatcctgc cccggcactt cgcccaatag cagccagtcc
     4861 cttcccgctt cagtgacaac gtcgagcaca gctgcgcaag gaacgcccgt cgtggccagc
     4921 cacgatagcc gcgctgcctc gtcctgcagt tcattcaggg caccggacag gtcggtcttg
     4981 acaaaaagaa ccgggcgccc ctgcgctgac agccggaaca cggcggcatc agagcagccg
     5041 attgtctgtt gtgcccagtc atagccgaat agcctctcca cccaagcggc cggagaacct
     5101 gcgtgcaatc catcttgttc aatcatgcga aacgatcctc atcctgtctc ttgatcagat
     5161 ccgaaaatgg atatacaagc tcccgggagc tttttgcaaa agcctaggcc tccaaaaaag
     5221 cctcctcact acttctggaa tagctcagag gcagaggcgg cctcggcctc tgcataaata
     5281 aaaaaaatta gtcagccatg gggcggagaa tgggcggaac tgggcggagt taggggcggg
     5341 atgggcggag ttaggggcgg gactatggtt gctgactaat tgagatgcat gctttgcata
     5401 cttctgcctg ctggggagcc tggggacttt ccacacctgg ttgctgacta attgagatgc
     5461 atgctttgca tacttctgcc tgctggggag cctggggact ttccacaccc taactgacac
     5521 acattccaca gaattaattc gcgttaaatt tttgttaaat cagctcattt tttaaccaat
     5581 aggccgaaat cggcaaaatc ccttataaat caaaagaata gaccgagata gggttgagtg
     5641 ttgttccagt ttggaacaag agtccactat taaagaacgt ggactccaac gtcaaagggc
     5701 gaaaaaccgt ctatcagggc gatggcccac tacgtgaacc atcaccctaa tcaagttttt
     5761 tggggtcgag gtgccgtaaa gcactaaatc ggaaccctaa agggagcccc cgatttagag
     5821 cttgacgggg aaagccggcg aacgtggcga gaaaggaagg gaagaaagcg aaaggagcgg
     5881 gcgctagggc gctggcaagt gtagcggtca cgctgcgcgt aaccaccaca cccgccgcgc
     5941 ttaatgcgcc gctacagggc gcgtggggat accccctaga gccccagctg gttctttccg
     6001 cctcagaagc catagagccc accgcatccc cagcatgcct gctattgtct tcccaatcct
     6061 cccccttgct gtcctgcccc accccacccc ccagaataga atgacaccta ctcagacaat
     6121 gcgatgcaat ttcctcattt tattaggaaa ggac
//
