LOCUS       sequence-430431-        6106 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  For overexpression of Gal4-DBD NGN2 WT.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6106)
  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 6106)
  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-NGN2-WT
FEATURES             Location/Qualifiers
     source          1..6106
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(join(86..877,878..946))
                     /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     709..728
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(947..1051)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(1126..1145)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        1317..1696
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        1697..1900
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     1851..1871
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     1945..1964
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1945..1963
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             2026..2466
                     /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"
     promoter        complement(2741..2759)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(2742..2759)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(2779..2796)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2785..3009
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      3055..3483
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3142..3161)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3352..3373
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3492..3512)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3497..3826
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3677..3812
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     3739..3758
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             3893..4687
                     /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(3947..3966)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4557..4576
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    4861..4982
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4898..4917)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4952..4971
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5031..5047)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5031..5047)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5044..5066)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5055..5071
                     /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)."
     promoter        complement(join(5079..5085,5086..5103,5104..5109))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5124..5145
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5262..5279)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5433..6021)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5513..5532)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 aaaggatctt cacctagatc cttttaaatt aaaaatgaag ttttaaatca atctaaagta
       61 tatatgagta aacttggtct gacagttacc aatgcttaat cagtgaggca cctatctcag
      121 cgatctgtct atttcgttca tccatagttg cctgacttcc cgtcgtgtag ataactacga
      181 tacgggaggg cttaccatct ggccccagtg ctgcaatgat accgcgagac ccacgctcac
      241 cggctccaga tttatcagca ataaaccagc cagccggaag ggccgagcgc agaagtggtc
      301 ctgcaacttt atccgcctcc atccagtcta ttaattgttg ccgggaagct agagtaagta
      361 gttcgccagt taatagtttg cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac
      421 gctcgtcgtt tggtatggct tcattcagct ccggttccca acgatcaagg cgagttacat
      481 gatcccccat gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc gttgtcagaa
      541 gtaagttggc cgcagtgtta tcactcatgg ttatggcagc actgcataat tctcttactg
      601 tcatgccatc cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag tcattctgag
      661 aatagtgtat gcggcgaccg agttgctctt gcccggcgtc aatacgggat aataccgcgc
      721 cacatagcag aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct
      781 caaggatctt accgctgttg agatccagtt cgatgtaacc cactcgtgca cccaactgat
      841 cttcagcatc ttttactttc accagcgttt ctgggtgagc aaaaacagga aggcaaaatg
      901 ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat actcatactc ttcctttttc
      961 aatattattg aagcatttat cagggttatt gtctcatgag cggatacata tttgaatgta
     1021 tttagaaaaa taaacaaata ggggttccgc gcacatttcc ccgaaaagtg ccacctgacg
     1081 tcgacggatc gggagatctc ccgatcccct atggtgcact ctcagtacaa tctgctctga
     1141 tgccgcatag ttaagccagt atctgctccc tgcttgtgtg ttggaggtcg ctgagtagtg
     1201 cgcgagcaaa atttaagcta caacaaggca aggcttgacc gacaattgca tgaagaatct
     1261 gcttagggtt aggcgttttg cgctgcttcg cgatgtacgg gccagatata cgcgttgaca
     1321 ttgattattg actagttatt aatagtaatc aattacgggg tcattagttc atagcccata
     1381 tatggagttc cgcgttacat aacttacggt aaatggcccg cctggctgac cgcccaacga
     1441 cccccgccca ttgacgtcaa taatgacgta tgttcccata gtaacgccaa tagggacttt
     1501 ccattgacgt caatgggtgg agtatttacg gtaaactgcc cacttggcag tacatcaagt
     1561 gtatcatatg ccaagtacgc cccctattga cgtcaatgac ggtaaatggc ccgcctggca
     1621 ttatgcccag tacatgacct tatgggactt tcctacttgg cagtacatct acgtattagt
     1681 catcgctatt accatggtga tgcggttttg gcagtacatc aatgggcgtg gatagcggtt
     1741 tgactcacgg ggatttccaa gtctccaccc cattgacgtc aatgggagtt tgttttggca
     1801 ccaaaatcaa cgggactttc caaaatgtcg taacaactcc gccccattga cgcaaatggg
     1861 cggtaggcgt gtacggtggg aggtctatat aagcagagct ctctggctaa ctagagaacc
     1921 cactgcttac tggcttatcg aaattaatac gactcactat agggagaccc aagcttggta
     1981 ccgagctcgg atccactagt aacggccgcc agtgtgctgg aattcatgaa gctactgtct
     2041 tctatcgaac aagcatgcga tatttgccga cttaaaaagc tcaagtgctc caaagaaaaa
     2101 ccgaagtgcg ccaagtgtct gaagaacaac tgggagtgtc gctactctcc caaaaccaaa
     2161 aggtctccgc tgactagggc acatctgaca gaagtggaat caaggctaga aagactggaa
     2221 cagctatttc tactgatttt tcctcgagaa gaccttgaca tgattttgaa aatggattct
     2281 ttacaggata taaaagcatt gttaacagga ttatttgtac aagataatgt gaataaagat
     2341 gccgtcacag atagattggc ttcagtggag actgatatgc ctctaacatt gagacagcat
     2401 agaataagtg cgacatcatc atcggaagag agtagtaaca aaggtcaaag acagttgact
     2461 gtatcggcga tagcggtcct cttgtcaccc ggcggggcat ccgcagcttt gtcctcttcc
     2521 ggcgatagtc catcccctgc ttctacttgg tcatgtacga attctccagc cccatctagc
     2581 agtgtcagct ctaactctac gagtccttat tcctgtacac tgtccccagc gtcccccgcg
     2641 ggctctgata tggattactg gcaacctcct ccccctgaca agcaccgcta tgcacctcac
     2701 ctccccatag ccagggattg tatcgtctag agggccctat tctatagtgt cacctaaatg
     2761 ctagagctcg ctgatcagcc tcgactgtgc cttctagttg ccagccatct gttgtttgcc
     2821 cctcccccgt gccttccttg accctggaag gtgccactcc cactgtcctt tcctaataaa
     2881 atgaggaaat tgcatcgcat tgtctgagta ggtgtcattc tattctgggg ggtggggtgg
     2941 ggcaggacag caagggggag gattgggaag acaatagcag gcatgctggg gatgcggtgg
     3001 gctctatggc ttctgaggcg gaaagaacca gctggggctc tagggggtat ccccacgcgc
     3061 cctgtagcgg cgcattaagc gcggcgggtg tggtggttac gcgcagcgtg accgctacac
     3121 ttgccagcgc cctagcgccc gctcctttcg ctttcttccc ttcctttctc gccacgttcg
     3181 ccggctttcc ccgtcaagct ctaaatcggg ggctcccttt agggttccga tttagtgctt
     3241 tacggcacct cgaccccaaa aaacttgatt agggtgatgg ttcacgtagt gggccatcgc
     3301 cctgatagac ggtttttcgc cctttgacgt tggagtccac gttctttaat agtggactct
     3361 tgttccaaac tggaacaaca ctcaacccta tctcggtcta ttcttttgat ttataaggga
     3421 ttttgccgat ttcggcctat tggttaaaaa atgagctgat ttaacaaaaa tttaacgcga
     3481 attaattctg tggaatgtgt gtcagttagg gtgtggaaag tccccaggct ccccagcagg
     3541 cagaagtatg caaagcatgc atctcaatta gtcagcaacc aggtgtggaa agtccccagg
     3601 ctccccagca ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa ccatagtccc
     3661 gcccctaact ccgcccatcc cgcccctaac tccgcccagt tccgcccatt ctccgcccca
     3721 tggctgacta atttttttta tttatgcaga ggccgaggcc gcctctgcct ctgagctatt
     3781 ccagaagtag tgaggaggct tttttggagg cctaggcttt tgcaaaaagc tcccgggagc
     3841 ttgtatatcc attttcggat ctgatcaaga gacaggatga ggatcgtttc gcatgattga
     3901 acaagatgga ttgcacgcag gttctccggc cgcttgggtg gagaggctat tcggctatga
     3961 ctgggcacaa cagacaatcg gctgctctga tgccgccgtg ttccggctgt cagcgcaggg
     4021 gcgcccggtt ctttttgtca agaccgacct gtccggtgcc ctgaatgaac tgcaggacga
     4081 ggcagcgcgg ctatcgtggc tggccacgac gggcgttcct tgcgcagctg tgctcgacgt
     4141 tgtcactgaa gcgggaaggg actggctgct attgggcgaa gtgccggggc aggatctcct
     4201 gtcatctcac cttgctcctg ccgagaaagt atccatcatg gctgatgcaa tgcggcggct
     4261 gcatacgctt gatccggcta cctgcccatt cgaccaccaa gcgaaacatc gcatcgagcg
     4321 agcacgtact cggatggaag ccggtcttgt cgatcaggat gatctggacg aagagcatca
     4381 ggggctcgcg ccagccgaac tgttcgccag gctcaaggcg cgcatgcccg acggcgagga
     4441 tctcgtcgtg acccatggcg atgcctgctt gccgaatatc atggtggaaa atggccgctt
     4501 ttctggattc atcgactgtg gccggctggg tgtggcggac cgctatcagg acatagcgtt
     4561 ggctacccgt gatattgctg aagagcttgg cggcgaatgg gctgaccgct tcctcgtgct
     4621 ttacggtatc gccgctcccg attcgcagcg catcgccttc tatcgccttc ttgacgagtt
     4681 cttctgagcg ggactctggg gttcgaaatg accgaccaag cgacgcccaa cctgccatca
     4741 cgagatttcg attccaccgc cgccttctat gaaaggttgg gcttcggaat cgttttccgg
     4801 gacgccggct ggatgatcct ccagcgcggg gatctcatgc tggagttctt cgcccacccc
     4861 aacttgttta ttgcagctta taatggttac aaataaagca atagcatcac aaatttcaca
     4921 aataaagcat ttttttcact gcattctagt tgtggtttgt ccaaactcat caatgtatct
     4981 tatcatgtct gtataccgtc gacctctagc tagagcttgg cgtaatcatg gtcatagctg
     5041 tttcctgtgt gaaattgtta tccgctcaca attccacaca acatacgagc cggaagcata
     5101 aagtgtaaag cctggggtgc ctaatgagtg agctaactca cattaattgc gttgcgctca
     5161 ctgcccgctt tccagtcggg aaacctgtcg tgccagctgc attaatgaat cggccaacgc
     5221 gcggggagag gcggtttgcg tattgggcgc tcttccgctt cctcgctcac tgactcgctg
     5281 cgctcggtcg ttcggctgcg gcgagcggta tcagctcact caaaggcggt aatacggtta
     5341 tccacagaat caggggataa cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc
     5401 aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc ccctgacgag
     5461 catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact ataaagatac
     5521 caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct gccgcttacc
     5581 ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcatag ctcacgctgt
     5641 aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc
     5701 gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggtaaga
     5761 cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc gaggtatgta
     5821 ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag aagaacagta
     5881 tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg tagctcttga
     5941 tccggcaaac aaaccaccgc tggtagcggt ggtttttttg tttgcaagca gcagattacg
     6001 cgcagaaaaa aaggatctca agaagatcct ttgatctttt ctacggggtc tgacgctcag
     6061 tggaacgaaa actcacgtta agggattttg gtcatgagat tatcaa
//
