LOCUS       sequence-437917-        6604 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  For overexpression of Gal4-DBD CEBPa AroPERFECT IS15+2.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6604)
  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 6604)
  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-CEBPa-AroPERFECT_IS15+2
FEATURES             Location/Qualifiers
     source          1..6604
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     10..29
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     263..280
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    397..418
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(433..438,439..456,457..463)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    471..487
                     /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     476..498
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     495..511
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     495..511
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     polyA_signal    560..681
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(571..590)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     625..644
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     CDS             complement(855..1649)
                     /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(966..985)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     1576..1595
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     promoter        complement(1716..2045)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      1730..1865
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(1784..1803)
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     primer_bind     2030..2050
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      complement(2059..2487)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2169..2190)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2381..2400
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     polyA_signal    2533..2757
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     2746..2763
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     promoter        2783..2801
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     2783..2800
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     CDS             complement(3574..4014)
                     /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(4076..4095)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(4077..4095)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     promoter        complement(4140..4343)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(4169..4189)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        4344..4723
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     primer_bind     4895..4914
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        4989..5093
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5094..5162,5163..5954)
                     /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(5312..5331)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      join(6125..6604,1..109)
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
ORIGIN
        1 gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact
       61 tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa
      121 cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgt tctttcctgc
      181 gttatcccct gattctgtgg ataaccgtat taccgccttt gagtgagctg ataccgctcg
      241 ccgcagccga acgaccgagc gcagcgagtc agtgagcgag gaagcggaag agcgcccaat
      301 acgcaaaccg cctctccccg cgcgttggcc gattcattaa tgcagctggc acgacaggtt
      361 tcccgactgg aaagcgggca gtgagcgcaa cgcaattaat gtgagttagc tcactcatta
      421 ggcaccccag gctttacact ttatgcttcc ggctcgtatg ttgtgtggaa ttgtgagcgg
      481 ataacaattt cacacaggaa acagctatga ccatgattac gccaagctct agctagaggt
      541 cgacggtata cagacatgat aagatacatt gatgagtttg gacaaaccac aactagaatg
      601 cagtgaaaaa aatgctttat ttgtgaaatt tgtgatgcta ttgctttatt tgtaaccatt
      661 ataagctgca ataaacaagt tggggtgggc gaagaactcc agcatgagat ccccgcgctg
      721 gaggatcatc cagccggcgt cccggaaaac gattccgaag cccaaccttt catagaaggc
      781 ggcggtggaa tcgaaatctc gtgatggcag gttgggcgtc gcttggtcgg tcatttcgaa
      841 ccccagagtc ccgctcagaa gaactcgtca agaaggcgat agaaggcgat gcgctgcgaa
      901 tcgggagcgg cgataccgta aagcacgagg aagcggtcag cccattcgcc gccaagctct
      961 tcagcaatat cacgggtagc caacgctatg tcctgatagc ggtccgccac acccagccgg
     1021 ccacagtcga tgaatccaga aaagcggcca ttttccacca tgatattcgg caagcaggca
     1081 tcgccatggg tcacgacgag atcctcgccg tcgggcatgc gcgccttgag cctggcgaac
     1141 agttcggctg gcgcgagccc ctgatgctct tcgtccagat catcctgatc gacaagaccg
     1201 gcttccatcc gagtacgtgc tcgctcgatg cgatgtttcg cttggtggtc gaatgggcag
     1261 gtagccggat caagcgtatg cagccgccgc attgcatcag ccatgatgga tactttctcg
     1321 gcaggagcaa ggtgagatga caggagatcc tgccccggca cttcgcccaa tagcagccag
     1381 tcccttcccg cttcagtgac aacgtcgagc acagctgcgc aaggaacgcc cgtcgtggcc
     1441 agccacgata gccgcgctgc ctcgtcctgc agttcattca gggcaccgga caggtcggtc
     1501 ttgacaaaaa gaaccgggcg cccctgcgct gacagccgga acacggcggc atcagagcag
     1561 ccgattgtct gttgtgccca gtcatagccg aatagcctct ccacccaagc ggccggagaa
     1621 cctgcgtgca atccatcttg ttcaatcatg cgaaacgatc ctcatcctgt ctcttgatca
     1681 gatccgaaaa tggatataca agctcccggg agctttttgc aaaagcctag gcctccaaaa
     1741 aagcctcctc actacttctg gaatagctca gaggcagagg cggcctcggc ctctgcataa
     1801 ataaaaaaaa ttagtcagcc atggggcgga gaatgggcgg aactgggcgg agttaggggc
     1861 gggatgggcg gagttagggg cgggactatg gttgctgact aattgagatg catgctttgc
     1921 atacttctgc ctgctgggga gcctggggac tttccacacc tggttgctga ctaattgaga
     1981 tgcatgcttt gcatacttct gcctgctggg gagcctgggg actttccaca ccctaactga
     2041 cacacattcc acagaattaa ttcgcgttaa atttttgtta aatcagctca ttttttaacc
     2101 aataggccga aatcggcaaa atcccttata aatcaaaaga atagaccgag atagggttga
     2161 gtgttgttcc agtttggaac aagagtccac tattaaagaa cgtggactcc aacgtcaaag
     2221 ggcgaaaaac cgtctatcag ggcgatggcc cactacgtga accatcaccc taatcaagtt
     2281 ttttggggtc gaggtgccgt aaagcactaa atcggaaccc taaagggagc ccccgattta
     2341 gagcttgacg gggaaagccg gcgaacgtgg cgagaaagga agggaagaaa gcgaaaggag
     2401 cgggcgctag ggcgctggca agtgtagcgg tcacgctgcg cgtaaccacc acacccgccg
     2461 cgcttaatgc gccgctacag ggcgcgtggg gataccccct agagccccag ctggttcttt
     2521 ccgcctcaga agccatagag cccaccgcat ccccagcatg cctgctattg tcttcccaat
     2581 cctccccctt gctgtcctgc cccaccccac cccccagaat agaatgacac ctactcagac
     2641 aatgcgatgc aatttcctca ttttattagg aaaggacagt gggagtggca ccttccaggg
     2701 tcaaggaagg cacgggggag gggcaaacaa cagatggctg gcaactagaa ggcacagtcg
     2761 aggctgatca gcgagctcta gcatttaggt gacactatag aatagggccc tctagacttg
     2821 tccacaatga aaggctattt gttgcaaatg aggagcagcg agatgagcag ggggtggata
     2881 atgtggatga ggatggcttg gtggtggtgg gggtggtggt tgtggaaaca gaccagccaa
     2941 tgcaagttgt ttagcctcgt cttcttcccg tggatattct tgtttaatga ccaagggtcg
     3001 aagagcaggg gcacccaccc gatattccaa aggttccaac cgcccatcga gacctgcagc
     3061 ggcacaacca taacctggtg gtggtccatg agccccgcct ggcattacag ccccgccata
     3121 aggtccggct ggagcacctg gatcatctcc gcccccccca ccgccgaagg ttggtcctac
     3181 agcggctttt gctttctctt gttggcggga atgaaattgg agatctgcga gttcatcatt
     3241 tgctgcagga tcaatagcac taaatatatc gatactggtt tcatgttcac aaattccgcc
     3301 gagaggttca taaggcgccg cgggtggagc aggtggttga gcaggtccgg cccctctaaa
     3361 aggtcccgct gcagaggaag gagcatgagg tggggattga agatgaaatg atgacattgg
     3421 cggacgaggt tctgcttcat ctgctgattc catgaaagga gaatttgaat tttccctaca
     3481 gggagaaccc cgccgtccgc catatgcttg ggctggccgt cttcgccgcc cacccggaga
     3541 ccctgctcgg aacccgcgcc cgcgcatgat cgccgataca gtcaactgtc tttgaccttt
     3601 gttactactc tcttccgatg atgatgtcgc acttattcta tgctgtctca atgttagagg
     3661 catatcagtc tccactgaag ccaatctatc tgtgacggca tctttattca cattatcttg
     3721 tacaaataat cctgttaaca atgcttttat atcctgtaaa gaatccattt tcaaaatcat
     3781 gtcaaggtct tctcgaggaa aaatcagtag aaatagctgt tccagtcttt ctagccttga
     3841 ttccacttct gtcagatgtg ccctagtcag cggagacctt ttggttttgg gagagtagcg
     3901 acactcccag ttgttcttca gacacttggc gcacttcggt ttttctttgg agcacttgag
     3961 ctttttaagt cggcaaatat cgcatgcttg ttcgatagaa gacagtagct tcatgaattc
     4021 cagcacactg gcggccgtta ctagtggatc cgagctcggt accaagcttg ggtctcccta
     4081 tagtgagtcg tattaatttc gataagccag taagcagtgg gttctctagt tagccagaga
     4141 gctctgctta tatagacctc ccaccgtaca cgcctaccgc ccatttgcgt caatggggcg
     4201 gagttgttac gacattttgg aaagtcccgt tgattttggt gccaaaacaa actcccattg
     4261 acgtcaatgg ggtggagact tggaaatccc cgtgagtcaa accgctatcc acgcccattg
     4321 atgtactgcc aaaaccgcat caccatggta atagcgatga ctaatacgta gatgtactgc
     4381 caagtaggaa agtcccataa ggtcatgtac tgggcataat gccaggcggg ccatttaccg
     4441 tcattgacgt caataggggg cgtacttggc atatgataca cttgatgtac tgccaagtgg
     4501 gcagtttacc gtaaatactc cacccattga cgtcaatgga aagtccctat tggcgttact
     4561 atgggaacat acgtcattat tgacgtcaat gggcgggggt cgttgggcgg tcagccaggc
     4621 gggccattta ccgtaagtta tgtaacgcgg aactccatat atgggctatg aactaatgac
     4681 cccgtaattg attactatta ataactagtc aataatcaat gtcaacgcgt atatctggcc
     4741 cgtacatcgc gaagcagcgc aaaacgccta accctaagca gattcttcat gcaattgtcg
     4801 gtcaagcctt gccttgttgt agcttaaatt ttgctcgcgc actactcagc gacctccaac
     4861 acacaagcag ggagcagata ctggcttaac tatgcggcat cagagcagat tgtactgaga
     4921 gtgcaccata ggggatcggg agatctcccg atccgtcgac gtcaggtggc acttttcggg
     4981 gaaatgtgcg cggaacccct atttgtttat ttttctaaat acattcaaat atgtatccgc
     5041 tcatgagaca ataaccctga taaatgcttc aataatattg aaaaaggaag agtatgagta
     5101 ttcaacattt ccgtgtcgcc cttattccct tttttgcggc attttgcctt cctgtttttg
     5161 ctcacccaga aacgctggtg aaagtaaaag atgctgaaga tcagttgggt gcacgagtgg
     5221 gttacatcga actggatctc aacagcggta agatccttga gagttttcgc cccgaagaac
     5281 gttttccaat gatgagcact tttaaagttc tgctatgtgg cgcggtatta tcccgtattg
     5341 acgccgggca agagcaactc ggtcgccgca tacactattc tcagaatgac ttggttgagt
     5401 actcaccagt cacagaaaag catcttacgg atggcatgac agtaagagaa ttatgcagtg
     5461 ctgccataac catgagtgat aacactgcgg ccaacttact tctgacaacg atcggaggac
     5521 cgaaggagct aaccgctttt ttgcacaaca tgggggatca tgtaactcgc cttgatcgtt
     5581 gggaaccgga gctgaatgaa gccataccaa acgacgagcg tgacaccacg atgcctgtag
     5641 caatggcaac aacgttgcgc aaactattaa ctggcgaact acttactcta gcttcccggc
     5701 aacaattaat agactggatg gaggcggata aagttgcagg accacttctg cgctcggccc
     5761 ttccggctgg ctggtttatt gctgataaat ctggagccgg tgagcgtggg tctcgcggta
     5821 tcattgcagc actggggcca gatggtaagc cctcccgtat cgtagttatc tacacgacgg
     5881 gaagtcaggc aactatggat gaacgaaata gacagatcgc tgagataggt gcctcactga
     5941 ttaagcattg gtaactgtca gaccaagttt actcatatat actttagatt gatttaaaac
     6001 ttcattttta atttaaaagg atctaggtga agatcctttt tgataatctc atgaccaaaa
     6061 tcccttaacg tgagttttcg ttccactgag cgtcagaccc cgtagaaaag atcaaaggat
     6121 cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc
     6181 taccagcggt ggtttgtttg ccggatcaag agctaccaac tctttttccg aaggtaactg
     6241 gcttcagcag agcgcagata ccaaatactg ttcttctagt gtagccgtag ttaggccacc
     6301 acttcaagaa ctctgtagca ccgcctacat acctcgctct gctaatcctg ttaccagtgg
     6361 ctgctgccag tggcgataag tcgtgtctta ccgggttgga ctcaagacga tagttaccgg
     6421 ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac acagcccagc ttggagcgaa
     6481 cgacctacac cgaactgaga tacctacagc gtgagctatg agaaagcgcc acgcttcccg
     6541 aagggagaaa ggcggacagg tatccggtaa gcggcagggt cggaacagga gagcgcacga
     6601 ggga
//
