LOCUS       sequence-443026-        6232 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  For overexpression of Gal4-DBD EGR1 AroSCRAMBLED.
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-AroSCRAMBLED
FEATURES             Location/Qualifiers
     source          1..6232
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      103..531
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(190..209)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     400..421
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(540..560)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        545..874
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      725..860
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     787..806
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             941..1735
                     /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(995..1014)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     1605..1624
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    1909..2030
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1946..1965)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2000..2019
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(2079..2095)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2079..2095)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2092..2114)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2103..2119
                     /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(2127..2133,2134..2151,2152..2157))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2172..2193
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2310..2327)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2481..3069)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2561..2580)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3240..4031,4032..4100))
                     /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     3863..3882
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4101..4205)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(4280..4299)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        4471..4850
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        4851..5054
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     5005..5025
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     5099..5118
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        5099..5117
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             5180..5620
                     /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(6021..6039)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(6022..6039)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(6059..6076)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    join(6065..6232,1..57)
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
ORIGIN
        1 agggggagga ttgggaagac aatagcaggc atgctgggga tgcggtgggc tctatggctt
       61 ctgaggcgga aagaaccagc tggggctcta gggggtatcc ccacgcgccc tgtagcggcg
      121 cattaagcgc ggcgggtgtg gtggttacgc gcagcgtgac cgctacactt gccagcgccc
      181 tagcgcccgc tcctttcgct ttcttccctt cctttctcgc cacgttcgcc ggctttcccc
      241 gtcaagctct aaatcggggg ctccctttag ggttccgatt tagtgcttta cggcacctcg
      301 accccaaaaa acttgattag ggtgatggtt cacgtagtgg gccatcgccc tgatagacgg
      361 tttttcgccc tttgacgttg gagtccacgt tctttaatag tggactcttg ttccaaactg
      421 gaacaacact caaccctatc tcggtctatt cttttgattt ataagggatt ttgccgattt
      481 cggcctattg gttaaaaaat gagctgattt aacaaaaatt taacgcgaat taattctgtg
      541 gaatgtgtgt cagttagggt gtggaaagtc cccaggctcc ccagcaggca gaagtatgca
      601 aagcatgcat ctcaattagt cagcaaccag gtgtggaaag tccccaggct ccccagcagg
      661 cagaagtatg caaagcatgc atctcaatta gtcagcaacc atagtcccgc ccctaactcc
      721 gcccatcccg cccctaactc cgcccagttc cgcccattct ccgccccatg gctgactaat
      781 tttttttatt tatgcagagg ccgaggccgc ctctgcctct gagctattcc agaagtagtg
      841 aggaggcttt tttggaggcc taggcttttg caaaaagctc ccgggagctt gtatatccat
      901 tttcggatct gatcaagaga caggatgagg atcgtttcgc atgattgaac aagatggatt
      961 gcacgcaggt tctccggccg cttgggtgga gaggctattc ggctatgact gggcacaaca
     1021 gacaatcggc tgctctgatg ccgccgtgtt ccggctgtca gcgcaggggc gcccggttct
     1081 ttttgtcaag accgacctgt ccggtgccct gaatgaactg caggacgagg cagcgcggct
     1141 atcgtggctg gccacgacgg gcgttccttg cgcagctgtg ctcgacgttg tcactgaagc
     1201 gggaagggac tggctgctat tgggcgaagt gccggggcag gatctcctgt catctcacct
     1261 tgctcctgcc gagaaagtat ccatcatggc tgatgcaatg cggcggctgc atacgcttga
     1321 tccggctacc tgcccattcg accaccaagc gaaacatcgc atcgagcgag cacgtactcg
     1381 gatggaagcc ggtcttgtcg atcaggatga tctggacgaa gagcatcagg ggctcgcgcc
     1441 agccgaactg ttcgccaggc tcaaggcgcg catgcccgac ggcgaggatc tcgtcgtgac
     1501 ccatggcgat gcctgcttgc cgaatatcat ggtggaaaat ggccgctttt ctggattcat
     1561 cgactgtggc cggctgggtg tggcggaccg ctatcaggac atagcgttgg ctacccgtga
     1621 tattgctgaa gagcttggcg gcgaatgggc tgaccgcttc ctcgtgcttt acggtatcgc
     1681 cgctcccgat tcgcagcgca tcgccttcta tcgccttctt gacgagttct tctgagcggg
     1741 actctggggt tcgaaatgac cgaccaagcg acgcccaacc tgccatcacg agatttcgat
     1801 tccaccgccg ccttctatga aaggttgggc ttcggaatcg ttttccggga cgccggctgg
     1861 atgatcctcc agcgcgggga tctcatgctg gagttcttcg cccaccccaa cttgtttatt
     1921 gcagcttata atggttacaa ataaagcaat agcatcacaa atttcacaaa taaagcattt
     1981 ttttcactgc attctagttg tggtttgtcc aaactcatca atgtatctta tcatgtctgt
     2041 ataccgtcga cctctagcta gagcttggcg taatcatggt catagctgtt tcctgtgtga
     2101 aattgttatc cgctcacaat tccacacaac atacgagccg gaagcataaa gtgtaaagcc
     2161 tggggtgcct aatgagtgag ctaactcaca ttaattgcgt tgcgctcact gcccgctttc
     2221 cagtcgggaa acctgtcgtg ccagctgcat taatgaatcg gccaacgcgc ggggagaggc
     2281 ggtttgcgta ttgggcgctc ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt
     2341 cggctgcggc gagcggtatc agctcactca aaggcggtaa tacggttatc cacagaatca
     2401 ggggataacg caggaaagaa catgtgagca aaaggccagc aaaaggccag gaaccgtaaa
     2461 aaggccgcgt tgctggcgtt tttccatagg ctccgccccc ctgacgagca tcacaaaaat
     2521 cgacgctcaa gtcagaggtg gcgaaacccg acaggactat aaagatacca ggcgtttccc
     2581 cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc
     2641 gcctttctcc cttcgggaag cgtggcgctt tctcatagct cacgctgtag gtatctcagt
     2701 tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt tcagcccgac
     2761 cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggtaagaca cgacttatcg
     2821 ccactggcag cagccactgg taacaggatt agcagagcga ggtatgtagg cggtgctaca
     2881 gagttcttga agtggtggcc taactacggc tacactagaa gaacagtatt tggtatctgc
     2941 gctctgctga agccagttac cttcggaaaa agagttggta gctcttgatc cggcaaacaa
     3001 accaccgctg gtagcggtgg tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa
     3061 ggatctcaag aagatccttt gatcttttct acggggtctg acgctcagtg gaacgaaaac
     3121 tcacgttaag ggattttggt catgagatta tcaaaaagga tcttcaccta gatcctttta
     3181 aattaaaaat gaagttttaa atcaatctaa agtatatatg agtaaacttg gtctgacagt
     3241 taccaatgct taatcagtga ggcacctatc tcagcgatct gtctatttcg ttcatccata
     3301 gttgcctgac ttcccgtcgt gtagataact acgatacggg agggcttacc atctggcccc
     3361 agtgctgcaa tgataccgcg agacccacgc tcaccggctc cagatttatc agcaataaac
     3421 cagccagccg gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag
     3481 tctattaatt gttgccggga agctagagta agtagttcgc cagttaatag tttgcgcaac
     3541 gttgttgcca ttgctacagg catcgtggtg tcacgctcgt cgtttggtat ggcttcattc
     3601 agctccggtt cccaacgatc aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg
     3661 gttagctcct tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt gttatcactc
     3721 atggttatgg cagcactgca taattctctt actgtcatgc catccgtaag atgcttttct
     3781 gtgactggtg agtactcaac caagtcattc tgagaatagt gtatgcggcg accgagttgc
     3841 tcttgcccgg cgtcaatacg ggataatacc gcgccacata gcagaacttt aaaagtgctc
     3901 atcattggaa aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc
     3961 agttcgatgt aacccactcg tgcacccaac tgatcttcag catcttttac tttcaccagc
     4021 gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca
     4081 cggaaatgtt gaatactcat actcttcctt tttcaatatt attgaagcat ttatcagggt
     4141 tattgtctca tgagcggata catatttgaa tgtatttaga aaaataaaca aataggggtt
     4201 ccgcgcacat ttccccgaaa agtgccacct gacgtcgacg gatcgggaga tctcccgatc
     4261 ccctatggtg cactctcagt acaatctgct ctgatgccgc atagttaagc cagtatctgc
     4321 tccctgcttg tgtgttggag gtcgctgagt agtgcgcgag caaaatttaa gctacaacaa
     4381 ggcaaggctt gaccgacaat tgcatgaaga atctgcttag ggttaggcgt tttgcgctgc
     4441 ttcgcgatgt acgggccaga tatacgcgtt gacattgatt attgactagt tattaatagt
     4501 aatcaattac ggggtcatta gttcatagcc catatatgga gttccgcgtt acataactta
     4561 cggtaaatgg cccgcctggc tgaccgccca acgacccccg cccattgacg tcaataatga
     4621 cgtatgttcc catagtaacg ccaataggga ctttccattg acgtcaatgg gtggagtatt
     4681 tacggtaaac tgcccacttg gcagtacatc aagtgtatca tatgccaagt acgcccccta
     4741 ttgacgtcaa tgacggtaaa tggcccgcct ggcattatgc ccagtacatg accttatggg
     4801 actttcctac ttggcagtac atctacgtat tagtcatcgc tattaccatg gtgatgcggt
     4861 tttggcagta catcaatggg cgtggatagc ggtttgactc acggggattt ccaagtctcc
     4921 accccattga cgtcaatggg agtttgtttt ggcaccaaaa tcaacgggac tttccaaaat
     4981 gtcgtaacaa ctccgcccca ttgacgcaaa tgggcggtag gcgtgtacgg tgggaggtct
     5041 atataagcag agctctctgg ctaactagag aacccactgc ttactggctt atcgaaatta
     5101 atacgactca ctatagggag acccaagctt ggtaccgagc tcggatccac tagtaacggc
     5161 cgccagtgtg ctggaattca tgaagctact gtcttctatc gaacaagcat gcgatatttg
     5221 ccgacttaaa aagctcaagt gctccaaaga aaaaccgaag tgcgccaagt gtctgaagaa
     5281 caactgggag tgtcgctact ctcccaaaac caaaaggtct ccgctgacta gggcacatct
     5341 gacagaagtg gaatcaaggc tagaaagact ggaacagcta tttctactga tttttcctcg
     5401 agaagacctt gacatgattt tgaaaatgga ttctttacag gatataaaag cattgttaac
     5461 aggattattt gtacaagata atgtgaataa agatgccgtc acagatagat tggcttcagt
     5521 ggagactgat atgcctctaa cattgagaca gcatagaata agtgcgacat catcatcgga
     5581 agagagtagt aacaaaggtc aaagacagtt gactgtatcg gcgatattgc ggcagaagga
     5641 caagaaagca gacaaaagtg ttgtggcctc ttcggccacc tcctctctct cttcctaccc
     5701 gtccccggtt gctttcacct actctccgtc cccggttact acctctccat ccccgtatgc
     5761 cacctacacc tcaccatccc ctgtgcccac ctcctcctct ttccccggct cctcgtactt
     5821 caccccatcc cctgtgcaca gtggccccta ctccccgtcg gtggccacca cgtcctctgt
     5881 tccccctgct ccggcccagg tcagcagccc ttcctcagct gtcttcacca actccttcag
     5941 cgcctccaca gggtttcttt cggacatgac agcaacctct cccaggacaa ttgaaatttg
     6001 ctgagtctag agggccctat tctatagtgt cacctaaatg ctagagctcg ctgatcagcc
     6061 tcgactgtgc cttctagttg ccagccatct gttgtttgcc cctcccccgt gccttccttg
     6121 accctggaag gtgccactcc cactgtcctt tcctaataaa atgaggaaat tgcatcgcat
     6181 tgtctgagta ggtgtcattc tattctgggg ggtggggtgg ggcaggacag ca
//
