LOCUS       sequence-460788-        6685 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Bacterial expression of N-terminally 6His tagged Gcn4 Med15.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6685)
  AUTHORS   Bremer A, Lang WH, Kempen RP, Sweta K, Taylor AB, Borgia MB, Ansari 
            AZ, Mittag T
  TITLE     Reconciling competing models on the roles of condensates and soluble
            complexes in transcription factor function.
  JOURNAL   bioRxiv [Preprint]. 2024 Nov 22:2024.11.21.624739. doi: 
            10.1101/2024.11.21.624739.
  PUBMED    39605529
REFERENCE   2  (bases 1 to 6685)
  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"; journalName: "bioRxiv 
            [Preprint]. 2024 Nov 22:2024.11.21.624739. doi: 
            10.1101/2024.11.21.624739."
COMMENT     Sequence Label: pDest17_Med15
FEATURES             Location/Qualifiers
     source          1..6685
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     21..40
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        21..39
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     RBS             71..93
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             113..130
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     protein_bind    140..164
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             167..187
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     protein_bind    complement(2154..2178)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     primer_bind     complement(2231..2249)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      2245..2292
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     promoter        complement(join(2658..2663,2664..2680,2681..2686))
                     /label=tet promoter
                     /note="E. coli promoter for tetracycline efflux protein 
                     gene"
     primer_bind     complement(2713..2731)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        2799..2903
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2904..2972,2973..3764)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(3122..3141)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3935..4523
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4424..4443
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4677..4694
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    4709..4848
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(4774..4793)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4934..4956
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(4950..5141)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     primer_bind     6603..6622
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
ORIGIN
        1 agatctcgat cccgcgaaat taatacgact cactataggg agaccacaac ggtttccctc
       61 tagaaataat tttgtttaac tttaagaagg agatatacat atgtcgtact accatcacca
      121 tcaccatcac ctcgaatcaa caagtttgta caaaaaagca ggcttcgaga acctgtattt
      181 tcagggtagc caggataaag ataccctgag caatgcagaa cgtgccaaaa atgttaatgg
      241 tctgctgcag gttctgatgg atattaacac cctgaatggt ggtagcagcg ataccgcaga
      301 taaaattcgt attcacgcca aaaactttga agcagcactg tttgcaaaaa gcagcagcaa
      361 aaaagaatac atggatagca tgaacgaaaa ggttgccgtt atgcgcaata cctataacac
      421 ccgtaaaaat gcagttaccg cagcagcagc caataataac attaaaccgg ttgaacagca
      481 ccacatcaac aatctgaaaa atagcggtaa tagcgccaac aacatgaacg tgaatatgaa
      541 tctgaatccg cagatgtttc tgaatcagca ggcacaggca cgtcagcagg ttgcacagca
      601 gctgcgtaat cagcagcaac aacaacagca acagcaacaa cagcagcgtc gccagctgac
      661 tcctcaacag cagcagctgg tgaaccagat gaaagttgca ccgattccga aacaactgct
      721 gcagcgtatt ccgaatattc cgcctaacat taatacctgg cagcaggtta ccgcactggc
      781 acagcagaaa ctgctgacac cgcaggatat ggaagcagcc aaagaagtgt acaaaattca
      841 tcagcaactg ctgtttaaag cacgtctgca gcagcagcag gcccaggctc aggcacaagc
      901 gaataacaat aacaatggcc tgccgcagaa tggcaatatc aacaacaata ttaacatccc
      961 gcagcaacag cagatgcagc ctccgaatag cagcgcaaat aacaatccgc ttcagcagca
     1021 gagcagtcag aataccgttc cgaatgtgct gaatcagatt aaccagattt ttagccctga
     1081 agaacagcgt agcctgctgc aagaagcaat tgaaacctgc aaaaacttcg aaaaaacaca
     1141 gctgggtagc accatgaccg aaccggtgaa acagagcttt attcgcaaat acattaatca
     1201 gaaagccctg cgtaaaattc aggcactgcg tgatgtgaaa aataacaaca atgccaacaa
     1261 taacggcagc aatctgcagc gtgcacagaa tgttccgatg aacattattc agcagcagca
     1321 gcaacagaac accaataata acgataccat tgcaaccagc gcaaccccga atgcagcagc
     1381 ctttagccag caacaaaatg caagcagtaa actgtatcag atgcaacagc agcaacaggc
     1441 acaagcacag gcgcaggccc aagcgcaagc tcaagcacaa gcccaggcac aggcgcaggc
     1501 agcacaagca gcacaggcgc aagcgcaagc ccaggcgcag gctcaagccc aggcacaagc
     1561 ccaagctcaa gcgcaggcgc aagcacaggc tcaggcccag gcacaggcac aggcgcatgc
     1621 acagcatcag ccgagccagc agcctcagca agcccagcag caaccgaatc cgctgcatgg
     1681 tctgaccccg accgcaaaag atgttgaagt tattaaacag ctgagtctgg atgccagcaa
     1741 aaccaatctg cgtctgaccg atgttaccaa tagcctgtca aatgaagaaa aagaaaaaat
     1801 caaaatgaaa ctgaaacagg gccagaaact gtttgttcag gttagcaatt ttgcaccgca
     1861 ggtttatatc atcaccaaaa acgagaactt tctgaaagag gtttttcagc tgcgcatctt
     1921 cgttaaagaa atcctggaaa aatgtgccga aggcatcttt gttgttaaac tggataccgt
     1981 tgatcgcctg atcatcaaat atcagaaata ttgggaaagc atgcgcattc agatcctgcg
     2041 tcgtcaggca attctgcgtc aacaacaaca aatggcaaat aacaacggta atccgggtac
     2101 aaccagcacc ggtaacaaca ataacattgc aacccagcag aacatgcagt aagacccagc
     2161 tttcttgtac aaagtggttg attcgaggct gctaacaaag cccgaaagga agctgagttg
     2221 gctgctgcca ccgctgagca ataactagca taaccccttg gggcctctaa acgggtcttg
     2281 aggggttttt tgctgaaagg aggaactata tccggatatc cacaggacgg gtgtggtcgc
     2341 catgatcgcg tagtcgatag tggctccaag tagcgaagcg agcaggactg ggcggcggcc
     2401 aaagcggtcg gacagtgctc cgagaacggg tgcgcataga aattgcatca acgcatatag
     2461 cgctagcagc acgccatagt gactggcgat gctgtcggaa tggacgatat cccgcaagag
     2521 gcccggcagt accggcataa ccaagcctat gcctacagca tccagggtga cggtgccgag
     2581 gatgacgatg agcgcattgt tagatttcat acacggtgcc tgactgcgtt agcaatttaa
     2641 ctgtgataaa ctaccgcatt aaagcttatc gatgataagc tgtcaaacat gagaattctt
     2701 gaagacgaaa gggcctcgtg atacgcctat ttttataggt taatgtcatg ataataatgg
     2761 tttcttagac gtcaggtggc acttttcggg gaaatgtgcg cggaacccct atttgtttat
     2821 ttttctaaat acattcaaat atgtatccgc tcatgagaca ataaccctga taaatgcttc
     2881 aataatattg aaaaaggaag agtatgagta ttcaacattt ccgtgtcgcc cttattccct
     2941 tttttgcggc attttgcctt cctgtttttg ctcacccaga aacgctggtg aaagtaaaag
     3001 atgctgaaga tcagttgggt gcacgagtgg gttacatcga actggatctc aacagcggta
     3061 agatccttga gagttttcgc cccgaagaac gttttccaat gatgagcact tttaaagttc
     3121 tgctatgtgg cgcggtatta tcccgtgttg acgccgggca agagcaactc ggtcgccgca
     3181 tacactattc tcagaatgac ttggttgagt actcaccagt cacagaaaag catcttacgg
     3241 atggcatgac agtaagagaa ttatgcagtg ctgccataac catgagtgat aacactgcgg
     3301 ccaacttact tctgacaacg atcggaggac cgaaggagct aaccgctttt ttgcacaaca
     3361 tgggggatca tgtaactcgc cttgatcgtt gggaaccgga gctgaatgaa gccataccaa
     3421 acgacgagcg tgacaccacg atgcctgcag caatggcaac aacgttgcgc aaactattaa
     3481 ctggcgaact acttactcta gcttcccggc aacaattaat agactggatg gaggcggata
     3541 aagttgcagg accacttctg cgctcggccc ttccggctgg ctggtttatt gctgataaat
     3601 ctggagccgg tgagcgtggg tctcgcggta tcattgcagc actggggcca gatggtaagc
     3661 cctcccgtat cgtagttatc tacacgacgg ggagtcaggc aactatggat gaacgaaata
     3721 gacagatcgc tgagataggt gcctcactga ttaagcattg gtaactgtca gaccaagttt
     3781 actcatatat actttagatt gatttaaaac ttcattttta atttaaaagg atctaggtga
     3841 agatcctttt tgataatctc atgaccaaaa tcccttaacg tgagttttcg ttccactgag
     3901 cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa
     3961 tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg ccggatcaag
     4021 agctaccaac tctttttccg aaggtaactg gcttcagcag agcgcagata ccaaatactg
     4081 tccttctagt gtagccgtag ttaggccacc acttcaagaa ctctgtagca ccgcctacat
     4141 acctcgctct gctaatcctg ttaccagtgg ctgctgccag tggcgataag tcgtgtctta
     4201 ccgggttgga ctcaagacga tagttaccgg ataaggcgca gcggtcgggc tgaacggggg
     4261 gttcgtgcac acagcccagc ttggagcgaa cgacctacac cgaactgaga tacctacagc
     4321 gtgagctatg agaaagcgcc acgcttcccg aagggagaaa ggcggacagg tatccggtaa
     4381 gcggcagggt cggaacagga gagcgcacga gggagcttcc agggggaaac gcctggtatc
     4441 tttatagtcc tgtcgggttt cgccacctct gacttgagcg tcgatttttg tgatgctcgt
     4501 caggggggcg gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct
     4561 tttgctggcc ttttgctcac atgttctttc ctgcgttatc ccctgattct gtggataacc
     4621 gtattaccgc ctttgagtga gctgataccg ctcgccgcag ccgaacgacc gagcgcagcg
     4681 agtcagtgag cgaggaagcg gaagagcgcc tgatgcggta ttttctcctt acgcatctgt
     4741 gcggtatttc acaccgcaat ggtgcactct cagtacaatc tgctctgatg ccgcatagtt
     4801 aagccagtat acactccgct atcgctacgt gactgggtca tggctgcgcc ccgacacccg
     4861 ccaacacccg ctgacgcgcc ctgacgggct tgtctgctcc cggcatccgc ttacagacaa
     4921 gctgtgaccg tctccgggag ctgcatgtgt cagaggtttt caccgtcatc accgaaacgc
     4981 gcgaggcagc tgcggtaaag ctcatcagcg tggtcgtgaa gcgattcaca gatgtctgcc
     5041 tgttcatccg cgtccagctc gttgagtttc tccagaagcg ttaatgtctg gcttctgata
     5101 aagcgggcca tgttaagggc ggttttttcc tgtttggtca ctgatgcctc cgtgtaaggg
     5161 ggatttctgt tcatgggggt aatgataccg atgaaacgag agaggatgct cacgatacgg
     5221 gttactgatg atgaacatgc ccggttactg gaacgttgtg agggtaaaca actggcggta
     5281 tggatgcggc gggaccagag aaaaatcact cagggtcaat gccagcgctt cgttaataca
     5341 gatgtaggtg ttccacaggg tagccagcag catcctgcga tgcagatccg gaacataatg
     5401 gtgcagggcg ctgacttccg cgtttccaga ctttacgaaa cacggaaacc gaagaccatt
     5461 catgttgttg ctcaggtcgc agacgttttg cagcagcagt cgcttcacgt tcgctcgcgt
     5521 atcggtgatt cattctgcta accagtaagg caaccccgcc agcctagccg ggtcctcaac
     5581 gacaggagca cgatcatgcg cacccgtggc caggacccaa cgctgcccga gatgcgccgc
     5641 gtgcggctgc tggagatggc ggacgcgatg gatatgttct gccaagggtt ggtttgcgca
     5701 ttcacagttc tccgcaagaa ttgattggct ccaattcttg gagtggtgaa tccgttagcg
     5761 aggtgccgcc ggcttccatt caggtcgagg tggcccggct ccatgcaccg cgacgcaacg
     5821 cggggaggca gacaaggtat agggcggcgc ctacaatcca tgccaacccg ttccatgtgc
     5881 tcgccgaggc ggcataaatc gccgtgacga tcagcggtcc aatgatcgaa gttaggctgg
     5941 taagagccgc gagcgatcct tgaagctgtc cctgatggtc gtcatctacc tgcctggaca
     6001 gcatggcctg caacgcgggc atcccgatgc cgccggaagc gagaagaatc ataatgggga
     6061 aggccatcca gcctcgcgtc gcgaacgcca gcaagacgta gcccagcgcg tcggccgcca
     6121 tgccggcgat aatggcctgc ttctcgccga aacgtttggt ggcgggacca gtgacgaagg
     6181 cttgagcgag ggcgtgcaag attccgaata ccgcaagcga caggccgatc atcgtcgcgc
     6241 tccagcgaaa gcggtcctcg ccgaaaatga cccagagcgc tgccggcacc tgtcctacga
     6301 gttgcatgat aaagaagaca gtcataagtg cggcgacgat agtcatgccc cgcgcccacc
     6361 ggaaggagct gactgggttg aaggctctca agggcatcgg tcgatcgacg ctctccctta
     6421 tgcgactcct gcattaggaa gcagcccagt agtaggttga ggccgttgag caccgccgcc
     6481 gcaaggaatg gtgcatgcaa ggagatggcg cccaacagtc ccccggccac ggggcctgcc
     6541 accataccca cgccgaaaca agcgctcatg agcccgaagt ggcgagcccg atcttcccca
     6601 tcggtgatgt cggcgatata ggcgccagca accgcacctg tggcgccggt gatgccggcc
     6661 acgatgcgtc cggcgtagag gatcg
//
