LOCUS       sequence-473985-        5918 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  The wHTH DNA binding domain used to understand the allosteric 
            mechanism.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5918)
  AUTHORS   Cannistraci E, Srinivasu BY, Chavez Orozco J Jr, Gozzi K, Wales TE, 
            Schumacher MA
  TITLE     Allosteric activation mechanism of DriD, a WYL-domain containing 
            transcription regulator.
  JOURNAL   Commun Biol. 2025 Apr 29;8(1):679. doi: 10.1038/s42003-025-08111-x.
  PUBMED    40301632
REFERENCE   2  (bases 1 to 5918)
  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: "Commun 
            Biol."; date: "2025-04-29"; volume: "8(1):679. doi"; pages: " 
            10.1038/s42003-025-08111-x"
COMMENT     Sequence Label: DriD-wHTH-DNABD
FEATURES             Location/Qualifiers
     source          1..5918
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     RBS             36..58
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             77..94
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             104..121
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
     primer_bind     complement(396..414)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      410..457
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     promoter        complement(join(632..637,638..654,655..660))
                     /label=tet promoter
                     /note="E. coli promoter for tetracycline efflux protein 
                     gene"
     primer_bind     complement(687..705)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        773..877
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(878..946,947..1738)
                     /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(1096..1115)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      1909..2497
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2398..2417
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2651..2668
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    2683..2822
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(2748..2767)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     2908..2930
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(2924..3115)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(4427..5509)
                     /codon_start=1
                     /gene="lacI"
                     /product="lac repressor"
                     /label=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)."
                     /translation="MKPVTLYDVAEYAGVSYQTVSRVVNQASHVSAKTREKVEAAMAEL
                     NYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSRADQLGASVVVSMVERSGV
                     EACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSIIFSH
                     EDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREGDWSA
                     MSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTEDSSC
                     YIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR
                     ALADSLMQLARQVSRLESGQ"
     primer_bind     5471..5490
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(5510..5587)
                     /gene="lacI"
                     /label=lacI promoter
     primer_bind     5793..5812
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     5896..5915
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        5896..5914
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    join(5915..5918,1..21)
                     /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)."
ORIGIN
        1 ttgtgagcgg ataacaattc ccctctagaa ataattttgt ttaactttaa gaaggagata
       61 taccatgggc agcagccatc atcatcatca tcacagcagc ggcctggtgc cgcgcggcag
      121 ccatatgcgt cacgaaaaag cgacccgtct gctggacctg gcgcgtatgc tggcgggtag
      181 cgcggagggt ctgaccctgg atgaaatggc ggcggcgctg ggtgtgggtc gtcgtaccgc
      241 ggagcgtatg cgtgatgcgg tttgggcggc gttcccgcag atggaagcga tcgacgatcc
      301 gccgaccaaa cgttttcgta ttccgagcgg tctggattaa ctcgaggatc cggctgctaa
      361 caaagcccga aaggaagctg agttggctgc tgccaccgct gagcaataac tagcataacc
      421 ccttggggcc tctaaacggg tcttgagggg ttttttgctg aaaggaggaa ctatatccgg
      481 atatcccgca agaggcccgg cagtaccggc ataaccaagc ctatgcctac agcatccagg
      541 gtgacggtgc cgaggatgac gatgagcgca ttgttagatt tcatacacgg tgcctgactg
      601 cgttagcaat ttaactgtga taaactaccg cattaaagct tatcgatgat aagctgtcaa
      661 acatgagaat tcttgaagac gaaagggcct cgtgatacgc ctatttttat aggttaatgt
      721 catgataata atggtttctt agacgtcagg tggcactttt cggggaaatg tgcgcggaac
      781 ccctatttgt ttatttttct aaatacattc aaatatgtat ccgctcatga gacaataacc
      841 ctgataaatg cttcaataat attgaaaaag gaagagtatg agtattcaac atttccgtgt
      901 cgcccttatt cccttttttg cggcattttg ccttcctgtt tttgctcacc cagaaacgct
      961 ggtgaaagta aaagatgctg aagatcagtt gggtgcacga gtgggttaca tcgaactgga
     1021 tctcaacagc ggtaagatcc ttgagagttt tcgccccgaa gaacgttttc caatgatgag
     1081 cacttttaaa gttctgctat gtggcgcggt attatcccgt gttgacgccg ggcaagagca
     1141 actcggtcgc cgcatacact attctcagaa tgacttggtt gagtactcac cagtcacaga
     1201 aaagcatctt acggatggca tgacagtaag agaattatgc agtgctgcca taaccatgag
     1261 tgataacact gcggccaact tacttctgac aacgatcgga ggaccgaagg agctaaccgc
     1321 ttttttgcac aacatggggg atcatgtaac tcgccttgat cgttgggaac cggagctgaa
     1381 tgaagccata ccaaacgacg agcgtgacac cacgatgcct gcagcaatgg caacaacgtt
     1441 gcgcaaacta ttaactggcg aactacttac tctagcttcc cggcaacaat taatagactg
     1501 gatggaggcg gataaagttg caggaccact tctgcgctcg gcccttccgg ctggctggtt
     1561 tattgctgat aaatctggag ccggtgagcg tgggtctcgc ggtatcattg cagcactggg
     1621 gccagatggt aagccctccc gtatcgtagt tatctacacg acggggagtc aggcaactat
     1681 ggatgaacga aatagacaga tcgctgagat aggtgcctca ctgattaagc attggtaact
     1741 gtcagaccaa gtttactcat atatacttta gattgattta aaacttcatt tttaatttaa
     1801 aaggatctag gtgaagatcc tttttgataa tctcatgacc aaaatccctt aacgtgagtt
     1861 ttcgttccac tgagcgtcag accccgtaga aaagatcaaa ggatcttctt gagatccttt
     1921 ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg
     1981 tttgccggat caagagctac caactctttt tccgaaggta actggcttca gcagagcgca
     2041 gataccaaat actgtccttc tagtgtagcc gtagttaggc caccacttca agaactctgt
     2101 agcaccgcct acatacctcg ctctgctaat cctgttacca gtggctgctg ccagtggcga
     2161 taagtcgtgt cttaccgggt tggactcaag acgatagtta ccggataagg cgcagcggtc
     2221 gggctgaacg gggggttcgt gcacacagcc cagcttggag cgaacgacct acaccgaact
     2281 gagataccta cagcgtgagc tatgagaaag cgccacgctt cccgaaggga gaaaggcgga
     2341 caggtatccg gtaagcggca gggtcggaac aggagagcgc acgagggagc ttccaggggg
     2401 aaacgcctgg tatctttata gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt
     2461 tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac gccagcaacg cggccttttt
     2521 acggttcctg gccttttgct ggccttttgc tcacatgttc tttcctgcgt tatcccctga
     2581 ttctgtggat aaccgtatta ccgcctttga gtgagctgat accgctcgcc gcagccgaac
     2641 gaccgagcgc agcgagtcag tgagcgagga agcggaagag cgcctgatgc ggtattttct
     2701 ccttacgcat ctgtgcggta tttcacaccg caatggtgca ctctcagtac aatctgctct
     2761 gatgccgcat agttaagcca gtatacactc cgctatcgct acgtgactgg gtcatggctg
     2821 cgccccgaca cccgccaaca cccgctgacg cgccctgacg ggcttgtctg ctcccggcat
     2881 ccgcttacag acaagctgtg accgtctccg ggagctgcat gtgtcagagg ttttcaccgt
     2941 catcaccgaa acgcgcgagg cagctgcggt aaagctcatc agcgtggtcg tgaagcgatt
     3001 cacagatgtc tgcctgttca tccgcgtcca gctcgttgag tttctccaga agcgttaatg
     3061 tctggcttct gataaagcgg gccatgttaa gggcggtttt ttcctgtttg gtcactgatg
     3121 cctccgtgta agggggattt ctgttcatgg gggtaatgat accgatgaaa cgagagagga
     3181 tgctcacgat acgggttact gatgatgaac atgcccggtt actggaacgt tgtgagggta
     3241 aacaactggc ggtatggatg cggcgggacc agagaaaaat cactcagggt caatgccagc
     3301 gcttcgttaa tacagatgta ggtgttccac agggtagcca gcagcatcct gcgatgcaga
     3361 tccggaacat aatggtgcag ggcgctgact tccgcgtttc cagactttac gaaacacgga
     3421 aaccgaagac cattcatgtt gttgctcagg tcgcagacgt tttgcagcag cagtcgcttc
     3481 acgttcgctc gcgtatcggt gattcattct gctaaccagt aaggcaaccc cgccagccta
     3541 gccgggtcct caacgacagg agcacgatca tgcgcacccg tggccaggac ccaacgctgc
     3601 ccgagatgcg ccgcgtgcgg ctgctggaga tggcggacgc gatggatatg ttctgccaag
     3661 ggttggtttg cgcattcaca gttctccgca agaattgatt ggctccaatt cttggagtgg
     3721 tgaatccgtt agcgaggtgc cgccggcttc cattcaggtc gaggtggccc ggctccatgc
     3781 accgcgacgc aacgcgggga ggcagacaag gtatagggcg gcgcctacaa tccatgccaa
     3841 cccgttccat gtgctcgccg aggcggcata aatcgccgtg acgatcagcg gtccaatgat
     3901 cgaagttagg ctggtaagag ccgcgagcga tccttgaagc tgtccctgat ggtcgtcatc
     3961 tacctgcctg gacagcatgg cctgcaacgc gggcatcccg atgccgccgg aagcgagaag
     4021 aatcataatg gggaaggcca tccagcctcg cgtcgcgaac gccagcaaga cgtagcccag
     4081 cgcgtcggcc gccatgccgg cgataatggc ctgcttctcg ccgaaacgtt tggtggcggg
     4141 accagtgacg aaggcttgag cgagggcgtg caagattccg aataccgcaa gcgacaggcc
     4201 gatcatcgtc gcgctccagc gaaagcggtc ctcgccgaaa atgacccaga gcgctgccgg
     4261 cacctgtcct acgagttgca tgataaagaa gacagtcata agtgcggcga cgatagtcat
     4321 gccccgcgcc caccggaagg agctgactgg gttgaaggct ctcaagggca tcggtcgaga
     4381 tcccggtgcc taatgagtga gctaacttac attaattgcg ttgcgctcac tgcccgcttt
     4441 ccagtcggga aacctgtcgt gccagctgca ttaatgaatc ggccaacgcg cggggagagg
     4501 cggtttgcgt attgggcgcc agggtggttt ttcttttcac cagtgagacg ggcaacagct
     4561 gattgccctt caccgcctgg ccctgagaga gttgcagcaa gcggtccacg ctggtttgcc
     4621 ccagcaggcg aaaatcctgt ttgatggtgg ttaacggcgg gatataacat gagctgtctt
     4681 cggtatcgtc gtatcccact accgagatat ccgcaccaac gcgcagcccg gactcggtaa
     4741 tggcgcgcat tgcgcccagc gccatctgat cgttggcaac cagcatcgca gtgggaacga
     4801 tgccctcatt cagcatttgc atggtttgtt gaaaaccgga catggcactc cagtcgcctt
     4861 cccgttccgc tatcggctga atttgattgc gagtgagata tttatgccag ccagccagac
     4921 gcagacgcgc cgagacagaa cttaatgggc ccgctaacag cgcgatttgc tggtgaccca
     4981 atgcgaccag atgctccacg cccagtcgcg taccgtcttc atgggagaaa ataatactgt
     5041 tgatgggtgt ctggtcagag acatcaagaa ataacgccgg aacattagtg caggcagctt
     5101 ccacagcaat ggcatcctgg tcatccagcg gatagttaat gatcagccca ctgacgcgtt
     5161 gcgcgagaag attgtgcacc gccgctttac aggcttcgac gccgcttcgt tctaccatcg
     5221 acaccaccac gctggcaccc agttgatcgg cgcgagattt aatcgccgcg acaatttgcg
     5281 acggcgcgtg cagggccaga ctggaggtgg caacgccaat cagcaacgac tgtttgcccg
     5341 ccagttgttg tgccacgcgg ttgggaatgt aattcagctc cgccatcgcc gcttccactt
     5401 tttcccgcgt tttcgcagaa acgtggctgg cctggttcac cacgcgggaa acggtctgat
     5461 aagagacacc ggcatactct gcgacatcgt ataacgttac tggtttcaca ttcaccaccc
     5521 tgaattgact ctcttccggg cgctatcatg ccataccgcg aaaggttttg cgccattcga
     5581 tggtgtccgg gatctcgacg ctctccctta tgcgactcct gcattaggaa gcagcccagt
     5641 agtaggttga ggccgttgag caccgccgcc gcaaggaatg gtgcatgcaa ggagatggcg
     5701 cccaacagtc ccccggccac ggggcctgcc accataccca cgccgaaaca agcgctcatg
     5761 agcccgaagt ggcgagcccg atcttcccca tcggtgatgt cggcgatata ggcgccagca
     5821 accgcacctg tggcgccggt gatgccggcc acgatgcgtc cggcgtagag gatcgagatc
     5881 tcgatcccgc gaaattaata cgactcacta taggggaa
//
