LOCUS       sequence-473988-        6458 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  DriD missing the wHTH DNABD used to crystallize the apo protein.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6458)
  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 6458)
  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_78-327
FEATURES             Location/Qualifiers
     source          1..6458
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(55..73)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      69..116
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     promoter        complement(join(291..296,297..313,314..319))
                     /label=tet promoter
                     /note="E. coli promoter for tetracycline efflux protein 
                     gene"
     primer_bind     complement(346..364)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        432..536
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(537..605,606..1397)
                     /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(755..774)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      1568..2156
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2057..2076
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2310..2327
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    2342..2481
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(2407..2426)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     2567..2589
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(2583..2774)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(4086..5168)
                     /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     5130..5149
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(5169..5246)
                     /gene="lacI"
                     /label=lacI promoter
     primer_bind     5452..5471
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     5555..5574
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        5555..5573
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    5574..5598
                     /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)."
     RBS             5613..5635
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             5654..5671
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             5681..5698
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
ORIGIN
        1 tcgaggatcc ggctgctaac aaagcccgaa aggaagctga gttggctgct gccaccgctg
       61 agcaataact agcataaccc cttggggcct ctaaacgggt cttgaggggt tttttgctga
      121 aaggaggaac tatatccgga tatcccgcaa gaggcccggc agtaccggca taaccaagcc
      181 tatgcctaca gcatccaggg tgacggtgcc gaggatgacg atgagcgcat tgttagattt
      241 catacacggt gcctgactgc gttagcaatt taactgtgat aaactaccgc attaaagctt
      301 atcgatgata agctgtcaaa catgagaatt cttgaagacg aaagggcctc gtgatacgcc
      361 tatttttata ggttaatgtc atgataataa tggtttctta gacgtcaggt ggcacttttc
      421 ggggaaatgt gcgcggaacc cctatttgtt tatttttcta aatacattca aatatgtatc
      481 cgctcatgag acaataaccc tgataaatgc ttcaataata ttgaaaaagg aagagtatga
      541 gtattcaaca tttccgtgtc gcccttattc ccttttttgc ggcattttgc cttcctgttt
      601 ttgctcaccc agaaacgctg gtgaaagtaa aagatgctga agatcagttg ggtgcacgag
      661 tgggttacat cgaactggat ctcaacagcg gtaagatcct tgagagtttt cgccccgaag
      721 aacgttttcc aatgatgagc acttttaaag ttctgctatg tggcgcggta ttatcccgtg
      781 ttgacgccgg gcaagagcaa ctcggtcgcc gcatacacta ttctcagaat gacttggttg
      841 agtactcacc agtcacagaa aagcatctta cggatggcat gacagtaaga gaattatgca
      901 gtgctgccat aaccatgagt gataacactg cggccaactt acttctgaca acgatcggag
      961 gaccgaagga gctaaccgct tttttgcaca acatggggga tcatgtaact cgccttgatc
     1021 gttgggaacc ggagctgaat gaagccatac caaacgacga gcgtgacacc acgatgcctg
     1081 cagcaatggc aacaacgttg cgcaaactat taactggcga actacttact ctagcttccc
     1141 ggcaacaatt aatagactgg atggaggcgg ataaagttgc aggaccactt ctgcgctcgg
     1201 cccttccggc tggctggttt attgctgata aatctggagc cggtgagcgt gggtctcgcg
     1261 gtatcattgc agcactgggg ccagatggta agccctcccg tatcgtagtt atctacacga
     1321 cggggagtca ggcaactatg gatgaacgaa atagacagat cgctgagata ggtgcctcac
     1381 tgattaagca ttggtaactg tcagaccaag tttactcata tatactttag attgatttaa
     1441 aacttcattt ttaatttaaa aggatctagg tgaagatcct ttttgataat ctcatgacca
     1501 aaatccctta acgtgagttt tcgttccact gagcgtcaga ccccgtagaa aagatcaaag
     1561 gatcttcttg agatcctttt tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac
     1621 cgctaccagc ggtggtttgt ttgccggatc aagagctacc aactcttttt ccgaaggtaa
     1681 ctggcttcag cagagcgcag ataccaaata ctgtccttct agtgtagccg tagttaggcc
     1741 accacttcaa gaactctgta gcaccgccta catacctcgc tctgctaatc ctgttaccag
     1801 tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac
     1861 cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc agcttggagc
     1921 gaacgaccta caccgaactg agatacctac agcgtgagct atgagaaagc gccacgcttc
     1981 ccgaagggag aaaggcggac aggtatccgg taagcggcag ggtcggaaca ggagagcgca
     2041 cgagggagct tccaggggga aacgcctggt atctttatag tcctgtcggg tttcgccacc
     2101 tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg
     2161 ccagcaacgc ggccttttta cggttcctgg ccttttgctg gccttttgct cacatgttct
     2221 ttcctgcgtt atcccctgat tctgtggata accgtattac cgcctttgag tgagctgata
     2281 ccgctcgccg cagccgaacg accgagcgca gcgagtcagt gagcgaggaa gcggaagagc
     2341 gcctgatgcg gtattttctc cttacgcatc tgtgcggtat ttcacaccgc aatggtgcac
     2401 tctcagtaca atctgctctg atgccgcata gttaagccag tatacactcc gctatcgcta
     2461 cgtgactggg tcatggctgc gccccgacac ccgccaacac ccgctgacgc gccctgacgg
     2521 gcttgtctgc tcccggcatc cgcttacaga caagctgtga ccgtctccgg gagctgcatg
     2581 tgtcagaggt tttcaccgtc atcaccgaaa cgcgcgaggc agctgcggta aagctcatca
     2641 gcgtggtcgt gaagcgattc acagatgtct gcctgttcat ccgcgtccag ctcgttgagt
     2701 ttctccagaa gcgttaatgt ctggcttctg ataaagcggg ccatgttaag ggcggttttt
     2761 tcctgtttgg tcactgatgc ctccgtgtaa gggggatttc tgttcatggg ggtaatgata
     2821 ccgatgaaac gagagaggat gctcacgata cgggttactg atgatgaaca tgcccggtta
     2881 ctggaacgtt gtgagggtaa acaactggcg gtatggatgc ggcgggacca gagaaaaatc
     2941 actcagggtc aatgccagcg cttcgttaat acagatgtag gtgttccaca gggtagccag
     3001 cagcatcctg cgatgcagat ccggaacata atggtgcagg gcgctgactt ccgcgtttcc
     3061 agactttacg aaacacggaa accgaagacc attcatgttg ttgctcaggt cgcagacgtt
     3121 ttgcagcagc agtcgcttca cgttcgctcg cgtatcggtg attcattctg ctaaccagta
     3181 aggcaacccc gccagcctag ccgggtcctc aacgacagga gcacgatcat gcgcacccgt
     3241 ggccaggacc caacgctgcc cgagatgcgc cgcgtgcggc tgctggagat ggcggacgcg
     3301 atggatatgt tctgccaagg gttggtttgc gcattcacag ttctccgcaa gaattgattg
     3361 gctccaattc ttggagtggt gaatccgtta gcgaggtgcc gccggcttcc attcaggtcg
     3421 aggtggcccg gctccatgca ccgcgacgca acgcggggag gcagacaagg tatagggcgg
     3481 cgcctacaat ccatgccaac ccgttccatg tgctcgccga ggcggcataa atcgccgtga
     3541 cgatcagcgg tccaatgatc gaagttaggc tggtaagagc cgcgagcgat ccttgaagct
     3601 gtccctgatg gtcgtcatct acctgcctgg acagcatggc ctgcaacgcg ggcatcccga
     3661 tgccgccgga agcgagaaga atcataatgg ggaaggccat ccagcctcgc gtcgcgaacg
     3721 ccagcaagac gtagcccagc gcgtcggccg ccatgccggc gataatggcc tgcttctcgc
     3781 cgaaacgttt ggtggcggga ccagtgacga aggcttgagc gagggcgtgc aagattccga
     3841 ataccgcaag cgacaggccg atcatcgtcg cgctccagcg aaagcggtcc tcgccgaaaa
     3901 tgacccagag cgctgccggc acctgtccta cgagttgcat gataaagaag acagtcataa
     3961 gtgcggcgac gatagtcatg ccccgcgccc accggaagga gctgactggg ttgaaggctc
     4021 tcaagggcat cggtcgagat cccggtgcct aatgagtgag ctaacttaca ttaattgcgt
     4081 tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg ccagctgcat taatgaatcg
     4141 gccaacgcgc ggggagaggc ggtttgcgta ttgggcgcca gggtggtttt tcttttcacc
     4201 agtgagacgg gcaacagctg attgcccttc accgcctggc cctgagagag ttgcagcaag
     4261 cggtccacgc tggtttgccc cagcaggcga aaatcctgtt tgatggtggt taacggcggg
     4321 atataacatg agctgtcttc ggtatcgtcg tatcccacta ccgagatatc cgcaccaacg
     4381 cgcagcccgg actcggtaat ggcgcgcatt gcgcccagcg ccatctgatc gttggcaacc
     4441 agcatcgcag tgggaacgat gccctcattc agcatttgca tggtttgttg aaaaccggac
     4501 atggcactcc agtcgccttc ccgttccgct atcggctgaa tttgattgcg agtgagatat
     4561 ttatgccagc cagccagacg cagacgcgcc gagacagaac ttaatgggcc cgctaacagc
     4621 gcgatttgct ggtgacccaa tgcgaccaga tgctccacgc ccagtcgcgt accgtcttca
     4681 tgggagaaaa taatactgtt gatgggtgtc tggtcagaga catcaagaaa taacgccgga
     4741 acattagtgc aggcagcttc cacagcaatg gcatcctggt catccagcgg atagttaatg
     4801 atcagcccac tgacgcgttg cgcgagaaga ttgtgcaccg ccgctttaca ggcttcgacg
     4861 ccgcttcgtt ctaccatcga caccaccacg ctggcaccca gttgatcggc gcgagattta
     4921 atcgccgcga caatttgcga cggcgcgtgc agggccagac tggaggtggc aacgccaatc
     4981 agcaacgact gtttgcccgc cagttgttgt gccacgcggt tgggaatgta attcagctcc
     5041 gccatcgccg cttccacttt ttcccgcgtt ttcgcagaaa cgtggctggc ctggttcacc
     5101 acgcgggaaa cggtctgata agagacaccg gcatactctg cgacatcgta taacgttact
     5161 ggtttcacat tcaccaccct gaattgactc tcttccgggc gctatcatgc cataccgcga
     5221 aaggttttgc gccattcgat ggtgtccggg atctcgacgc tctcccttat gcgactcctg
     5281 cattaggaag cagcccagta gtaggttgag gccgttgagc accgccgccg caaggaatgg
     5341 tgcatgcaag gagatggcgc ccaacagtcc cccggccacg gggcctgcca ccatacccac
     5401 gccgaaacaa gcgctcatga gcccgaagtg gcgagcccga tcttccccat cggtgatgtc
     5461 ggcgatatag gcgccagcaa ccgcacctgt ggcgccggtg atgccggcca cgatgcgtcc
     5521 ggcgtagagg atcgagatct cgatcccgcg aaattaatac gactcactat aggggaattg
     5581 tgagcggata acaattcccc tctagaaata attttgttta actttaagaa ggagatatac
     5641 catgggcagc agccatcatc atcatcatca cagcagcggc ctggtgccgc gcggcagcca
     5701 tatgaccgcg gaggaactgg cggcgctgcg tgttgcggcg gacagcctgc gtgcgagcgg
     5761 tgcggatgcg cgtgcgagca gcctgtatgc gctggaggcg aaactgctga gcgcgctgcg
     5821 tggcagcgcg cgtcgtcgtg ttgcgccgga tgttgaggcg ctggtgcagg cggaaaccat
     5881 tgcggttcat gcgggtccgc gtccgtacga agatcaagcg gtgctgggtg cgatccgtgc
     5941 ggcgattaaa ggcctgcaag cgctgagctt tcgttatgag ggtggcagca ccccgggtcg
     6001 tacccgtgaa gttaccccgc tgggtgtgct gttcggccgt agcaactacc tggttgcgct
     6061 ggagggcaag ggtggcaaac cgcgtagctg gcgtctggac cgtatgagcg atctgaaagt
     6121 gctggacaaa ccggcgccgc cgccgcagga tttcagcctg caagcgtttg cggacgagag
     6181 cttcggcatc tatcacgacg aaattcaaga tgtggttctg cgtatccaca aaagccgtgc
     6241 ggaggatgcg ctgcgttggc gttttcacgc gacccagcaa gtgaccccgg aagcggacgg
     6301 tagcgtgctg gttacctttc gtgcgggtgg catgcgtgag ctgagctggc acctgttcac
     6361 ctggggcgat gcggtggaaa ttgttgcgcc gcaggttctg aaagacatga tggtgcaaga
     6421 actgcgtgaa gcgggtcgtg cgcacggcgc gtggtaac
//
