LOCUS       sequence-457026-        4992 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses RssB with R115A mutation.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4992)
  AUTHORS   Dorich V, Brugger C, Tripathi A, Hoskins JR, Tong S, Suhanovsky MM, 
            Sastry A, Wickner S, Gottesman S, Deaconescu AM
  TITLE     Structural basis for inhibition of a response regulator of sigma(S) 
            stability by a ClpXP antiadaptor.
  JOURNAL   Genes Dev. 2019 Jun 1;33(11-12):718-732. doi: 
            10.1101/gad.320168.118. Epub 2019 Apr 11.
  PUBMED    30975721
REFERENCE   2  (bases 1 to 4992)
  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: "Genes 
            Dev."; date: "2019-06-1"; volume: "33(11-12):718-732. doi"; pages: "
            10.1101/gad.320168.118. Epub 2019 Apr 11"
COMMENT     Sequence Label: pVF30
FEATURES             Location/Qualifiers
     source          1..4992
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     6..25
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(244..348)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     416..434
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     637..656
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        637..655
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    656..680
                     /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             695..717
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             1785..1802
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(1855..1873)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      1869..1916
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      1953..2333
                     /direction=RIGHT
                     /label=M13 ori
                     /note="M13 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2040..2059)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2250..2271
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2981..3000
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     3062..3081
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     primer_bind     complement(3531..3548)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3665..4210)
                     /direction=LEFT
                     /label=p15A ori
                     /note="Plasmids containing the medium-copy-number p15A 
                     origin of replication can be propagated in E. coli cells 
                     that contain a second plasmid with the ColE1 origin.
                     "
     primer_bind     complement(3745..3764)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4375..4992,1..174,175..243))
                     /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"
ORIGIN
        1 acgggataat accgcgccac atagcagaac tttaaaagtg ctcatcattg gaaaacgttc
       61 ttcggggcga aaactctcaa ggatcttacc gctgttgaga tccagttcga tgtaacccac
      121 tcgtgcaccc aactgatctt cagcatcttt tactttcacc agcgtttctg ggtgagcaaa
      181 aacaggaagg caaaatgccg caaaaaaggg aataagggcg acacggaaat gttgaatact
      241 catactcttc ctttttcaat attattgaag catttatcag ggttattgtc tcatgagcgg
      301 atacatattt gaatgtattt agaaaaataa acaaataggg gttccgcgca catttccccg
      361 aaaagtgcca cctgacgtct aagaaaccat tattatcatg acattaacct ataaaaatag
      421 gcgtatcacg aggccctttc gtcttcaaga attttataaa ccgtggagcg ggcaatactg
      481 agctgatgag caatttccgt tgcaccagtg cccttctgat gaagcgtcag cacgacgttc
      541 ctgtccacgg tacgcctgcg gccaaatttg attcctttca gctttgcttc ctgtcggccc
      601 tcattcgtgc gctctaggat ctcgatcccg cgaaattaat acgactcact ataggggaat
      661 tgtgagcgga taacaattcc cctctagaaa taattttgtt taactttaag aaggagatat
      721 acatatgacg cagccattgg tcggaaaaca gattctcatt gttgaagatg agcaggtatt
      781 tcgctcgctt ctggattcat ggttttcctc attgggagcg acaacggtac tggcggctga
      841 tggggtggat gcccttgagt tgctgggagg tttcactcca gacctgatga tatgtgatat
      901 cgcgatgcca cgaatgaacg ggcttaaact gctggagcat atacgtaaca gaggcgacca
      961 gaccccagtt ctggtgatat ctgccactga aaatatggca gatattgcca aagcgttacg
     1021 tctgggcgtt gaagatgttt tgctgaaacc agttaaagat ctgaatgcct tgcgcgagat
     1081 ggtttttgcc tgtctctatc ccagcatgtt taattcgcgc gttgaggaag aggaaaggct
     1141 ttttcgcgac tgggatgcaa tggttgataa ccctgccgca gcggcgaaat tattacagga
     1201 actacaaccg ccggttcagc aggtgatttc ccattgccgg gttaattatc gtcaattggt
     1261 tgccgcggac aaacccggcc tggtgcttga tattgccgca ctttcggaaa acgatctggc
     1321 attttattgc cttgatgtca cccgagctgg acataatggc gtacttgctg ccttgttatt
     1381 acgcgcattg tttaacggat tattacagga acagcttgca caccaaaatc aacggttgcc
     1441 agagttgggc gcgttattga agcaggtaaa ccatttactt cgtcaggcca atctgccggg
     1501 gcagtttccg ctattagttg gctattatca tcgcgaactg aaaaatctca ttctggtttc
     1561 tgcgggtctg aatgcgacgt taaataccgg cgaacaccag gtgcaaatca gtaatggtgt
     1621 tccgttaggc actttaggta acgcttattt gaatcaattg agccagcgat gcgatgcctg
     1681 gcaatgccaa atatggggaa ccggtggtcg actgcgcttg atgttgtctg cagaatgagg
     1741 atccgaattc gagctccgtc gacaagcttg cggccgcact cgagcaccac caccaccacc
     1801 actgagatcc ggctgctaac aaagcccgaa aggaagctga gttggctgct gccaccgctg
     1861 agcaataact agcataaccc cttggggcct ctaaacgggt cttgaggggt tttttgctga
     1921 aaggaggaac tatatccgga ttggcgaatg ggacgcgccc tgtagcggcg cattaagcgc
     1981 ggcgggtgtg gtggttacgc gcagcgtgac cgctacactt gccagcgccc tagcgcccgc
     2041 tcctttcgct ttcttccctt cctttctcgc cacgttcgcc ggctttcccc gtcaagctct
     2101 aaatcggggg ctccctttag ggttccgatt tagtgcttta cggcacctcg accccaaaaa
     2161 acttgattag ggtgatggtt cacgtagtgg gccatcgccc tgatagacgg tttttcgccc
     2221 tttgacgttg gagtccacgt tctttaatag tggactcttg ttccaaactg gaacaacact
     2281 caaccctatc tcggtctatt cttttgattt ataagggatt ttgccgattt cggcctattg
     2341 gttaaaaaat gagctgattt aacaaaaatt tttataggtt aatgtcatga taataatggt
     2401 ttcttagacg tcaggtggca cttttcgggg aaatgtgcgc ggaaccccta tttgtttatt
     2461 tttctaaata cattcaaata tgtatccgct catgaattaa ttcttagaaa aactcatcga
     2521 gcatcaaatg aaactgcaat ttattcatat caggattatc aataccatat ttttgaaaaa
     2581 gccgtttctg taatgaagga gaaaactcac cgaggcagtt ccataggatg gcaagatcct
     2641 ggtatcggtc tgcgattccg actcgtccaa catcaataca acctattaat ttcccctcgt
     2701 caaaaataag gttatcaagt gagaaatcac catgagtgac gactgaatcc ggtgagaatg
     2761 gcaacccggg gatcgcagtg gtgagtaacc atgcatcatc aggagtacgg ataaaatgct
     2821 tgatggtcgg aagaggcata aattccgtca gccagtttag tctgaccatc tcatctgtaa
     2881 catcattggc aacgctacct ttgccatgtt tcagaaacaa ctctggcgca tcgggcttcc
     2941 catacaatcg atagattgtc gcacctgatt gcccgacatt atcgcgagcc catttatacc
     3001 catataaatc agcatccatg ttggaattta atcgcggcag ataagacgtt tcccgttgaa
     3061 tatggctcat aacacccctt gtattactgt ttatgtaagc agacagtttt attgttcatg
     3121 atgatatatt tttatcttgt gcaatgtaac atcagagatt ttgagacaca acgtggcttt
     3181 gttgaataaa tcgaactttt gctgagttga aggatcagat cacgcatctt cccgacaacg
     3241 cagaccgttc cgtggcaaag caaaagttca aaatcaccaa ctggtccacc tacaacaaag
     3301 ctctcatcaa ccgtggctcc ctcactttct ggctggatga tggggcgatt caggcctgga
     3361 atgagtcagc aacaccttct tcacgaggca gacctcagcg ctagcggagt gtatactggc
     3421 ttactatgtt ggcactgatg agggtgtcag tgaagtgctt catgtggcag gagaaaaaag
     3481 gctgcaccgg tgcgtcagca gaatatgtga tacaggatat attccgcttc ctcgctcact
     3541 gactcgctac gctcggtcgt tcgactgcgg cgagcggaaa tggcttacga acggggcgga
     3601 gatttcctgg aagatgccag gaagatactt aacagggaag tgagagggcc gcggcaaagc
     3661 cgtttttcca taggctccgc ccccctgaca agcatcacga aatctgacgc tcaaatcagt
     3721 ggtggcgaaa cccgacagga ctataaagat accaggcgtt tccccctggc ggctccctcg
     3781 tgcgctctcc tgttcctgcc tttcggttta ccggtgtcat tccgctgtta tggccgcgtt
     3841 tgtctcattc cacgcctgac actcagttcc gggtaggcag ttcgctccaa gctggactgt
     3901 atgcacgaac cccccgttca gtccgaccgc tgcgccttat ccggtaacta tcgtcttgag
     3961 tccaacccgg aaagacatgc aaaagcacca ctggcagcag ccactggtaa ttgatttaga
     4021 ggagttagtc ttgaagtcat gagccggtta aggctaaact gaaaggacaa gttttggtga
     4081 ctgcgctcct ccaagccagt tacctcggtt caaagagttg gtagctcaga gaaccttcga
     4141 aaaaccgccc tgcaaggcgg ttttttcgtt ttcagagcaa gagattacgc gcagaccaaa
     4201 acgatctcaa gaagatcatc ttattaaggg gtctgacgct cagtggaacg aaaactcacg
     4261 ttaagggatt ttggtcatga gattatcaaa aaggatcttc acctagatcc ttttaaatta
     4321 aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa acttggtctg acagttacca
     4381 atgcttaatc agtgaggcac ctatctcagc gatctgtcta tttcgttcat ccatagttgc
     4441 ctgactcccc gtcgtgtaga taactacgat acgggagggc ttaccatctg gccccagtgc
     4501 tgcaatgata ccgcgagacc cacgctcacc ggctccagat ttatcagcaa taaaccagcc
     4561 agccggaagg gccgagcgca gaagtggtcc tgcaacttta tccgcctcca tccagtctat
     4621 taattgttgc cgggaagcta gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt
     4681 tgccattgct gcaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc
     4741 cggttcccaa cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa aagcggttag
     4801 ctccttcggt cctccgatcg ttgtcagaag taagttggcc gcagtgttat cactcatggt
     4861 tatggcagca ctgcataatt ctcttactgt catgccatcc gtaagatgct tttctgtgac
     4921 tggtgagtac tcaaccaagt cattctgaga atagtgtatg cggcgaccga gttgctcttg
     4981 cccggcgtca ac
//
