LOCUS       sequence-380748-        3432 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of Rok from the hns promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3432)
  AUTHORS   Erkelens AM, Qin L, van Erp B, Miguel-Arribas A, Abia D, Keek HGJ, 
            Markus D, Cajili MKM, Schwab S, Meijer WJJ, Dame RT
  TITLE     The B. subtilis Rok protein is an atypical H-NS-like protein 
            irresponsive to physico-chemical cues.
  JOURNAL   Nucleic Acids Res. 2022 Nov 21. pii: 6835367. doi: 
            10.1093/nar/gkac1064.
  PUBMED    36408910
REFERENCE   2  (bases 1 to 3432)
  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: "Nucleic 
            Acids Res. 2022 Nov 21. pii: 6835367. doi: 10.1093/nar/gkac1064."
COMMENT     Sequence Label: pRD424
FEATURES             Location/Qualifiers
     source          1..3432
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(28..50)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     150..169
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     363..385
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     377..394
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     378..394
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1210..1226)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1210..1226)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1223..1245)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1234..1250
                     /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(1258..1264,1265..1282,1283..1288))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1303..1324
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(1441..1458)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1612..2200)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1692..1711)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2371..3162,3163..3231))
                     /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     2994..3013
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3232..3336)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     3404..3422
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
ORIGIN
        1 cgcgcgtttc ggtgatgacg gtgaaaacct ctgacacatg cagctcccgg agacggtcac
       61 agcttgtctg taagcggatg ccgggagcag acaagcccgt cagggcgcgt cagcgggtgt
      121 tggcgggtgt cggggctggc ttaactatgc ggcatcagag cagattgtac tgagagtgca
      181 ccatatgcgg tgtgaaatac cgcacagatg cgtaaggaga aaataccgca tcaggcgcca
      241 ttcgccattc aggctgcgca actgttggga agggcgatcg gtgcgggcct cttcgctatt
      301 acgccagctg gcgaaagggg gatgtgctgc aaggcgatta agttgggtaa cgccagggtt
      361 ttcccagtca cgacgttgta aaacgacggc cagtgaattc gagctcggta cccggggatc
      421 ttctggctaa ttttatgaaa agatatttat tggcggcaca aaataaagaa caattttgaa
      481 ttccttacat tcctggctat tgcacaactg aatttaaggc tctattatta cctcaacaaa
      541 ccaccccaat ataagtttga gattactaca atgtttaatg aaagagaagc tttgcgcttg
      601 cggttagaac aattaaatga agctgaagtg aaagtcattc gtgaatatca aattgaacgt
      661 gataaaatat acgcaaaatt aagagagctg gacagaaatg gaagcccttc cgaaatcaaa
      721 aaggatttcc gttctgaaaa aaaacctgac tctctgccgg ttctcgctga gcttgcggct
      781 caggaaatca gaagctatca gccgcaatca cagcagcagt ctgttcagcc tcagcttcaa
      841 tctatttcct ctcttcctgc cggtatacca gacggaacaa cacgaagaag aagaggaacg
      901 gcaagaccgg gatcaaaagc agctaagctg cgtgaagctg ctattaaaac attaaaacgc
      961 cacaacgcgg cgattaagag ctcggagctt caaaaagaga ttgaaaaaga aagcggtctt
     1021 gaaatcccta atatgacaac gtttatgcaa agcttaatta aaatgtaccc ggaagtgaaa
     1081 aagccatatc gcgggcagta cattttagag ggtgaaatcg aatctgcaga atcagcaaac
     1141 gaataaatat aaagaaaaac tgcttgctct agagtcgacc tgcaggcatg caagcttggc
     1201 gtaatcatgg tcatagctgt ttcctgtgtg aaattgttat ccgctcacaa ttccacacaa
     1261 catacgagcc ggaagcataa agtgtaaagc ctggggtgcc taatgagtga gctaactcac
     1321 attaattgcg ttgcgctcac tgcccgcttt ccagtcggga aacctgtcgt gccagctgca
     1381 ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt attgggcgct cttccgcttc
     1441 ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat cagctcactc
     1501 aaaggcggta atacggttat ccacagaatc aggggataac gcaggaaaga acatgtgagc
     1561 aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag
     1621 gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc
     1681 gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt
     1741 tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct
     1801 ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg
     1861 ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct
     1921 tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat
     1981 tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg
     2041 ctacactaga agaacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa
     2101 aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt
     2161 ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc
     2221 tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt
     2281 atcaaaaagg atcttcacct agatcctttt aaattaaaaa tgaagtttta aatcaatcta
     2341 aagtatatat gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg aggcacctat
     2401 ctcagcgatc tgtctatttc gttcatccat agttgcctga ctccccgtcg tgtagataac
     2461 tacgatacgg gagggcttac catctggccc cagtgctgca atgataccgc gagacccacg
     2521 ctcaccggct ccagatttat cagcaataaa ccagccagcc ggaagggccg agcgcagaag
     2581 tggtcctgca actttatccg cctccatcca gtctattaat tgttgccggg aagctagagt
     2641 aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt
     2701 gtcacgctcg tcgtttggta tggcttcatt cagctccggt tcccaacgat caaggcgagt
     2761 tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt
     2821 cagaagtaag ttggccgcag tgttatcact catggttatg gcagcactgc ataattctct
     2881 tactgtcatg ccatccgtaa gatgcttttc tgtgactggt gagtactcaa ccaagtcatt
     2941 ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac gggataatac
     3001 cgcgccacat agcagaactt taaaagtgct catcattgga aaacgttctt cggggcgaaa
     3061 actctcaagg atcttaccgc tgttgagatc cagttcgatg taacccactc gtgcacccaa
     3121 ctgatcttca gcatctttta ctttcaccag cgtttctggg tgagcaaaaa caggaaggca
     3181 aaatgccgca aaaaagggaa taagggcgac acggaaatgt tgaatactca tactcttcct
     3241 ttttcaatat tattgaagca tttatcaggg ttattgtctc atgagcggat acatatttga
     3301 atgtatttag aaaaataaac aaataggggt tccgcgcaca tttccccgaa aagtgccacc
     3361 tgacgtctaa gaaaccatta ttatcatgac attaacctat aaaaataggc gtatcacgag
     3421 gccctttcgt ct
//
