LOCUS       sequence-373854-        3270 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of H-NS from its natural promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3270)
  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 3270)
  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: pRD408
FEATURES             Location/Qualifiers
     source          1..3270
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     42..60
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(98..120)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     220..239
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     433..455
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     447..464
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     448..464
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1118..1134)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1118..1134)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1131..1153)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1142..1158
                     /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(1166..1172,1173..1190,1191..1196))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1211..1232
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(1349..1366)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1520..2108)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1600..1619)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2279..3070,3071..3139))
                     /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     2902..2921
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3140..3244)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 acgtctaaga aaccattatt atcatgacat taacctataa aaataggcgt atcacgaggc
       61 cctttcgtct cgcgcgtttc ggtgatgacg gtgaaaacct ctgacacatg cagctcccgg
      121 agacggtcac agcttgtctg taagcggatg ccgggagcag acaagcccgt cagggcgcgt
      181 cagcgggtgt tggcgggtgt cggggctggc ttaactatgc ggcatcagag cagattgtac
      241 tgagagtgca ccatatgcgg tgtgaaatac cgcacagatg cgtaaggaga aaataccgca
      301 tcaggcgcca ttcgccattc aggctgcgca actgttggga agggcgatcg gtgcgggcct
      361 cttcgctatt acgccagctg gcgaaagggg gatgtgctgc aaggcgatta agttgggtaa
      421 cgccagggtt ttcccagtca cgacgttgta aaacgacggc cagtgaattc gagctcggta
      481 cccggggatc ttctggctaa ttttatgaaa agatatttat tggcggcaca aaataaagaa
      541 caattttgaa ttccttacat tcctggctat tgcacaactg aatttaaggc tctattatta
      601 cctcaacaaa ccaccccaat ataagtttga gattactaca atgagcgaag cacttaaaat
      661 tctgaacaac atccgtactc ttcgtgcgca ggcaagagaa tgtacacttg aaacgctgga
      721 agaaatgctg gaaaaattag aagttgttgt taacgaacgt cgcgaagaag aaagcgcggc
      781 tgctgctgaa gttgaagagc gcactcgtaa actgcagcaa tatcgcgaaa tgctgatcgc
      841 tgacggtatt gacccgaacg aactgctgaa tagccttgct gccgttaaat ctggcaccaa
      901 agctaaacgt gctcagcgtc cggcaaaata tagctacgtt gacgaaaacg gcgaaactaa
      961 aacctggact ggccaaggcc gtactccagc tgtaatcaaa aaagcaatgg atgagcaagg
     1021 taaatccctc gacgatttcc tgatcaagca ataatctttt gtagattgca cttgctctag
     1081 agtcgacctg caggcatgca agcttggcgt aatcatggtc atagctgttt cctgtgtgaa
     1141 attgttatcc gctcacaatt ccacacaaca tacgagccgg aagcataaag tgtaaagcct
     1201 ggggtgccta atgagtgagc taactcacat taattgcgtt gcgctcactg cccgctttcc
     1261 agtcgggaaa cctgtcgtgc cagctgcatt aatgaatcgg ccaacgcgcg gggagaggcg
     1321 gtttgcgtat tgggcgctct tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc
     1381 ggctgcggcg agcggtatca gctcactcaa aggcggtaat acggttatcc acagaatcag
     1441 gggataacgc aggaaagaac atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa
     1501 aggccgcgtt gctggcgttt ttccataggc tccgcccccc tgacgagcat cacaaaaatc
     1561 gacgctcaag tcagaggtgg cgaaacccga caggactata aagataccag gcgtttcccc
     1621 ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga tacctgtccg
     1681 cctttctccc ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg tatctcagtt
     1741 cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga accccccgtt cagcccgacc
     1801 gctgcgcctt atccggtaac tatcgtcttg agtccaaccc ggtaagacac gacttatcgc
     1861 cactggcagc agccactggt aacaggatta gcagagcgag gtatgtaggc ggtgctacag
     1921 agttcttgaa gtggtggcct aactacggct acactagaag aacagtattt ggtatctgcg
     1981 ctctgctgaa gccagttacc ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa
     2041 ccaccgctgg tagcggtggt ttttttgttt gcaagcagca gattacgcgc agaaaaaaag
     2101 gatctcaaga agatcctttg atcttttcta cggggtctga cgctcagtgg aacgaaaact
     2161 cacgttaagg gattttggtc atgagattat caaaaaggat cttcacctag atccttttaa
     2221 attaaaaatg aagttttaaa tcaatctaaa gtatatatga gtaaacttgg tctgacagtt
     2281 accaatgctt aatcagtgag gcacctatct cagcgatctg tctatttcgt tcatccatag
     2341 ttgcctgact ccccgtcgtg tagataacta cgatacggga gggcttacca tctggcccca
     2401 gtgctgcaat gataccgcga gacccacgct caccggctcc agatttatca gcaataaacc
     2461 agccagccgg aagggccgag cgcagaagtg gtcctgcaac tttatccgcc tccatccagt
     2521 ctattaattg ttgccgggaa gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg
     2581 ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg gcttcattca
     2641 gctccggttc ccaacgatca aggcgagtta catgatcccc catgttgtgc aaaaaagcgg
     2701 ttagctcctt cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg ttatcactca
     2761 tggttatggc agcactgcat aattctctta ctgtcatgcc atccgtaaga tgcttttctg
     2821 tgactggtga gtactcaacc aagtcattct gagaatagtg tatgcggcga ccgagttgct
     2881 cttgcccggc gtcaatacgg gataataccg cgccacatag cagaacttta aaagtgctca
     2941 tcattggaaa acgttcttcg gggcgaaaac tctcaaggat cttaccgctg ttgagatcca
     3001 gttcgatgta acccactcgt gcacccaact gatcttcagc atcttttact ttcaccagcg
     3061 tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata agggcgacac
     3121 ggaaatgttg aatactcata ctcttccttt ttcaatatta ttgaagcatt tatcagggtt
     3181 attgtctcat gagcggatac atatttgaat gtatttagaa aaataaacaa ataggggttc
     3241 cgcgcacatt tccccgaaaa gtgccacctg
//
