LOCUS       sequence-350249-        5599 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  sRok overexpression.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5599)
  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 5599)
  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: pRD411
FEATURES             Location/Qualifiers
     source          1..5599
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(121..1203)
                     /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
                     YIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTAPPR
                     ALADSLMQLVRQVSRLESGQ"
     primer_bind     1165..1184
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(1204..1281)
                     /gene="lacI"
                     /label=lacI promoter
     primer_bind     1487..1506
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     1594..1613
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1594..1612
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    1613..1637
                     /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             1652..1674
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     primer_bind     complement(2123..2141)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      2137..2184
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      2221..2676
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2308..2327)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2518..2539
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(2768..3583)
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     CDS             complement(2768..3583)
                     /codon_start=1
                     /gene="aph(3')-Ia"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     3491..3510
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     3575..3594
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     rep_origin      3705..4293
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4194..4213
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4447..4464
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    4479..4618
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(4544..4563)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4704..4726
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(4720..4911)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
ORIGIN
        1 gcgacgatag tcatgccccg cgcccaccgg aaggagctga ctgggttgaa ggctctcaag
       61 ggcatcggtc gagatcccgg tgcctaatga gtgagctaac ttacattaat tgcgttgcac
      121 tcactgcccg ctttccagtc gggaaacctg tcgtaccagc tgcattaatg aatcggccaa
      181 cgcgcgggga ggggcggttt gcgtattggg cgccagggtg gtttttcttt tcaccagtga
      241 gacgggcaac agctgattgc ccttcaccgc ctggccctga gagagttgca gcaagcggtc
      301 cacgctggtt tgccccagca ggcgaaaatc ctgtttgatg gtggttaacg gcgggatata
      361 acatgagctg tcttcggtat cgtcgtatcc cactaccgag atgtccgcac caacgcgcag
      421 cccggactcg gtaatggcgc gcattgcgcc cagcgccatc tgatcgttgg caaccagcat
      481 cgcagtggga acgatgccct cattcagcat ttgcatggtt tgttgaaaac cggacatggc
      541 actccagtcg ccttcccgtt ccgctatcgg ctgaatttga ttgcgagtga gatatttatg
      601 ccagccagcc agacgcagac gcgccgagac agaacttaat gggcccgcta acagcgcgat
      661 ttgctggtga cccaatgcga ccagatgctc cacgcccagt cgcgtaccgt cttcatggga
      721 gaaaataata ctgttgatgg gtgtctggtc agagacatca agaaataacg ccggaacatt
      781 agtgcaggca gcttccacag caatggcatc ctggtcatcc agcggatagt taatgatcag
      841 cccactgacg cgttgcgcga gaagattgtg caccgccgct ttacaggctt cgacgccgct
      901 tcgttctacc atcgacacca ccacgctggc acccagttga tcggcgcgag atttaatcgc
      961 cgcgacaatt tgcgacggcg cgtgtagggc cagactggag gtggcaacgc caatcagcaa
     1021 cgactgtttg cccgccagtt gttgtgccac gcggttggga atgtaattca gctccgccat
     1081 cgccgcttcc actttttccc gcgttttcgc agaaacgtgg ctggcctggt tcaccacgcg
     1141 ggaaacggtc tgataagaga caccggcata ctctgcgaca tcgtataacg ttactggttt
     1201 cacattcacc accctgaatt gactctcttc cgggcgctat catgccatac cgcgaaaggt
     1261 tttgcgccat tcgatggtgt ccgggatctc gacgctctcc cttatgcgac tcctgcatta
     1321 ggaagcagcc cagtagtagg ttgaggccgt tgagcaccgc cgccgcaagg aatggtgcat
     1381 gcaaggagat ggcgcccaac agtcccccgg ccacggggcc tgccaccata cccacgccga
     1441 aacaagcgct catgagcccg aagtggcgag cccgatcttc cccatcggtg atgtcggcga
     1501 tataggcgcc agcaaccgca cctgtggcgc cggtgatgcc ggccacgatg cgtccggcgt
     1561 agaggatcga gatcgatctc gatcccgcga aattaatacg actcactata ggggaattgt
     1621 gagcggataa caattcccct ctagaaataa ttttgtttaa ctttaagaag gagatataca
     1681 tatgctgacc gaacgtcagg cactgcagga tcgtctggaa aaaattgata aagatgagat
     1741 caccctgatc aaagagtatc agaaacagcg caaccagatt tttgaacgtc tgcgtgaaat
     1801 tgatcgcgaa gagtataaaa acctgccgga tctgaaacag ctggcaagcc tggaaattca
     1861 tcagaaaagc aaaccggaac gtgatattcg taaacatgtt gccgtgaaca ttctgaaagt
     1921 taatccggat ggtctgagcg cagatgaact gcgtagcaaa atcgaaaaag aaaccaacat
     1981 gcagatcctg aacatgacca attttatgcg cagcatcatg gaaaaaaatc cgagcgttaa
     2041 aaaaccgcgt cgtggttatt atcgctttga agaaatgtaa ctgctaacaa agcccgaaag
     2101 gaagctgagt tggctgctgc caccgctgag caataactag cataacccct tggggcctct
     2161 aaacgggtct tgaggggttt tttgctgaaa ggaggaacta tatccggatt ggcgaatggg
     2221 acgcgccctg tagcggcgca ttaagcgcgg cgggtgtggt ggttacgcgc agcgtgaccg
     2281 ctacacttgc cagcgcccta gcgcccgctc ctttcgcttt cttcccttcc tttctcgcca
     2341 cgttcgccgg ctttccccgt caagctctaa atcgggggct ccctttaggg ttccgattta
     2401 gtgctttacg gcacctcgac cccaaaaaac ttgattaggg tgatggttca cgtagtgggc
     2461 catcgccctg atagacggtt tttcgccctt tgacgttgga gtccacgttc tttaatagtg
     2521 gactcttgtt ccaaactgga acaacactca accctatctc ggtctattct tttgatttat
     2581 aagggatttt gccgatttcg gcctattggt taaaaaatga gctgatttaa caaaaattta
     2641 acgcgaattt taacaaaata ttaacgctta caatttaggt ggcacttttc ggggaaatgt
     2701 gcgcggaacc cctatttgtt tatttttcta aatacattca aatatgtatc cgctcatgaa
     2761 ttaattctta gaaaaactca tcgagcatca aatgaaactg caatttattc atatcaggat
     2821 tatcaatacc atatttttga aaaagccgtt tctgtaatga aggagaaaac tcaccgaggc
     2881 agttccatag gatggcaaga tcctggtatc ggtctgcgat tccgactcgt ccaacatcaa
     2941 tacaacctat taatttcccc tcgtcaaaaa taaggttatc aagtgagaaa tcaccatgag
     3001 tgacgactga atccggtgag aatggcaaaa gtttatgcat ttctttccag acttgttcaa
     3061 caggccagcc attacgctcg tcatcaaaat cactcgcatc aaccaaaccg ttattcattc
     3121 gtgattgcgc ctgagcgaga cgaaatacgc gatcgctgtt aaaaggacaa ttacaaacag
     3181 gaatcgaatg caaccggcgc aggaacactg ccagcgcatc aacaatattt tcacctgaat
     3241 caggatattc ttctaatacc tggaatgctg ttttcccggg gatcgcagtg gtgagtaacc
     3301 atgcatcatc aggagtacgg ataaaatgct tgatggtcgg aagaggcata aattccgtca
     3361 gccagtttag tctgaccatc tcatctgtaa catcattggc aacgctacct ttgccatgtt
     3421 tcagaaacaa ctctggcgca tcgggcttcc catacaatcg atagattgtc gcacctgatt
     3481 gcccgacatt atcgcgagcc catttatacc catataaatc agcatccatg ttggaattta
     3541 atcgcggcct agagcaagac gtttcccgtt gaatatggct cataacaccc cttgtattac
     3601 tgtttatgta agcagacagt tttattgttc atgaccaaaa tcccttaacg tgagttttcg
     3661 ttccactgag cgtcagaccc cgtagaaaag atcaaaggat cttcttgaaa tccttttttt
     3721 ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg
     3781 ccggatcaag agctaccaac tctttttccg aaggtaactg gcttcagcag agcgcagata
     3841 ccaaatactg tccttctagt gtagccgtag ttaggccacc acttcaagaa ctctgtagca
     3901 ccgcctacat acctcgctct gctaatcctg ttaccagtgg ctgctgccag tggcgataag
     3961 tcgtgtctta ccgggttgga ctcaagacga tagttaccgg ataaggcgca gcggtcgggc
     4021 tgaacggggg gttcgtgcac acagcccagc ttggagcgaa cgacctacac cgaactgaga
     4081 tacctacagc gtgagctatg agaaagcgcc acgcttcccg aagggagaaa ggcggacagg
     4141 tatccggtaa gcggcagggt cggaacagga gagcgcacga gggagcttcc agggggaaac
     4201 gcctggtatc tttatagtcc tgtcgggttt cgccacctct gacttgagcg tcgatttttg
     4261 tgatgctcgt caggggggcg gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg
     4321 ttcctggcct tttgctggcc ttttgctcac atgttctttc ctgcgttatc ccctgattct
     4381 gtggataacc gtattaccgc ctttgagtga gctgataccg ctcgccgcag ccgaacgacc
     4441 gagcgcagcg agtcagtgag cgaggaagcg gaagagcgcc tgatgcggta ttttctcctt
     4501 acgcatctgt gcggtatttc acaccgcaat ggtgcactct cagtacaatc tgctctgatg
     4561 ccgcatagtt aagccagtat acactccgct atcgctacgt gactgggtca tggctgcgcc
     4621 ccgacacccg ccaacacccg ctgacgcgcc ctgacgggct tgtctgctcc cggcatccgc
     4681 ttacagacaa gctgtgaccg tctccgggag ctgcatgtgt cagaggtttt caccgtcatc
     4741 accgaaacgc gcgaggcagc tgcggtaaag ctcatcagcg tggtcgtgaa gcgattcaca
     4801 gatgtctgcc tgttcatccg cgtccagctc gttgagtttc tccagaagcg ttaatgtctg
     4861 gcttctgata aagcgggcca tgttaagggc ggttttttcc tgtttggtca ctgatgcctc
     4921 cgtgtaaggg ggatttctgt tcatgggggt aatgataccg atgaaacgag agaggatgct
     4981 cacgatacgg gttactgatg atgaacatgc ccggttactg gaacgttgtg agggtaaaca
     5041 actggcggta tggatgcggc gggaccagag aaaaatcact cagggtcaat gccagcgctt
     5101 cgttaataca gatgtaggtg ttccacaggg tagccagcag catcctgcga tgcagatccg
     5161 gaacataatg gtgcagggcg ctgacttccg cgtttccaga ctttacgaaa cacggaaacc
     5221 gaagaccatt catgttgttg ctcaggtcgc agacgttttg cagcagcagt cgcttcacgt
     5281 tcgctcgcgt atcggtgatt cattctgcta accagtaagg caaccccgcc agcctagccg
     5341 ggtcctcaac gacaggagca cgatcatgcg cacccgtggg gccgccatgc cggcgataat
     5401 ggcctgcttc tcgccgaaac gtttggtggc gggaccagtg acgaaggctt gagcgagggc
     5461 gtgcaagatt ccgaataccg caagcgacag gccgatcatc gtcgcgctcc agcgaaagcg
     5521 gtcctcgccg aaaatgaccc agagcgctgc cggcacctgt cctacgagtt gcatgataaa
     5581 gaagacagtc ataagtgcg
//
