LOCUS       sequence-305085-        6062 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Controller plasmid for TS circuit and cL and cR controls.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6062)
  AUTHORS   Santos-Moreno J, Tasiudi E, Stelling J, Schaerli Y
  TITLE     Multistable and dynamic CRISPRi-based synthetic circuits.
  JOURNAL   Nat Commun. 2020 Jun 2;11(1):2746. doi: 10.1038/s41467-020-16574-1.
  PUBMED    32488086
REFERENCE   2  (bases 1 to 6062)
  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: "Nat 
            Commun."; date: "2020-06-2"; volume: "11(1):2746. doi"; pages: " 
            10.1038/s41467-020-16574-1"
COMMENT     Sequence Label: pJ2077.2
FEATURES             Location/Qualifiers
     source          1..6062
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    112..131
                     /label=Csy4 Site
                     /bound_moiety="Pseudomonas aeruginosa Csy4 (Cas6f)"
                     /note="cleavage site for the Csy4 endoribonuclease (Cermak 
                     et al. 2017)"
     protein_bind    238..257
                     /label=Csy4 Site
                     /bound_moiety="Pseudomonas aeruginosa Csy4 (Cas6f)"
                     /note="cleavage site for the Csy4 endoribonuclease (Cermak 
                     et al. 2017)"
     protein_bind    651..667
                     /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)."
     protein_bind    742..761
                     /label=Csy4 Site
                     /bound_moiety="Pseudomonas aeruginosa Csy4 (Cas6f)"
                     /note="cleavage site for the Csy4 endoribonuclease (Cermak 
                     et al. 2017)"
     protein_bind    868..887
                     /label=Csy4 Site
                     /bound_moiety="Pseudomonas aeruginosa Csy4 (Cas6f)"
                     /note="cleavage site for the Csy4 endoribonuclease (Cermak 
                     et al. 2017)"
     primer_bind     complement(1413..1431)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     CDS             complement(1950..2900)
                     /codon_start=1
                     /gene="rep101"
                     /product="RepA protein needed for replication with the 
                     pSC101 origin"
                     /label=Rep101
                     /translation="MSELVVFKANELAISRYDLTEHETKLILCCVALLNPTIENPTRKE
                     RTVSFTYNQYAQMMNISRENAYGVLAKATRELMTRTVEIRNPLVKGFEIFQWTNYAKFS
                     SEKLELVFSEEILPYLFQLKKFIKYNLEHVKSFENKYSMRIYEWLLKELTQKKTHKANI
                     EISLDEFKFMLMLENNYHEFKRLNQWVLKPISKDLNTYSNMKLVVDKRGRPTDTLIFQV
                     ELDRQMDLVTELENNQIKMNGDKIPTTITSDSYLHNGLRKTLHDALTAKIQLTSFEAKF
                     LSDMQSKYDLNGSFSWLTQKQRTTLENILAKYGRI"
     rep_origin      2948..3171
                     /label=pSC101 ori
                     /note="low-copy replication origin that requires the Rep101
                     protein"
     terminator      complement(3663..3757)
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             complement(join(3783..4574,4575..4643))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDKRNRQIAEIGAS
                     LIKHW"
     primer_bind     4406..4425
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4644..4748)
                     /gene="bla"
                     /label=AmpR promoter
     CDS             complement(4924..5802)
                     /codon_start=1
                     /gene="araC"
                     /product="L-arabinose regulatory protein"
                     /label=araC
                     /translation="MAEAQNDPLLPGYSFNAHLVAGLTPIEANGYLDFFIDRPLGMKGY
                     ILNLTIRGQGVVKNQGREFVCRPGDILLFPPGEIHHYGRHPEAREWYHQWVYFRPRAYW
                     HEWLNWPSIFANTGFFRPDEAHQPHFSDLFGQIINAGQGEGRYSELLAINLLEQLLLRR
                     MEAINESLHPPMDNRVREACQYISDHLADSNFDIASVAQHVCLSPSRLSHLFRQQLGIS
                     VLSWREDQRISQAKLLLSTTRMPIATVGRNVGFDDQLYFSRVFKKCTGASPSEFRAGCE
                     EKVNDVAVKLS"
     promoter        join(5829..6062,1..51)
                     /gene="araBAD"
                     /label=araBAD promoter
                     /note="promoter of the L-arabinose operon of E. coli; the 
                     araC regulatory gene is transcribed in the opposite 
                     direction (Guzman et al., 1995)"
     primer_bind     6036..6055
                     /label=pBAD Forward
                     /note="For vectors with E. coli araBAD promoter, forward 
                     primer"
ORIGIN
        1 tagcggttcc tacctgacgc tttttatcgc aactctctac tgtttctcca taccgaattc
       61 ataggataga ttctggaaac tttaccgtcc gagctccagc ctgcggtccg ggttcactgc
      121 cgtataggca gtctcaagct agactctagt ggtttcagag ctatgctgga aacagcatag
      181 caagttgaaa taaggctagt ccgttatcaa cttgaaaaag tggcaccgag tcggtgcgtt
      241 cactgccgta taggcagtcg ctgggacgcc cgctcgagca ataaacagtt gatagggctt
      301 ctccgttacc atggttcagc caaaaaactt aagaccgccg gtcttgtcca ctaccttgca
      361 gtaatgcggt ggacaggatc ggcggttttc ttttctcttc tcaattcttc tgacctgtaa
      421 cgaataatag atagtaaagt agtctccgat tgagttttct ctgccgagtc ccacccagtt
      481 ctgtgatttc agtaagttgg taattgatac actgttgcga gaactgctgc ctggtagtag
      541 ataggttgtt attgagtaag aaggtaaagt gaacgaaatc cctgaaactg agactgtaga
      601 aaataagctt cagcctgcgg tccggacctg taggatcgta catgctgaca ttgtgagcgg
      661 ataacaatac tgtttcttca atcgctggga cgcccggtag tgcttatcag acccaatact
      721 gttgaacagc ctgcggtccg ggttcactgc cgtataggca ggacacatct tagagtatgt
      781 agtttcagag ctatgctgga aacagcatag caagttgaaa taaggctagt ccgttatcaa
      841 cttgaaaaag tggcaccgag tcggtgcgtt cactgccgta taggcagtcg ctgggacgcc
      901 cgggatccaa gagatttcta cacgattgag cactgtctcc tgcaggctcg gtaccaaatt
      961 ccagaaaaga ggcctcccga aaggggggcc ttttttcgtt ttggtcctaa tagataaagg
     1021 ataggtctgg tagtgttgtt cgttctcgca ggtaaatcaa taatactcag cagttccgta
     1081 gacttttcag tgggacaggg tagcgataac agatagattg taataagaca cagtaggtgc
     1141 tcgtagttgc gtgaagagaa ccgctcagga aatccagtca gaagtattgg taatcgttga
     1201 aaactcagtc gacgcactta ctgaagacgt cctattacac tcgtcgttgg aaactgaaga
     1261 tgcggccgcg gaaacacaga aaaaagcccg cacctgacag tgcgggcttt ttttttcgac
     1321 caaaggtagc gaacgacgag tcactgttga ggataaatac tttctctact aggcgcctgt
     1381 tacacaggtc ctcagcggcg cgcctgctgc caccgctgag caataactag cgtacgggtt
     1441 ttgctgcccg caaacgggct gttctggtgt tgctagtttg ttatcagaat cgcagatccg
     1501 gcttcagccg gtttgccggc tgaaagcgct atttcttcca gaattgccat gattttttcc
     1561 ccacgggagg cgtcactggc tcccgtgttg tcggcagctt tgattcgata agcagcatcg
     1621 cctgtttcag gctgtctatg tgtgactgtt gagctgtaac aagttgtctc aggtgttcaa
     1681 tttcatgttc tagttgcttt gttttactgg tttcacctgt tctattaggt gttacatgct
     1741 gttcatctgt tacattgtcg atctgttcat ggtgaacagc tttgaatgca ccaaaaactc
     1801 gtaaaagctc tgatgtatct atctttttta caccgttttc atctgtgcat atggacagtt
     1861 ttccctttga tatgtaacgg tgaacagttg ttctactttt gtttgttagt cttgatgctt
     1921 cactgataga tacaagagcc ataagaacct cagatccttc cgtatttagc cagtatgttc
     1981 tctagtgtgg ttcgttgttt ttgcgtgagc catgagaacg aaccattgag atcatactta
     2041 ctttgcatgt cactcaaaaa ttttgcctca aaactggtga gctgaatttt tgcagttaaa
     2101 gcatcgtgta gtgtttttct tagtccgtta tgtaggtagg aatctgatgt aatggttgtt
     2161 ggtattttgt caccattcat ttttatctgg ttgttctcaa gttcggttac gagatccatt
     2221 tgtctatcta gttcaacttg gaaaatcaac gtatcagtcg ggcggcctcg cttatcaacc
     2281 accaatttca tattgctgta agtgtttaaa tctttactta ttggtttcaa aacccattgg
     2341 ttaagccttt taaactcatg gtagttattt tcaagcatta acatgaactt aaattcatca
     2401 aggctaatct ctatatttgc cttgtgagtt ttcttttgtg ttagttcttt taataaccac
     2461 tcataaatcc tcatagagta tttgttttca aaagacttaa catgttccag attatatttt
     2521 atgaattttt ttaactggaa aagataaggc aatatctctt cactaaaaac taattctaat
     2581 ttttcgcttg agaacttggc atagtttgtc cactggaaaa tctcaaagcc tttaaccaaa
     2641 ggattcctga tttccacagt tctcgtcatc agctctctgg ttgctttagc taatacacca
     2701 taagcatttt ccctactgat gttcatcatc tgagcgtatt ggttataagt gaacgatacc
     2761 gtccgttctt tccttgtagg gttttcaatc gtggggttga gtagtgccac acagcataaa
     2821 attagcttgg tttcatgctc cgttaagtca tagcgactaa tcgctagttc atttgctttg
     2881 aaaacaacta attcagacat acatctcaat tggtctaggt gattttaatc actataccaa
     2941 ttgagatggg ctagtcaatg ataattacta gtccttttcc tttgagttgt gggtatctgt
     3001 aaattctgct agacctttgc tggaaaactt gtaaattctg ctagaccctc tgtaaattcc
     3061 gctagacctt tgtgtgtttt tttttgttta tattcaagtg gttataattt atagaataaa
     3121 gaaagaataa aaaaagataa aaagaataga tcccagccct gtgtataact cactacttta
     3181 gtcagttccg cagtattaca aaaggatgtc gcaaacgctg tttgctcctc tacaaaacag
     3241 accttaaaac cctaaaggct taagtagcac cctcgcaagc tcgggcaaat cgctgaatat
     3301 tccttttgtc tccgaccatc aggcacctga gtcgctgtct ttttcgtgac attcagttcg
     3361 ctgcgctcac ggctctggca gtgaatgggg gtaaatggca ctacaggctc cttttatgga
     3421 ttcatgcaag gaaactaccc ataatacaag aaaagcccgt cacgggcttc tcagggcgtt
     3481 ttatggcggg tctgctatgt ggtgctatct gactttttgc tgttcagcag ttcctgccct
     3541 ctgattttcc agtctgacca cttcggatta tcccgtgaca ggtcattcag actggctaat
     3601 gcacccagta aggcagcggt atcatcaaca ggcttacccg tcttactgtc cctagtgctt
     3661 ggattctcac caataaaaaa cgcccggcgg caaccgagcg ttctgaacaa atccagatgg
     3721 agttctgagg tcattactgg atctatcaac aggagtccaa gcgcgtaaac ttggtctgac
     3781 agttaccaat gcttaatcag tgaggcacct atctcagcga tctgtctatt tcgtttatcc
     3841 atagttgcct gactccccgt cgtgtagata actacgatac gggagggctt accatctggc
     3901 cccagtgctg caatgatacc gcgagaccca cgctcaccgg ctccagattt atcagcaata
     3961 aaccagccag ccggaagggc cgagcgcaga agtggtcctg caactttatc cgcctccatc
     4021 cagtctatta attgttgccg ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc
     4081 aacgttgttg ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca
     4141 ttcagctccg gttcccaacg atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa
     4201 gcggttagct ccttcggtcc tccgatcgtt gtcagaagta agttggccgc agtgttatca
     4261 ctcatggtta tggcagcact gcataattct cttactgtca tgccatccgt aagatgcttt
     4321 tctgtgactg gtgagtactc aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt
     4381 tgctcttgcc cggcgtcaat acgggataat accgcgccac atagcagaac tttaaaagtg
     4441 ctcatcattg gaaaacgttc ttcggggcga aaactctcaa ggatcttacc gctgttgaga
     4501 tccagttcga tgtaacccac tcgtgcaccc aactgatctt cagcatcttt tactttcacc
     4561 agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg aataagggcg
     4621 acacggaaat gttgaatact catactcttc ctttttcaat attattgaag catttatcag
     4681 ggttattgtc tcatgagcgg atacatattt gaatgtattt agaaaaataa acaaataggg
     4741 gttccgcgca catttccccg aaaagtgcca ccttcactct agatttcagt gcaatttatc
     4801 tcttcaaatg tagcacctga agtcagcccc atacgatata agttgtaatt ctcatgttag
     4861 tcatgccccg cgcccaccgg aaggagctga ctgggttgct cctagggtct gattcgttac
     4921 caattatgac aacttgacgg ctacatcatt cactttttct tcacaaccgg cacggaactc
     4981 gctcgggctg gccccggtgc attttttaaa tacccgcgag aaatagagtt gatcgtcaaa
     5041 accaacattg cgaccgacgg tggcgatagg catccgggtg gtgctcaaaa gcagcttcgc
     5101 ctggctgata cgttggtcct cgcgccagct taagacgcta atccctaact gctggcggaa
     5161 aagatgtgac agacgcgacg gcgacaagca aacatgctgt gcgacgctgg cgatatcaaa
     5221 attgctgtct gccaggtgat cgctgatgta ctgacaagcc tcgcgtaccc gattatccat
     5281 cggtggatgg agcgactcgt taatcgcttc catgcgccgc agtaacaatt gctcaagcag
     5341 atttatcgcc agcagctccg aatagcgccc ttccccttgc ccggcgttaa tgatttgccc
     5401 aaacaggtcg ctgaaatgcg gctggtgcgc ttcatccggg cgaaagaacc ccgtattggc
     5461 aaatattgac ggccagttaa gccattcatg ccagtaggcg cgcggacgaa agtaaaccca
     5521 ctggtgatac cattcgcgag cctccggatg acgaccgtag tgatgaatct ctcctggcgg
     5581 gaacagcaaa atatcacccg gtcggcaaac aaattctcgt ccctgatttt tcaccacccc
     5641 ctgaccgcga atggtgagat tgagaatata acctttcatt cccagcggtc ggtcgataaa
     5701 aaaatcgaga taaccgttgg cctcaatcgg cgttaaaccc gccaccagat gggcattaaa
     5761 cgagtatccc ggcagcaggg gatcattttg cgcttcagcc atacttttca tactcccgcc
     5821 attcagagaa gaaaccaatt gtccatattg catcagacat tgccgtcact gcgtctttta
     5881 ctggctcttc tcgctaacca aaccggtaac cccgcttatt aaaagcattc tgtaacaaag
     5941 cgggaccaaa gccatgacaa aaacgcgtaa caaaagtgtc tataatcacg gcagaaaagt
     6001 ccacattgat tatttgcacg gcgtcacact ttgctatgcc atagcatttt tatccataag
     6061 at
//
