LOCUS       sequence-257709-        3007 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Engineered sgRNA-LEA4 (2 BoxB aptamers) generator circuit including 
            promoter Plux2.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3007)
  AUTHORS   Liu Y, Wan X, Wang B
  TITLE     Engineered CRISPRa enables programmable eukaryote-like gene 
            activation in bacteria.
  JOURNAL   Nat Commun. 2019 Aug 26;10(1):3693. doi: 10.1038/s41467-019-11479-0.
  PUBMED    31451697
REFERENCE   2  (bases 1 to 3007)
  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: "2019-08-26"; volume: "10(1):3693. doi"; pages: " 
            10.1038/s41467-019-11479-0"
COMMENT     Sequence Label: pLY106
FEATURES             Location/Qualifiers
     source          1..3007
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      complement(116..143)
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     promoter        151..185
                     /label=BBa_J23117
                     /note="Anderson constitutive promoter 
                     (https://parts.igem.org/Promoters/Catalog/Anderson). 
                     Measured strength relative to BBa_J223100: 0.06"
     RBS             197..208
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     promoter        1147..1249
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     CDS             1250..1909
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(1316..1335)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     terminator      1930..2024
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     rep_origin      2138..2683
                     /direction=RIGHT
                     /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     2584..2603
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2800..2817
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    2941..2959
                     /label=boxB
                     /bound_moiety="lambda-N peptide"
                     /note="lambda N peptide binding site consisting of an RNA 
                     hairpin within the N-utliization (nut) site of the lambda 
                     bacteriophage transcript (Tan and Frankel, 1995)"
     protein_bind    join(2996..3007,1..7)
                     /label=boxB
                     /bound_moiety="lambda-N peptide"
                     /note="lambda N peptide binding site consisting of an RNA 
                     hairpin within the N-utliization (nut) site of the lambda 
                     bacteriophage transcript (Tan and Frankel, 1995)"
ORIGIN
        1 agggccctag ctctcaaacg ggaacgaagc gagataactc tctagtttta ttcgaaagta
       61 acaaaccatt ttcttgcgta aacctgtacg atcctacagg tctctagtat ataaacgcag
      121 aaaggcccac ccgaaggtga gccagtgtga ttgacagcta gctcagtcct agggattgtg
      181 ctagctacta gagattaaag aggagaaata ctagatgaaa aacataaatg ccgacgacac
      241 atacagaata attaataaaa ttaaagcttg tagaagcaat aatgatatta atcaatgctt
      301 atctgatatg actaaaatgg tacattgtga atattattta ctcgcgatca tttatcctca
      361 ttctatggtt aaatctgata tttcaatcct agataattac cctaaaaaat ggaggcaata
      421 ttatgatgac gctaatttaa taaaatatga tcctatagta gattattcta actccaatca
      481 ttcaccaatt aattggaata tatttgaaaa caatgctgta aataaaaaat ctccaaatgt
      541 aattaaagaa gcgaaaacat caggtcttat cactgggttt agtttcccta ttcatacggc
      601 taacaatggc ttcggaatgc ttagttttgc acattcagaa aaagacaact atatagatag
      661 tttattttta catgcgtgta tgaacatacc attaattgtt ccttctctag ttgataatta
      721 tcgaaaaata aatatagcaa ataataaatc aaacaacgat ttaaccaaaa gagaaaaaga
      781 atgtttagcg tgggcatgcg aaggaaaaag ctcttgggat atttcaaaaa tattaggttg
      841 cagtgagcgt actgtcactt tccatttaac caatgcgcaa atgaaactca atacaacaaa
      901 ccgctgccaa agtatttcta aagcaatttt aacaggagca attgattgcc catactttaa
      961 aaattaataa cactgatagt gctagtgtag atcactacta gagctcggta ccaaagacga
     1021 acaataagac gctgaaaagc gtcttttttc gttttggtcc ctctagtatt aattttgtct
     1081 tcatgcatga agacaactag gctctagaag cggccgcgaa ttagatatct ggcgaaaatg
     1141 agacgttgat cggcacgtaa gaggttccaa ctttcaccat aatgaaataa gatcactacc
     1201 gggcgtattt tttgagttat cgagattttc aggagctaag gaagctaaaa tggagaaaaa
     1261 aatcactgga tataccaccg ttgatatatc ccaatggcat cgtaaagaac attttgaggc
     1321 atttcagtca gttgctcaat gtacctataa ccagaccgtt cagctggata ttacggcctt
     1381 tttaaagacc gtaaagaaaa ataagcacaa gttttatccg gcctttattc acattcttgc
     1441 ccgcctgatg aatgctcatc cggaatttcg tatggcaatg aaagacggtg agctggtgat
     1501 atgggatagt gttcaccctt gttacaccgt tttccatgag caaactgaaa cgttttcatc
     1561 gctctggagt gaataccacg acgatttccg gcagtttcta cacatatatt cgcaagatgt
     1621 ggcgtgttac ggtgaaaacc tggcctattt ccctaaaggg tttattgaga atatgttttt
     1681 cgtctcagcc aatccctggg tgagtttcac cagttttgat ttaaacgtgg ccaatatgga
     1741 caacttcttc gcccccgttt tcaccatggg caaatattat acgcaaggcg acaaggtgct
     1801 gatgccgctg gcgattcagg ttcatcatgc cgtttgtgat ggcttccatg tcggcagaat
     1861 gcttaatgaa ttacaacagt actgcgatga gtggcagggc ggggcgtaat ttgatatcga
     1921 gctcgcttgg actcctgttg atagatccag taatgacctc agaactccat ctggatttgt
     1981 tcagaacgct cggttgccgc cgggcgtttt ttattggtga gaatccaagc actagtaaca
     2041 acttatatcg tatggggctg acttcaggtg ctacatttga agagataaat tgcactgaaa
     2101 tctagaaata ttttatctga ttaataagat gatcttcttg agatcgtttt ggtctgcgcg
     2161 taatctcttg ctctgaaaac gaaaaaaccg ccttgcaggg cggtttttcg aaggttctct
     2221 gagctaccaa ctctttgaac cgaggtaact ggcttggagg agcgcagtca ccaaaacttg
     2281 tcctttcagt ttagccttaa ccggcgcatg acttcaagac taactcctct aaatcaatta
     2341 ccagtggctg ctgccagtgg tgcttttgca tgtctttccg ggttggactc aagacgatag
     2401 ttaccggata aggcgcagcg gtcggactga acggggggtt cgtgcataca gtccagcttg
     2461 gagcgaactg cctacccgga actgagtgtc aggcgtggaa tgagacaaac gcggccataa
     2521 cagcggaatg acaccggtaa accgaaaggc aggaacagga gagcgcacga gggagccgcc
     2581 agggggaaac gcctggtatc tttatagtcc tgtcgggttt cgccaccact gatttgagcg
     2641 tcagatttcg tgatgcttgt caggggggcg gagcctatgg aaaaacggct ttgccgcggc
     2701 cctctcactt ccctgttaag tatcttcctg gcatcttcca ggaaatctcc gccccgttcg
     2761 taagccattt ccgctcgccg cagtcgaacg accgagcgta gcgagtcagt gagcgaggaa
     2821 gcggaatata tccctaggcc tagtaggacc aaaacgaaaa aaggcccccc tttcgggagg
     2881 cctcttttct ggaatttggt accgagaagc ttcaaaaaaa gcaccgactc ggtgccactt
     2941 gggcccttct tcagggccca agttgataac ggactagcct tatttgaact tgctagggcc
     3001 cttcttc
//
