LOCUS       sequence-258310-        3004 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Engineered sgRNA-LEB1 (2 BoxB aptamers) generator circuit including 
            promoter Plux2.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3004)
  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 3004)
  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: pLY108
FEATURES             Location/Qualifiers
     source          1..3004
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    4..22
                     /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    59..77
                     /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)"
     terminator      complement(183..210)
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     promoter        218..252
                     /label=BBa_J23117
                     /note="Anderson constitutive promoter 
                     (https://parts.igem.org/Promoters/Catalog/Anderson). 
                     Measured strength relative to BBa_J223100: 0.06"
     RBS             264..275
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     promoter        1214..1316
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     CDS             1317..1976
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(1383..1402)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     terminator      1997..2091
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     rep_origin      2205..2750
                     /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     2651..2670
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2867..2884
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 cttgggccct tcttcagggc ccaagttgat aacggactag ccttatttga acttgctagg
       61 gcccttcttc agggccctag ctctcaaaca tcttccagac ctcagactct agttttattc
      121 gaaagtaaca aaccattttc ttgcgtaaac ctgtacgatc ctacaggtct ctagtatata
      181 aacgcagaaa ggcccacccg aaggtgagcc agtgtgattg acagctagct cagtcctagg
      241 gattgtgcta gctactagag attaaagagg agaaatacta gatgaaaaac ataaatgccg
      301 acgacacata cagaataatt aataaaatta aagcttgtag aagcaataat gatattaatc
      361 aatgcttatc tgatatgact aaaatggtac attgtgaata ttatttactc gcgatcattt
      421 atcctcattc tatggttaaa tctgatattt caatcctaga taattaccct aaaaaatgga
      481 ggcaatatta tgatgacgct aatttaataa aatatgatcc tatagtagat tattctaact
      541 ccaatcattc accaattaat tggaatatat ttgaaaacaa tgctgtaaat aaaaaatctc
      601 caaatgtaat taaagaagcg aaaacatcag gtcttatcac tgggtttagt ttccctattc
      661 atacggctaa caatggcttc ggaatgctta gttttgcaca ttcagaaaaa gacaactata
      721 tagatagttt atttttacat gcgtgtatga acataccatt aattgttcct tctctagttg
      781 ataattatcg aaaaataaat atagcaaata ataaatcaaa caacgattta accaaaagag
      841 aaaaagaatg tttagcgtgg gcatgcgaag gaaaaagctc ttgggatatt tcaaaaatat
      901 taggttgcag tgagcgtact gtcactttcc atttaaccaa tgcgcaaatg aaactcaata
      961 caacaaaccg ctgccaaagt atttctaaag caattttaac aggagcaatt gattgcccat
     1021 actttaaaaa ttaataacac tgatagtgct agtgtagatc actactagag ctcggtacca
     1081 aagacgaaca ataagacgct gaaaagcgtc ttttttcgtt ttggtccctc tagtattaat
     1141 tttgtcttca tgcatgaaga caactaggct ctagaagcgg ccgcgaatta gatatctggc
     1201 gaaaatgaga cgttgatcgg cacgtaagag gttccaactt tcaccataat gaaataagat
     1261 cactaccggg cgtatttttt gagttatcga gattttcagg agctaaggaa gctaaaatgg
     1321 agaaaaaaat cactggatat accaccgttg atatatccca atggcatcgt aaagaacatt
     1381 ttgaggcatt tcagtcagtt gctcaatgta cctataacca gaccgttcag ctggatatta
     1441 cggccttttt aaagaccgta aagaaaaata agcacaagtt ttatccggcc tttattcaca
     1501 ttcttgcccg cctgatgaat gctcatccgg aatttcgtat ggcaatgaaa gacggtgagc
     1561 tggtgatatg ggatagtgtt cacccttgtt acaccgtttt ccatgagcaa actgaaacgt
     1621 tttcatcgct ctggagtgaa taccacgacg atttccggca gtttctacac atatattcgc
     1681 aagatgtggc gtgttacggt gaaaacctgg cctatttccc taaagggttt attgagaata
     1741 tgtttttcgt ctcagccaat ccctgggtga gtttcaccag ttttgattta aacgtggcca
     1801 atatggacaa cttcttcgcc cccgttttca ccatgggcaa atattatacg caaggcgaca
     1861 aggtgctgat gccgctggcg attcaggttc atcatgccgt ttgtgatggc ttccatgtcg
     1921 gcagaatgct taatgaatta caacagtact gcgatgagtg gcagggcggg gcgtaatttg
     1981 atatcgagct cgcttggact cctgttgata gatccagtaa tgacctcaga actccatctg
     2041 gatttgttca gaacgctcgg ttgccgccgg gcgtttttta ttggtgagaa tccaagcact
     2101 agtaacaact tatatcgtat ggggctgact tcaggtgcta catttgaaga gataaattgc
     2161 actgaaatct agaaatattt tatctgatta ataagatgat cttcttgaga tcgttttggt
     2221 ctgcgcgtaa tctcttgctc tgaaaacgaa aaaaccgcct tgcagggcgg tttttcgaag
     2281 gttctctgag ctaccaactc tttgaaccga ggtaactggc ttggaggagc gcagtcacca
     2341 aaacttgtcc tttcagttta gccttaaccg gcgcatgact tcaagactaa ctcctctaaa
     2401 tcaattacca gtggctgctg ccagtggtgc ttttgcatgt ctttccgggt tggactcaag
     2461 acgatagtta ccggataagg cgcagcggtc ggactgaacg gggggttcgt gcatacagtc
     2521 cagcttggag cgaactgcct acccggaact gagtgtcagg cgtggaatga gacaaacgcg
     2581 gccataacag cggaatgaca ccggtaaacc gaaaggcagg aacaggagag cgcacgaggg
     2641 agccgccagg gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc caccactgat
     2701 ttgagcgtca gatttcgtga tgcttgtcag gggggcggag cctatggaaa aacggctttg
     2761 ccgcggccct ctcacttccc tgttaagtat cttcctggca tcttccagga aatctccgcc
     2821 ccgttcgtaa gccatttccg ctcgccgcag tcgaacgacc gagcgtagcg agtcagtgag
     2881 cgaggaagcg gaatatatcc ctaggcctag taggaccaaa acgaaaaaag gccccccttt
     2941 cgggaggcct cttttctgga atttggtacc gagaagcttc aaaaaaagca ccgactcggt
     3001 gcca
//
