LOCUS       sequence-258926-        3004 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Engineered sgRNA-LEB3 (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: pLY110
FEATURES             Location/Qualifiers
     source          1..3004
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    48..66
                     /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(172..199)
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     promoter        207..241
                     /label=BBa_J23117
                     /note="Anderson constitutive promoter 
                     (https://parts.igem.org/Promoters/Catalog/Anderson). 
                     Measured strength relative to BBa_J223100: 0.06"
     RBS             253..264
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     promoter        1203..1305
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     CDS             1306..1965
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(1372..1391)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     terminator      1986..2080
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     rep_origin      2194..2739
                     /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     2640..2659
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2856..2873
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    join(2997..3004,1..11)
                     /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 cttcagggcc caagttgata acggactagc cttatttgaa cttgctaggg cccttcttca
       61 gggccctagc tctcaaacat gggaaacgaa ctatgctcta gttttattcg aaagtaacaa
      121 accattttct tgcgtaaacc tgtacgatcc tacaggtctc tagtatataa acgcagaaag
      181 gcccacccga aggtgagcca gtgtgattga cagctagctc agtcctaggg attgtgctag
      241 ctactagaga ttaaagagga gaaatactag atgaaaaaca taaatgccga cgacacatac
      301 agaataatta ataaaattaa agcttgtaga agcaataatg atattaatca atgcttatct
      361 gatatgacta aaatggtaca ttgtgaatat tatttactcg cgatcattta tcctcattct
      421 atggttaaat ctgatatttc aatcctagat aattacccta aaaaatggag gcaatattat
      481 gatgacgcta atttaataaa atatgatcct atagtagatt attctaactc caatcattca
      541 ccaattaatt ggaatatatt tgaaaacaat gctgtaaata aaaaatctcc aaatgtaatt
      601 aaagaagcga aaacatcagg tcttatcact gggtttagtt tccctattca tacggctaac
      661 aatggcttcg gaatgcttag ttttgcacat tcagaaaaag acaactatat agatagttta
      721 tttttacatg cgtgtatgaa cataccatta attgttcctt ctctagttga taattatcga
      781 aaaataaata tagcaaataa taaatcaaac aacgatttaa ccaaaagaga aaaagaatgt
      841 ttagcgtggg catgcgaagg aaaaagctct tgggatattt caaaaatatt aggttgcagt
      901 gagcgtactg tcactttcca tttaaccaat gcgcaaatga aactcaatac aacaaaccgc
      961 tgccaaagta tttctaaagc aattttaaca ggagcaattg attgcccata ctttaaaaat
     1021 taataacact gatagtgcta gtgtagatca ctactagagc tcggtaccaa agacgaacaa
     1081 taagacgctg aaaagcgtct tttttcgttt tggtccctct agtattaatt ttgtcttcat
     1141 gcatgaagac aactaggctc tagaagcggc cgcgaattag atatctggcg aaaatgagac
     1201 gttgatcggc acgtaagagg ttccaacttt caccataatg aaataagatc actaccgggc
     1261 gtattttttg agttatcgag attttcagga gctaaggaag ctaaaatgga gaaaaaaatc
     1321 actggatata ccaccgttga tatatcccaa tggcatcgta aagaacattt tgaggcattt
     1381 cagtcagttg ctcaatgtac ctataaccag accgttcagc tggatattac ggccttttta
     1441 aagaccgtaa agaaaaataa gcacaagttt tatccggcct ttattcacat tcttgcccgc
     1501 ctgatgaatg ctcatccgga atttcgtatg gcaatgaaag acggtgagct ggtgatatgg
     1561 gatagtgttc acccttgtta caccgttttc catgagcaaa ctgaaacgtt ttcatcgctc
     1621 tggagtgaat accacgacga tttccggcag tttctacaca tatattcgca agatgtggcg
     1681 tgttacggtg aaaacctggc ctatttccct aaagggttta ttgagaatat gtttttcgtc
     1741 tcagccaatc cctgggtgag tttcaccagt tttgatttaa acgtggccaa tatggacaac
     1801 ttcttcgccc ccgttttcac catgggcaaa tattatacgc aaggcgacaa ggtgctgatg
     1861 ccgctggcga ttcaggttca tcatgccgtt tgtgatggct tccatgtcgg cagaatgctt
     1921 aatgaattac aacagtactg cgatgagtgg cagggcgggg cgtaatttga tatcgagctc
     1981 gcttggactc ctgttgatag atccagtaat gacctcagaa ctccatctgg atttgttcag
     2041 aacgctcggt tgccgccggg cgttttttat tggtgagaat ccaagcacta gtaacaactt
     2101 atatcgtatg gggctgactt caggtgctac atttgaagag ataaattgca ctgaaatcta
     2161 gaaatatttt atctgattaa taagatgatc ttcttgagat cgttttggtc tgcgcgtaat
     2221 ctcttgctct gaaaacgaaa aaaccgcctt gcagggcggt ttttcgaagg ttctctgagc
     2281 taccaactct ttgaaccgag gtaactggct tggaggagcg cagtcaccaa aacttgtcct
     2341 ttcagtttag ccttaaccgg cgcatgactt caagactaac tcctctaaat caattaccag
     2401 tggctgctgc cagtggtgct tttgcatgtc tttccgggtt ggactcaaga cgatagttac
     2461 cggataaggc gcagcggtcg gactgaacgg ggggttcgtg catacagtcc agcttggagc
     2521 gaactgccta cccggaactg agtgtcaggc gtggaatgag acaaacgcgg ccataacagc
     2581 ggaatgacac cggtaaaccg aaaggcagga acaggagagc gcacgaggga gccgccaggg
     2641 ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc accactgatt tgagcgtcag
     2701 atttcgtgat gcttgtcagg ggggcggagc ctatggaaaa acggctttgc cgcggccctc
     2761 tcacttccct gttaagtatc ttcctggcat cttccaggaa atctccgccc cgttcgtaag
     2821 ccatttccgc tcgccgcagt cgaacgaccg agcgtagcga gtcagtgagc gaggaagcgg
     2881 aatatatccc taggcctagt aggaccaaaa cgaaaaaagg cccccctttc gggaggcctc
     2941 ttttctggaa tttggtaccg agaagcttca aaaaaagcac cgactcggtg ccacttgggc
     3001 cctt
//
