LOCUS       sequence-258320-        3004 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Engineered sgRNA-LEB5 (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: pLY112
FEATURES             Location/Qualifiers
     source          1..3004
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        599..701
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     CDS             702..1361
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(768..787)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     terminator      1382..1476
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     rep_origin      1590..2135
                     /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     2036..2055
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2252..2269
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    2393..2411
                     /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    2448..2466
                     /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(2572..2599)
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     promoter        2607..2641
                     /label=BBa_J23117
                     /note="Anderson constitutive promoter 
                     (https://parts.igem.org/Promoters/Catalog/Anderson). 
                     Measured strength relative to BBa_J223100: 0.06"
     RBS             2653..2664
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
ORIGIN
        1 aagcgaaaac atcaggtctt atcactgggt ttagtttccc tattcatacg gctaacaatg
       61 gcttcggaat gcttagtttt gcacattcag aaaaagacaa ctatatagat agtttatttt
      121 tacatgcgtg tatgaacata ccattaattg ttccttctct agttgataat tatcgaaaaa
      181 taaatatagc aaataataaa tcaaacaacg atttaaccaa aagagaaaaa gaatgtttag
      241 cgtgggcatg cgaaggaaaa agctcttggg atatttcaaa aatattaggt tgcagtgagc
      301 gtactgtcac tttccattta accaatgcgc aaatgaaact caatacaaca aaccgctgcc
      361 aaagtatttc taaagcaatt ttaacaggag caattgattg cccatacttt aaaaattaat
      421 aacactgata gtgctagtgt agatcactac tagagctcgg taccaaagac gaacaataag
      481 acgctgaaaa gcgtcttttt tcgttttggt ccctctagta ttaattttgt cttcatgcat
      541 gaagacaact aggctctaga agcggccgcg aattagatat ctggcgaaaa tgagacgttg
      601 atcggcacgt aagaggttcc aactttcacc ataatgaaat aagatcacta ccgggcgtat
      661 tttttgagtt atcgagattt tcaggagcta aggaagctaa aatggagaaa aaaatcactg
      721 gatataccac cgttgatata tcccaatggc atcgtaaaga acattttgag gcatttcagt
      781 cagttgctca atgtacctat aaccagaccg ttcagctgga tattacggcc tttttaaaga
      841 ccgtaaagaa aaataagcac aagttttatc cggcctttat tcacattctt gcccgcctga
      901 tgaatgctca tccggaattt cgtatggcaa tgaaagacgg tgagctggtg atatgggata
      961 gtgttcaccc ttgttacacc gttttccatg agcaaactga aacgttttca tcgctctgga
     1021 gtgaatacca cgacgatttc cggcagtttc tacacatata ttcgcaagat gtggcgtgtt
     1081 acggtgaaaa cctggcctat ttccctaaag ggtttattga gaatatgttt ttcgtctcag
     1141 ccaatccctg ggtgagtttc accagttttg atttaaacgt ggccaatatg gacaacttct
     1201 tcgcccccgt tttcaccatg ggcaaatatt atacgcaagg cgacaaggtg ctgatgccgc
     1261 tggcgattca ggttcatcat gccgtttgtg atggcttcca tgtcggcaga atgcttaatg
     1321 aattacaaca gtactgcgat gagtggcagg gcggggcgta atttgatatc gagctcgctt
     1381 ggactcctgt tgatagatcc agtaatgacc tcagaactcc atctggattt gttcagaacg
     1441 ctcggttgcc gccgggcgtt ttttattggt gagaatccaa gcactagtaa caacttatat
     1501 cgtatggggc tgacttcagg tgctacattt gaagagataa attgcactga aatctagaaa
     1561 tattttatct gattaataag atgatcttct tgagatcgtt ttggtctgcg cgtaatctct
     1621 tgctctgaaa acgaaaaaac cgccttgcag ggcggttttt cgaaggttct ctgagctacc
     1681 aactctttga accgaggtaa ctggcttgga ggagcgcagt caccaaaact tgtcctttca
     1741 gtttagcctt aaccggcgca tgacttcaag actaactcct ctaaatcaat taccagtggc
     1801 tgctgccagt ggtgcttttg catgtctttc cgggttggac tcaagacgat agttaccgga
     1861 taaggcgcag cggtcggact gaacgggggg ttcgtgcata cagtccagct tggagcgaac
     1921 tgcctacccg gaactgagtg tcaggcgtgg aatgagacaa acgcggccat aacagcggaa
     1981 tgacaccggt aaaccgaaag gcaggaacag gagagcgcac gagggagccg ccagggggaa
     2041 acgcctggta tctttatagt cctgtcgggt ttcgccacca ctgatttgag cgtcagattt
     2101 cgtgatgctt gtcagggggg cggagcctat ggaaaaacgg ctttgccgcg gccctctcac
     2161 ttccctgtta agtatcttcc tggcatcttc caggaaatct ccgccccgtt cgtaagccat
     2221 ttccgctcgc cgcagtcgaa cgaccgagcg tagcgagtca gtgagcgagg aagcggaata
     2281 tatccctagg cctagtagga ccaaaacgaa aaaaggcccc cctttcggga ggcctctttt
     2341 ctggaatttg gtaccgagaa gcttcaaaaa aagcaccgac tcggtgccac ttgggccctt
     2401 cttcagggcc caagttgata acggactagc cttatttgaa cttgctaggg cccttcttca
     2461 gggccctagc tctcaaacca cttatatatt catccctcta gttttattcg aaagtaacaa
     2521 accattttct tgcgtaaacc tgtacgatcc tacaggtctc tagtatataa acgcagaaag
     2581 gcccacccga aggtgagcca gtgtgattga cagctagctc agtcctaggg attgtgctag
     2641 ctactagaga ttaaagagga gaaatactag atgaaaaaca taaatgccga cgacacatac
     2701 agaataatta ataaaattaa agcttgtaga agcaataatg atattaatca atgcttatct
     2761 gatatgacta aaatggtaca ttgtgaatat tatttactcg cgatcattta tcctcattct
     2821 atggttaaat ctgatatttc aatcctagat aattacccta aaaaatggag gcaatattat
     2881 gatgacgcta atttaataaa atatgatcct atagtagatt attctaactc caatcattca
     2941 ccaattaatt ggaatatatt tgaaaacaat gctgtaaata aaaaatctcc aaatgtaatt
     3001 aaag
//
