LOCUS       sequence-258316-        3007 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Engineered sgRNA-LEA5 (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: pLY107
FEATURES             Location/Qualifiers
     source          1..3007
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     RBS             188..199
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     promoter        complement(211..245)
                     /label=BBa_J23117
                     /note="Anderson constitutive promoter 
                     (https://parts.igem.org/Promoters/Catalog/Anderson). 
                     Measured strength relative to BBa_J223100: 0.06"
     terminator      253..280
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     protein_bind    389..407
                     /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    444..462
                     /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)"
     primer_bind     complement(586..603)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(720..1265)
                     /direction=LEFT
                     /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     complement(800..819)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     terminator      complement(1379..1473)
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             complement(1494..2153)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     2068..2087
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     promoter        complement(2154..2256)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
ORIGIN
        1 tatctaggat tgaaatatca gatttaacca tagaatgagg ataaatgatc gcgagtaaat
       61 aatattcaca atgtaccatt ttagtcatat cagataagca ttgattaata tcattattgc
      121 ttctacaagc tttaatttta ttaattattc tgtatgtgtc gtcggcattt atgtttttca
      181 tctagtattt ctcctcttta atctctagta gctagcacaa tccctaggac tgagctagct
      241 gtcaatcaca ctggctcacc ttcgggtggg cctttctgcg tttatatact agagacctgt
      301 aggatcgtac aggtttacgc aagaaaatgg tttgttactt tcgaataaaa ctagagagtt
      361 atatgttggt aactacgttt gagagctagg gccctgaaga agggccctag caagttcaaa
      421 taaggctagt ccgttatcaa cttgggccct gaagaagggc ccaagtggca ccgagtcggt
      481 gctttttttg aagcttctcg gtaccaaatt ccagaaaaga ggcctcccga aaggggggcc
      541 ttttttcgtt ttggtcctac taggcctagg gatatattcc gcttcctcgc tcactgactc
      601 gctacgctcg gtcgttcgac tgcggcgagc ggaaatggct tacgaacggg gcggagattt
      661 cctggaagat gccaggaaga tacttaacag ggaagtgaga gggccgcggc aaagccgttt
      721 ttccataggc tccgcccccc tgacaagcat cacgaaatct gacgctcaaa tcagtggtgg
      781 cgaaacccga caggactata aagataccag gcgtttcccc ctggcggctc cctcgtgcgc
      841 tctcctgttc ctgcctttcg gtttaccggt gtcattccgc tgttatggcc gcgtttgtct
      901 cattccacgc ctgacactca gttccgggta ggcagttcgc tccaagctgg actgtatgca
      961 cgaacccccc gttcagtccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa
     1021 cccggaaaga catgcaaaag caccactggc agcagccact ggtaattgat ttagaggagt
     1081 tagtcttgaa gtcatgcgcc ggttaaggct aaactgaaag gacaagtttt ggtgactgcg
     1141 ctcctccaag ccagttacct cggttcaaag agttggtagc tcagagaacc ttcgaaaaac
     1201 cgccctgcaa ggcggttttt tcgttttcag agcaagagat tacgcgcaga ccaaaacgat
     1261 ctcaagaaga tcatcttatt aatcagataa aatatttcta gatttcagtg caatttatct
     1321 cttcaaatgt agcacctgaa gtcagcccca tacgatataa gttgttacta gtgcttggat
     1381 tctcaccaat aaaaaacgcc cggcggcaac cgagcgttct gaacaaatcc agatggagtt
     1441 ctgaggtcat tactggatct atcaacagga gtccaagcga gctcgatatc aaattacgcc
     1501 ccgccctgcc actcatcgca gtactgttgt aattcattaa gcattctgcc gacatggaag
     1561 ccatcacaaa cggcatgatg aacctgaatc gccagcggca tcagcacctt gtcgccttgc
     1621 gtataatatt tgcccatggt gaaaacgggg gcgaagaagt tgtccatatt ggccacgttt
     1681 aaatcaaaac tggtgaaact cacccaggga ttggctgaga cgaaaaacat attctcaata
     1741 aaccctttag ggaaataggc caggttttca ccgtaacacg ccacatcttg cgaatatatg
     1801 tgtagaaact gccggaaatc gtcgtggtat tcactccaga gcgatgaaaa cgtttcagtt
     1861 tgctcatgga aaacggtgta acaagggtga acactatccc atatcaccag ctcaccgtct
     1921 ttcattgcca tacgaaattc cggatgagca ttcatcaggc gggcaagaat gtgaataaag
     1981 gccggataaa acttgtgctt atttttcttt acggtcttta aaaaggccgt aatatccagc
     2041 tgaacggtct ggttataggt acattgagca actgactgaa atgcctcaaa atgttcttta
     2101 cgatgccatt gggatatatc aacggtggta tatccagtga tttttttctc cattttagct
     2161 tccttagctc ctgaaaatct cgataactca aaaaatacgc ccggtagtga tcttatttca
     2221 ttatggtgaa agttggaacc tcttacgtgc cgatcaacgt ctcattttcg ccagatatct
     2281 aattcgcggc cgcttctaga gcctagttgt cttcatgcat gaagacaaaa ttaatactag
     2341 agggaccaaa acgaaaaaag acgcttttca gcgtcttatt gttcgtcttt ggtaccgagc
     2401 tctagtagtg atctacacta gcactatcag tgttattaat ttttaaagta tgggcaatca
     2461 attgctcctg ttaaaattgc tttagaaata ctttggcagc ggtttgttgt attgagtttc
     2521 atttgcgcat tggttaaatg gaaagtgaca gtacgctcac tgcaacctaa tatttttgaa
     2581 atatcccaag agctttttcc ttcgcatgcc cacgctaaac attctttttc tcttttggtt
     2641 aaatcgttgt ttgatttatt atttgctata tttatttttc gataattatc aactagagaa
     2701 ggaacaatta atggtatgtt catacacgca tgtaaaaata aactatctat atagttgtct
     2761 ttttctgaat gtgcaaaact aagcattccg aagccattgt tagccgtatg aatagggaaa
     2821 ctaaacccag tgataagacc tgatgttttc gcttctttaa ttacatttgg agatttttta
     2881 tttacagcat tgttttcaaa tatattccaa ttaattggtg aatgattgga gttagaataa
     2941 tctactatag gatcatattt tattaaatta gcgtcatcat aatattgcct ccatttttta
     3001 gggtaat
//
