LOCUS       sequence-258927-        3004 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Engineered sgRNA-LEB4 (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: pLY111
FEATURES             Location/Qualifiers
     source          1..3004
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        778..880
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     CDS             881..1540
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(947..966)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     terminator      1561..1655
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     rep_origin      1769..2314
                     /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     2215..2234
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2431..2448
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    2572..2590
                     /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    2627..2645
                     /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(2751..2778)
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     promoter        2786..2820
                     /label=BBa_J23117
                     /note="Anderson constitutive promoter 
                     (https://parts.igem.org/Promoters/Catalog/Anderson). 
                     Measured strength relative to BBa_J223100: 0.06"
     RBS             2832..2843
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
ORIGIN
        1 taaatctgat atttcaatcc tagataatta ccctaaaaaa tggaggcaat attatgatga
       61 cgctaattta ataaaatatg atcctatagt agattattct aactccaatc attcaccaat
      121 taattggaat atatttgaaa acaatgctgt aaataaaaaa tctccaaatg taattaaaga
      181 agcgaaaaca tcaggtctta tcactgggtt tagtttccct attcatacgg ctaacaatgg
      241 cttcggaatg cttagttttg cacattcaga aaaagacaac tatatagata gtttattttt
      301 acatgcgtgt atgaacatac cattaattgt tccttctcta gttgataatt atcgaaaaat
      361 aaatatagca aataataaat caaacaacga tttaaccaaa agagaaaaag aatgtttagc
      421 gtgggcatgc gaaggaaaaa gctcttggga tatttcaaaa atattaggtt gcagtgagcg
      481 tactgtcact ttccatttaa ccaatgcgca aatgaaactc aatacaacaa accgctgcca
      541 aagtatttct aaagcaattt taacaggagc aattgattgc ccatacttta aaaattaata
      601 acactgatag tgctagtgta gatcactact agagctcggt accaaagacg aacaataaga
      661 cgctgaaaag cgtctttttt cgttttggtc cctctagtat taattttgtc ttcatgcatg
      721 aagacaacta ggctctagaa gcggccgcga attagatatc tggcgaaaat gagacgttga
      781 tcggcacgta agaggttcca actttcacca taatgaaata agatcactac cgggcgtatt
      841 ttttgagtta tcgagatttt caggagctaa ggaagctaaa atggagaaaa aaatcactgg
      901 atataccacc gttgatatat cccaatggca tcgtaaagaa cattttgagg catttcagtc
      961 agttgctcaa tgtacctata accagaccgt tcagctggat attacggcct ttttaaagac
     1021 cgtaaagaaa aataagcaca agttttatcc ggcctttatt cacattcttg cccgcctgat
     1081 gaatgctcat ccggaatttc gtatggcaat gaaagacggt gagctggtga tatgggatag
     1141 tgttcaccct tgttacaccg ttttccatga gcaaactgaa acgttttcat cgctctggag
     1201 tgaataccac gacgatttcc ggcagtttct acacatatat tcgcaagatg tggcgtgtta
     1261 cggtgaaaac ctggcctatt tccctaaagg gtttattgag aatatgtttt tcgtctcagc
     1321 caatccctgg gtgagtttca ccagttttga tttaaacgtg gccaatatgg acaacttctt
     1381 cgcccccgtt ttcaccatgg gcaaatatta tacgcaaggc gacaaggtgc tgatgccgct
     1441 ggcgattcag gttcatcatg ccgtttgtga tggcttccat gtcggcagaa tgcttaatga
     1501 attacaacag tactgcgatg agtggcaggg cggggcgtaa tttgatatcg agctcgcttg
     1561 gactcctgtt gatagatcca gtaatgacct cagaactcca tctggatttg ttcagaacgc
     1621 tcggttgccg ccgggcgttt tttattggtg agaatccaag cactagtaac aacttatatc
     1681 gtatggggct gacttcaggt gctacatttg aagagataaa ttgcactgaa atctagaaat
     1741 attttatctg attaataaga tgatcttctt gagatcgttt tggtctgcgc gtaatctctt
     1801 gctctgaaaa cgaaaaaacc gccttgcagg gcggtttttc gaaggttctc tgagctacca
     1861 actctttgaa ccgaggtaac tggcttggag gagcgcagtc accaaaactt gtcctttcag
     1921 tttagcctta accggcgcat gacttcaaga ctaactcctc taaatcaatt accagtggct
     1981 gctgccagtg gtgcttttgc atgtctttcc gggttggact caagacgata gttaccggat
     2041 aaggcgcagc ggtcggactg aacggggggt tcgtgcatac agtccagctt ggagcgaact
     2101 gcctacccgg aactgagtgt caggcgtgga atgagacaaa cgcggccata acagcggaat
     2161 gacaccggta aaccgaaagg caggaacagg agagcgcacg agggagccgc cagggggaaa
     2221 cgcctggtat ctttatagtc ctgtcgggtt tcgccaccac tgatttgagc gtcagatttc
     2281 gtgatgcttg tcaggggggc ggagcctatg gaaaaacggc tttgccgcgg ccctctcact
     2341 tccctgttaa gtatcttcct ggcatcttcc aggaaatctc cgccccgttc gtaagccatt
     2401 tccgctcgcc gcagtcgaac gaccgagcgt agcgagtcag tgagcgagga agcggaatat
     2461 atccctaggc ctagtaggac caaaacgaaa aaaggccccc ctttcgggag gcctcttttc
     2521 tggaatttgg taccgagaag cttcaaaaaa agcaccgact cggtgccact tgggcccttc
     2581 ttcagggccc aagttgataa cggactagcc ttatttgaac ttgctagggc ccttcttcag
     2641 ggccctagct ctcaaactta tttccgaacg atcactctag ttttattcga aagtaacaaa
     2701 ccattttctt gcgtaaacct gtacgatcct acaggtctct agtatataaa cgcagaaagg
     2761 cccacccgaa ggtgagccag tgtgattgac agctagctca gtcctaggga ttgtgctagc
     2821 tactagagat taaagaggag aaatactaga tgaaaaacat aaatgccgac gacacataca
     2881 gaataattaa taaaattaaa gcttgtagaa gcaataatga tattaatcaa tgcttatctg
     2941 atatgactaa aatggtacat tgtgaatatt atttactcgc gatcatttat cctcattcta
     3001 tggt
//
