LOCUS       sequence-258928-        3006 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Engineered sgRNA-G6 (2 BoxB aptamers) generator circuit including 
            promoter Plux2.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3006)
  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 3006)
  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: pLY74
FEATURES             Location/Qualifiers
     source          1..3006
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    116..134
                     /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    171..189
                     /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(297..324)
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     promoter        332..366
                     /label=BBa_J23117
                     /note="Anderson constitutive promoter 
                     (https://parts.igem.org/Promoters/Catalog/Anderson). 
                     Measured strength relative to BBa_J223100: 0.06"
     RBS             378..389
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     promoter        1328..1430
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     CDS             1431..2090
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(1497..1516)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     terminator      2111..2205
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     rep_origin      2319..2864
                     /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     2765..2784
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2981..2998
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 atatatccct aggcctagta ggaccaaaac gaaaaaaggc ccccctttcg ggaggcctct
       61 tttctggaat ttggtaccga gaagcttcaa aaaaagcacc gactcggtgc cacttgggcc
      121 cttcttcagg gcccaagttg ataacggact agccttattt gaacttgcta gggcccttct
      181 tcagggccct agctctcaaa cttttttgcc acaacagctt ttctagtttt attcgaaagt
      241 aacaaaccat tttcttgcgt aaacctgtac gatcctacag gtctctagta tataaacgca
      301 gaaaggccca cccgaaggtg agccagtgtg attgacagct agctcagtcc tagggattgt
      361 gctagctact agagattaaa gaggagaaat actagatgaa aaacataaat gccgacgaca
      421 catacagaat aattaataaa attaaagctt gtagaagcaa taatgatatt aatcaatgct
      481 tatctgatat gactaaaatg gtacattgtg aatattattt actcgcgatc atttatcctc
      541 attctatggt taaatctgat atttcaatcc tagataatta ccctaaaaaa tggaggcaat
      601 attatgatga cgctaattta ataaaatatg atcctatagt agattattct aactccaatc
      661 attcaccaat taattggaat atatttgaaa acaatgctgt aaataaaaaa tctccaaatg
      721 taattaaaga agcgaaaaca tcaggtctta tcactgggtt tagtttccct attcatacgg
      781 ctaacaatgg cttcggaatg cttagttttg cacattcaga aaaagacaac tatatagata
      841 gtttattttt acatgcgtgt atgaacatac cattaattgt tccttctcta gttgataatt
      901 atcgaaaaat aaatatagca aataataaat caaacaacga tttaaccaaa agagaaaaag
      961 aatgtttagc gtgggcatgc gaaggaaaaa gctcttggga tatttcaaaa atattaggtt
     1021 gcagtgagcg tactgtcact ttccatttaa ccaatgcgca aatgaaactc aatacaacaa
     1081 accgctgcca aagtatttct aaagcaattt taacaggagc aattgattgc ccatacttta
     1141 aaaattaata acactgatag tgctagtgta gatcactact agagctcggt accaaagacg
     1201 aacaataaga cgctgaaaag cgtctttttt cgttttggtc cctctagtat taattttgtc
     1261 ttcatgcatg aagacaacta ggctctagaa gcggccgcga attagatatc tggcgaaaat
     1321 gagacgttga tcggcacgta agaggttcca actttcacca taatgaaata agatcactac
     1381 cgggcgtatt ttttgagtta tcgagatttt caggagctaa ggaagctaaa atggagaaaa
     1441 aaatcactgg atataccacc gttgatatat cccaatggca tcgtaaagaa cattttgagg
     1501 catttcagtc agttgctcaa tgtacctata accagaccgt tcagctggat attacggcct
     1561 ttttaaagac cgtaaagaaa aataagcaca agttttatcc ggcctttatt cacattcttg
     1621 cccgcctgat gaatgctcat ccggaatttc gtatggcaat gaaagacggt gagctggtga
     1681 tatgggatag tgttcaccct tgttacaccg ttttccatga gcaaactgaa acgttttcat
     1741 cgctctggag tgaataccac gacgatttcc ggcagtttct acacatatat tcgcaagatg
     1801 tggcgtgtta cggtgaaaac ctggcctatt tccctaaagg gtttattgag aatatgtttt
     1861 tcgtctcagc caatccctgg gtgagtttca ccagttttga tttaaacgtg gccaatatgg
     1921 acaacttctt cgcccccgtt ttcaccatgg gcaaatatta tacgcaaggc gacaaggtgc
     1981 tgatgccgct ggcgattcag gttcatcatg ccgtttgtga tggcttccat gtcggcagaa
     2041 tgcttaatga attacaacag tactgcgatg agtggcaggg cggggcgtaa tttgatatcg
     2101 agctcgcttg gactcctgtt gatagatcca gtaatgacct cagaactcca tctggatttg
     2161 ttcagaacgc tcggttgccg ccgggcgttt tttattggtg agaatccaag cactagtaac
     2221 aacttatatc gtatggggct gacttcaggt gctacatttg aagagataaa ttgcactgaa
     2281 atctagaaat attttatctg attaataaga tgatcttctt gagatcgttt tggtctgcgc
     2341 gtaatctctt gctctgaaaa cgaaaaaacc gccttgcagg gcggtttttc gaaggttctc
     2401 tgagctacca actctttgaa ccgaggtaac tggcttggag gagcgcagtc accaaaactt
     2461 gtcctttcag tttagcctta accggcgcat gacttcaaga ctaactcctc taaatcaatt
     2521 accagtggct gctgccagtg gtgcttttgc atgtctttcc gggttggact caagacgata
     2581 gttaccggat aaggcgcagc ggtcggactg aacggggggt tcgtgcatac agtccagctt
     2641 ggagcgaact gcctacccgg aactgagtgt caggcgtgga atgagacaaa cgcggccata
     2701 acagcggaat gacaccggta aaccgaaagg caggaacagg agagcgcacg agggagccgc
     2761 cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt tcgccaccac tgatttgagc
     2821 gtcagatttc gtgatgcttg tcaggggggc ggagcctatg gaaaaacggc tttgccgcgg
     2881 ccctctcact tccctgttaa gtatcttcct ggcatcttcc aggaaatctc cgccccgttc
     2941 gtaagccatt tccgctcgcc gcagtcgaac gaccgagcgt agcgagtcag tgagcgagga
     3001 agcgga
//
