LOCUS       sequence-258923-        3007 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Engineered sgRNA-LEA1 (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: pLY103
FEATURES             Location/Qualifiers
     source          1..3007
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    20..38
                     /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(147..174)
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     promoter        182..216
                     /label=BBa_J23117
                     /note="Anderson constitutive promoter 
                     (https://parts.igem.org/Promoters/Catalog/Anderson). 
                     Measured strength relative to BBa_J223100: 0.06"
     RBS             228..239
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     promoter        1178..1280
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     CDS             1281..1940
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(1347..1366)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     terminator      1961..2055
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     rep_origin      2169..2714
                     /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     2615..2634
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2831..2848
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    2972..2990
                     /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 gccttatttg aacttgctag ggcccttctt cagggcccta gctctcaaac gacggcgcgt
       61 cgggataact ctctagtttt attcgaaagt aacaaaccat tttcttgcgt aaacctgtac
      121 gatcctacag gtctctagta tataaacgca gaaaggccca cccgaaggtg agccagtgtg
      181 attgacagct agctcagtcc tagggattgt gctagctact agagattaaa gaggagaaat
      241 actagatgaa aaacataaat gccgacgaca catacagaat aattaataaa attaaagctt
      301 gtagaagcaa taatgatatt aatcaatgct tatctgatat gactaaaatg gtacattgtg
      361 aatattattt actcgcgatc atttatcctc attctatggt taaatctgat atttcaatcc
      421 tagataatta ccctaaaaaa tggaggcaat attatgatga cgctaattta ataaaatatg
      481 atcctatagt agattattct aactccaatc attcaccaat taattggaat atatttgaaa
      541 acaatgctgt aaataaaaaa tctccaaatg taattaaaga agcgaaaaca tcaggtctta
      601 tcactgggtt tagtttccct attcatacgg ctaacaatgg cttcggaatg cttagttttg
      661 cacattcaga aaaagacaac tatatagata gtttattttt acatgcgtgt atgaacatac
      721 cattaattgt tccttctcta gttgataatt atcgaaaaat aaatatagca aataataaat
      781 caaacaacga tttaaccaaa agagaaaaag aatgtttagc gtgggcatgc gaaggaaaaa
      841 gctcttggga tatttcaaaa atattaggtt gcagtgagcg tactgtcact ttccatttaa
      901 ccaatgcgca aatgaaactc aatacaacaa accgctgcca aagtatttct aaagcaattt
      961 taacaggagc aattgattgc ccatacttta aaaattaata acactgatag tgctagtgta
     1021 gatcactact agagctcggt accaaagacg aacaataaga cgctgaaaag cgtctttttt
     1081 cgttttggtc cctctagtat taattttgtc ttcatgcatg aagacaacta ggctctagaa
     1141 gcggccgcga attagatatc tggcgaaaat gagacgttga tcggcacgta agaggttcca
     1201 actttcacca taatgaaata agatcactac cgggcgtatt ttttgagtta tcgagatttt
     1261 caggagctaa ggaagctaaa atggagaaaa aaatcactgg atataccacc gttgatatat
     1321 cccaatggca tcgtaaagaa cattttgagg catttcagtc agttgctcaa tgtacctata
     1381 accagaccgt tcagctggat attacggcct ttttaaagac cgtaaagaaa aataagcaca
     1441 agttttatcc ggcctttatt cacattcttg cccgcctgat gaatgctcat ccggaatttc
     1501 gtatggcaat gaaagacggt gagctggtga tatgggatag tgttcaccct tgttacaccg
     1561 ttttccatga gcaaactgaa acgttttcat cgctctggag tgaataccac gacgatttcc
     1621 ggcagtttct acacatatat tcgcaagatg tggcgtgtta cggtgaaaac ctggcctatt
     1681 tccctaaagg gtttattgag aatatgtttt tcgtctcagc caatccctgg gtgagtttca
     1741 ccagttttga tttaaacgtg gccaatatgg acaacttctt cgcccccgtt ttcaccatgg
     1801 gcaaatatta tacgcaaggc gacaaggtgc tgatgccgct ggcgattcag gttcatcatg
     1861 ccgtttgtga tggcttccat gtcggcagaa tgcttaatga attacaacag tactgcgatg
     1921 agtggcaggg cggggcgtaa tttgatatcg agctcgcttg gactcctgtt gatagatcca
     1981 gtaatgacct cagaactcca tctggatttg ttcagaacgc tcggttgccg ccgggcgttt
     2041 tttattggtg agaatccaag cactagtaac aacttatatc gtatggggct gacttcaggt
     2101 gctacatttg aagagataaa ttgcactgaa atctagaaat attttatctg attaataaga
     2161 tgatcttctt gagatcgttt tggtctgcgc gtaatctctt gctctgaaaa cgaaaaaacc
     2221 gccttgcagg gcggtttttc gaaggttctc tgagctacca actctttgaa ccgaggtaac
     2281 tggcttggag gagcgcagtc accaaaactt gtcctttcag tttagcctta accggcgcat
     2341 gacttcaaga ctaactcctc taaatcaatt accagtggct gctgccagtg gtgcttttgc
     2401 atgtctttcc gggttggact caagacgata gttaccggat aaggcgcagc ggtcggactg
     2461 aacggggggt tcgtgcatac agtccagctt ggagcgaact gcctacccgg aactgagtgt
     2521 caggcgtgga atgagacaaa cgcggccata acagcggaat gacaccggta aaccgaaagg
     2581 caggaacagg agagcgcacg agggagccgc cagggggaaa cgcctggtat ctttatagtc
     2641 ctgtcgggtt tcgccaccac tgatttgagc gtcagatttc gtgatgcttg tcaggggggc
     2701 ggagcctatg gaaaaacggc tttgccgcgg ccctctcact tccctgttaa gtatcttcct
     2761 ggcatcttcc aggaaatctc cgccccgttc gtaagccatt tccgctcgcc gcagtcgaac
     2821 gaccgagcgt agcgagtcag tgagcgagga agcggaatat atccctaggc ctagtaggac
     2881 caaaacgaaa aaaggccccc ctttcgggag gcctcttttc tggaatttgg taccgagaag
     2941 cttcaaaaaa agcaccgact cggtgccact tgggcccttc ttcagggccc aagttgataa
     3001 cggacta
//
