LOCUS       sequence-376130-        3044 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  sgRNA with 3 RNA aptemers BoxB at the 3'-end for CRISPRa.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3044)
  AUTHORS   Liu Y, Pinto F, Wan X, Yang Z, Peng S, Li M, Cooper JM, Xie Z, 
            French CE, Wang B
  TITLE     Reprogrammed tracrRNAs enable repurposing of RNAs as crRNAs and 
            sequence-specific RNA biosensors.
  JOURNAL   Nat Commun. 2022 Apr 11;13(1):1937. doi: 10.1038/s41467-022-29604-x.
  PUBMED    35410423
REFERENCE   2  (bases 1 to 3044)
  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: "2022-04-11"; volume: "13(1):1937. doi"; pages: " 
            10.1038/s41467-022-29604-x"
COMMENT     Sequence Label: pLY171
FEATURES             Location/Qualifiers
     source          1..3044
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    102..120
                     /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    124..142
                     /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    150..168
                     /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)"
     misc_RNA        complement(169..244)
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     terminator      complement(340..367)
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     promoter        375..409
                     /label=BBa_J23117
                     /note="Anderson constitutive promoter 
                     (https://parts.igem.org/Promoters/Catalog/Anderson). 
                     Measured strength relative to BBa_J223100: 0.06"
     RBS             421..432
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     promoter        1371..1473
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     CDS             1474..2133
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(1540..1559)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     terminator      2154..2248
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     rep_origin      2362..2907
                     /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     2808..2827
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3024..3041
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 gcggaatata tccctaggcc tagtaggacc aaaacgaaaa aaggcccccc tttcgggagg
       61 cctcttttct ggaatttggt accgagctct agtaaaaaaa agggcccttc ttcagggccc
      121 tgcgggccct tcttcagggc cctagccttg ggcccttctt cagggcccgc accgactcgg
      181 tgccactttt tcaagttgat aacggactag ccttatttga acttgctatt tctagctctc
      241 aaacgaccga actatataat aacttctagt tttattcgaa agtaacaaac cattttcttg
      301 cgtaaacctg tacgatccta caggtctcta gtatataaac gcagaaaggc ccacccgaag
      361 gtgagccagt gtgattgaca gctagctcag tcctagggat tgtgctagct actagagatt
      421 aaagaggaga aatactagat gaaaaacata aatgccgacg acacatacag aataattaat
      481 aaaattaaag cttgtagaag caataatgat attaatcaat gcttatctga tatgactaaa
      541 atggtacatt gtgaatatta tttactcgcg atcatttatc ctcattctat ggttaaatct
      601 gatatttcaa tcctagataa ttaccctaaa aaatggaggc aatattatga tgacgctaat
      661 ttaataaaat atgatcctat agtagattat tctaactcca atcattcacc aattaattgg
      721 aatatatttg aaaacaatgc tgtaaataaa aaatctccaa atgtaattaa agaagcgaaa
      781 acatcaggtc ttatcactgg gtttagtttc cctattcata cggctaacaa tggcttcgga
      841 atgcttagtt ttgcacattc agaaaaagac aactatatag atagtttatt tttacatgcg
      901 tgtatgaaca taccattaat tgttccttct ctagttgata attatcgaaa aataaatata
      961 gcaaataata aatcaaacaa cgatttaacc aaaagagaaa aagaatgttt agcgtgggca
     1021 tgcgaaggaa aaagctcttg ggatatttca aaaatattag gttgcagtga gcgtactgtc
     1081 actttccatt taaccaatgc gcaaatgaaa ctcaatacaa caaaccgctg ccaaagtatt
     1141 tctaaagcaa ttttaacagg agcaattgat tgcccatact ttaaaaatta ataacactga
     1201 tagtgctagt gtagatcact actagagctc ggtaccaaag acgaacaata agacgctgaa
     1261 aagcgtcttt tttcgttttg gtccctctag tattaatttt gtcttcatgc atgaagacaa
     1321 ctaggctcta gaagcggccg cgaattagat atctggcgaa aatgagacgt tgatcggcac
     1381 gtaagaggtt ccaactttca ccataatgaa ataagatcac taccgggcgt attttttgag
     1441 ttatcgagat tttcaggagc taaggaagct aaaatggaga aaaaaatcac tggatatacc
     1501 accgttgata tatcccaatg gcatcgtaaa gaacattttg aggcatttca gtcagttgct
     1561 caatgtacct ataaccagac cgttcagctg gatattacgg cctttttaaa gaccgtaaag
     1621 aaaaataagc acaagtttta tccggccttt attcacattc ttgcccgcct gatgaatgct
     1681 catccggaat ttcgtatggc aatgaaagac ggtgagctgg tgatatggga tagtgttcac
     1741 ccttgttaca ccgttttcca tgagcaaact gaaacgtttt catcgctctg gagtgaatac
     1801 cacgacgatt tccggcagtt tctacacata tattcgcaag atgtggcgtg ttacggtgaa
     1861 aacctggcct atttccctaa agggtttatt gagaatatgt ttttcgtctc agccaatccc
     1921 tgggtgagtt tcaccagttt tgatttaaac gtggccaata tggacaactt cttcgccccc
     1981 gttttcacca tgggcaaata ttatacgcaa ggcgacaagg tgctgatgcc gctggcgatt
     2041 caggttcatc atgccgtttg tgatggcttc catgtcggca gaatgcttaa tgaattacaa
     2101 cagtactgcg atgagtggca gggcggggcg taatttgata tcgagctcgc ttggactcct
     2161 gttgatagat ccagtaatga cctcagaact ccatctggat ttgttcagaa cgctcggttg
     2221 ccgccgggcg ttttttattg gtgagaatcc aagcactagt aacaacttat atcgtatggg
     2281 gctgacttca ggtgctacat ttgaagagat aaattgcact gaaatctaga aatattttat
     2341 ctgattaata agatgatctt cttgagatcg ttttggtctg cgcgtaatct cttgctctga
     2401 aaacgaaaaa accgccttgc agggcggttt ttcgaaggtt ctctgagcta ccaactcttt
     2461 gaaccgaggt aactggcttg gaggagcgca gtcaccaaaa cttgtccttt cagtttagcc
     2521 ttaaccggcg catgacttca agactaactc ctctaaatca attaccagtg gctgctgcca
     2581 gtggtgcttt tgcatgtctt tccgggttgg actcaagacg atagttaccg gataaggcgc
     2641 agcggtcgga ctgaacgggg ggttcgtgca tacagtccag cttggagcga actgcctacc
     2701 cggaactgag tgtcaggcgt ggaatgagac aaacgcggcc ataacagcgg aatgacaccg
     2761 gtaaaccgaa aggcaggaac aggagagcgc acgagggagc cgccaggggg aaacgcctgg
     2821 tatctttata gtcctgtcgg gtttcgccac cactgatttg agcgtcagat ttcgtgatgc
     2881 ttgtcagggg ggcggagcct atggaaaaac ggctttgccg cggccctctc acttccctgt
     2941 taagtatctt cctggcatct tccaggaaat ctccgccccg ttcgtaagcc atttccgctc
     3001 gccgcagtcg aacgaccgag cgtagcgagt cagtgagcga ggaa
//
