LOCUS       sequence-377034-        5264 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  short tracrRNA generator with WT anti-repeat region.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5264)
  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 5264)
  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: pLY232
FEATURES             Location/Qualifiers
     source          1..5264
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     1..23
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     misc_feature    48..69
                     /label=BioBrick prefix
                     /note="BioBrick prefix for parts that do not start with 
                     ""ATG"""
     CDS             complement(176..1054)
                     /codon_start=1
                     /gene="araC"
                     /product="L-arabinose regulatory protein"
                     /label=araC
                     /translation="MAEAQNDPLLPGYSFNAHLVAGLTPIEANGYLDFFIDRPLGMKGY
                     ILNLTIRGQGVVKNQGREFVCRPGDILLFPPGEIHHYGRHPEAREWYHQWVYFRPRAYW
                     HEWLNWPSIFANTGFFRPDEAHQPHFSDLFGQIINAGQGEGRYSELLAINLLEQLLLRR
                     MEAINESLHPPMDNRVREACQYISDHLADSNFDIASVAQHVCLSPSRLSHLFRQQLGIS
                     VLSWREDQRISQAKLLLSTTRMPIATVGRNVGFDDQLYFSRVFKKCTGASPSEFRAGCE
                     EKVNDVAVKLS"
     promoter        1081..1365
                     /gene="araBAD"
                     /label=araBAD promoter
                     /note="promoter of the L-arabinose operon of E. coli; the 
                     araC regulatory gene is transcribed in the opposite 
                     direction (Guzman et al., 1995)"
     primer_bind     1288..1307
                     /label=pBAD Forward
                     /note="For vectors with E. coli araBAD promoter, forward 
                     primer"
     protein_bind    1434..1452
                     /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    1460..1478
                     /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    1482..1500
                     /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      1588..1682
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     primer_bind     complement(1803..1822)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             1814..2629
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     oriT            2796..2904
                     /note="incP origin of transfer"
     CDS             complement(2965..4113)
                     /codon_start=1
                     /product="trans-acting replication protein that binds to 
                     and activates oriV"
                     /label=trfA
                     /translation="MNRTFDRKAYRQELIDAGFSAEDAETIASRTVMRAPRETFQSVGS
                     IVQQATAKIERDSVQLAPPALPAPSAAVERSRRLEQEAAGLAKSMTIDTRGTMTTKKRK
                     TAGEDLAKQVSEAKQAALLKHTKQQIKEMQLSLFDIAPWPDTMRAMPNDTARSALFTTR
                     NKKIPREALQNKVIFHVNKDVKITYTGVELRADDDELVWQQVLEYAKRTPIGEPITFTF
                     YELCQDLGWSINGRYYTKAEECLSRLQATAMGFTSDRVGHLESVSLLHRFRVLDRGKKT
                     SRCQVLIDEEIVVLFAGDHYTKFIWEKYRKLSPTARRMFDYFSSHREPYPLKLETFRLM
                     CGSDSTRVKKWREQVGEACEELRGSGLVEHAWVNDDLVHCKR"
     rep_origin      4506..5037
                     /label=oriV
                     /note="incP origin of replication"
     terminator      complement(5164..5250)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
ORIGIN
        1 agcggataac aatttcacac aggaggccgc ctaggccgcg gccgcgcgaa ttcgcggccg
       61 cttctagagc ctagttgtct tcatgcatga agacaaaatt aatactagag ggaccaaaac
      121 gaaaaaagac gctcgaaagc gtctcttttc tggaatttgg taccgagctc tagtattatg
      181 acaacttgac ggctacatca ttcacttttt cttcacaacc ggcacggaac tcgctcgggc
      241 tggccccggt gcatttttta aatacccgcg agaaatagag ttgatcgtca aaaccaacat
      301 tgcgaccgac ggtggcgata ggcatccggg tggtgctcaa aagcagcttc gcctggctga
      361 tacgttggtc ctcgcgccag cttaagacgc taatccctaa ctgctggcgg aaaagatgtg
      421 acagacgcga cggcgacaag caaacatgct gtgcgacgct ggcgatatca aaattgctgt
      481 ctgccaggtg atcgctgatg tactgacaag cctcgcgtac ccgattatcc atcggtggat
      541 ggagcgactc gttaatcgct tccatgcgcc gcagtaacaa ttgctcaagc agatttatcg
      601 ccagcagctc cgaatagcgc ccttcccctt gcccggcgtt aatgatttgc ccaaacaggt
      661 cgctgaaatg cggctggtgc gcttcatccg ggcgaaagaa ccccgtattg gcaaatattg
      721 acggccagtt aagccattca tgccagtagg cgcgcggacg aaagtaaacc cactggtgat
      781 accattcgcg agcctccgga tgacgaccgt agtgatgaat ctctcctggc gggaacagca
      841 aaatatcacc cggtcggcaa acaaattctc gtccctgatt tttcaccacc ccctgaccgc
      901 gaatggtgag attgagaata taacctttca ttcccagcgg tcggtcgata aaaaaatcga
      961 gataaccgtt ggcctcaatc ggcgttaaac ccgccaccag atgggcatta aacgagtatc
     1021 ccggcagcag gggatcattt tgcgcttcag ccatactttt catactcccg ccattcagag
     1081 aagaaaccaa ttgtccatat tgcatcagac attgccgtca ctgcgtcttt tactggctct
     1141 tctcgctaac caaaccggta accccgctta ttaaaagcat tctgtaacaa agcgggacca
     1201 aagccatgac aaaaacgcgt aacaaaagtg tctataatca cggcagaaaa gtccacattg
     1261 attatttgca cggcgtcaca ctttgctatg ccatagcatt tttatccata agattagcgg
     1321 atcctacctg acgcttttta tcgcaactct ctactgtttc tccatactag gcatagcaag
     1381 ttaaaataag gctagtccgt tatcaacttg aaaaagtggc accgagtcgg tgcgggccct
     1441 gaagaagggc ccaaggctag ggccctgaag aagggcccgc agggccctga agaagggccc
     1501 ttttttttac tagagctcgg taccaaattc cagaaaagag gcctcccgaa aggggggcct
     1561 tttttcgttt tggtcctact agtcttggac tcctgttgat agatccagta atgacctcag
     1621 aactccatct ggatttgttc agaacgctcg gttgccgccg ggcgtttttt attggtgaga
     1681 atccaggggt ccccaataat tacgatttaa atttgtgtct caaaatctct gatgttacat
     1741 tgcacaagat aaaaatatat catcatgaac aataaaactg tctgcttaca taaacagtaa
     1801 tacaaggggt gttatgagcc atattcagcg tgaaacgagc tgtagccgtc cgcgtctgaa
     1861 cagcaacatg gatgcggatc tgtatggcta taaatgggcg cgtgataacg tgggtcagag
     1921 cggcgcgacc atttatcgtc tgtatggcaa accggatgcg ccggaactgt ttctgaaaca
     1981 tggcaaaggc agcgtggcga acgatgtgac cgatgaaatg gtgcgtctga actggctgac
     2041 cgaatttatg ccgctgccga ccattaaaca ttttattcgc accccggatg atgcgtggct
     2101 gctgaccacc gcgattccgg gcaaaaccgc gtttcaggtg ctggaagaat atccggatag
     2161 cggcgaaaac attgtggatg cgctggccgt gtttctgcgt cgtctgcata gcattccggt
     2221 gtgcaactgc ccgtttaaca gcgatcgtgt gtttcgtctg gcccaggcgc agagccgtat
     2281 gaacaacggc ctggtggatg cgagcgattt tgatgatgaa cgtaacggct ggccggtgga
     2341 acaggtgtgg aaagaaatgc ataaactgct gccgtttagc ccggatagcg tggtgaccca
     2401 cggcgatttt agcctggata acctgatttt cgatgaaggc aaactgattg gctgcattga
     2461 tgtgggccgt gtgggcattg cggatcgtta tcaggatctg gccattctgt ggaactgcct
     2521 gggcgaattt agcccgagcc tgcaaaaacg tctgtttcag aaatatggca ttgataatcc
     2581 ggatatgaac aaactgcaat ttcatctgat gctggatgaa tttttctaat aattaattgg
     2641 accgcggtcc gcgcgttgtc cttttccgct gcataaccct gcttcggggt cattatagcg
     2701 attttttcgg tatatccatc ctttttcgca cgatatacag gattttgcca aagggttcgt
     2761 gtagactttc cttggtgtat ccaacggcgt cagccgggca ggataggtga agtaggccca
     2821 cccgcgagcg ggtgttcctt cttcactgtc ccttattcgc acctggcggt gctcaacggg
     2881 aatcctgctc tgcgaggctg gccgtaggcc ggccgcgatg caggtggctg ctgaaccccc
     2941 agccggaact gaccccacaa ggccctagcg tttgcaatgc accaggtcat cattgaccca
     3001 ggcgtgttcc accaggccgc tgcctcgcaa ctcttcgcag gcttcgccga cctgctcgcg
     3061 ccacttcttc acgcgggtgg aatccgatcc gcacatgagg cggaaggttt ccagcttgag
     3121 cgggtacggc tcccggtgcg agctgaaata gtcgaacatc cgtcgggccg tcggcgacag
     3181 cttgcggtac ttctcccata tgaatttcgt gtagtggtcg ccagcaaaca gcacgacgat
     3241 ttcctcgtcg atcaggacct ggcaacggga cgttttcttg ccacggtcca ggacgcggaa
     3301 gcggtgcagc agcgacaccg attccaggtg cccaacgcgg tcggacgtga agcccatcgc
     3361 cgtcgcctgt aggcgcgaca ggcattcctc ggccttcgtg taataccggc cattgatcga
     3421 ccagcccagg tcctggcaaa gctcgtagaa cgtgaaggtg atcggctcgc cgataggggt
     3481 gcgcttcgcg tactccaaca cctgctgcca caccagttcg tcatcgtcgg cccgcagctc
     3541 gacgccggtg taggtgatct tcacgtcctt gttgacgtgg aaaatgacct tgttttgcag
     3601 cgcctcgcgc gggattttct tgttgcgcgt ggtgaacagg gcagagcggg ccgtgtcgtt
     3661 tggcatcgct cgcatcgtgt ccggccacgg cgcaatatcg aacaaggaaa gctgcatttc
     3721 cttgatctgc tgcttcgtgt gtttcagcaa cgcggcctgc ttggcttcgc tgacctgttt
     3781 tgccaggtcc tcgccggcgg tttttcgctt cttggtcgtc atagttcctc gcgtgtcgat
     3841 ggtcatcgac ttcgccaaac ctgccgcctc ctgttcgaga cgacgcgaac gctccacggc
     3901 ggccgatggc gcgggcaggg cagggggagc cagttgcacg ctgtcgcgct cgatcttggc
     3961 cgtagcttgc tggactatcg agccgacgga ctggaaggtt tcgcggggcg cacgcatgac
     4021 ggtgcggctt gcgatggttt cggcatcctc ggcggaaaac cccgcgtcga tcagttcttg
     4081 cctgtatgcc ttccggtcaa acgtccgatt cattcaccct ccttgcggga ttgccccgga
     4141 attaattccc cggatcgatc cgtcgatctt gatcccctgc gccatcagat ccttggcggc
     4201 aagaaagcca tccagtttac tttgcagggc ttcccaacct taccagaggg cgccccagct
     4261 ggcaattccg gttcgcttgc tgtccataaa accgcccagt ctagctatcg ccatgtaagc
     4321 ccactgcaag ctacctgctt tctctttgcg cttgcgtttt cccttgtcca gatagcccag
     4381 tagctgacat tcatccgggg tcagcaccgt ttctgcggac tggctttcta cgtggctgcc
     4441 atttttgggg tgaggccgtt cgcggccgag gggcgcagcc cctgggggga tgggaggccc
     4501 gcgttagcgg gccgggaggg ttcgagaagg gggggcaccc cccttcggcg tgcgcggtca
     4561 cgcgcacagg gcgcagccct ggttaaaaac aaggtttata aatattggtt taaaagcagg
     4621 ttaaaagaca ggttagcggt ggccgaaaaa cgggcggaaa cccttgcaaa tgctggattt
     4681 tctgcctgtg gacagcccct caaatgtcaa taggtgcgcc cctcatctgt cagcactctg
     4741 cccctcaagt gtcaaggatc gcgcccctca tctgtcagta gtcgcgcccc tcaagtgtca
     4801 ataccgcagg gcacttatcc ccaggcttgt ccacatcatc tgtgggaaac tcgcgtaaaa
     4861 tcaggcgttt tcgccgattt gcgaggctgg ccagctccac gtcgccggcc gaaatcgagc
     4921 ctgcccctca tctgtcaacg ccgcgccggg tgagtcggcc cctcaagtgt caacgtccgc
     4981 ccctcatctg tcagtgaggg ccaagttttc cgcgaggtat ccacaacgcc ggcggcccta
     5041 catggctctg ctgtagtgag tgggttgcgc tccggcagcg gtcctgatcc cccgcagaaa
     5101 aaaaggatct caagaagatc ctttgatctt ttctacggcg cgcccagctg tctagggcgg
     5161 cggatttgtc ctactcagga gagcgttcac cgacaaacaa cagataaaac gaaaggccca
     5221 gtctttcgac tgagcctttc gttttatttg atgcctttaa ttaa
//
