LOCUS       sequence-376090-        5266 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Reprogrammed short tracrRNA for hijacking mRNA of arsenic responsive
            gene cluster (site 2).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5266)
  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 5266)
  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: pLY251
FEATURES             Location/Qualifiers
     source          1..5266
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    51..69
                     /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    77..95
                     /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    99..117
                     /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      205..299
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     primer_bind     complement(420..439)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             431..1246
                     /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            1413..1521
                     /note="incP origin of transfer"
     CDS             complement(1582..2730)
                     /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      3123..3654
                     /label=oriV
                     /note="incP origin of replication"
     terminator      complement(3781..3867)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     3882..3904
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     misc_feature    3929..3950
                     /label=BioBrick prefix
                     /note="BioBrick prefix for parts that do not start with 
                     ""ATG"""
     CDS             complement(4057..4935)
                     /codon_start=1
                     /gene="araC"
                     /product="L-arabinose regulatory protein"
                     /label=araC
                     /translation="MAEAQNDPLLPGYSFNAHLVAGLTPIEANGYLDFFIDRPLGMKGY
                     ILNLTIRGQGVVKNQGREFVCRPGDILLFPPGEIHHYGRHPEAREWYHQWVYFRPRAYW
                     HEWLNWPSIFANTGFFRPDEAHQPHFSDLFGQIINAGQGEGRYSELLAINLLEQLLLRR
                     MEAINESLHPPMDNRVREACQYISDHLADSNFDIASVAQHVCLSPSRLSHLFRQQLGIS
                     VLSWREDQRISQAKLLLSTTRMPIATVGRNVGFDDQLYFSRVFKKCTGASPSEFRAGCE
                     EKVNDVAVKLS"
     promoter        4962..5246
                     /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     5169..5188
                     /label=pBAD Forward
                     /note="For vectors with E. coli araBAD promoter, forward 
                     primer"
ORIGIN
        1 atctaaggct agtccgttat caacttgaaa aagtggcacc gagtcggtgc gggccctgaa
       61 gaagggccca aggctagggc cctgaagaag ggcccgcagg gccctgaaga agggcccttt
      121 tttttactag agctcggtac caaattccag aaaagaggcc tcccgaaagg ggggcctttt
      181 ttcgttttgg tcctactagt cttggactcc tgttgataga tccagtaatg acctcagaac
      241 tccatctgga tttgttcaga acgctcggtt gccgccgggc gttttttatt ggtgagaatc
      301 caggggtccc caataattac gatttaaatt tgtgtctcaa aatctctgat gttacattgc
      361 acaagataaa aatatatcat catgaacaat aaaactgtct gcttacataa acagtaatac
      421 aaggggtgtt atgagccata ttcagcgtga aacgagctgt agccgtccgc gtctgaacag
      481 caacatggat gcggatctgt atggctataa atgggcgcgt gataacgtgg gtcagagcgg
      541 cgcgaccatt tatcgtctgt atggcaaacc ggatgcgccg gaactgtttc tgaaacatgg
      601 caaaggcagc gtggcgaacg atgtgaccga tgaaatggtg cgtctgaact ggctgaccga
      661 atttatgccg ctgccgacca ttaaacattt tattcgcacc ccggatgatg cgtggctgct
      721 gaccaccgcg attccgggca aaaccgcgtt tcaggtgctg gaagaatatc cggatagcgg
      781 cgaaaacatt gtggatgcgc tggccgtgtt tctgcgtcgt ctgcatagca ttccggtgtg
      841 caactgcccg tttaacagcg atcgtgtgtt tcgtctggcc caggcgcaga gccgtatgaa
      901 caacggcctg gtggatgcga gcgattttga tgatgaacgt aacggctggc cggtggaaca
      961 ggtgtggaaa gaaatgcata aactgctgcc gtttagcccg gatagcgtgg tgacccacgg
     1021 cgattttagc ctggataacc tgattttcga tgaaggcaaa ctgattggct gcattgatgt
     1081 gggccgtgtg ggcattgcgg atcgttatca ggatctggcc attctgtgga actgcctggg
     1141 cgaatttagc ccgagcctgc aaaaacgtct gtttcagaaa tatggcattg ataatccgga
     1201 tatgaacaaa ctgcaatttc atctgatgct ggatgaattt ttctaataat taattggacc
     1261 gcggtccgcg cgttgtcctt ttccgctgca taaccctgct tcggggtcat tatagcgatt
     1321 ttttcggtat atccatcctt tttcgcacga tatacaggat tttgccaaag ggttcgtgta
     1381 gactttcctt ggtgtatcca acggcgtcag ccgggcagga taggtgaagt aggcccaccc
     1441 gcgagcgggt gttccttctt cactgtccct tattcgcacc tggcggtgct caacgggaat
     1501 cctgctctgc gaggctggcc gtaggccggc cgcgatgcag gtggctgctg aacccccagc
     1561 cggaactgac cccacaaggc cctagcgttt gcaatgcacc aggtcatcat tgacccaggc
     1621 gtgttccacc aggccgctgc ctcgcaactc ttcgcaggct tcgccgacct gctcgcgcca
     1681 cttcttcacg cgggtggaat ccgatccgca catgaggcgg aaggtttcca gcttgagcgg
     1741 gtacggctcc cggtgcgagc tgaaatagtc gaacatccgt cgggccgtcg gcgacagctt
     1801 gcggtacttc tcccatatga atttcgtgta gtggtcgcca gcaaacagca cgacgatttc
     1861 ctcgtcgatc aggacctggc aacgggacgt tttcttgcca cggtccagga cgcggaagcg
     1921 gtgcagcagc gacaccgatt ccaggtgccc aacgcggtcg gacgtgaagc ccatcgccgt
     1981 cgcctgtagg cgcgacaggc attcctcggc cttcgtgtaa taccggccat tgatcgacca
     2041 gcccaggtcc tggcaaagct cgtagaacgt gaaggtgatc ggctcgccga taggggtgcg
     2101 cttcgcgtac tccaacacct gctgccacac cagttcgtca tcgtcggccc gcagctcgac
     2161 gccggtgtag gtgatcttca cgtccttgtt gacgtggaaa atgaccttgt tttgcagcgc
     2221 ctcgcgcggg attttcttgt tgcgcgtggt gaacagggca gagcgggccg tgtcgtttgg
     2281 catcgctcgc atcgtgtccg gccacggcgc aatatcgaac aaggaaagct gcatttcctt
     2341 gatctgctgc ttcgtgtgtt tcagcaacgc ggcctgcttg gcttcgctga cctgttttgc
     2401 caggtcctcg ccggcggttt ttcgcttctt ggtcgtcata gttcctcgcg tgtcgatggt
     2461 catcgacttc gccaaacctg ccgcctcctg ttcgagacga cgcgaacgct ccacggcggc
     2521 cgatggcgcg ggcagggcag ggggagccag ttgcacgctg tcgcgctcga tcttggccgt
     2581 agcttgctgg actatcgagc cgacggactg gaaggtttcg cggggcgcac gcatgacggt
     2641 gcggcttgcg atggtttcgg catcctcggc ggaaaacccc gcgtcgatca gttcttgcct
     2701 gtatgccttc cggtcaaacg tccgattcat tcaccctcct tgcgggattg ccccggaatt
     2761 aattccccgg atcgatccgt cgatcttgat cccctgcgcc atcagatcct tggcggcaag
     2821 aaagccatcc agtttacttt gcagggcttc ccaaccttac cagagggcgc cccagctggc
     2881 aattccggtt cgcttgctgt ccataaaacc gcccagtcta gctatcgcca tgtaagccca
     2941 ctgcaagcta cctgctttct ctttgcgctt gcgttttccc ttgtccagat agcccagtag
     3001 ctgacattca tccggggtca gcaccgtttc tgcggactgg ctttctacgt ggctgccatt
     3061 tttggggtga ggccgttcgc ggccgagggg cgcagcccct ggggggatgg gaggcccgcg
     3121 ttagcgggcc gggagggttc gagaaggggg ggcacccccc ttcggcgtgc gcggtcacgc
     3181 gcacagggcg cagccctggt taaaaacaag gtttataaat attggtttaa aagcaggtta
     3241 aaagacaggt tagcggtggc cgaaaaacgg gcggaaaccc ttgcaaatgc tggattttct
     3301 gcctgtggac agcccctcaa atgtcaatag gtgcgcccct catctgtcag cactctgccc
     3361 ctcaagtgtc aaggatcgcg cccctcatct gtcagtagtc gcgcccctca agtgtcaata
     3421 ccgcagggca cttatcccca ggcttgtcca catcatctgt gggaaactcg cgtaaaatca
     3481 ggcgttttcg ccgatttgcg aggctggcca gctccacgtc gccggccgaa atcgagcctg
     3541 cccctcatct gtcaacgccg cgccgggtga gtcggcccct caagtgtcaa cgtccgcccc
     3601 tcatctgtca gtgagggcca agttttccgc gaggtatcca caacgccggc ggccctacat
     3661 ggctctgctg tagtgagtgg gttgcgctcc ggcagcggtc ctgatccccc gcagaaaaaa
     3721 aggatctcaa gaagatcctt tgatcttttc tacggcgcgc ccagctgtct agggcggcgg
     3781 atttgtccta ctcaggagag cgttcaccga caaacaacag ataaaacgaa aggcccagtc
     3841 tttcgactga gcctttcgtt ttatttgatg cctttaatta aagcggataa caatttcaca
     3901 caggaggccg cctaggccgc ggccgcgcga attcgcggcc gcttctagag cctagttgtc
     3961 ttcatgcatg aagacaaaat taatactaga gggaccaaaa cgaaaaaaga cgctcgaaag
     4021 cgtctctttt ctggaatttg gtaccgagct ctagtattat gacaacttga cggctacatc
     4081 attcactttt tcttcacaac cggcacggaa ctcgctcggg ctggccccgg tgcatttttt
     4141 aaatacccgc gagaaataga gttgatcgtc aaaaccaaca ttgcgaccga cggtggcgat
     4201 aggcatccgg gtggtgctca aaagcagctt cgcctggctg atacgttggt cctcgcgcca
     4261 gcttaagacg ctaatcccta actgctggcg gaaaagatgt gacagacgcg acggcgacaa
     4321 gcaaacatgc tgtgcgacgc tggcgatatc aaaattgctg tctgccaggt gatcgctgat
     4381 gtactgacaa gcctcgcgta cccgattatc catcggtgga tggagcgact cgttaatcgc
     4441 ttccatgcgc cgcagtaaca attgctcaag cagatttatc gccagcagct ccgaatagcg
     4501 cccttcccct tgcccggcgt taatgatttg cccaaacagg tcgctgaaat gcggctggtg
     4561 cgcttcatcc gggcgaaaga accccgtatt ggcaaatatt gacggccagt taagccattc
     4621 atgccagtag gcgcgcggac gaaagtaaac ccactggtga taccattcgc gagcctccgg
     4681 atgacgaccg tagtgatgaa tctctcctgg cgggaacagc aaaatatcac ccggtcggca
     4741 aacaaattct cgtccctgat ttttcaccac cccctgaccg cgaatggtga gattgagaat
     4801 ataacctttc attcccagcg gtcggtcgat aaaaaaatcg agataaccgt tggcctcaat
     4861 cggcgttaaa cccgccacca gatgggcatt aaacgagtat cccggcagca ggggatcatt
     4921 ttgcgcttca gccatacttt tcatactccc gccattcaga gaagaaacca attgtccata
     4981 ttgcatcaga cattgccgtc actgcgtctt ttactggctc ttctcgctaa ccaaaccggt
     5041 aaccccgctt attaaaagca ttctgtaaca aagcgggacc aaagccatga caaaaacgcg
     5101 taacaaaagt gtctataatc acggcagaaa agtccacatt gattatttgc acggcgtcac
     5161 actttgctat gccatagcat ttttatccat aagattagcg gatcctacct gacgcttttt
     5221 atcgcaactc tctactgttt ctccataatg gtaatgttgc aagtat
//
