LOCUS       sequence-376088-        5266 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Reprogrammed short tracrRNA for hijacking mRNA of zraP.
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: pLY273
FEATURES             Location/Qualifiers
     source          1..5266
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     11..33
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     misc_feature    58..79
                     /label=BioBrick prefix
                     /note="BioBrick prefix for parts that do not start with 
                     ""ATG"""
     CDS             complement(186..1064)
                     /codon_start=1
                     /gene="araC"
                     /product="L-arabinose regulatory protein"
                     /label=araC
                     /translation="MAEAQNDPLLPGYSFNAHLVAGLTPIEANGYLDFFIDRPLGMKGY
                     ILNLTIRGQGVVKNQGREFVCRPGDILLFPPGEIHHYGRHPEAREWYHQWVYFRPRAYW
                     HEWLNWPSIFANTGFFRPDEAHQPHFSDLFGQIINAGQGEGRYSELLAINLLEQLLLRR
                     MEAINESLHPPMDNRVREACQYISDHLADSNFDIASVAQHVCLSPSRLSHLFRQQLGIS
                     VLSWREDQRISQAKLLLSTTRMPIATVGRNVGFDDQLYFSRVFKKCTGASPSEFRAGCE
                     EKVNDVAVKLS"
     promoter        1091..1375
                     /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     1298..1317
                     /label=pBAD Forward
                     /note="For vectors with E. coli araBAD promoter, forward 
                     primer"
     protein_bind    1446..1464
                     /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    1472..1490
                     /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    1494..1512
                     /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      1600..1694
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     primer_bind     complement(1815..1834)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             1826..2641
                     /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            2808..2916
                     /note="incP origin of transfer"
     CDS             complement(2977..4125)
                     /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      4518..5049
                     /label=oriV
                     /note="incP origin of replication"
     terminator      complement(5176..5262)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
ORIGIN
        1 ctttaattaa agcggataac aatttcacac aggaggccgc ctaggccgcg gccgcgcgaa
       61 ttcgcggccg cttctagagc ctagttgtct tcatgcatga agacaaaatt aatactagag
      121 ggaccaaaac gaaaaaagac gctcgaaagc gtctcttttc tggaatttgg taccgagctc
      181 tagtattatg acaacttgac ggctacatca ttcacttttt cttcacaacc ggcacggaac
      241 tcgctcgggc tggccccggt gcatttttta aatacccgcg agaaatagag ttgatcgtca
      301 aaaccaacat tgcgaccgac ggtggcgata ggcatccggg tggtgctcaa aagcagcttc
      361 gcctggctga tacgttggtc ctcgcgccag cttaagacgc taatccctaa ctgctggcgg
      421 aaaagatgtg acagacgcga cggcgacaag caaacatgct gtgcgacgct ggcgatatca
      481 aaattgctgt ctgccaggtg atcgctgatg tactgacaag cctcgcgtac ccgattatcc
      541 atcggtggat ggagcgactc gttaatcgct tccatgcgcc gcagtaacaa ttgctcaagc
      601 agatttatcg ccagcagctc cgaatagcgc ccttcccctt gcccggcgtt aatgatttgc
      661 ccaaacaggt cgctgaaatg cggctggtgc gcttcatccg ggcgaaagaa ccccgtattg
      721 gcaaatattg acggccagtt aagccattca tgccagtagg cgcgcggacg aaagtaaacc
      781 cactggtgat accattcgcg agcctccgga tgacgaccgt agtgatgaat ctctcctggc
      841 gggaacagca aaatatcacc cggtcggcaa acaaattctc gtccctgatt tttcaccacc
      901 ccctgaccgc gaatggtgag attgagaata taacctttca ttcccagcgg tcggtcgata
      961 aaaaaatcga gataaccgtt ggcctcaatc ggcgttaaac ccgccaccag atgggcatta
     1021 aacgagtatc ccggcagcag gggatcattt tgcgcttcag ccatactttt catactcccg
     1081 ccattcagag aagaaaccaa ttgtccatat tgcatcagac attgccgtca ctgcgtcttt
     1141 tactggctct tctcgctaac caaaccggta accccgctta ttaaaagcat tctgtaacaa
     1201 agcgggacca aagccatgac aaaaacgcgt aacaaaagtg tctataatca cggcagaaaa
     1261 gtccacattg attatttgca cggcgtcaca ctttgctatg ccatagcatt tttatccata
     1321 agattagcgg atcctacctg acgcttttta tcgcaactct ctactgtttc tccattttcg
     1381 tgttccgtta agtatggtta aggctagtcc gttatcaact tgaaaaagtg gcaccgagtc
     1441 ggtgcgggcc ctgaagaagg gcccaaggct agggccctga agaagggccc gcagggccct
     1501 gaagaagggc cctttttttt actagagctc ggtaccaaat tccagaaaag aggcctcccg
     1561 aaaggggggc cttttttcgt tttggtccta ctagtcttgg actcctgttg atagatccag
     1621 taatgacctc agaactccat ctggatttgt tcagaacgct cggttgccgc cgggcgtttt
     1681 ttattggtga gaatccaggg gtccccaata attacgattt aaatttgtgt ctcaaaatct
     1741 ctgatgttac attgcacaag ataaaaatat atcatcatga acaataaaac tgtctgctta
     1801 cataaacagt aatacaaggg gtgttatgag ccatattcag cgtgaaacga gctgtagccg
     1861 tccgcgtctg aacagcaaca tggatgcgga tctgtatggc tataaatggg cgcgtgataa
     1921 cgtgggtcag agcggcgcga ccatttatcg tctgtatggc aaaccggatg cgccggaact
     1981 gtttctgaaa catggcaaag gcagcgtggc gaacgatgtg accgatgaaa tggtgcgtct
     2041 gaactggctg accgaattta tgccgctgcc gaccattaaa cattttattc gcaccccgga
     2101 tgatgcgtgg ctgctgacca ccgcgattcc gggcaaaacc gcgtttcagg tgctggaaga
     2161 atatccggat agcggcgaaa acattgtgga tgcgctggcc gtgtttctgc gtcgtctgca
     2221 tagcattccg gtgtgcaact gcccgtttaa cagcgatcgt gtgtttcgtc tggcccaggc
     2281 gcagagccgt atgaacaacg gcctggtgga tgcgagcgat tttgatgatg aacgtaacgg
     2341 ctggccggtg gaacaggtgt ggaaagaaat gcataaactg ctgccgttta gcccggatag
     2401 cgtggtgacc cacggcgatt ttagcctgga taacctgatt ttcgatgaag gcaaactgat
     2461 tggctgcatt gatgtgggcc gtgtgggcat tgcggatcgt tatcaggatc tggccattct
     2521 gtggaactgc ctgggcgaat ttagcccgag cctgcaaaaa cgtctgtttc agaaatatgg
     2581 cattgataat ccggatatga acaaactgca atttcatctg atgctggatg aatttttcta
     2641 ataattaatt ggaccgcggt ccgcgcgttg tccttttccg ctgcataacc ctgcttcggg
     2701 gtcattatag cgattttttc ggtatatcca tcctttttcg cacgatatac aggattttgc
     2761 caaagggttc gtgtagactt tccttggtgt atccaacggc gtcagccggg caggataggt
     2821 gaagtaggcc cacccgcgag cgggtgttcc ttcttcactg tcccttattc gcacctggcg
     2881 gtgctcaacg ggaatcctgc tctgcgaggc tggccgtagg ccggccgcga tgcaggtggc
     2941 tgctgaaccc ccagccggaa ctgaccccac aaggccctag cgtttgcaat gcaccaggtc
     3001 atcattgacc caggcgtgtt ccaccaggcc gctgcctcgc aactcttcgc aggcttcgcc
     3061 gacctgctcg cgccacttct tcacgcgggt ggaatccgat ccgcacatga ggcggaaggt
     3121 ttccagcttg agcgggtacg gctcccggtg cgagctgaaa tagtcgaaca tccgtcgggc
     3181 cgtcggcgac agcttgcggt acttctccca tatgaatttc gtgtagtggt cgccagcaaa
     3241 cagcacgacg atttcctcgt cgatcaggac ctggcaacgg gacgttttct tgccacggtc
     3301 caggacgcgg aagcggtgca gcagcgacac cgattccagg tgcccaacgc ggtcggacgt
     3361 gaagcccatc gccgtcgcct gtaggcgcga caggcattcc tcggccttcg tgtaataccg
     3421 gccattgatc gaccagccca ggtcctggca aagctcgtag aacgtgaagg tgatcggctc
     3481 gccgataggg gtgcgcttcg cgtactccaa cacctgctgc cacaccagtt cgtcatcgtc
     3541 ggcccgcagc tcgacgccgg tgtaggtgat cttcacgtcc ttgttgacgt ggaaaatgac
     3601 cttgttttgc agcgcctcgc gcgggatttt cttgttgcgc gtggtgaaca gggcagagcg
     3661 ggccgtgtcg tttggcatcg ctcgcatcgt gtccggccac ggcgcaatat cgaacaagga
     3721 aagctgcatt tccttgatct gctgcttcgt gtgtttcagc aacgcggcct gcttggcttc
     3781 gctgacctgt tttgccaggt cctcgccggc ggtttttcgc ttcttggtcg tcatagttcc
     3841 tcgcgtgtcg atggtcatcg acttcgccaa acctgccgcc tcctgttcga gacgacgcga
     3901 acgctccacg gcggccgatg gcgcgggcag ggcaggggga gccagttgca cgctgtcgcg
     3961 ctcgatcttg gccgtagctt gctggactat cgagccgacg gactggaagg tttcgcgggg
     4021 cgcacgcatg acggtgcggc ttgcgatggt ttcggcatcc tcggcggaaa accccgcgtc
     4081 gatcagttct tgcctgtatg ccttccggtc aaacgtccga ttcattcacc ctccttgcgg
     4141 gattgccccg gaattaattc cccggatcga tccgtcgatc ttgatcccct gcgccatcag
     4201 atccttggcg gcaagaaagc catccagttt actttgcagg gcttcccaac cttaccagag
     4261 ggcgccccag ctggcaattc cggttcgctt gctgtccata aaaccgccca gtctagctat
     4321 cgccatgtaa gcccactgca agctacctgc tttctctttg cgcttgcgtt ttcccttgtc
     4381 cagatagccc agtagctgac attcatccgg ggtcagcacc gtttctgcgg actggctttc
     4441 tacgtggctg ccatttttgg ggtgaggccg ttcgcggccg aggggcgcag cccctggggg
     4501 gatgggaggc ccgcgttagc gggccgggag ggttcgagaa gggggggcac cccccttcgg
     4561 cgtgcgcggt cacgcgcaca gggcgcagcc ctggttaaaa acagggttta taaatattgg
     4621 tttaaaagca ggttaaaaga caggttagcg gtggccgaaa aacgggcgga aacccttgca
     4681 aatgctggat tttctgcctg tggacagccc ctcaaatgtc aataggtgcg cccctcatct
     4741 gtcagcactc tgcccctcaa gtgtcaagga tcgcgcccct catctgtcag tagtcgcgcc
     4801 cctcaagtgt caataccgca gggcacttat ccccaggctt gtccacatca tctgtgggaa
     4861 actcgcgtaa aatcaggcgt tttcgccgat ttgcgaggct ggccagctcc acgtcgccgg
     4921 ccgaaatcga gcctgcccct catctgtcaa cgccgcgccg ggtgagtcgg cccctcaagt
     4981 gtcaacgtcc gcccctcatc tgtcagtgag ggccaagttt tccgcgaggt atccacaacg
     5041 ccggcggccc tacatggctc tgctgtagtg agtgggttgc gctccggcag cggtcctgat
     5101 cccccgcaga aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg cgcgcccagc
     5161 tgtctagggc ggcggatttg tcctactcag gagagcgttc accgacaaac aacagataaa
     5221 acgaaaggcc cagtctttcg actgagcctt tcgttttatt tgatgc
//
