LOCUS       sequence-361921-        3971 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  arabinose-induced synthesis of a hybrid RNA containing a single MS2 
            hairpin followed by E coli ChiX inserted between XmaI and HindIII 
            sites. Insert can be replaced with RNA bait of interest..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3971)
  AUTHORS   Pandey S, Gravel CM, Stockert OM, Wang CD, Hegner CL, LeBlanc H, 
            Berry KE
  TITLE     Genetic identification of the functional surface for RNA binding by 
            Escherichia coli ProQ.
  JOURNAL   Nucleic Acids Res. 2020 May 7;48(8):4507-4520. doi: 
            10.1093/nar/gkaa144.
  PUBMED    32170306
REFERENCE   2  (bases 1 to 3971)
  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"; doi: "10"; journalName: 
            "Nucleic Acids Res"; date: "2020-05-7- 7"; volume: "48"; issue: "8";
            pages: "4507-4520"
COMMENT     Sequence Label: pCH6 - pBait-ChiX
FEATURES             Location/Qualifiers
     source          1..3971
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(165..956)
                     /codon_start=1
                     /gene="aadA"
                     /product="aminoglycoside adenylyltransferase (Murphy, 
                     1985)"
                     /label=SmR
                     /note="confers resistance to spectinomycin and 
                     streptomycin"
                     /translation="MREAVIAEVSTQLSEVVGVIERHLEPTLLAVHLYGSAVDGGLKPH
                     SDIDLLVTVTVRLDETTRRALINDLLETSASPGESEILRAVEVTIVVHDDIIPWRYPAK
                     RELQFGEWQRNDILAGIFEPATIDIDLAILLTKAREHSVALVGPAAEELFDPVPEQDLF
                     EALNETLTLWNSPPDWAGDERNVVLTLSRIWYSAVTGKIAPKDVAADWAMERLPAQYQP
                     VILEARQAYLGQEEDRLASRADQLEEFVHYVKGEITKVVGK"
     promoter        complement(957..1048)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(1096..1834)
                     /direction=LEFT
                     /label=CloDF13 ori
                     /note="Plasmids containing the CloDF13 (CDF) origin of 
                     replication can be propagated in E. coli cells that contain
                     additional plasmids with compatible origins."
     CDS             complement(2233..3111)
                     /codon_start=1
                     /gene="araC"
                     /product="L-arabinose regulatory protein"
                     /label=araC
                     /translation="MAEAQNDPLLPGYSFNAHLVAGLTPIEANGYLDFFIDRPLGMKGY
                     ILNLTIRGQGVVKNQGREFVCRPGDILLFPPGEIHHYGRHPEAREWYHQWVYFRPRAYW
                     HEWLNWPSIFANTGFFRPDEAHQPHFSDLFGQIINAGQGEGRYSELLAINLLEQLLLRR
                     MEAINESLHPPMDNRVREACQYISDHLADSNFDIASVAQHVCLSPSRLSHLFRQQLGIS
                     VLSWREDQRISQAKLLLSTTRMPIATVGRNVGFDDQLYFSRVFKKCTGASPSEFRAGCE
                     EKVNDVAVKLS"
     primer_bind     3345..3364
                     /label=pBAD Forward
                     /note="For vectors with E. coli araBAD promoter, forward 
                     primer"
     misc_RNA        3428..3446
                     /label=MS2 stem loop
                     /note="stem loop that binds the bacteriophage MS2 coat 
                     protein"
     primer_bind     complement(3581..3598)
                     /label=pBAD Reverse
                     /note="For vectors with E. coli araBAD promoter, reverse 
                     primer"
     terminator      3751..3837
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      3929..3956
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
ORIGIN
        1 cctcaggcat ttgagaagca cacggtcaca ctgcttccgg tagtcaataa accggtaaac
       61 cagcaataga cataagcggc tatttaacga ccctgccctg aaccgacgac cgggtcatcg
      121 tggccggatc ttgcggcccc tcggcttgaa cgaattgtta gacattattt gccgactacc
      181 ttggtgatct cgcctttcac gtagtggaca aattcttcca actgatctgc gcgcgaggcc
      241 aagcgatctt cttcttgtcc aagataagcc tgtctagctt caagtatgac gggctgatac
      301 tgggccggca ggcgctccat tgcccagtcg gcagcgacat ccttcggcgc gattttgccg
      361 gttactgcgc tgtaccaaat gcgggacaac gtaagcacta catttcgctc atcgccagcc
      421 cagtcgggcg gcgagttcca tagcgttaag gtttcattta gcgcctcaaa tagatcctgt
      481 tcaggaaccg gatcaaagag ttcctccgcc gctggaccta ccaaggcaac gctatgttct
      541 cttgcttttg tcagcaagat agccagatca atgtcgatcg tggctggctc gaagatacct
      601 gcaagaatgt cattgcgctg ccattctcca aattgcagtt cgcgcttagc tggataacgc
      661 cacggaatga tgtcgtcgtg cacaacaatg gtgacttcta cagcgcggag aatctcgctc
      721 tctccagggg aagccgaagt ttccaaaagg tcgttgatca aagctcgccg cgttgtttca
      781 tcaagcctta cggtcaccgt aaccagcaaa tcaatatcac tgtgtggctt caggccgcca
      841 tccactgcgg agccgtacaa atgtacggcc agcaacgtcg gttcgagatg gcgctcgatg
      901 acgccaacta cctctgatag ttgagtcgat acttcggcga tcaccgcttc cctcatactc
      961 ttcctttttc aatattattg aagcatttat cagggttatt gtctcatgag cggatacata
     1021 tttgaatgta tttagaaaaa taaacaaata gctagctcac tcggtcgcta cgctccgggc
     1081 gtgagactgc ggcgggcgct gcggacacat acaaagttac ccacagattc cgtggataag
     1141 caggggacta acatgtgagg caaaacagca gggccgcgcc ggtggcgttt ttccataggc
     1201 tccgccctcc tgccagagtt cacataaaca gacgcttttc cggtgcatct gtgggagccg
     1261 tgaggctcaa ccatgaatct gacagtacgg gcgaaacccg acaggactta aagatcccca
     1321 ccgtttccgg cgggtcgctc cctcttgcgc tctcctgttc cgaccctgcc gtttaccgga
     1381 tacctgttcc gcctttctcc cttacgggaa gtgtggcgct ttctcatagc tcacacactg
     1441 gtatctcggc tcggtgtagg tcgttcgctc caagctgggc tgtaagcaag aactccccgt
     1501 tcagcccgac tgctgcgcct tatccggtaa ctgttcactt gagtccaacc cggaaaagca
     1561 cggtaaaacg ccactggcag cagccattgg taactgggag ttcgcagagg atttgtttag
     1621 ctaaacacgc ggttgctctt gaagtgtgcg ccaaagtccg gctacactgg aaggacagat
     1681 ttggttgctg tgctctgcga aagccagtta ccacggttaa gcagttcccc aactgactta
     1741 accttcgatc aaaccacctc cccaggtggt tttttcgttt acagggcaaa agattacgcg
     1801 cagaaaaaaa ggatctcaag aagatccttt gatcttttct actgaaccgc tctagatttc
     1861 agtgcaattt atctcttcaa atgtagcacc tgaagtcagc cccatacgat ataagttgta
     1921 attctcatgt tagtcatgcc ccgcgcccac cggaaggagc tgactgggtt gaaggctctc
     1981 aagggcatcg gtcgagatcc cggtgcctaa tgagtgagct aacttacatt aattgcgttg
     2041 cgctcactgc ccgctttcca gtcgggaaac ctgtcgtgcc agctgcatta atgaatcggc
     2101 caacgcgcgg ggagaggcgg tttgcgtatt gggcgccatc gatgcataat gtgcctgtca
     2161 aatggacgaa gcagggattc tgcaaaccct atgctactcc gtcaagccgt caattgtctg
     2221 attcgttacc aattatgaca acttgacggc tacatcattc actttttctt cacaaccggc
     2281 acggaactcg ctcgggctgg ccccggtgca ttttttaaat acccgcgaga aatagagttg
     2341 atcgtcaaaa ccaacattgc gaccgacggt ggcgataggc atccgggtgg tgctcaaaag
     2401 cagcttcgcc tggctgatac gttggtcctc gcgccagctt aagacgctaa tccctaactg
     2461 ctggcggaaa agatgtgaca gacgcgacgg cgacaagcaa acatgctgtg cgacgctggc
     2521 gatatcaaaa ttgctgtctg ccaggtgatc gctgatgtac tgacaagcct cgcgtacccg
     2581 attatccatc ggtggatgga gcgactcgtt aatcgcttcc atgcgccgca gtaacaattg
     2641 ctcaagcaga tttatcgcca gcagctccga atagcgccct tccccttgcc cggcgttaat
     2701 gatttgccca aacaggtcgc tgaaatgcgg ctggtgcgct tcatccgggc gaaagaaccc
     2761 cgtattggca aatattgacg gccagttaag ccattcatgc cagtaggcgc gcggacgaaa
     2821 gtaaacccac tggtgatacc attcgcgagc ctccggatga cgaccgtagt gatgaatctc
     2881 tcctggcggg aacagcaaaa tatcacccgg tcggcaaaca aattctcgtc cctgattttt
     2941 caccaccccc tgaccgcgaa tggtgagatt gagaatataa cctttcattc ccagcggtcg
     3001 gtcgataaaa aaatcgagat aaccgttggc ctcaatcggc gttaaacccg ccaccagatg
     3061 ggcattaaac gagtatcccg gcagcagggg atcattttgc gcttcagcca tacttttcat
     3121 actcccgcca ttcagagaag aaaccaattg tccatattgc atcagacatt gccgtcactg
     3181 cgtcttttac tggctcttct cgctaaccaa accggtaacc ccgcttatta aaagcattct
     3241 gtaacaaagc gggaccaaag ccatgacaaa aacgcgtaac aaaagtgtct ataatcacgg
     3301 cagaaaagtc cacattgatt atttgcacgg cgtcacactt tgctatgcca tagcattttt
     3361 atccataaga ttagcggatc ctacctgacg ctttttatcg caactctcta ctgtttgaat
     3421 tcagaaaaca tgaggatcac ccatgtctgc agcccgggac accgtcgctt aaagtgacgg
     3481 cataataata aaaaaatgaa attcctcttt gacgggccaa tagcgatatt ggccattttt
     3541 ttaagcttgg ctgttttggc ggatgagaga agattttcag cctgatacag attaaatcag
     3601 aacgcagaag cggtctgata aaacagaatt tgcctggcgg cagtagcgcg gtggtcccac
     3661 ctgaccccat gccgaactca gaagtgaaac gccgtagcgc cgatggtagt gtggggtctc
     3721 cccatgcgag agtagggaac tgccaggcat caaataaaac gaaaggctca gtcgaaagac
     3781 tgggcctttc gttttatctg ttgtttgtcg gtgaacgctc tcctgagtag gacaaatccg
     3841 ccgggagcgg atttgaacgt tgcgaagcaa cggcccggag ggtggcgggc aggacgcccg
     3901 ccataaactg ccaggcatca aattaagcag aaggccatcc tgacggatgg cctttttgcg
     3961 tttctacaaa c
//
