LOCUS       sequence-451847-        4012 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  arabinose-induced synthesis of an RNA containing an MS2 hairpin, 7GC
            clamp and trpA terminator. Used as a three-hybrid control. For 
            cloning purposes, see plasmid #222404..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4012)
  AUTHORS   
  TITLE     Improved constructs for bait RNA display in a bacterial three-hybrid
            assay
  JOURNAL   Sci Rep 15, 3820 (2025)
REFERENCE   2  (bases 1 to 4012)
  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: "Sci Rep 15,
            3820 (2025)"
COMMENT     Sequence Label: pSS2 - pBait-1xMS2-7GC[empty]-TtrpA
FEATURES             Location/Qualifiers
     source          1..4012
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(1194..2072)
                     /codon_start=1
                     /gene="araC"
                     /product="L-arabinose regulatory protein"
                     /label=araC
                     /translation="MAEAQNDPLLPGYSFNAHLVAGLTPIEANGYLDFFIDRPLGMKGY
                     ILNLTIRGQGVVKNQGREFVCRPGDILLFPPGEIHHYGRHPEAREWYHQWVYFRPRAYW
                     HEWLNWPSIFANTGFFRPDEAHQPHFSDLFGQIINAGQGEGRYSELLAINLLEQLLLRR
                     MEAINESLHPPMDNRVREACQYISDHLADSNFDIASVAQHVCLSPSRLSHLFRQQLGIS
                     VLSWREDQRISQAKLLLSTTRMPIATVGRNVGFDDQLYFSRVFKKCTGASPSEFRAGCE
                     EKVNDVAVKLS"
     primer_bind     2306..2325
                     /label=pBAD Forward
                     /note="For vectors with E. coli araBAD promoter, forward 
                     primer"
     misc_RNA        2389..2407
                     /label=MS2 stem loop
                     /note="stem loop that binds the bacteriophage MS2 coat 
                     protein"
     primer_bind     complement(2518..2535)
                     /label=pBAD Reverse
                     /note="For vectors with E. coli araBAD promoter, reverse 
                     primer"
     terminator      2688..2774
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      2866..2893
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     CDS             complement(3073..3864)
                     /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(3865..3956)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 acacatacaa agttacccac agattccgtg gataagcagg ggactaacat gtgaggcaaa
       61 acagcagggc cgcgccggtg gcgtttttcc ataggctccg ccctcctgcc agagttcaca
      121 taaacagacg cttttccggt gcatctgtgg gagccgtgag gctcaaccat gaatctgaca
      181 gtacgggcga aacccgacag gacttaaaga tccccaccgt ttccggcggg tcgctccctc
      241 ttgcgctctc ctgttccgac cctgccgttt accggatacc tgttccgcct ttctccctta
      301 cgggaagtgt ggcgctttct catagctcac acactggtat ctcggctcgg tgtaggtcgt
      361 tcgctccaag ctgggctgta agcaagaact ccccgttcag cccgactgct gcgccttatc
      421 cggtaactgt tcacttgagt ccaacccgga aaagcacggt aaaacgccac tggcagcagc
      481 cattggtaac tgggagttcg cagaggattt gtttagctaa acacgcggtt gctcttgaag
      541 tgtgcgccaa agtccggcta cactggaagg acagatttgg ttgctgtgct ctgcgaaagc
      601 cagttaccac ggttaagcag ttccccaact gacttaacct tcgatcaaac cacctcccca
      661 ggtggttttt tcgaaggtta agtcagttgg ggaactgctt aaccttcgat caaaccacct
      721 ccccaggtgg ttttttcgtt tacagggcaa aagattacgc gcagaaaaaa aggatctcaa
      781 gaagatcctt tgatcttttc tactgaaccg ctctagattt cagtgcaatt tatctcttca
      841 aatgtagcac ctgaagtcag ccccatacga tataagttgt aattctcatg ttagtcatgc
      901 cccgcgccca ccggaaggag ctgactgggt tgaaggctct caagggcatc ggtcgagatc
      961 ccggtgccta atgagtgagc taacttacat taattgcgtt gcgctcactg cccgctttcc
     1021 agtcgggaaa cctgtcgtgc cagctgcatt aatgaatcgg ccaacgcgcg gggagaggcg
     1081 gtttgcgtat tgggcgccat cgatgcataa tgtgcctgtc aaatggacga agcagggatt
     1141 ctgcaaaccc tatgctactc cgtcaagccg tcaattgtct gattcgttac caattatgac
     1201 aacttgacgg ctacatcatt cactttttct tcacaaccgg cacggaactc gctcgggctg
     1261 gccccggtgc attttttaaa tacccgcgag aaatagagtt gatcgtcaaa accaacattg
     1321 cgaccgacgg tggcgatagg catccgggtg gtgctcaaaa gcagcttcgc ctggctgata
     1381 cgttggtcct cgcgccagct taagacgcta atccctaact gctggcggaa aagatgtgac
     1441 agacgcgacg gcgacaagca aacatgctgt gcgacgctgg cgatatcaaa attgctgtct
     1501 gccaggtgat cgctgatgta ctgacaagcc tcgcgtaccc gattatccat cggtggatgg
     1561 agcgactcgt taatcgcttc catgcgccgc agtaacaatt gctcaagcag atttatcgcc
     1621 agcagctccg aatagcgccc ttccccttgc ccggcgttaa tgatttgccc aaacaggtcg
     1681 ctgaaatgcg gctggtgcgc ttcatccggg cgaaagaacc ccgtattggc aaatattgac
     1741 ggccagttaa gccattcatg ccagtaggcg cgcggacgaa agtaaaccca ctggtgatac
     1801 cattcgcgag cctccggatg acgaccgtag tgatgaatct ctcctggcgg gaacagcaaa
     1861 atatcacccg gtcggcaaac aaattctcgt ccctgatttt tcaccacccc ctgaccgcga
     1921 atggtgagat tgagaatata acctttcatt cccagcggtc ggtcgataaa aaaatcgaga
     1981 taaccgttgg cctcaatcgg cgttaaaccc gccaccagat gggcattaaa cgagtatccc
     2041 ggcagcaggg gatcattttg cgcttcagcc atacttttca tactcccgcc attcagagaa
     2101 gaaaccaatt gtccatattg catcagacat tgccgtcact gcgtctttta ctggctcttc
     2161 tcgctaacca aaccggtaac cccgcttatt aaaagcattc tgtaacaaag cgggaccaaa
     2221 gccatgacaa aaacgcgtaa caaaagtgtc tataatcacg gcagaaaagt ccacattgat
     2281 tatttgcacg gcgtcacact ttgctatgcc atagcatttt tatccataag attagcggat
     2341 cctacctgac gctttttatc gcaactctct actgtttgaa ttcagaaaac atgaggatca
     2401 cccatgtctg cagggcccgg gacctgcagg catgcaagct ttaaccgggc cagcccgcct
     2461 aatgagcggg cttttttttc agcttggctg ttttggcgga tgagagaaga ttttcagcct
     2521 gatacagatt aaatcagaac gcagaagcgg tctgataaaa cagaatttgc ctggcggcag
     2581 tagcgcggtg gtcccacctg accccatgcc gaactcagaa gtgaaacgcc gtagcgccga
     2641 tggtagtgtg gggtctcccc atgcgagagt agggaactgc caggcatcaa ataaaacgaa
     2701 aggctcagtc gaaagactgg gcctttcgtt ttatctgttg tttgtcggtg aacgctctcc
     2761 tgagtaggac aaatccgccg ggagcggatt tgaacgttgc gaagcaacgg cccggagggt
     2821 ggcgggcagg acgcccgcca taaactgcca ggcatcaaat taagcagaag gccatcctga
     2881 cggatggcct ttttgcgttt ctacaaaccc tcaggcattt gagaagcaca cggtcacact
     2941 gcttccggta gtcaataaac cggtaaacca gcaatagaca taagcggcta tttaacgacc
     3001 ctgccctgaa ccgacgaccg ggtcatcgtg gccggatctt gcggcccctc ggcttgaacg
     3061 aattgttaga cattatttgc cgactacctt ggtgatctcg cctttcacgt agtggacaaa
     3121 ttcttccaac tgatctgcgc gcgaggccaa gcgatcttct tcttgtccaa gataagcctg
     3181 tctagcttca agtatgacgg gctgatactg ggccggcagg cgctccattg cccagtcggc
     3241 agcgacatcc ttcggcgcga ttttgccggt tactgcgctg taccaaatgc gggacaacgt
     3301 aagcactaca tttcgctcat cgccagccca gtcgggcggc gagttccata gcgttaaggt
     3361 ttcatttagc gcctcaaata gatcctgttc aggaaccgga tcaaagagtt cctccgccgc
     3421 tggacctacc aaggcaacgc tatgttctct tgcttttgtc agcaagatag ccagatcaat
     3481 gtcgatcgtg gctggctcga agatacctgc aagaatgtca ttgcgctgcc attctccaaa
     3541 ttgcagttcg cgcttagctg gataacgcca cggaatgatg tcgtcgtgca caacaatggt
     3601 gacttctaca gcgcggagaa tctcgctctc tccaggggaa gccgaagttt ccaaaaggtc
     3661 gttgatcaaa gctcgccgcg ttgtttcatc aagccttacg gtcaccgtaa ccagcaaatc
     3721 aatatcactg tgtggcttca ggccgccatc cactgcggag ccgtacaaat gtacggccag
     3781 caacgtcggt tcgagatggc gctcgatgac gccaactacc tctgatagtt gagtcgatac
     3841 ttcggcgatc accgcttccc tcatactctt cctttttcaa tattattgaa gcatttatca
     3901 gggttattgt ctcatgagcg gatacatatt tgaatgtatt tagaaaaata aacaaatagc
     3961 tagctcactc ggtcgctacg ctccgggcgt gagactgcgg cgggcgctgc gg
//
