LOCUS       sequence-451849-        4051 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  arabinose-induced synthesis of a hybrid RNA with an MS2 hairpin 
            surrounded by a 5bp GC clamp with E coli McaS 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 4051)
  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 4051)
  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: pLN34 - pBait-5GC[1xMS2]-McaS
FEATURES             Location/Qualifiers
     source          1..4051
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        894..985
                     /gene="bla"
                     /label=AmpR promoter
     CDS             986..1777
                     /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"
     terminator      complement(1957..1984)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(2076..2162)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     2315..2332
                     /label=pBAD Reverse
                     /note="For vectors with E. coli araBAD promoter, reverse 
                     primer"
     misc_RNA        complement(2476..2494)
                     /label=MS2 stem loop
                     /note="stem loop that binds the bacteriophage MS2 coat 
                     protein"
     primer_bind     complement(2564..2583)
                     /label=pBAD Forward
                     /note="For vectors with E. coli araBAD promoter, forward 
                     primer"
     CDS             2817..3695
                     /codon_start=1
                     /gene="araC"
                     /product="L-arabinose regulatory protein"
                     /label=araC
                     /translation="MAEAQNDPLLPGYSFNAHLVAGLTPIEANGYLDFFIDRPLGMKGY
                     ILNLTIRGQGVVKNQGREFVCRPGDILLFPPGEIHHYGRHPEAREWYHQWVYFRPRAYW
                     HEWLNWPSIFANTGFFRPDEAHQPHFSDLFGQIINAGQGEGRYSELLAINLLEQLLLRR
                     MEAINESLHPPMDNRVREACQYISDHLADSNFDIASVAQHVCLSPSRLSHLFRQQLGIS
                     VLSWREDQRISQAKLLLSTTRMPIATVGRNVGFDDQLYFSRVFKKCTGASPSEFRAGCE
                     EKVNDVAVKLS"
ORIGIN
        1 agagataaat tgcactgaaa tctagagcgg ttcagtagaa aagatcaaag gatcttcttg
       61 agatcctttt tttctgcgcg taatcttttg ccctgtaaac gaaaaaacca cctggggagg
      121 tggtttgatc gaaggttaag cagttcccca actgacttaa ccttcgaaaa aaccacctgg
      181 ggaggtggtt tgatcgaagg ttaagtcagt tggggaactg cttaaccgtg gtaactggct
      241 ttcgcagagc acagcaacca aatctgtcct tccagtgtag ccggactttg gcgcacactt
      301 caagagcaac cgcgtgttta gctaaacaaa tcctctgcga actcccagtt accaatggct
      361 gctgccagtg gcgttttacc gtgcttttcc gggttggact caagtgaaca gttaccggat
      421 aaggcgcagc agtcgggctg aacggggagt tcttgcttac agcccagctt ggagcgaacg
      481 acctacaccg agccgagata ccagtgtgtg agctatgaga aagcgccaca cttcccgtaa
      541 gggagaaagg cggaacaggt atccggtaaa cggcagggtc ggaacaggag agcgcaagag
      601 ggagcgaccc gccggaaacg gtggggatct ttaagtcctg tcgggtttcg cccgtactgt
      661 cagattcatg gttgagcctc acggctccca cagatgcacc ggaaaagcgt ctgtttatgt
      721 gaactctggc aggagggcgg agcctatgga aaaacgccac cggcgcggcc ctgctgtttt
      781 gcctcacatg ttagtcccct gcttatccac ggaatctgtg ggtaactttg tatgtgtccg
      841 cagcgcccgc cgcagtctca cgcccggagc gtagcgaccg agtgagctag ctatttgttt
      901 atttttctaa atacattcaa atatgtatcc gctcatgaga caataaccct gataaatgct
      961 tcaataatat tgaaaaagga agagtatgag ggaagcggtg atcgccgaag tatcgactca
     1021 actatcagag gtagttggcg tcatcgagcg ccatctcgaa ccgacgttgc tggccgtaca
     1081 tttgtacggc tccgcagtgg atggcggcct gaagccacac agtgatattg atttgctggt
     1141 tacggtgacc gtaaggcttg atgaaacaac gcggcgagct ttgatcaacg accttttgga
     1201 aacttcggct tcccctggag agagcgagat tctccgcgct gtagaagtca ccattgttgt
     1261 gcacgacgac atcattccgt ggcgttatcc agctaagcgc gaactgcaat ttggagaatg
     1321 gcagcgcaat gacattcttg caggtatctt cgagccagcc acgatcgaca ttgatctggc
     1381 tatcttgctg acaaaagcaa gagaacatag cgttgccttg gtaggtccag cggcggagga
     1441 actctttgat ccggttcctg aacaggatct atttgaggcg ctaaatgaaa ccttaacgct
     1501 atggaactcg ccgcccgact gggctggcga tgagcgaaat gtagtgctta cgttgtcccg
     1561 catttggtac agcgcagtaa ccggcaaaat cgcgccgaag gatgtcgctg ccgactgggc
     1621 aatggagcgc ctgccggccc agtatcagcc cgtcatactt gaagctagac aggcttatct
     1681 tggacaagaa gaagatcgct tggcctcgcg cgcagatcag ttggaagaat ttgtccacta
     1741 cgtgaaaggc gagatcacca aggtagtcgg caaataatgt ctaacaattc gttcaagccg
     1801 aggggccgca agatccggcc acgatgaccc ggtcgtcggt tcagggcagg gtcgttaaat
     1861 agccgcttat gtctattgct ggtttaccgg tttattgact accggaagca gtgtgaccgt
     1921 gtgcttctca aatgcctgag ggtttgtaga aacgcaaaaa ggccatccgt caggatggcc
     1981 ttctgcttaa tttgatgcct ggcagtttat ggcgggcgtc ctgcccgcca ccctccgggc
     2041 cgttgcttcg caacgttcaa atccgctccc ggcggatttg tcctactcag gagagcgttc
     2101 accgacaaac aacagataaa acgaaaggcc cagtctttcg actgagcctt tcgttttatt
     2161 tgatgcctgg cagttcccta ctctcgcatg gggagacccc acactaccat cggcgctacg
     2221 gcgtttcact tctgagttcg gcatggggtc aggtgggacc accgcgctac tgccgccagg
     2281 caaattctgt tttatcagac cgcttctgcg ttctgattta atctgtatca ggctgaaaat
     2341 cttctctcat ccgccaaaac agccaagctt aaaaaataga gtctgtcgac atccgccaga
     2401 ctctacagta cacacagcag tgcatccgcg tcttaaatcc ggcattgtct cctctgcgcc
     2461 ggtcccgggc tgcagacatg ggtgatcctc atgttttctc ccggtgaatt caaacagtag
     2521 agagttgcga taaaaagcgt caggtaggat ccgctaatct tatggataaa aatgctatgg
     2581 catagcaaag tgtgacgccg tgcaaataat caatgtggac ttttctgccg tgattataga
     2641 cacttttgtt acgcgttttt gtcatggctt tggtcccgct ttgttacaga atgcttttaa
     2701 taagcggggt taccggtttg gttagcgaga agagccagta aaagacgcag tgacggcaat
     2761 gtctgatgca atatggacaa ttggtttctt ctctgaatgg cgggagtatg aaaagtatgg
     2821 ctgaagcgca aaatgatccc ctgctgccgg gatactcgtt taatgcccat ctggtggcgg
     2881 gtttaacgcc gattgaggcc aacggttatc tcgatttttt tatcgaccga ccgctgggaa
     2941 tgaaaggtta tattctcaat ctcaccattc gcggtcaggg ggtggtgaaa aatcagggac
     3001 gagaatttgt ttgccgaccg ggtgatattt tgctgttccc gccaggagag attcatcact
     3061 acggtcgtca tccggaggct cgcgaatggt atcaccagtg ggtttacttt cgtccgcgcg
     3121 cctactggca tgaatggctt aactggccgt caatatttgc caatacgggg ttctttcgcc
     3181 cggatgaagc gcaccagccg catttcagcg acctgtttgg gcaaatcatt aacgccgggc
     3241 aaggggaagg gcgctattcg gagctgctgg cgataaatct gcttgagcaa ttgttactgc
     3301 ggcgcatgga agcgattaac gagtcgctcc atccaccgat ggataatcgg gtacgcgagg
     3361 cttgtcagta catcagcgat cacctggcag acagcaattt tgatatcgcc agcgtcgcac
     3421 agcatgtttg cttgtcgccg tcgcgtctgt cacatctttt ccgccagcag ttagggatta
     3481 gcgtcttaag ctggcgcgag gaccaacgta tcagccaggc gaagctgctt ttgagcacca
     3541 cccggatgcc tatcgccacc gtcggtcgca atgttggttt tgacgatcaa ctctatttct
     3601 cgcgggtatt taaaaaatgc accggggcca gcccgagcga gttccgtgcc ggttgtgaag
     3661 aaaaagtgaa tgatgtagcc gtcaagttgt cataattggt aacgaatcag acaattgacg
     3721 gcttgacgga gtagcatagg gtttgcagaa tccctgcttc gtccatttga caggcacatt
     3781 atgcatcgat ggcgcccaat acgcaaaccg cctctccccg cgcgttggcc gattcattaa
     3841 tgcagctggc acgacaggtt tcccgactgg aaagcgggca gtgagcgcaa cgcaattaat
     3901 gtaagttagc tcactcatta ggcaccggga tctcgaccga tgcccttgag agccttcaac
     3961 ccagtcagct ccttccggtg ggcgcggggc atgactaaca tgagaattac aacttatatc
     4021 gtatggggct gacttcaggt gctacatttg a
//
