LOCUS       sequence-451848-        4108 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  arabinose-induced synthesis of a hybrid RNA containing a 5' MS2 
            hairpin, E. coli chiP 5'UTR insert between XmaI and HindIII sites, 
            surrounded by a 7bp GC clamp, and followed by a 3' trpA terminator..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4108)
  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 4108)
  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: pLN43 - pBait-1xMS2-7GC[chiP-5']-TtrpA
FEATURES             Location/Qualifiers
     source          1..4108
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     278..297
                     /label=pBAD Forward
                     /note="For vectors with E. coli araBAD promoter, forward 
                     primer"
     misc_RNA        361..379
                     /label=MS2 stem loop
                     /note="stem loop that binds the bacteriophage MS2 coat 
                     protein"
     primer_bind     complement(586..603)
                     /label=pBAD Reverse
                     /note="For vectors with E. coli araBAD promoter, reverse 
                     primer"
     terminator      756..842
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      934..961
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     CDS             complement(1141..1932)
                     /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(1933..2024)
                     /gene="bla"
                     /label=AmpR promoter
     CDS             complement(join(3274..4108,1..44))
                     /codon_start=1
                     /gene="araC"
                     /product="L-arabinose regulatory protein"
                     /label=araC
                     /translation="MAEAQNDPLLPGYSFNAHLVAGLTPIEANGYLDFFIDRPLGMKGY
                     ILNLTIRGQGVVKNQGREFVCRPGDILLFPPGEIHHYGRHPEAREWYHQWVYFRPRAYW
                     HEWLNWPSIFANTGFFRPDEAHQPHFSDLFGQIINAGQGEGRYSELLAINLLEQLLLRR
                     MEAINESLHPPMDNRVREACQYISDHLADSNFDIASVAQHVCLSPSRLSHLFRQQLGIS
                     VLSWREDQRISQAKLLLSTTRMPIATVGRNVGFDDQLYFSRVFKKCTGASPSEFRAGCE
                     EKVNDVAVKLS"
ORIGIN
        1 aacgagtatc ccggcagcag gggatcattt tgcgcttcag ccatactttt catactcccg
       61 ccattcagag aagaaaccaa ttgtccatat tgcatcagac attgccgtca ctgcgtcttt
      121 tactggctct tctcgctaac caaaccggta accccgctta ttaaaagcat tctgtaacaa
      181 agcgggacca aagccatgac aaaaacgcgt aacaaaagtg tctataatca cggcagaaaa
      241 gtccacattg attatttgca cggcgtcaca ctttgctatg ccatagcatt tttatccata
      301 agattagcgg atcctacctg acgcttttta tcgcaactct ctactgtttg aattcagaaa
      361 acatgaggat cacccatgtc tgcagggccc ggggtagtca gcgagacttt tctcaacgct
      421 acttttttaa tttttatttt ttcgctgttc acctttggtg cagcaattta tacgtcaaag
      481 aggattaacc catgcgtacg tttaagcttt aaccgggcca gcccgcctaa tgagcgggct
      541 tttttttcag cttggctgtt ttggcggatg agagaagatt ttcagcctga tacagattaa
      601 atcagaacgc agaagcggtc tgataaaaca gaatttgcct ggcggcagta gcgcggtggt
      661 cccacctgac cccatgccga actcagaagt gaaacgccgt agcgccgatg gtagtgtggg
      721 gtctccccat gcgagagtag ggaactgcca ggcatcaaat aaaacgaaag gctcagtcga
      781 aagactgggc ctttcgtttt atctgttgtt tgtcggtgaa cgctctcctg agtaggacaa
      841 atccgccggg agcggatttg aacgttgcga agcaacggcc cggagggtgg cgggcaggac
      901 gcccgccata aactgccagg catcaaatta agcagaaggc catcctgacg gatggccttt
      961 ttgcgtttct acaaaccctc aggcatttga gaagcacacg gtcacactgc ttccggtagt
     1021 caataaaccg gtaaaccagc aatagacata agcggctatt taacgaccct gccctgaacc
     1081 gacgaccggg tcatcgtggc cggatcttgc ggcccctcgg cttgaacgaa ttgttagaca
     1141 ttatttgccg actaccttgg tgatctcgcc tttcacgtag tggacaaatt cttccaactg
     1201 atctgcgcgc gaggccaagc gatcttcttc ttgtccaaga taagcctgtc tagcttcaag
     1261 tatgacgggc tgatactggg ccggcaggcg ctccattgcc cagtcggcag cgacatcctt
     1321 cggcgcgatt ttgccggtta ctgcgctgta ccaaatgcgg gacaacgtaa gcactacatt
     1381 tcgctcatcg ccagcccagt cgggcggcga gttccatagc gttaaggttt catttagcgc
     1441 ctcaaataga tcctgttcag gaaccggatc aaagagttcc tccgccgctg gacctaccaa
     1501 ggcaacgcta tgttctcttg cttttgtcag caagatagcc agatcaatgt cgatcgtggc
     1561 tggctcgaag atacctgcaa gaatgtcatt gcgctgccat tctccaaatt gcagttcgcg
     1621 cttagctgga taacgccacg gaatgatgtc gtcgtgcaca acaatggtga cttctacagc
     1681 gcggagaatc tcgctctctc caggggaagc cgaagtttcc aaaaggtcgt tgatcaaagc
     1741 tcgccgcgtt gtttcatcaa gccttacggt caccgtaacc agcaaatcaa tatcactgtg
     1801 tggcttcagg ccgccatcca ctgcggagcc gtacaaatgt acggccagca acgtcggttc
     1861 gagatggcgc tcgatgacgc caactacctc tgatagttga gtcgatactt cggcgatcac
     1921 cgcttccctc atactcttcc tttttcaata ttattgaagc atttatcagg gttattgtct
     1981 catgagcgga tacatatttg aatgtattta gaaaaataaa caaatagcta gctcactcgg
     2041 tcgctacgct ccgggcgtga gactgcggcg ggcgctgcgg acacatacaa agttacccac
     2101 agattccgtg gataagcagg ggactaacat gtgaggcaaa acagcagggc cgcgccggtg
     2161 gcgtttttcc ataggctccg ccctcctgcc agagttcaca taaacagacg cttttccggt
     2221 gcatctgtgg gagccgtgag gctcaaccat gaatctgaca gtacgggcga aacccgacag
     2281 gacttaaaga tccccaccgt ttccggcggg tcgctccctc ttgcgctctc ctgttccgac
     2341 cctgccgttt accggatacc tgttccgcct ttctccctta cgggaagtgt ggcgctttct
     2401 catagctcac acactggtat ctcggctcgg tgtaggtcgt tcgctccaag ctgggctgta
     2461 agcaagaact ccccgttcag cccgactgct gcgccttatc cggtaactgt tcacttgagt
     2521 ccaacccgga aaagcacggt aaaacgccac tggcagcagc cattggtaac tgggagttcg
     2581 cagaggattt gtttagctaa acacgcggtt gctcttgaag tgtgcgccaa agtccggcta
     2641 cactggaagg acagatttgg ttgctgtgct ctgcgaaagc cagttaccac ggttaagcag
     2701 ttccccaact gacttaacct tcgatcaaac cacctcccca ggtggttttt tcgaaggtta
     2761 agtcagttgg ggaactgctt aaccttcgat caaaccacct ccccaggtgg ttttttcgtt
     2821 tacagggcaa aagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc
     2881 tactgaaccg ctctagattt cagtgcaatt tatctcttca aatgtagcac ctgaagtcag
     2941 ccccatacga tataagttgt aattctcatg ttagtcatgc cccgcgccca ccggaaggag
     3001 ctgactgggt tgaaggctct caagggcatc ggtcgagatc ccggtgccta atgagtgagc
     3061 taacttacat taattgcgtt gcgctcactg cccgctttcc agtcgggaaa cctgtcgtgc
     3121 cagctgcatt aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat tgggcgccat
     3181 cgatgcataa tgtgcctgtc aaatggacga agcagggatt ctgcaaaccc tatgctactc
     3241 cgtcaagccg tcaattgtct gattcgttac caattatgac aacttgacgg ctacatcatt
     3301 cactttttct tcacaaccgg cacggaactc gctcgggctg gccccggtgc attttttaaa
     3361 tacccgcgag aaatagagtt gatcgtcaaa accaacattg cgaccgacgg tggcgatagg
     3421 catccgggtg gtgctcaaaa gcagcttcgc ctggctgata cgttggtcct cgcgccagct
     3481 taagacgcta atccctaact gctggcggaa aagatgtgac agacgcgacg gcgacaagca
     3541 aacatgctgt gcgacgctgg cgatatcaaa attgctgtct gccaggtgat cgctgatgta
     3601 ctgacaagcc tcgcgtaccc gattatccat cggtggatgg agcgactcgt taatcgcttc
     3661 catgcgccgc agtaacaatt gctcaagcag atttatcgcc agcagctccg aatagcgccc
     3721 ttccccttgc ccggcgttaa tgatttgccc aaacaggtcg ctgaaatgcg gctggtgcgc
     3781 ttcatccggg cgaaagaacc ccgtattggc aaatattgac ggccagttaa gccattcatg
     3841 ccagtaggcg cgcggacgaa agtaaaccca ctggtgatac cattcgcgag cctccggatg
     3901 acgaccgtag tgatgaatct ctcctggcgg gaacagcaaa atatcacccg gtcggcaaac
     3961 aaattctcgt ccctgatttt tcaccacccc ctgaccgcga atggtgagat tgagaatata
     4021 acctttcatt cccagcggtc ggtcgataaa aaaatcgaga taaccgttgg cctcaatcgg
     4081 cgttaaaccc gccaccagat gggcatta
//
