LOCUS       sequence-451850-        3972 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  arabinose-induced synthesis of an RNA containing an MS2 hairpin 
            flanked by a 5GC clamp. Used as a three-hybrid control. For cloning 
            purposes, see plasmid #222403..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3972)
  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 3972)
  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: pSS1 - pBait-5GC[1xMS2]-empty
FEATURES             Location/Qualifiers
     source          1..3972
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      167..194
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     CDS             complement(374..1165)
                     /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(1166..1257)
                     /gene="bla"
                     /label=AmpR promoter
     CDS             complement(2507..3385)
                     /codon_start=1
                     /gene="araC"
                     /product="L-arabinose regulatory protein"
                     /label=araC
                     /translation="MAEAQNDPLLPGYSFNAHLVAGLTPIEANGYLDFFIDRPLGMKGY
                     ILNLTIRGQGVVKNQGREFVCRPGDILLFPPGEIHHYGRHPEAREWYHQWVYFRPRAYW
                     HEWLNWPSIFANTGFFRPDEAHQPHFSDLFGQIINAGQGEGRYSELLAINLLEQLLLRR
                     MEAINESLHPPMDNRVREACQYISDHLADSNFDIASVAQHVCLSPSRLSHLFRQQLGIS
                     VLSWREDQRISQAKLLLSTTRMPIATVGRNVGFDDQLYFSRVFKKCTGASPSEFRAGCE
                     EKVNDVAVKLS"
     primer_bind     3619..3638
                     /label=pBAD Forward
                     /note="For vectors with E. coli araBAD promoter, forward 
                     primer"
     misc_RNA        3708..3726
                     /label=MS2 stem loop
                     /note="stem loop that binds the bacteriophage MS2 coat 
                     protein"
     primer_bind     complement(3791..3808)
                     /label=pBAD Reverse
                     /note="For vectors with E. coli araBAD promoter, reverse 
                     primer"
     terminator      join(3961..3972,1..75)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
ORIGIN
        1 aaggctcagt cgaaagactg ggcctttcgt tttatctgtt gtttgtcggt gaacgctctc
       61 ctgagtagga caaatccgcc gggagcggat ttgaacgttg cgaagcaacg gcccggaggg
      121 tggcgggcag gacgcccgcc ataaactgcc aggcatcaaa ttaagcagaa ggccatcctg
      181 acggatggcc tttttgcgtt tctacaaacc ctcaggcatt tgagaagcac acggtcacac
      241 tgcttccggt agtcaataaa ccggtaaacc agcaatagac ataagcggct atttaacgac
      301 cctgccctga accgacgacc gggtcatcgt ggccggatct tgcggcccct cggcttgaac
      361 gaattgttag acattatttg ccgactacct tggtgatctc gcctttcacg tagtggacaa
      421 attcttccaa ctgatctgcg cgcgaggcca agcgatcttc ttcttgtcca agataagcct
      481 gtctagcttc aagtatgacg ggctgatact gggccggcag gcgctccatt gcccagtcgg
      541 cagcgacatc cttcggcgcg attttgccgg ttactgcgct gtaccaaatg cgggacaacg
      601 taagcactac atttcgctca tcgccagccc agtcgggcgg cgagttccat agcgttaagg
      661 tttcatttag cgcctcaaat agatcctgtt caggaaccgg atcaaagagt tcctccgccg
      721 ctggacctac caaggcaacg ctatgttctc ttgcttttgt cagcaagata gccagatcaa
      781 tgtcgatcgt ggctggctcg aagatacctg caagaatgtc attgcgctgc cattctccaa
      841 attgcagttc gcgcttagct ggataacgcc acggaatgat gtcgtcgtgc acaacaatgg
      901 tgacttctac agcgcggaga atctcgctct ctccagggga agccgaagtt tccaaaaggt
      961 cgttgatcaa agctcgccgc gttgtttcat caagccttac ggtcaccgta accagcaaat
     1021 caatatcact gtgtggcttc aggccgccat ccactgcgga gccgtacaaa tgtacggcca
     1081 gcaacgtcgg ttcgagatgg cgctcgatga cgccaactac ctctgatagt tgagtcgata
     1141 cttcggcgat caccgcttcc ctcatactct tcctttttca atattattga agcatttatc
     1201 agggttattg tctcatgagc ggatacatat ttgaatgtat ttagaaaaat aaacaaatag
     1261 ctagctcact cggtcgctac gctccgggcg tgagactgcg gcgggcgctg cggacacata
     1321 caaagttacc cacagattcc gtggataagc aggggactaa catgtgaggc aaaacagcag
     1381 ggccgcgccg gtggcgtttt tccataggct ccgccctcct gccagagttc acataaacag
     1441 acgcttttcc ggtgcatctg tgggagccgt gaggctcaac catgaatctg acagtacggg
     1501 cgaaacccga caggacttaa agatccccac cgtttccggc gggtcgctcc ctcttgcgct
     1561 ctcctgttcc gaccctgccg tttaccggat acctgttccg cctttctccc ttacgggaag
     1621 tgtggcgctt tctcatagct cacacactgg tatctcggct cggtgtaggt cgttcgctcc
     1681 aagctgggct gtaagcaaga actccccgtt cagcccgact gctgcgcctt atccggtaac
     1741 tgttcacttg agtccaaccc ggaaaagcac ggtaaaacgc cactggcagc agccattggt
     1801 aactgggagt tcgcagagga tttgtttagc taaacacgcg gttgctcttg aagtgtgcgc
     1861 caaagtccgg ctacactgga aggacagatt tggttgctgt gctctgcgaa agccagttac
     1921 cacggttaag cagttcccca actgacttaa ccttcgatca aaccacctcc ccaggtggtt
     1981 ttttcgaagg ttaagtcagt tggggaactg cttaaccttc gatcaaacca cctccccagg
     2041 tggttttttc gtttacaggg caaaagatta cgcgcagaaa aaaaggatct caagaagatc
     2101 ctttgatctt ttctactgaa ccgctctaga tttcagtgca atttatctct tcaaatgtag
     2161 cacctgaagt cagccccata cgatataagt tgtaattctc atgttagtca tgccccgcgc
     2221 ccaccggaag gagctgactg ggttgaaggc tctcaagggc atcggtcgag atcccggtgc
     2281 ctaatgagtg agctaactta cattaattgc gttgcgctca ctgcccgctt tccagtcggg
     2341 aaacctgtcg tgccagctgc attaatgaat cggccaacgc gcggggagag gcggtttgcg
     2401 tattgggcgc catcgatgca taatgtgcct gtcaaatgga cgaagcaggg attctgcaaa
     2461 ccctatgcta ctccgtcaag ccgtcaattg tctgattcgt taccaattat gacaacttga
     2521 cggctacatc attcactttt tcttcacaac cggcacggaa ctcgctcggg ctggccccgg
     2581 tgcatttttt aaatacccgc gagaaataga gttgatcgtc aaaaccaaca ttgcgaccga
     2641 cggtggcgat aggcatccgg gtggtgctca aaagcagctt cgcctggctg atacgttggt
     2701 cctcgcgcca gcttaagacg ctaatcccta actgctggcg gaaaagatgt gacagacgcg
     2761 acggcgacaa gcaaacatgc tgtgcgacgc tggcgatatc aaaattgctg tctgccaggt
     2821 gatcgctgat gtactgacaa gcctcgcgta cccgattatc catcggtgga tggagcgact
     2881 cgttaatcgc ttccatgcgc cgcagtaaca attgctcaag cagatttatc gccagcagct
     2941 ccgaatagcg cccttcccct tgcccggcgt taatgatttg cccaaacagg tcgctgaaat
     3001 gcggctggtg cgcttcatcc gggcgaaaga accccgtatt ggcaaatatt gacggccagt
     3061 taagccattc atgccagtag gcgcgcggac gaaagtaaac ccactggtga taccattcgc
     3121 gagcctccgg atgacgaccg tagtgatgaa tctctcctgg cgggaacagc aaaatatcac
     3181 ccggtcggca aacaaattct cgtccctgat ttttcaccac cccctgaccg cgaatggtga
     3241 gattgagaat ataacctttc attcccagcg gtcggtcgat aaaaaaatcg agataaccgt
     3301 tggcctcaat cggcgttaaa cccgccacca gatgggcatt aaacgagtat cccggcagca
     3361 ggggatcatt ttgcgcttca gccatacttt tcatactccc gccattcaga gaagaaacca
     3421 attgtccata ttgcatcaga cattgccgtc actgcgtctt ttactggctc ttctcgctaa
     3481 ccaaaccggt aaccccgctt attaaaagca ttctgtaaca aagcgggacc aaagccatga
     3541 caaaaacgcg taacaaaagt gtctataatc acggcagaaa agtccacatt gattatttgc
     3601 acggcgtcac actttgctat gccatagcat ttttatccat aagattagcg gatcctacct
     3661 gacgcttttt atcgcaactc tctactgttt gaattcaccg ggagaaaaca tgaggatcac
     3721 ccatgtctgc agcccgggac ctgcaggcat gcaagcttgg ctgttttggc ggatgagaga
     3781 agattttcag cctgatacag attaaatcag aacgcagaag cggtctgata aaacagaatt
     3841 tgcctggcgg cagtagcgcg gtggtcccac ctgaccccat gccgaactca gaagtgaaac
     3901 gccgtagcgc cgatggtagt gtggggtctc cccatgcgag agtagggaac tgccaggcat
     3961 caaataaaac ga
//
