LOCUS       sequence-78838-e        5932 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Gene encoding E. coli RNaseIII cloned as a N terminal translational 
            fussion with superfolder GFP. Expression driven by arabinose 
            (backbone is pBAD24)..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5932)
  AUTHORS   Malagon F
  TITLE     RNase III is required for localization to the nucleoid of the 5' 
            pre-rRNA leader and for optimal induction of rRNA synthesis in E. 
            coli.
  JOURNAL   RNA. 2013 Sep;19(9):1200-7. doi: 10.1261/rna.038588.113. Epub 2013 
            Jul 26.
  PUBMED    23893733
REFERENCE   2  (bases 1 to 5932)
  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: 
            "10.1261/rna.038588.113"; journalName: "RNA"; date: "2013-09"; 
            volume: "19"; issue: "9"; pages: "1200-7"
COMMENT     Sequence Label: pBAD24-rnc-sfGFP
FEATURES             Location/Qualifiers
     source          1..5932
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(96..974)
                     /codon_start=1
                     /gene="araC"
                     /product="L-arabinose regulatory protein"
                     /label=araC
                     /translation="MAEAQNDPLLPGYSFNAHLVAGLTPIEANGYLDFFIDRPLGMKGY
                     ILNLTIRGQGVVKNQGREFVCRPGDILLFPPGEIHHYGRHPEAREWYHQWVYFRPRAYW
                     HEWLNWPSIFANTGFFRPDEAHQPHFSDLFGQIINAGQGEGRYSELLAINLLEQLLLRR
                     MEAINESLHPPMDNRVREACQYISDHLADSNFDIASVAQHVCLSPSRLSHLFRQQLGIS
                     VLSWREDQRISQAKLLLSTTRMPIATVGRNVGFDDQLYFSRVFKKCTGASPSEFRAGCE
                     EKVNDVAVKLS"
     promoter        1001..1285
                     /gene="araBAD"
                     /label=araBAD promoter
                     /note="promoter of the L-arabinose operon of E. coli; the 
                     araC regulatory gene is transcribed in the opposite 
                     direction (Guzman et al., 1995)"
     primer_bind     1208..1227
                     /label=pBAD Forward
                     /note="For vectors with E. coli araBAD promoter, forward 
                     primer"
     CDS             2014..2724
                     /codon_start=1
                     /product="GFP variant that folds robustly even when fused 
                     to poorly folded proteins (Pedelacq et al., 2006)"
                     /label=superfolder GFP
                     /translation="RKGEELFTGVVPILVELDGDVNGHKFSVRGEGEGDATNGKLTLKF
                     ICTTGKLPVPWPTLVTTLTYGVQCFARYPDHMKQHDFFKSAMPEGYVQERTISFKDDGT
                     YKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNFNSHNVYITADKQKNGIKAN
                     FKIRHNVEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSVLSKDPNEKRDHMVLLEF
                     VTAAGITHGMDELYK"
     primer_bind     complement(2789..2806)
                     /label=pBAD Reverse
                     /note="For vectors with E. coli araBAD promoter, reverse 
                     primer"
     terminator      2959..3045
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      3137..3164
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     promoter        3183..3274
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3275..3343,3344..4135)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(3493..3512)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4177..4632
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4264..4283)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     rep_origin      4743..5331
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5232..5251
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5485..5502
                     /label=L4440
                     /note="L4440 vector, forward primer"
     primer_bind     5668..5690
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     5829..5848
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
ORIGIN
        1 atcgatgcat aatgtgcctg tcaaatggac gaagcaggga ttctgcaaac cctatgctac
       61 tccgtcaagc cgtcaattgt ctgattcgtt accaattatg acaacttgac ggctacatca
      121 ttcacttttt cttcacaacc ggcacggaac tcgctcgggc tggccccggt gcatttttta
      181 aatacccgcg agaaatagag ttgatcgtca aaaccaacat tgcgaccgac ggtggcgata
      241 ggcatccggg tggtgctcaa aagcagcttc gcctggctga tacgttggtc ctcgcgccag
      301 cttaagacgc taatccctaa ctgctggcgg aaaagatgtg acagacgcga cggcgacaag
      361 caaacatgct gtgcgacgct ggcgatatca aaattgctgt ctgccaggtg atcgctgatg
      421 tactgacaag cctcgcgtac ccgattatcc atcggtggat ggagcgactc gttaatcgct
      481 tccatgcgcc gcagtaacaa ttgctcaagc agatttatcg ccagcagctc cgaatagcgc
      541 ccttcccctt gcccggcgtt aatgatttgc ccaaacaggt cgctgaaatg cggctggtgc
      601 gcttcatccg ggcgaaagaa ccccgtattg gcaaatattg acggccagtt aagccattca
      661 tgccagtagg cgcgcggacg aaagtaaacc cactggtgat accattcgcg agcctccgga
      721 tgacgaccgt agtgatgaat ctctcctggc gggaacagca aaatatcacc cggtcggcaa
      781 acaaattctc gtccctgatt tttcaccacc ccctgaccgc gaatggtgag attgagaata
      841 taacctttca ttcccagcgg tcggtcgata aaaaaatcga gataaccgtt ggcctcaatc
      901 ggcgttaaac ccgccaccag atgggcatta aacgagtatc ccggcagcag gggatcattt
      961 tgcgcttcag ccatactttt catactcccg ccattcagag aagaaaccaa ttgtccatat
     1021 tgcatcagac attgccgtca ctgcgtcttt tactggctct tctcgctaac caaaccggta
     1081 accccgctta ttaaaagcat tctgtaacaa agcgggacca aagccatgac aaaaacgcgt
     1141 aacaaaagtg tctataatca cggcagaaaa gtccacattg attatttgca cggcgtcaca
     1201 ctttgctatg ccatagcatt tttatccata agattagcgg atcctacctg acgcttttta
     1261 tcgcaactct ctactgtttc tccatacccg tttttttggg ctagcaggag gaattcgcca
     1321 atgaacccca tcgtaattaa tcggcttcaa cggaagctgg gctacacttt taatcatcag
     1381 gaactgttgc agcaggcatt aactcatcgt agtgccagca gtaaacataa cgagcgttta
     1441 gaatttttag gcgactctat tctgagctac gttatcgcca atgcgcttta tcaccgtttc
     1501 cctcgtgtgg atgaaggcga tatgagccgg atgcgcgcca cgctggtccg tggcaatacg
     1561 ctggcggaac tggcgcgcga atttgagtta ggcgagtgct tacgtttagg gccaggtgaa
     1621 cttaaaagcg gtggatttcg tcgtgagtca attctcgccg acaccgtcga agcattaatt
     1681 ggtggcgtat tcctcgacag tgatattcaa accgtcgaga aattaatcct caactggtat
     1741 caaactcgtt tggacgaaat tagcccaggc gataaacaaa aagatccgaa aacgcgcttg
     1801 caagaatatt tgcagggtcg ccatctgccg ctgccgactt atctggtagt ccaggtacgt
     1861 ggcgaagcgc acgatcagga atttactatc cactgccagg tcagcggcct gagtgaaccg
     1921 gtggttggca caggttcaag ccgtcgtaag gctgagcagg ctgccgccga acaggcgttg
     1981 aaaaaactgg agctggaacc tcgagcccgg gcgcgtaaag gcgaagagct gttcactggt
     2041 gtcgtcccca ttctggtgga actggatggt gatgtcaacg gtcataagtt ttccgtgcgt
     2101 ggcgagggtg aaggtgacgc aactaatggt aaactgacgc tgaagttcat ctgtactact
     2161 ggtaaactgc cggtaccttg gccgactctg gtaacgacgc tgacttatgg tgttcagtgc
     2221 tttgctcgtt atccggacca tatgaagcag catgacttct tcaagtccgc catgccggaa
     2281 ggctatgtgc aggaacgcac gatttccttt aaggatgacg gcacgtacaa aacgcgtgcg
     2341 gaagtgaaat ttgaaggcga taccctggta aaccgcattg agctgaaagg cattgacttt
     2401 aaagaagacg gcaatatcct gggccataag ctggaataca attttaacag ccacaatgtt
     2461 tacatcaccg ccgataaaca aaaaaatggc attaaagcga attttaaaat tcgccacaac
     2521 gtggaggatg gcagcgtgca gctggctgat cactaccagc aaaacactcc aatcggtgat
     2581 ggtcctgttc tgctgccaga caatcactat ctgagcacgc aaagcgttct gtctaaagat
     2641 ccgaacgaga aacgcgatca tatggttctg ctggagttcg taaccgcagc gggcatcacg
     2701 catggtatgg atgaactgta caaaggtcta gagtcgacct gcaggcatgc aagcttggct
     2761 gttttggcgg atgagagaag attttcagcc tgatacagat taaatcagaa cgcagaagcg
     2821 gtctgataaa acagaatttg cctggcggca gtagcgcggt ggtcccacct gaccccatgc
     2881 cgaactcaga agtgaaacgc cgtagcgccg atggtagtgt ggggtctccc catgcgagag
     2941 tagggaactg ccaggcatca aataaaacga aaggctcagt cgaaagactg ggcctttcgt
     3001 tttatctgtt gtttgtcggt gaacgctctc ctgagtagga caaatccgcc gggagcggat
     3061 ttgaacgttg cgaagcaacg gcccggaggg tggcgggcag gacgcccgcc ataaactgcc
     3121 aggcatcaaa ttaagcagaa ggccatcctg acggatggcc tttttgcgtt tctacaaact
     3181 cttttgttta tttttctaaa tacattcaaa tatgtatccg ctcatgagac aataaccctg
     3241 ataaatgctt caataatatt gaaaaaggaa gagtatgagt attcaacatt tccgtgtcgc
     3301 ccttattccc ttttttgcgg cattttgcct tcctgttttt gctcacccag aaacgctggt
     3361 gaaagtaaaa gatgctgaag atcagttggg tgcacgagtg ggttacatcg aactggatct
     3421 caacagcggt aagatccttg agagttttcg ccccgaagaa cgttttccaa tgatgagcac
     3481 ttttaaagtt ctgctatgtg gcgcggtatt atcccgtgtt gacgccgggc aagagcaact
     3541 cggtcgccgc atacactatt ctcagaatga cttggttgag tactcaccag tcacagaaaa
     3601 gcatcttacg gatggcatga cagtaagaga attatgcagt gctgccataa ccatgagtga
     3661 taacactgcg gccaacttac ttctgacaac gatcggagga ccgaaggagc taaccgcttt
     3721 tttgcacaac atgggggatc atgtaactcg ccttgatcgt tgggaaccgg agctgaatga
     3781 agccatacca aacgacgagc gtgacaccac gatgcctgta gcaatggcaa caacgttgcg
     3841 caaactatta actggcgaac tacttactct agcttcccgg caacaattaa tagactggat
     3901 ggaggcggat aaagttgcag gaccacttct gcgctcggcc cttccggctg gctggtttat
     3961 tgctgataaa tctggagccg gtgagcgtgg gtctcgcggt atcattgcag cactggggcc
     4021 agatggtaag ccctcccgta tcgtagttat ctacacgacg gggagtcagg caactatgga
     4081 tgaacgaaat agacagatcg ctgagatagg tgcctcactg attaagcatt ggtaactgtc
     4141 agaccaagtt tactcatata tactttagat tgatttacgc gccctgtagc ggcgcattaa
     4201 gcgcggcggg tgtggtggtt acgcgcagcg tgaccgctac acttgccagc gccctagcgc
     4261 ccgctccttt cgctttcttc ccttcctttc tcgccacgtt cgccggcttt ccccgtcaag
     4321 ctctaaatcg ggggctccct ttagggttcc gatttagtgc tttacggcac ctcgacccca
     4381 aaaaacttga tttgggtgat ggttcacgta gtgggccatc gccctgatag acggtttttc
     4441 gccctttgac gttggagtcc acgttcttta atagtggact cttgttccaa acttgaacaa
     4501 cactcaaccc tatctcgggc tattcttttg atttataagg gattttgccg atttcggcct
     4561 attggttaaa aaatgagctg atttaacaaa aatttaacgc gaattttaac aaaatattaa
     4621 cgtttacaat ttaaaaggat ctaggtgaag atcctttttg ataatctcat gaccaaaatc
     4681 ccttaacgtg agttttcgtt ccactgagcg tcagaccccg tagaaaagat caaaggatct
     4741 tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta
     4801 ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc tttttccgaa ggtaactggc
     4861 ttcagcagag cgcagatacc aaatactgtc cttctagtgt agccgtagtt aggccaccac
     4921 ttcaagaact ctgtagcacc gcctacatac ctcgctctgc taatcctgtt accagtggct
     4981 gctgccagtg gcgataagtc gtgtcttacc gggttggact caagacgata gttaccggat
     5041 aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac agcccagctt ggagcgaacg
     5101 acctacaccg aactgagata cctacagcgt gagctatgag aaagcgccac gcttcccgaa
     5161 gggagaaagg cggacaggta tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg
     5221 gagcttccag ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg ccacctctga
     5281 cttgagcgtc gatttttgtg atgctcgtca ggggggcgga gcctatggaa aaacgccagc
     5341 aacgcggcct ttttacggtt cctggccttt tgctggcctt ttgctcacat gttctttcct
     5401 gcgttatccc ctgattctgt ggataaccgt attaccgcct ttgagtgagc tgataccgct
     5461 cgccgcagcc gaacgaccga gcgcagcgag tcagtgagcg aggaagcgga agagcgcctg
     5521 atgcggtatt ttctccttac gcatctgtgc ggtatttcac accgcatagg gtcatggctg
     5581 cgccccgaca cccgccaaca cccgctgacg cgccctgacg ggcttgtctg ctcccggcat
     5641 ccgcttacag acaagctgtg accgtctccg ggagctgcat gtgtcagagg ttttcaccgt
     5701 catcaccgaa acgcgcgagg cagcaaggag atggcgccca acagtccccc ggccacgggg
     5761 cctgccacca tacccacgcc gaaacaagcg ctcatgagcc cgaagtggcg agcccgatct
     5821 tccccatcgg tgatgtcggc gatataggcg ccagcaaccg cacctgtggc gccggtgatg
     5881 ccggccacga tgcgtccggc gtagaggatc tgctcatgtt tgacagctta tc
//
