LOCUS       sequence-78831-e        5257 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Superfolder GFP ORF cloned into pBAD24 for expression in E. coli. It
            contains additional aminoacids in the N and C terminus (polilinker 
            sequences for cloning proteins in frame with GFP).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5257)
  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 5257)
  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-sfGFPx2
FEATURES             Location/Qualifiers
     source          1..5257
                     /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             1339..2049
                     /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
                     YKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLECNFNSHNVYITADKQKNGIKAN
                     FKIRHNVEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSVLSIDPNEKRDHMVLLEF
                     VTAAGITHGMDELYK"
     primer_bind     complement(2114..2131)
                     /label=pBAD Reverse
                     /note="For vectors with E. coli araBAD promoter, reverse 
                     primer"
     terminator      2284..2370
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      2462..2489
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     promoter        2508..2599
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2600..2668,2669..3460)
                     /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(2818..2837)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3502..3957
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3589..3608)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     rep_origin      4068..4656
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4557..4576
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4810..4827
                     /label=L4440
                     /note="L4440 vector, forward primer"
     primer_bind     4993..5015
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     5154..5173
                     /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 atggctcgag cccgggcgcg taaaggcgaa gagctgttca ctggtgtcgt ccccattctg
     1381 gtggaactgg atggtgatgt caacggtcat aagttttccg tgcgtggcga gggtgaaggt
     1441 gacgcaacta atggtaaact gacgctgaag ttcatctgta ctactggtaa actgccggta
     1501 ccttggccga ctctggtaac gacgctgact tatggtgttc agtgctttgc tcgttatccg
     1561 gaccatatga agcagcatga cttcttcaag tccgccatgc cggaaggcta tgtgcaggaa
     1621 cgcacgattt cctttaagga tgacggcacg tacaaaacgc gtgcggaagt gaaatttgaa
     1681 ggcgataccc tggtaaaccg cattgagctg aaaggcattg actttaaaga agacggcaat
     1741 atcctgggcc ataagctgga atgcaatttt aacagccaca atgtttacat caccgccgat
     1801 aaacaaaaaa atggcattaa agcgaatttt aaaattcgcc acaacgtgga ggatggcagc
     1861 gtgcagctgg ctgatcacta ccagcaaaac actccaatcg gtgatggtcc tgttctgctg
     1921 ccagacaatc actatctgag cacgcaaagc gttctgtcta tagatccgaa cgagaaacgc
     1981 gatcatatgg ttctgctgga gttcgtaacc gcagcgggca tcacgcatgg tatggatgaa
     2041 ctgtacaaag gtctagagtc gacctgcagg catgcaagct tggctgtttt ggcggatgag
     2101 agaagatttt cagcctgata cagattaaat cagaacgcag aagcggtctg ataaaacaga
     2161 atttgcctgg cggcagtagc gcggtggtcc cacctgaccc catgccgaac tcagaagtga
     2221 aacgccgtag cgccgatggt agtgtggggt ctccccatgc gagagtaggg aactgccagg
     2281 catcaaataa aacgaaaggc tcagtcgaaa gactgggcct ttcgttttat ctgttgtttg
     2341 tcggtgaacg ctctcctgag taggacaaat ccgccgggag cggatttgaa cgttgcgaag
     2401 caacggcccg gagggtggcg ggcaggacgc ccgccataaa ctgccaggca tcaaattaag
     2461 cagaaggcca tcctgacgga tggccttttt gcgtttctac aaactctttt gtttattttt
     2521 ctaaatacat tcaaatatgt atccgctcat gagacaataa ccctgataaa tgcttcaata
     2581 atattgaaaa aggaagagta tgagtattca acatttccgt gtcgccctta ttcccttttt
     2641 tgcggcattt tgccttcctg tttttgctca cccagaaacg ctggtgaaag taaaagatgc
     2701 tgaagatcag ttgggtgcac gagtgggtta catcgaactg gatctcaaca gcggtaagat
     2761 ccttgagagt tttcgccccg aagaacgttt tccaatgatg agcactttta aagttctgct
     2821 atgtggcgcg gtattatccc gtgttgacgc cgggcaagag caactcggtc gccgcataca
     2881 ctattctcag aatgacttgg ttgagtactc accagtcaca gaaaagcatc ttacggatgg
     2941 catgacagta agagaattat gcagtgctgc cataaccatg agtgataaca ctgcggccaa
     3001 cttacttctg acaacgatcg gaggaccgaa ggagctaacc gcttttttgc acaacatggg
     3061 ggatcatgta actcgccttg atcgttggga accggagctg aatgaagcca taccaaacga
     3121 cgagcgtgac accacgatgc ctgtagcaat ggcaacaacg ttgcgcaaac tattaactgg
     3181 cgaactactt actctagctt cccggcaaca attaatagac tggatggagg cggataaagt
     3241 tgcaggacca cttctgcgct cggcccttcc ggctggctgg tttattgctg ataaatctgg
     3301 agccggtgag cgtgggtctc gcggtatcat tgcagcactg gggccagatg gtaagccctc
     3361 ccgtatcgta gttatctaca cgacggggag tcaggcaact atggatgaac gaaatagaca
     3421 gatcgctgag ataggtgcct cactgattaa gcattggtaa ctgtcagacc aagtttactc
     3481 atatatactt tagattgatt tacgcgccct gtagcggcgc attaagcgcg gcgggtgtgg
     3541 tggttacgcg cagcgtgacc gctacacttg ccagcgccct agcgcccgct cctttcgctt
     3601 tcttcccttc ctttctcgcc acgttcgccg gctttccccg tcaagctcta aatcgggggc
     3661 tccctttagg gttccgattt agtgctttac ggcacctcga ccccaaaaaa cttgatttgg
     3721 gtgatggttc acgtagtggg ccatcgccct gatagacggt ttttcgccct ttgacgttgg
     3781 agtccacgtt ctttaatagt ggactcttgt tccaaacttg aacaacactc aaccctatct
     3841 cgggctattc ttttgattta taagggattt tgccgatttc ggcctattgg ttaaaaaatg
     3901 agctgattta acaaaaattt aacgcgaatt ttaacaaaat attaacgttt acaatttaaa
     3961 aggatctagg tgaagatcct ttttgataat ctcatgacca aaatccctta acgtgagttt
     4021 tcgttccact gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg agatcctttt
     4081 tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt
     4141 ttgccggatc aagagctacc aactcttttt ccgaaggtaa ctggcttcag cagagcgcag
     4201 ataccaaata ctgtccttct agtgtagccg tagttaggcc accacttcaa gaactctgta
     4261 gcaccgccta catacctcgc tctgctaatc ctgttaccag tggctgctgc cagtggcgat
     4321 aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac cggataaggc gcagcggtcg
     4381 ggctgaacgg ggggttcgtg cacacagccc agcttggagc gaacgaccta caccgaactg
     4441 agatacctac agcgtgagct atgagaaagc gccacgcttc ccgaagggag aaaggcggac
     4501 aggtatccgg taagcggcag ggtcggaaca ggagagcgca cgagggagct tccaggggga
     4561 aacgcctggt atctttatag tcctgtcggg tttcgccacc tctgacttga gcgtcgattt
     4621 ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc ggccttttta
     4681 cggttcctgg ccttttgctg gccttttgct cacatgttct ttcctgcgtt atcccctgat
     4741 tctgtggata accgtattac cgcctttgag tgagctgata ccgctcgccg cagccgaacg
     4801 accgagcgca gcgagtcagt gagcgaggaa gcggaagagc gcctgatgcg gtattttctc
     4861 cttacgcatc tgtgcggtat ttcacaccgc atagggtcat ggctgcgccc cgacacccgc
     4921 caacacccgc tgacgcgccc tgacgggctt gtctgctccc ggcatccgct tacagacaag
     4981 ctgtgaccgt ctccgggagc tgcatgtgtc agaggttttc accgtcatca ccgaaacgcg
     5041 cgaggcagca aggagatggc gcccaacagt cccccggcca cggggcctgc caccataccc
     5101 acgccgaaac aagcgctcat gagcccgaag tggcgagccc gatcttcccc atcggtgatg
     5161 tcggcgatat aggcgccagc aaccgcacct gtggcgccgg tgatgccggc cacgatgcgt
     5221 ccggcgtaga ggatctgctc atgtttgaca gcttatc
//
