LOCUS       sequence-78830-e        5228 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Superfolder GFP ORF cloned into pBAD24 for expression in E. coli.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5228)
  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 5228)
  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-sfGFPx1
FEATURES             Location/Qualifiers
     source          1..5228
                     /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             1321..2034
                     /codon_start=1
                     /product="GFP variant that folds robustly even when fused 
                     to poorly folded proteins (Pedelacq et al., 2006)"
                     /label=superfolder GFP
                     /translation="MRKGEELFTGVVPILVELDGDVNGHKFSVRGEGEGDATNGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFARYPDHMKQHDFFKSAMPEGYVQERTISFKDDG
                     TYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNFNSHNVYITADKQKNGIKA
                     NFKIRHNVEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSVLSKDPNEKRDHMVLLE
                     FVTAAGITHGMDELYK"
     primer_bind     complement(2085..2102)
                     /label=pBAD Reverse
                     /note="For vectors with E. coli araBAD promoter, reverse 
                     primer"
     terminator      2255..2341
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      2433..2460
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     promoter        2479..2570
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2571..2639,2640..3431)
                     /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(2789..2808)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3473..3928
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3560..3579)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     rep_origin      4039..4627
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4528..4547
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4781..4798
                     /label=L4440
                     /note="L4440 vector, forward primer"
     primer_bind     4964..4986
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     5125..5144
                     /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 atgcgtaaag gcgaagagct gttcactggt gtcgtcccta ttctggtgga actggatggt
     1381 gatgtcaacg gtcataagtt ttccgtgcgt ggcgagggtg aaggtgacgc aactaatggt
     1441 aaactgacgc tgaagttcat ctgtactact ggtaaactgc cggtaccttg gccgactctg
     1501 gtaacgacgc tgacttatgg tgttcagtgc tttgctcgtt atccggacca tatgaagcag
     1561 catgacttct tcaagtccgc catgccggaa ggctatgtgc aggaacgcac gatttccttt
     1621 aaggatgacg gcacgtacaa aacgcgtgcg gaagtgaaat ttgaaggcga taccctggta
     1681 aaccgcattg agctgaaagg cattgacttt aaagaagacg gcaatatcct gggccataag
     1741 ctggaataca attttaacag ccacaatgtt tacatcaccg ccgataaaca aaaaaatggc
     1801 attaaagcga attttaaaat tcgccacaac gtggaggatg gcagcgtgca gctggctgat
     1861 cactaccagc aaaacactcc aatcggtgat ggtcctgttc tgctgccaga caatcactat
     1921 ctgagcacgc aaagcgttct gtctaaagat ccgaacgaga aacgcgatca tatggttctg
     1981 ctggagttcg taaccgcagc gggcatcacg catggtatgg atgaactgta caaatgatga
     2041 cccgggaagc ttggctgttt tggcggatga gagaagattt tcagcctgat acagattaaa
     2101 tcagaacgca gaagcggtct gataaaacag aatttgcctg gcggcagtag cgcggtggtc
     2161 ccacctgacc ccatgccgaa ctcagaagtg aaacgccgta gcgccgatgg tagtgtgggg
     2221 tctccccatg cgagagtagg gaactgccag gcatcaaata aaacgaaagg ctcagtcgaa
     2281 agactgggcc tttcgtttta tctgttgttt gtcggtgaac gctctcctga gtaggacaaa
     2341 tccgccggga gcggatttga acgttgcgaa gcaacggccc ggagggtggc gggcaggacg
     2401 cccgccataa actgccaggc atcaaattaa gcagaaggcc atcctgacgg atggcctttt
     2461 tgcgtttcta caaactcttt tgtttatttt tctaaataca ttcaaatatg tatccgctca
     2521 tgagacaata accctgataa atgcttcaat aatattgaaa aaggaagagt atgagtattc
     2581 aacatttccg tgtcgccctt attccctttt ttgcggcatt ttgccttcct gtttttgctc
     2641 acccagaaac gctggtgaaa gtaaaagatg ctgaagatca gttgggtgca cgagtgggtt
     2701 acatcgaact ggatctcaac agcggtaaga tccttgagag ttttcgcccc gaagaacgtt
     2761 ttccaatgat gagcactttt aaagttctgc tatgtggcgc ggtattatcc cgtgttgacg
     2821 ccgggcaaga gcaactcggt cgccgcatac actattctca gaatgacttg gttgagtact
     2881 caccagtcac agaaaagcat cttacggatg gcatgacagt aagagaatta tgcagtgctg
     2941 ccataaccat gagtgataac actgcggcca acttacttct gacaacgatc ggaggaccga
     3001 aggagctaac cgcttttttg cacaacatgg gggatcatgt aactcgcctt gatcgttggg
     3061 aaccggagct gaatgaagcc ataccaaacg acgagcgtga caccacgatg cctgtagcaa
     3121 tggcaacaac gttgcgcaaa ctattaactg gcgaactact tactctagct tcccggcaac
     3181 aattaataga ctggatggag gcggataaag ttgcaggacc acttctgcgc tcggcccttc
     3241 cggctggctg gtttattgct gataaatctg gagccggtga gcgtgggtct cgcggtatca
     3301 ttgcagcact ggggccagat ggtaagccct cccgtatcgt agttatctac acgacgggga
     3361 gtcaggcaac tatggatgaa cgaaatagac agatcgctga gataggtgcc tcactgatta
     3421 agcattggta actgtcagac caagtttact catatatact ttagattgat ttacgcgccc
     3481 tgtagcggcg cattaagcgc ggcgggtgtg gtggttacgc gcagcgtgac cgctacactt
     3541 gccagcgccc tagcgcccgc tcctttcgct ttcttccctt cctttctcgc cacgttcgcc
     3601 ggctttcccc gtcaagctct aaatcggggg ctccctttag ggttccgatt tagtgcttta
     3661 cggcacctcg accccaaaaa acttgatttg ggtgatggtt cacgtagtgg gccatcgccc
     3721 tgatagacgg tttttcgccc tttgacgttg gagtccacgt tctttaatag tggactcttg
     3781 ttccaaactt gaacaacact caaccctatc tcgggctatt cttttgattt ataagggatt
     3841 ttgccgattt cggcctattg gttaaaaaat gagctgattt aacaaaaatt taacgcgaat
     3901 tttaacaaaa tattaacgtt tacaatttaa aaggatctag gtgaagatcc tttttgataa
     3961 tctcatgacc aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag accccgtaga
     4021 aaagatcaaa ggatcttctt gagatccttt ttttctgcgc gtaatctgct gcttgcaaac
     4081 aaaaaaacca ccgctaccag cggtggtttg tttgccggat caagagctac caactctttt
     4141 tccgaaggta actggcttca gcagagcgca gataccaaat actgtccttc tagtgtagcc
     4201 gtagttaggc caccacttca agaactctgt agcaccgcct acatacctcg ctctgctaat
     4261 cctgttacca gtggctgctg ccagtggcga taagtcgtgt cttaccgggt tggactcaag
     4321 acgatagtta ccggataagg cgcagcggtc gggctgaacg gggggttcgt gcacacagcc
     4381 cagcttggag cgaacgacct acaccgaact gagataccta cagcgtgagc tatgagaaag
     4441 cgccacgctt cccgaaggga gaaaggcgga caggtatccg gtaagcggca gggtcggaac
     4501 aggagagcgc acgagggagc ttccaggggg aaacgcctgg tatctttata gtcctgtcgg
     4561 gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct
     4621 atggaaaaac gccagcaacg cggccttttt acggttcctg gccttttgct ggccttttgc
     4681 tcacatgttc tttcctgcgt tatcccctga ttctgtggat aaccgtatta ccgcctttga
     4741 gtgagctgat accgctcgcc gcagccgaac gaccgagcgc agcgagtcag tgagcgagga
     4801 agcggaagag cgcctgatgc ggtattttct ccttacgcat ctgtgcggta tttcacaccg
     4861 catagggtca tggctgcgcc ccgacacccg ccaacacccg ctgacgcgcc ctgacgggct
     4921 tgtctgctcc cggcatccgc ttacagacaa gctgtgaccg tctccgggag ctgcatgtgt
     4981 cagaggtttt caccgtcatc accgaaacgc gcgaggcagc aaggagatgg cgcccaacag
     5041 tcccccggcc acggggcctg ccaccatacc cacgccgaaa caagcgctca tgagcccgaa
     5101 gtggcgagcc cgatcttccc catcggtgat gtcggcgata taggcgccag caaccgcacc
     5161 tgtggcgccg gtgatgccgg ccacgatgcg tccggcgtag aggatctgct catgtttgac
     5221 agcttatc
//
