LOCUS       sequence-127163-        3405 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  pbuE Sense Expression Plasmid.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3405)
  AUTHORS   Chappell J, Takahashi MK, Lucks JB
  TITLE     Creating small transcription activating RNAs.
  JOURNAL   Nat Chem Biol. 2015 Mar;11(3):214-20. doi: 10.1038/nchembio.1737. 
            Epub 2015 Feb 2.
  PUBMED    25643173
REFERENCE   2  (bases 1 to 3405)
  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: 
            "10.1038/nchembio.1737"; journalName: "Nat Chem Biol"; date: 
            "2015-03"; volume: "11"; issue: "3"; pages: "214-20"
COMMENT     Sequence Label: pJBL2184
FEATURES             Location/Qualifiers
     source          1..3405
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        16..50
                     /label=J23119(SpeI) promoter
                     /note="bacterial promoter (Registry of Standard Biological 
                     Parts BBa_J23119) modified to end with an SpeI site"
     CDS             join(144..146,147..860)
                     /codon_start=1
                     /product="GFP variant that folds robustly even when fused 
                     to poorly folded proteins (Pedelacq et al., 2006)"
                     /label=superfolder GFP
                     /note="mammalian codon-optimized"
                     /translation="MSKGEELFTGVVPILVELDGDVNGHKFSVRGEGEGDATNGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKRHDFFKSAMPEGYVQERTISFKDDG
                     TYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNFNSHNVYITADKQKNGIKA
                     NFKIRHNVEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSVLSKDPNEKRDHMVLLE
                     FVTAAGITHGMDELYK"
     CDS             144..857
                     /codon_start=1
                     /product="GFP variant that folds robustly even when fused 
                     to poorly folded proteins (Nager et al., 2011)"
                     /label=superfolder GFP
                     /translation="MSKGEELFTGVVPILVELDGDVNGHKFSVRGEGEGDATNGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKRHDFFKSAMPEGYVQERTISFKDDG
                     TYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNFNSHNVYITADKQKNGIKA
                     NFKIRHNVEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSVLSKDPNEKRDHMVLLE
                     FVTAAGITHGMDELYK"
     primer_bind     complement(174..202)
                     /label=GFP-R
                     /note="GFP, reverse primer. Does NOT anneal to EGFP"
     primer_bind     797..817
                     /label=GFP-F
                     /note="GFP, forward primer. Does NOT anneal to EGFP"
     CDS             880..909
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             925..942
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(998..1015)
                     /label=pBAD Reverse
                     /note="For vectors with E. coli araBAD promoter, reverse 
                     primer"
     terminator      1168..1214
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     CDS             1367..2026
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(1433..1452)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     terminator      2047..2141
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     primer_bind     complement(2548..2565)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2682..3227)
                     /direction=LEFT
                     /label=p15A ori
                     /note="Plasmids containing the medium-copy-number p15A 
                     origin of replication can be propagated in E. coli cells 
                     that contain a second plasmid with the ColE1 origin.
                     "
     primer_bind     complement(2762..2781)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 gaattctaaa gatctttgac agctagctca gtcctaggta taatactagt gtgtctacca
       61 ggaaccgtaa aatcctgatt acaaaatttg tttatgacat tttttgtaat caggattttt
      121 tttggatcta ggaggaagga tctatgagca aaggagaaga acttttcact ggagttgtcc
      181 caattcttgt tgaattagat ggtgatgtta atgggcacaa attttctgtc cgtggagagg
      241 gtgaaggtga tgctacaaac ggaaaactca cccttaaatt tatttgcact actggaaaac
      301 tacctgttcc gtggccaaca cttgtcacta ctctgaccta tggtgttcaa tgcttttccc
      361 gttatccgga tcacatgaaa cggcatgact ttttcaagag tgccatgccc gaaggttatg
      421 tacaggaacg cactatatct ttcaaagatg acgggaccta caagacgcgt gctgaagtca
      481 agtttgaagg tgataccctt gttaatcgta tcgagttaaa gggtattgat tttaaagaag
      541 atggaaacat tcttggacac aaactcgagt acaactttaa ctcacacaat gtatacatca
      601 cggcagacaa acaaaagaat ggaatcaaag ctaacttcaa aattcgccac aacgttgaag
      661 atggttccgt tcaactagca gaccattatc aacaaaatac tccaattggc gatggccctg
      721 tccttttacc agacaaccat tacctgtcga cacaatctgt cctttcgaaa gatcccaacg
      781 aaaagcgtga ccacatggtc cttcttgagt ttgtaactgc tgctgggatt acacatggca
      841 tggatgagct ctacaaataa ggatctgaag cttgggcccg aacaaaaact catctcagaa
      901 gaggatctga atagcgccgt cgaccatcat catcatcatc attgagttta aacggtctcc
      961 agcttggctg ttttggcgga tgagagaaga ttttcagcct gatacagatt aaatcagaac
     1021 gcagaagcgg tctgataaaa cagaatttgc ctggcggcag tagcgcggtg gtcccacctg
     1081 accccatgcc gaactcagaa gtgaaacgcc gtagcgccga tggtagtgtg gggtctcccc
     1141 atgcgagagt agggaactgc caggcatcaa ataaaacgaa aggctcagtc gaaagactgg
     1201 gcctttcgtt ttatctgttg tttgtcggtg aactggatcc ttactcgagt ctagactgca
     1261 gttgatcggg cacgtaagag gttccaactt tcaccataat gaaataagat cactaccggg
     1321 cgtatttttt gagttatcga gattttcagg agctaaggaa gctaaaatgg agaaaaaaat
     1381 cactggatat accaccgttg atatatccca atggcatcgt aaagaacatt ttgaggcatt
     1441 tcagtcagtt gctcaatgta cctataacca gaccgttcag ctggatatta cggccttttt
     1501 aaagaccgta aagaaaaata agcacaagtt ttatccggcc tttattcaca ttcttgcccg
     1561 cctgatgaat gctcatccgg aatttcgtat ggcaatgaaa gacggtgagc tggtgatatg
     1621 ggatagtgtt cacccttgtt acaccgtttt ccatgagcaa actgaaacgt tttcatcgct
     1681 ctggagtgaa taccacgacg atttccggca gtttctacac atatattcgc aagatgtggc
     1741 gtgttacggt gaaaacctgg cctatttccc taaagggttt attgagaata tgtttttcgt
     1801 ctcagccaat ccctgggtga gtttcaccag ttttgattta aacgtggcca atatggacaa
     1861 cttcttcgcc cccgttttca ccatgggcaa atattatacg caaggcgaca aggtgctgat
     1921 gccgctggcg attcaggttc atcatgccgt ttgtgatggc ttccatgtcg gcagaatgct
     1981 taatgaatta caacagtact gcgatgagtg gcagggcggg gcgtaatttg atatcgagct
     2041 cgcttggact cctgttgata gatccagtaa tgacctcaga actccatctg gatttgttca
     2101 gaacgctcgg ttgccgccgg gcgtttttta ttggtgagaa tccaagcctc cgatcaacgt
     2161 ctcattttcg ccaaaagttg gcccagggct tcccggtatc aacagggaca ccaggattta
     2221 tttattctgc gaagtgatct tccgtcacag gtatttattc ggcgcaaagt gcgtcgggtg
     2281 atgctgccaa cttactgatt tagtgtatga tggtgttttt gaggtgctcc agtggcttct
     2341 gtttctatca gctgtccctc ctgttcagct actgacgggg tggtgcgtaa cggcaaaagc
     2401 accgccggac atcagcgcta gcggagtgta tactggctta ctatgttggc actgatgagg
     2461 gtgtcagtga agtgcttcat gtggcaggag aaaaaaggct gcaccggtgc gtcagcagaa
     2521 tatgtgatac aggatatatt ccgcttcctc gctcactgac tcgctacgct cggtcgttcg
     2581 actgcggcga gcggaaatgg cttacgaacg gggcggagat ttcctggaag atgccaggaa
     2641 gatacttaac agggaagtga gagggccgcg gcaaagccgt ttttccatag gctccgcccc
     2701 cctgacaagc atcacgaaat ctgacgctca aatcagtggt ggcgaaaccc gacaggacta
     2761 taaagatacc aggcgtttcc ccctggcggc tccctcgtgc gctctcctgt tcctgccttt
     2821 cggtttaccg gtgtcattcc gctgttatgg ccgcgtttgt ctcattccac gcctgacact
     2881 cagttccggg taggcagttc gctccaagct ggactgtatg cacgaacccc ccgttcagtc
     2941 cgaccgctgc gccttatccg gtaactatcg tcttgagtcc aacccggaaa gacatgcaaa
     3001 agcaccactg gcagcagcca ctggtaattg atttagagga gttagtcttg aagtcatgcg
     3061 ccggttaagg ctaaactgaa aggacaagtt ttggtgactg cgctcctcca agccagttac
     3121 ctcggttcaa agagttggta gctcagagaa ccttcgaaaa accgccctgc aaggcggttt
     3181 tttcgttttc agagcaagag attacgcgca gaccaaaacg atctcaagaa gatcatctta
     3241 ttaatcagat aaaatatttc tagatttcag tgcaatttat ctcttcaaat gtagcacctg
     3301 aagtcagccc catacgatat aagttgtaat tctcatgttt gacagcttat catcgataag
     3361 cttccgatgg cgcgccgaga ggctttacac tttatgcttc cggct
//
